here - The International Organization for Mycoplasmology

Transcription

Phylum XVI. Tenericutes Murray 1984a, 356VP
(Effective publication: Murray 1984b, 33.)
Daniel R. Brown
Ten.er¢i.cutes. L. adj. tener tender; L. fem. n. cutis skin; N.L. fem. n. Tenericutes prokaryotes
of a soft pliable nature indicative of a lack of a rigid cell wall.
Members of the Tenericutes are wall-less bacteria that do not synthesize precursors of peptidoglycan.
Further descriptive information
The nomenclatural type by monotypy (Murray, 1984a) is the
class Mollicutes, which consists of very small prokaryotes that
are devoid of cell walls. Electron microscopic evidence for the
absence of a cell wall was mandatory for describing novel species
of mollicutes until very recently. Genes encoding the pathways
for peptidoglycan biosynthesis are absent from the genomes of
more than 15 species that have been annotated to date. Some
species do possess an extracellular glycocalyx. The absence of
a cell wall confers such mechanical plasticity that most mollicutes are readily filterable through 450 nm pores and many species have some cells in their populations that are able to pass
through 220 nm or even 100 nm filters. However, they may vary
in shape from coccoid to flask-shaped cells or helical filaments
that reflect flexible cytoskeletal elements.
Taxonomic comments
To provide greater definition and formal nomenclature for
vernacular names used in the 8th edition of Bergey’s Manual of
Determinative Bacteriology (Bergey VIII; Buchanan and Gibbons,
1974), Gibbons and Murray (1978) proposed that the higher
taxa of prokaryotes be subdivided primarily according to the
presence and character, or absence, of a rigid or semirigid cell
wall as reflected in the determinative Gram reaction. Similar
to the non-hierarchical groupings of Bergey VIII, which were
based on a few readily determined criteria, the “wall-deficient”
organisms grouped together in the first edition of The Prokaryotes included the mollicutes (Starr et al., 1981). While acknowledging the emerging 16S rRNA-based evidence that indicated
a phylogenetic relationship between mollicutes and certain
Gram-stain-positive bacteria in the division Firmicutes, Murray
(1984b) proposed the separate division Tenericutes for the stable
and distinctive group of wall-less species that are not simply an
obvious subset of the Firmicutes.
The approved divisional rank of Tenericutes and the assignment
of class Mollicutes as its nomenclatural type (Murray, 1984a) were
adopted by the International Committee on Systematic Bacteriology’s Subcommittee on the Taxonomy of Mollicutes (Tully,
1988) and subsequent valid taxonomic descriptions assigned
novel species of mollicutes to the Tenericutes. However, the second
(1992) and third (2007) editions of The Prokaryotes described the
mollicutes instead as Firmicutes with low G+C DNA. The Subcommittee considered this to be an unfortunate grouping: “While
the organisms are evolutionarily related to certain clostridia,
the absence of a cell wall cannot be equated with Gram reaction positivity or with other members of the Firmicutes. It is
unfortunate that workers involved in determinative bacteriology
have a reference in which wall-free prokaryotes are described as
Gram-positive bacteria” (Tully, 1993a). Despite numerous valid
assignments of novel species of mollicutes to the Tenericutes during the intervening years, the class Mollicutes was still included
in the phylum Firmicutes in the most recent revision of the Taxonomic Outline of Bacteria and Archaea (TOBA), which is based
solely on the phylogeny of 16S rRNA genes (Garrity et al., 2007).
The taxon Tenericutes is not recognized in the TOBA, although
paradoxically it is the phylum consisting of the Mollicutes in the
most current release of the Ribosomal Database Project (Cole
et al., 2009). Mollicutes are specifically excluded from the most
recently emended description of the Firmicutes in Bergey’s Manual
of Systematic Bacteriology (2nd edition, volume 3; De Vos et al.,
2009) on the grounds of their lack of rigid cell walls plus analyses
of strongly supported alternative universal phylogenetic markers,
including RNA polymerase subunit B, the chaperonin GroEL,
several different aminoacyl tRNA synthetases, and subunits of
F0F1-ATPase (Ludwig et al., 2009; Ludwig and Schleifer, 2005).
The taxonomic dignity of Tenericutes bestowed by its original
formal validation, and upheld by a quarter of a century of valid
descriptions of novel species of mollicutes, has therefore been
respected in this volume of Bergey’s Manual.
Type order: Mycoplasmatales Freundt 1955, 71AL emend. Tully,
Bové, Laigret and Whitcomb 1993b, 382.
References
Buchanan, R.E. and N.E. Gibbons (editors). 1974. Bergey’s Manual of
Determinative Bacteriology, 8th edn. Williams & Wilkins, Baltimore.
Cole, J.R., Q. Wang, E. Cardenas, J. Fish, B. Chai, R.J. Farris, A.S. KulamSyed-Mohideen, D.M. McGarrell, T. Marsh, G.M. Garrity and J.M.
Tiedje. 2009. The Ribosomal Database Project: improved alignments
and new tools for rRNA analysis. Nucleic Acids Res. 37: (Database
issue): D141–D145.
De Vos, P., G. Garrity, D. Jones, N.R. Krieg, W. Ludwig, F.A. Rainey, K.H.
Schleifer and W.B. Whitman. 2009. In Bergey’s Manual of Systematic
Bacteriology, 2nd edn, vol. 3. Springer, New York.
Freundt, E.A. 1955. The classification of the pleuropneumoniae group
of organisms (Borrelomycetales). Int. Bull. Bacteriol. Nomencl. Taxon.
5: 67–78.
Garrity, G.M., T.G. Lilburn, J.R. Cole, S.H. Harrison, J. Euzéby and B.J.
Tindall. 2007. The Taxonomic Outline of the Bacteria and Archaea,
Release 7.7, Part 11 – The Bacteria: Phyla Planctomycetes, Chlamydiae, Spirochaetes, Fibrobacteres, Acidobacteria, Bacteroidetes, Fusobacteria,
567
568
Phylum XVI. Tenericutes
errucomicrobia, Dictyoglomi, Gemmatomonadetes, and Lentisphaerae. pp.
V
540–595. (http://www.taxonomicoutline.org/).
Gibbons, N.E. and R.G.E. Murray. 1978. Proposals concerning the
higher taxa of bacteria. Int. J. Syst. Bacteriol. 28: 1–6.
Ludwig, W. and K.H. Schleifer. 2005. Molecular phylogeny of bacteria based on comparative sequence analysis of conserved genes. In
Microbial Phylogeny and Evolution, Concepts and Controversies,
(edited by Sapp). Oxford University Press, New York, pp. 70–98.
Ludwig, W., K.H. Schleifer and W.B. Whitman. 2009. Revised road map to
the phylum Firmicutes. In Bergey’s Manual of Systematic Bacteriology, 2nd
edn, vol. 3, The Firmicutes (edited by De Vos, Garrity, Jones, Krieg, Ludwig, Rainey, Schleifer and Whitman). Springer, New York, pp. 1–13.
Murray, R.G.E. 1984a. In Validation of the publication of new names
and new combinations previously effectively published outside the
IJSB. List no. 15. Int. J. Syst. Bacteriol. 34: 355–357.
Murray, R.G.E. 1984b. The higher taxa, or, a place for everything…? In
Bergey’s Manual of Systematic Bacteriology, vol. 1 (edited by Krieg
and Holt). Williams & Wilkins, Baltimore, pp. 31–34.
Starr, M.P., H. Stolp, H.G. Trüper, A. Balows and H.G. Schlegel (editors).
1981. The Prokaryotes. Springer, Berlin.
Tully, J.G. 1988. International Committee on Systematic Bacteriology, Subcommittee on the Taxonomy of Mollicutes, Minutes of the
Interim Meeting, 25 and 28 August 1986, Birmingham, Alabama. Int.
J. Syst. Bacteriol. 38: 226–230.
Tully, J.G. 1993a. International Committee on Systematic Bacteriology, Subcommittee on the Taxonomy of Mollicutes, Minutes of the
Interim Meetings, 1 and 2 August, 1992, Ames, Iowa. Int. J. Syst.
Bacteriol. 43: 394–397.
Tully, J.G., J.M. Bové, F. Laigret and R.F. Whitcomb. 1993b. Revised
taxonomy of the class Mollicutes – proposed elevation of a monophyletic cluster of arthropod-associated mollicutes to ordinal rank (Entomoplasmatales ord. nov.), with provision for familial rank to separate
species with nonhelical morphology (Entomoplasmataceae fam. nov.)
from helical species (Spiroplasmataceae), and emended descriptions
of the order Mycoplasmatales, family Mycoplasmataceae. Int. J. Syst.
Bacteriol. 43: 378–385.
Class I. Mollicutes Edward and Freundt 1967, 267AL
Daniel R. Brown, Meghan May, Janet M. Bradbury and Karl-Erik Johansson
Mol¢li.cutes or Mol.li.cu¢tes. L. adj. mollis soft, pliable; L. fem. n. cutis skin; N.L. fem. pl. n.
Mollicutes class with pliable cell boundary.
Very small prokaryotes totally devoid of cell walls. Bounded
by a plasma membrane only. Incapable of synthesis of peptidoglycan or its precursors. Consequently resistant to penicillin
and its derivatives and sensitive to lysis by osmotic shock, detergents, alcohols, and specific antibody plus complement. Gramstain-negative due to lack of cell wall, but constitute a distinct
phylogenetic lineage within the Gram-stain-positive bacteria
(Woese et al., 1980). Pleomorphic, varying from spherical or
flask-shaped structures to branched or helical filaments. The
coccoid and flask-shaped cells usually range from 200–500 nm
in diameter, although cells as large as 2000 nm have been seen.
Replicate by binary fission, but genome replication may precede cytoplasmic division, leading to the formation of multinucleated filaments. Colonies on solid media are very small,
usually much less than 1 mm in diameter. The organisms tend
to penetrate and grow inside the solid medium. Under suitable
conditions, almost all species form colonies that have a characteristic fried-egg appearance. Usually nonmotile, but some
species show gliding motility. Species that occur as helical filaments show rotary, flexional, and translational motility. No resting stages are known.
The species recognized so far can be grown on artificial cellfree media of varying complexity, although certain strains may
be more readily isolated by cell-culture procedures. Many “Candidatus” species have been proposed and characterized at the
molecular level, but not yet cultivated axenically. Most cultivable species require sterols and fatty acids for growth. However,
members of some genera can grow well in either serum-free
media or serum-free media supplemented with polyoxyethylene sorbitan. Most species are facultative anaerobes, but some
are obligate anaerobes that are killed by exposure to minute
quantities of oxygen. No tricarboxylic acid cycle enzymes, quinones, or cytochromes have been found.
All mollicutes are commensals or parasites, occurring in a wide
range of vertebrate, insect, and plant hosts. Many are significant
pathogens of humans, animals, insects, or plants. Genome sizes
range from 580 to 2200 kbp, among the smallest recorded in
prokaryotes. The genomes of more than 20 species have been
completely sequenced and annotated to date (Table 135). The
G+C content of the DNA is usually low, ~23–34 mol%, but in some
species is as high as ~40 mol% (Bd, Tm). Can be distinguished from
other bacteria in having only one or two rRNA operons (one species of Mesoplasma has three) and an RNA polymerase that is resistant to rifampin. The 5S rRNA contains fewer nucleotides than
that of other bacteria and there are fewer tRNA genes. In some
genera, instead of a stop, the UGA codon encodes tryptophan.
Plasmids and viruses (phage) occur in some species.
Type order: Mycoplasmatales Freundt 1955, 71AL emend. Tully,
Bové, Laigret and Whitcomb 1993, 382.
Table 136 summarizes the present classification of the Mollicutes
into families and genera and provides the major distinguishing
characteristics of these taxa. The trivial term mycoplasma has
been used to denote any species included in the class Mollicutes,
but the term mollicute(s) is now considered most appropriate
as the trivial name for all members of the class, so that the trivial name mycoplasma can be retained only for members of the
genus Mycoplasma. Hemotropic mycoplasmas are referred to by
the trivial name hemoplasmas. The trivial names ureaplasma,
entomoplasma, mesoplasma, spiroplasma, acholeplasma,
anaeroplasma, and asteroleplasma are commonly used when
reference is made to members of the corresponding genus.
Their 16S rRNA gene sequences usefully place the mollicutes into phylogenetic groups ( Johansson, 2002; Weisburg
et al., 1989) and an analysis of 16S rRNA gene sequences is
now mandatory for characterization of novel species (Brown
et al., 2007). 16S rRNA gene sequences have also shown that
certain hemotropic bacteria, previously considered to be
members of the Rickettsia, belong to the order Mycoplasmatales.
0.82
30
631
nd
80
Dybvig
et al.
(2008)
PG2T
0.88
29
742
50
87
SirandPugnet
et al.
(2007)
Strain
Size (mb)
DNA G+C
content
(mol%)
Open reading
frames
Hypothetical
genes (%)
Coding
density (%)
References
Mycoplasma arthritidis
Mycoplasma capricolum subsp. capricolum
J. Glass
and
others,
unpublished.
88
nd
812
ATCC
27343T
1.0
23
Mycoplasma conjunctivae
91
37
726
0.99
31
R
Calderon- Papazisi
Copete
et al.
et al.
(2003)
(2009)
90
45
727
0.85
28
HRC/581T
Mycoplasma genitalium
Fraser
et al.
(1995)
90
21
475
0.58
31
G-37T
Mycoplasma hominis
Mycoplasma hyopneumoniae b
87
nd
657
0.90
28
JT
Mycoplasma mobile
90
27
633
0.78
25
163KT
Mycoplasma mycoides subsp. mycoides SC
80
41
1016
1.21
24
PG1T
88
41
1037
1.36
26
HF-2
Mycoplasma penetrans
Pereyre Vasconcelos Jaffe Westberg Sasaki
et al.
et al.
et al.
et al.
et al.
(2009)
(2005)
(2004) (2004) (2002)
90
36
537
0.67
27
PG21T
Mycoplasma pneumoniae
Himmelreich
et al.
(1996)
87
38
689
0.82
40
M129
b
a
91
33
659
0.80
28
53
Mycoplasma synoviae
Chambaud Vasconcelos
et al. (2001)
et al.
(2005)
90
38
782
0.96
26
UAB CTIP
Mycoplasma pulmonis
nd, Not determined; na, not available.
Mycoplasma hyopneumoniae strains 232 and 7448 were sequenced by Minion et al. (2004) and Vasconcelos et al. (2005).
c
Data for Ureaplasma parvum and Ureaplasma urealyticum refer to serovars 3 and 10, respectively; serovars 1–14 were sequenced and deposited directly into GenBank.
d
AYWB, Aster yellows witches’ broom; the onion yellows strain was sequenced by Oshima et al. (2004).
158L3-1
Mycoplasma agalactiae
Species
Mycoplasma gallisepticum
Table 135. Characteristics of sequenced mollicute genomesa
Mesoplasma florum
Knight
et al.
(2004)
92
nd
687
0.79
27
L1T
Ureaplasma parvumc
Glass
et al.
(2000)
91
33
653
ATCC
700970
0.75
25
Ureaplasma urealyticumc
na
89
nd
692
ATCC
33699
0.87
25
Acholeplasma laidlawii
na
90
nd
1433
1.5
31
PG8T
Bai
et al.
(2006)
73
nd
671
0.71
26
AYWB
“Candidatus Phytoplasma asteris”d
Mycoplasmataceae
Mycoplasmataceae
Incertae sedis
Incertae sedis
Entomoplasmataceae
Entomoplasmataceae
Spiroplasmataceae
Acholeplasmataceae
Incertae sedis
Anaeroplasmataceae
Anaeroplasmataceae
Family
Numbers of species: valid, Candidatus, incertae sedis, invalid.
Mycoplasma
Ureaplasma
Eperythrozoon
Haemobartonella
Entomoplasma
Mesoplasma
Spiroplasma
Acholeplasma
“Candidatus Phytoplasma”
Anaeroplasma
Asteroleplasma
Genus
c
H, human; A, vertebrate animal; N, invertebrate animal; P, plant.
d
Affiliation of the constituent genera within the Mycoplasmatales has not been formalized.
b
a
nd, Not determined; PES, polyoxyethylene sorbitan.
I. Mycoplasmatales
I. Mycoplasmatales
I. Mycoplasmatalesd
I. Mycoplasmatalesd
II. Entomoplasmatales
III. Acholeplasmatales
III. Acholeplasmatales
IV. Anaeroplasmatales
IV. Anaeroplasmatales
Order
Table 136. Description of the class Mollicutes a
116, 9, 1, 4
7, 0, 0, 0
4, 0, 0, 0
1, 0, 0, 0
6, 0, 0, 0
11, 0, 0, 0
37, 0, 0, 0
18, 0, 0, 0
0, 27, 0, 0
4, 0, 0, 0
1, 0, 0, 0
Speciesb
580–1,350
760–1,140
Nd
Nd
870–900
825–930
780–2,220
1,500–1,650
530–1,350
1,500–1,600
1,500
Genome size range (kbp)
+
+
nd
nd
+
−
+
−
nd
+
−
Cholesterol
requirement
H, A
H, A
A
A
N, P
N, P
N, P
A, N, P
N, P
A
A
Habitatc
Not yet cultured in vitro
Strictly anaerobic
Strictly anaerobic
Growth with PES
Helical morphology
Urea hydrolysis
Hemotropic
Hemotropic
Defining features
570
The phytoplasmas, a large group of uncultivated mollicutes
occurring as agents that can cycle between plant and invertebrate hosts, have been given a provisional “Candidatus Phytoplasma” genus designation. The 16S rRNA gene sequences
from at least ten unique phylotypes, recently discovered among
the human microbial flora through global 16S rRNA gene PCR
(Eckburg et al., 2005), cluster distinctly enough to suggest the
existence of a yet-uncircumscribed order within the class (May
et al., 2009).
Non-helical mollicutes isolated from insects and plants have
been placed in the order Entomoplasmatales, the two genera
of which are distinguished by their requirement for cholesterol (Tully et al., 1993). Members of the genus Entomoplasma
require cholesterol; those of the genus Mesoplasma do not.
However, sterol requirements do not correlate well with phylogenetic analyses in other groups. At least four species of
spiroplasmas do not require sterol for growth, but they do not
form a phylogenetic group. Within the order including the
obligately anaerobic mollicutes Anaeroplasmatales, members
of the genus Anaeroplasma require sterols for growth, whereas
members of the genus Asteroleplasma do not (Robinson et al.,
1975; Robinson and Freundt, 1987). Thus, sterol requirement
is a useful phylogenetic marker only in the Acholeplasmatales
and Anaeroplasmatales.
In the past, there was some risk of confusing mollicutes with
wall-less “L (Lister)-phase” variants of certain other bacteria, but
simple PCR-based analyses of 16S rRNA or other gene sequences
now obviate that concern. Wall-less members of the genus Thermoplasma, previously assigned to the Mollicutes, are Archaea and
differ from all other members of this class in their 16S rRNA
nucleotide sequences plus a number of important features relating to their mode of life and metabolism. Thus, they are quite
unrelated to this class (Fox et al., 1980; Razin and Freundt, 1984;
Woese et al., 1980). Members of the Erysipelothrix line of descent,
also formerly assigned to the Mollicutes, are now assigned to the
class Erysipelotrichi in the phylum Firmicutes (Stackebrandt, 2009;
Verbarg et al., 2004).
Taxonomic comments
The origin of mollicutes and their relationships to other
prokaryotes was controversial for many years, especially since
their small genomes and comparative phenotypic simplicity
suggested that they might have descended from a primitive
organism. The first comparative phylogenetic analysis of the
origin of mollicutes was carried out by oligonucleotide mapping of 16S rRNA gene sequences by Woese et al. (1980). The
organisms then assigned to the genera Mycoplasma, Spiroplasma,
and Acholeplasma seemed to have arisen by reductive evolution
as a deep branch of the clostridial lineage leading to the genera
Bacillus and Lactobacillus. This relationship had been proposed
earlier (Neimark, 1979) because the low G+C mollicutes, streptococci, and lactic acid bacteria share characteristic enzymes.
In particular, acholeplasma and streptococcus aldolases show
high amino acid sequence similarity.
These findings were generally confirmed by studies of 5S
rRNA gene sequences (Rogers et al., 1985), which included a
number of acholeplasmas, anaeroplasmas, mycoplasmas, ureaplasmas, and Clostridium innocuum. Dendrograms constructed
from evolutionary distance matrices indicated that the mollicutes form a coherent phylogenetic group that developed as
571
a branch of the Firmicutes. The initial event in this evolution
was proposed to be the formation of the Acholeplasma branch,
although the position of the Anaeroplasma species (Anaeroplasma bactoclasticum and Anaeroplasma abactoclasticum) was not
definitely established within these dendrograms. Formation
of the acholeplasmas may have coincided with a reduction in
genome size to about 1500–1700 kb and loss of the cell wall.
With a genome size similar to the acholeplasmas, the spiroplasmas may have formed from the acholeplasmas. Later independent genome reductions to 500–1000 kb may have led to the
origins of the sterol-requiring mycoplasma and ureaplasma lineages. The more extensive phylogenetic analysis of Weisburg
et al. (1989) examined the 16S rRNA gene sequences of about
50 species of mollicutes and confirmed a number of these
observations and provided additional insights into mollicute
evolution. These results also indicated that the acholeplasmas
formed upon the initial divergence of mollicutes from clostridial ancestors. Further divergence of this stem led to the sterolrequiring, anaerobic Anaeroplasma and the non-sterol-requiring
Asteroleplasma branches. The Spiroplasma branch also appeared
to originate from within the acholeplasmas, with further evolution leading to a series of repeated and independent genome
reductions from nearly 2000 kb to 600–1200 kb to yield the
Mycoplasma and Ureaplasma lineages.
Based on the phylogeny of 16S rRNA genes, the class
Mollicutes was included in the phylum Firmicutes in the most
recent revision of the Taxonomic Outline of Bacteria and Archaea
(Garrity et al., 2007). However, the Mollicutes are excluded
from the most recently emended description of the Firmicutes
(De Vos et al., 2009) based on alternative phylogenetic markers,
including RNA polymerase subunit B, the chaperonin GroEL,
several different aminoacyl tRNA synthetases, and subunits of
F0F1-ATPase (Ludwig and Schleifer, 2005).
The Weisburg et al. (1989) study also proposed five additional phylogenetic groupings within the mollicutes, including
the anaeroplasma, asteroleplasma, spiroplasma, pneumoniae,
and hominis groups (Figure 105). Phytoplasmas are similar to
acholeplasmas in their 16S rRNA gene sequences and UGA
codon usage (IRPCM Phytoplasma/Spiroplasma Working
Team – Phytoplasma Taxonomy Group, 2004). They probably
diverged from acholeplasmas at about the same time as the split
of spiroplasmas into helical and non-helical lineages (Maniloff,
2002). The modern species concept for mollicutes is justified
principally by DNA–DNA hybridization, serology, and 16S rRNA
gene sequence similarity (Brown et al., 2007). A large number
of individual species have been assigned to phylogenetic groups,
clusters, and subclusters that also share other characteristics,
although the cluster boundaries are sometimes subjective
(Harasawa and Cassell, 1996; Johansson, 2002; Pettersson et al.,
2000, 2001).
Lastly, the type order Mycoplasmatales is assigned to the class
as this clearly appeared to be the intention of Edward and Freundt (1967) in their paper entitled “Proposal for Mollicutes as
name of the class established for the order Mycoplasmatales”.
Acknowledgements
The lifetime achievements in mycoplasmology and major contributions to the foundation of this material by Joseph G. Tully
are gratefully acknowledged. Daniel R. Brown and Meghan May
were supported by NIH grant 5R01GM076584.
572
Mycoplasma hominis
Ureaplasma urealyticum
*
Mycoplasma coccoides
Spiroplasma apis
Mycoplasma mycoides subsp. mycoides
Entomoplasma ellychniae
Mesoplasma florum
Spiroplasma citri
Spiroplasma ixodetis
‘Candidatus Phytoplasma’ strain OY-M
Anaeroplasma abactoclasticum
Asteroleplasma anaerobium
Clostridium innocuum
Scale:
0.1 substitutions / site
Figure 105. Phylogenetic grouping of the class Mollicutes. The phylogram was based on a Jukes–Cantor corrected distance matrix and weighted
neighbor-joining analysis of the 16S rRNA gene sequences of the type genera, plus representatives of other major clusters within the Mycoplasmatales and Entomoplasmatales and a phytoplasma. Clostridium innocuum was the outgroup. All bootstrap values (100 replicates) are >50% except
where indicated (asterisk).
Further reading
Barile, M.F., S. Razin, J.G. Tully and R.F. Whitcomb (Editors).
1979, 1985, 1989. The Mycoplasmas (five volumes). Academic
Press, New York.
Maniloff, J., R.N. McElhaney, L.R. Finch and J.B. Baseman (editors). 1992. Mycoplasmas: Molecular Biology and Pathogenesis. American Society for Microbiology, Washington, D.C.
Murray, R.G.E. 1984. The higher taxa, or, a place for everything…?. In Bergey’s Manual of Systematic Bacteriology, vol.
1 (edited by Krieg and Holt). Williams & Wilkins, Baltimore,
pp. 31–34.
Razin, S. and J.G.E. Tully. 1995. Molecular and Diagnostic Procedures in Mycoplasmology, vol. 1. Academic Press, San Diego.
References
Bai, X., J. Zhang, A. Ewing, S.A. Miller, A. Jancso Radek, D.V.
Shevchenko, K. Tsukerman, T. Walunas, A. Lapidus, J.W. Campbell
and S.A. Hogenhout. 2006. Living with genome instability: the adaptation of phytoplasmas to diverse environments of their insect and
plant hosts. J. Bacteriol. 188: 3682–3696.
Brown, D., R. Whitcomb and J. Bradbury. 2007. Revised minimal standards for description of new species of the class Mollicutes (division
Tenericutes). Int. J. Syst. Evol. Microbiol. 57: 2703–2719.
Calderon-Copete, S.P., G. Wigger, C. Wunderlin, T. Schmidheini, J.
Frey, M.A. Quail and L. Falquet. 2009. The Mycoplasma conjunctivae
genome sequencing, annotation and analysis. BMC Bioinformatics
10 Suppl 6: S7.
Chambaud, I., R. Heilig, S. Ferris, V. Barbe, D. Samson, F. Galisson, I. Moszer, K. Dybvig, H. Wroblewski, A. Viari, E.P. Rocha and
A. Blanchard. 2001. The complete genome sequence of the murine
respiratory pathogen Mycoplasma pulmonis. Nucleic Acids Res. 29:
2145–2153.
Taylor-Robinson, D. and J. Bradbury. 1998. Mycoplasma diseases.
In Topley and Wilson’s Principles and Practice of Microbiology, vol. 3 (edited by Hausler and Sussman). Edward Arnold,
London, pp. 1013–1037.
Taylor-Robinson, D. and J.G. Tully. 1998. Mycoplasmas, ureaplasmas, spiroplasmas, and related organisms. In Topley and
Wilson, Principles and Practice of Microbiology, 9th edn, vol.
2 (edited by Balows and Duerden). Arnold Publishers, London, pp. 799–827.
Tully, J.G. and S. Razin (editors). 1996. Molecular and Diagnostic
Procedures in Mycoplasmology, vol. 2. Academic Press,
San Diego, CA.
De Vos, P., G. Garrity, D. Jones, N.R. Krieg, W. Ludwig, F.A. Rainey,
K. H. Schleifer and W.B. Whitman. 2009. In Bergey’s Manual of
Systematic Bacteriology, 2nd edn, vol. 3. Springer, New York.
Dybvig, K., C. Zuhua, P. Lao, D.S. Jordan, C.T. French, A.H. Tu and
A.E. Loraine. 2008. Genome of Mycoplasma arthritidis. Infect. Immun.
76: 4000–4008.
Eckburg, P., E. Bik, C. Bernstein, E. Purdom, L. Dethlefsen, M. Sargent,
S. Gill, K. Nelson and D. Relman. 2005. Diversity of the human intestinal microbial flora. Science 308: 1635–1638.
Edward, D.G.ff. and E.A. Freundt. 1967. Proposal for Mollicutes as name
of the class established for the order Mycoplasmatales. Int. J. Syst.
Bacteriol. 17: 267–268.
Fox, G.E., E. Stackebrandt, R.B. Hespell, J. Gibson, J. Maniloff,
T.A. Dyer, R.S. Wolfe, W.E. Balch, R.S. Tanner, L.J. Magrum,
L.B. Zablen, R. Blakemore, R. Gupta, L. Bonen, B.J. Lewis, D.A. Stahl,
K.R. Luehrsen, K.N. Chen and C.R. Woese. 1980. The phylogeny of
prokaryotes. Science 209: 457–463.
Fraser, C.M., J.D. Gocayne, O. White, M.D. Adams, R.A. Clayton, R.D.
Fleischmann, C.J. Bult, A.R. Kerlavage, G. Sutton, J.M. Kelley and
et al. 1995. The minimal gene complement of Mycoplasma genitalium. Science 270 : 397–403.
5: 67–78.
Garrity, G.M., T.G. Lilburn, J.R. Cole, S.H. Harrison, J. Euzéby and
B.J. Tindall. 2007. The Taxonomic Outline of the Bacteria and
Archaea, Release 7.7, Part 11 – The Bacteria: phyla Planctomycetes,
Chlamydiae, Spirochaetes, Fibrobacteres, Acidobacteria, Bacteroidetes,
Fusobacteria, Verrucomicrobia, Dictyoglomi, Gemmatomonadetes, and
Lentisphaerae. pp. 540–595. (http://www.taxonomicoutline.org/).
Glass, J.I., E.J. Lefkowitz, J.S. Glass, C.R. Heiner, E.Y. Chen and
G.H. Cassell. 2000. The complete sequence of the mucosal pathogen
Ureaplasma urealyticum. Nature 407: 757–762.
Harasawa, R. and G.H. Cassell. 1996. Phylogenetic analysis of genes
coding for 16S rRNA in mammalian ureaplasmas. Int. J. Syst. Bacteriol. 46: 827–829.
Himmelreich, R., H. Hilbert, H. Plagens, E. Pirkl, B.C. Li and R. Herrmann.
1996. Complete sequence analysis of the genome of the bacterium
Mycoplasma pneumoniae. Nucleic Acids Res. 24: 4420–4449.
IRPCM Phytoplasma/Spiroplasma Working Team - Phytoplasma Taxonomy Group. 2004. Description of the genus ‘Candidatus Phytoplasma’,
a taxon for the wall-less non-helical prokaryotes that colonize plant
phloem and insects. Int. J. Syst. Evol. Microbiol. 54: 1243–1255.
Jaffe, J.D., N. Stange-Thomann, C. Smith, D. DeCaprio, S. Fisher, J.
Butler, S. Calvo, T. Elkins, M.G. Fitzgerald, N. Hafez, C.D. Kodira, J.
Major, S. Wang, J. Wilkinson, R. Nicol, C. Nusbaum, B. Birren, H.C.
Berg and G.M. Church. 2004. The complete genome and proteome
of Mycoplasma mobile. Genome Res. 14: 1447–1461.
Johansson, K.-E. 2002. Taxonomy of Mollicutes. In Molecular Biology
and Pathogenicity of Mycoplasmas (edited by Razin and Herrmann).
Kluwer Academic/Plenum Publishers, New York, pp. 1–29.
Knight, T.F., Jr. 2004. Reclassification of Mesoplasma pleciae as Acholeplasma pleciae comb. nov. on the basis of 16S rRNA and gyrB gene
sequence data. Int. J. Syst. Evol. Microbiol. 54: 1951–1952.
Ludwig, W. and K.H. Schleifer. 2005. Molecular phylogeny of bacteria based on comparative sequence analysis of conserved genes. In
Microbial Phylogeny and Evolution, Concepts and Controversies
(edited by Sapp). Oxford University Press, New York, pp. 70–98.
Maniloff, J. 2002. Phylogeny and Evolution. In Molecular Biology and
Pathogenicity of Mycoplasmas. Kluwer Academic/Plenum Publishers, pp. 31–43.
May, M., R.F. Whitcomb and D.R. Brown. 2009. Mycoplasma and related
organisms. In CRC Practical Handbook of Microbiology, 2nd edn.
(edited by Goldman and Green). Taylor & Francis, pp. 456–479.
Minion, F.C., E.J. Lefkowitz, M.L. Madsen, B.J. Cleary, S.M. Swartzell
and G.G. Mahairas. 2004. The genome sequence of Mycoplasma hyopneumoniae strain 232, the agent of swine mycoplasmosis. J. Bacteriol. 186: 7123–7133.
Neimark, H.C. 1979. Phylogenetic relationships between mycoplasmas
and other prokaryotes. In The Mycoplasmas, vol. 1 (edited by Barile
and Razin). Academic Press, New York, pp. 43–61.
Oshima, K., S. Kakizawa, H. Nishigawa, H.Y. Jung, W. Wei, S. Suzuki, R.
Arashida, D. Nakata, S. Miyata, M. Ugaki and S. Namba. 2004. Reductive evolution suggested from the complete genome sequence of a
plant-pathogenic phytoplasma. Nat. Genet. 36: 27–29.
Papazisi, L., T. Gorton, G. Kutish, P. Markham, G. Browning, D. Nguyen,
S. Swartzell, A. Madan, G. Mahairas and S. Geary. 2003. The complete genome sequence of the avian pathogen Mycoplasma gallisepticum strain R(low). Microbiology 149 : 2307–2316.
Pereyre, S., P. Sirand-Pugnet, L. Beven, A. Charron, H. Renaudin, A.
Barre, P. Avenaud, D. Jacob, A. Couloux, V. Barbe, A. de Daruvar,
A. Blanchard and C. Bebear. 2009. Life on arginine for Mycoplasma
hominis: clues from its minimal genome and comparison with other
human urogenital mycoplasmas. PLoS. Genet. 5: e1000677.
573
Pettersson, B., J.G. Tully, G. Bolske and K.E. Johansson. 2000. Updated
phylogenetic description of the Mycoplasma hominis cluster (Weisburg
et al. 1989) based on 16S rDNA sequences. Int. J. Syst. Evol. Microbiol. 50: 291–301.
Pettersson, B., J.G. Tully, G. Bolske and K.E. Johansson. 2001.
Re-evaluation of the classical Mycoplasma lipophilum cluster (Weisburg
et al. 1989) and description of two new clusters in the hominis group
based on 16S rDNA sequences. Int. J. Syst. Evol. Microbiol. 51:
633–643.
Razin, S. and E.A. Freundt. 1984. The Mollicutes, Mycoplasmatales, and
Mycoplasmataceae. In Bergey’s Manual of Systematic Bacteriology,
vol. 1 (edited by Krieg and Holt). Williams & Wilkins, Baltimore,
pp. 740–742.
Robinson, I.M., M.J. Allison and P.A. Hartman. 1975. Anaeroplasma abactoclasticum gen. nov., sp. nov., obligately anaerobic mycoplasma from
rumen. Int. J. Syst. Bacteriol. 25: 173–181.
Robinson, I.M. and E.A. Freundt. 1987. Proposal for an amended classification of anaerobic mollicutes. Int. J. Syst. Bacteriol. 37: 78–81.
Rogers, M.J., J. Simmons, R.T. Walker, W.G. Weisburg, C.R. Woese,
R.S. Tanner, I.M. Robinson, D.A. Stahl, G. Olsen, R.H. Leach and
J. Maniloff. 1985. Construction of the mycoplasma evolutionary
tree from 5S rRNA sequence data. Proc. Natl. Acad. Sci. U.S.A. 82:
1160–1164.
Sasaki, Y., J. Ishikawa, A. Yamashita, K. Oshima, T. Kenri, K. Furuya,
C. Yoshino, A. Horino, T. Shiba, T. Sasaki and M. Hattori. 2002.
The complete genomic sequence of Mycoplasma penetrans, an intracellular bacterial pathogen in humans. Nucleic Acids Res. 30 :
5293–5300.
Sirand-Pugnet, P., C. Lartigue, M. Marenda, D. Jacob, A. Barre, V. Barbe,
C. Schenowitz, S. Mangenot, A. Couloux, B. Segurens, A. de Daruvar, A. Blanchard and C. Citti. 2007. Being pathogenic, plastic, and
sexual while living with a nearly minimal bacterial genome. PLoS.
Genet. 3: e75.
Stackebrandt, E. 2009. Class III. Erysipelotrichia. In Bergey’s Manual
of Systematic Bacteriology, vol. 3 (edited by de Vos, Garrity, Jones,
Krieg, Ludwig, Rainey, Schleifer and Whitman). Springer, New York,
p. 1298.
Tully, J.G., J.M. Bove, F. Laigret and R.F. Whitcomb. 1993. Revised
taxonomy of the class Mollicutes - proposed elevation of a monophyletic cluster of arthropod-associated mollicutes to ordinal rank
(Entomoplasmatales ord. nov.), with provision for familial rank to
separate species with nonhelical morphology (Entomoplasmataceae
fam. nov.) from helical species (Spiroplasmataceae), and emended
descriptions of the order Mycoplasmatales, family Mycoplasmataceae.
Int. J. Syst. Bacteriol. 43: 378–385.
Vasconcelos, A.T. and a. coauthors. 2005. Swine and poultry pathogens: the complete genome sequences of two strains of Mycoplasma
hyopneumoniae and a strain of Mycoplasma synoviae. J. Bacteriol. 187:
5568–5577.
Verbarg, S., H. Rheims, S. Emus, A. Fruhling, R.M. Kroppenstedt, E.
Stackebrandt and P. Schumann. 2004. Erysipelothrix inopinata sp.
nov., isolated in the course of sterile filtration of vegetable peptone
broth, and description of Erysipelotrichaceae fam. nov. Int. J. Syst. Evol.
Microbiol. 54: 221–225.
Weisburg, W., J. Tully, D. Rose, J. Petzel, H. Oyaizu, D. Yang, L. Mandelco, J. Sechrest, T. Lawrence and J. Van Etten. 1989. A phylogenetic
analysis of the mycoplasmas: basis for their classification. J. Bacteriol.
171: 6455–6467.
Westberg, J., A. Persson, A. Holmberg, A. Goesmann, J. Lundeberg,
K.E. Johansson, B. Pettersson and M. Uhlen. 2004. The genome
sequence of Mycoplasma mycoides subsp. mycoides SC type strain PG1T,
the causative agent of contagious bovine pleuropneumonia (CBPP).
Genome Res. 14: 221–227.
Woese, C.R., J. Maniloff and L.B. Zablen. 1980. Phylogenetic analysis of
the mycoplasmas. Proc. Natl. Acad. Sci. U. S. A. 77: 494–498.
574
Order I. Mycoplasmatales Freundt 1955, 71AL emend. Tully, Bové, Laigret and Whitcomb 1993, 382
Daniel R. Brown, Meghan May, Janet M. Bradbury, Karl-Erik Johansson and Harold Neimark
My.co.plas.ma.ta¢les. N.L. neut. n. Mycoplasma, -atos type genus of the order; -ales ending
to denote an order; N.L. fem. pl. n. Mycoplasmatales the Mycoplasma order.
The first order in the class Mollicutes is assigned to a group of
sterol-requiring, wall-less prokaryotes that occur as commensals
or pathogens in a wide range of vertebrate hosts. The description of the order is essentially the same as for the class. A single
family Mycoplasmataceae with two genera, Mycoplasma and Urea
plasma, recognizes the prominent and distinct characteristics of
the assigned organisms, based on their sterol requirements for
growth, the capacity of some to hydrolyze urea, and conserved
16S rRNA gene sequences.
Type genus: Mycoplasma Nowak 1929, 1349 nom. cons. Jud.
Comm. Opin. 22, 1958, 166.
The entire class Mollicutes was encompassed initially by a single
order. The elevation of acholeplasmas to ordinal rank (Achole
plasmatales Freundt, Whitcomb, Barile, Razin and Tully 1984)
recognized their major distinctions in nutritional, biochemical,
physiological, and genetic characteristics from other members
of the class Mollicutes. Subsequently, additional orders were
proposed to recognize the anaerobic mollicutes and the wallless prokaryotes from plants and insects which were phylogenetically related to the remaining Mycoplasmatales. Thus, the
Anaeroplasmatales (Robinson and Freundt, 1987) recognized
the strictly anaerobic, wall-less prokaryotes first isolated from
the bovine and ovine rumen, and Entomoplasmatales (Tully
et al., 1993) provided a classification for a number of the mollicutes regularly associated with plant and insect hosts. On the
basis of 16S rRNA gene sequence similarities (Johansson and
References
Edward, D.G. 1971. Determination of sterol requirement for Mycoplas
matales. J. Gen. Microbiol. 69 : 205–210.
5: 67–78.
Freundt, E.A., R.F. Whitcomb, M.F. Barile, S. Razin and J.G. Tully. 1984.
Proposal for elevation of the family Acholeplasmataceae to ordinal
rank: Acholeplasmatales. Int. J. Syst. Bacteriol. 34: 346–349.
Johansson, K.E., Pettersson B. 2002. Taxonomy of Mollicutes. In Molecular biology and pathogenicity of mycoplasmas (edited by Razin and
Herrmann). Kluwer Academic, New York, pp. 1–30.
Judicial Commission. 1958. Opinion 22. Status of the generic name
Asterococcus and conservation of the generic name Mycoplasma. Int.
Bull. Bacteriol. Nomencl. Taxon. 8: 166–168.
Nowak, J. 1929. Morphologie, nature et cycle évolutif du microbe
de la péripneumonie des bovidés. Ann. Inst. Pasteur (Paris) 43:
1330–1352.
Pettersson, 2002), the Mycoplasmatales and Entomoplasmatales
represent a clade deeply split from the Acholeplasmatales and
Anaeroplasmatales.
A growth requirement for cholesterol or serum is shared by
the organisms assigned to the order Mycoplasmatales, as well as
most other organisms within the class Mollicutes. Therefore,
tests for cholesterol requirements are essential to classification.
Earlier assessments of the growth requirements for cholesterol
were based upon the capacity of organisms to grow in a number
of serum-free broth preparations to which various concentrations of cholesterol were added (Edward, 1971; Razin and Tully,
1970). In this test, species that do not require exogenous sterol usually show no significant growth response to increasing
cholesterol concentrations. Polyoxyethylene sorbitan (Tween
80) and palmitic acid should be included in the base medium
because acholeplasmas such as Acholeplasma axanthum and
Acholeplasma morum require additional fatty acids for adequate
growth. A modified method utilizing serial passage in selective
medium has been applied successfully to a large number of
mollicutes (Rose et al., 1993; Tully, 1995). The Acholeplasmatales
grow through end-point dilutions in serum-containing medium
and in serum-free preparations, or occasionally in serum-free
medium supplemented with Tween 80. Mesoplasmas from the
order Entomoplasmatales grow in serum-containing medium and
in serum-free medium supplemented only with Tween 80. Most
spiroplasmas, also from the Entomoplasmatales, and all members
of the order Mycoplasmatales grow only in serum-containing
medium.
Razin, S. and J.G. Tully. 1970. Cholesterol requirement of mycoplasmas.
J. Bacteriol. 102: 306–310.
Robinson, I.M. and E.A. Freundt. 1987. Proposal for an amended classification of anaerobic mollicutes. Int. J. Syst. Bacteriol. 37: 78–81.
Rose, D.L., J.G. Tully, J.M. Bove and R.F. Whitcomb. 1993. A test for
measuring growth responses of Mollicutes to serum and polyoxyethylene sorbitan. Int. J. Syst. Bacteriol. 43: 527–532.
Tully, J.G., J.M. Bové, F. Laigret and R.F. Whitcomb. 1993. Revised taxonomy of the class Mollicutes - proposed elevation of a monophyletic
cluster of arthropod-associated mollicutes to ordinal rank (Ento
moplasmatales ord. nov.), with provision for familial rank to separate
of the order Mycoplasmatales, family Mycoplasmataceae. Int. J. Syst. Bacteriol. 43: 378–385.
Tully, J.G. 1995. Determination of cholesterol and polyoxyethylene sorbitan growth requirements of mollicutes. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 1 (edited by Razin and Tully).
Academic Press, San Diego, pp. 381–389.
Genus I. Mycoplasma
575
Family I. Mycoplasmataceae Freundt 1955, 71AL emend. Tully, Bové, Laigret and Whitcomb 1993, 382
Daniel R. Brown, Meghan May, Janet M. Bradbury, Karl-Erik Johansson and Harold Neimark
My.co.plas.ma.ta.ce′ae. N.L. neut. n. Mycoplasma, -atos type genus of the family; -aceae ending to
denote a family; N.L. fem. pl. n. Mycoplasmataceae the Mycoplasma family.
Pleomorphic usually coccoid cells, 300–800 nm in diameter, to
slender branched filaments of uniform diameter. Some cells
have a terminal bleb or tip structure that mediates adhesion to
certain surfaces. Cells lack a cell wall and are bounded only by
a plasma membrane. Gram-stain-negative due to the absence of
a cell wall. Usually nonmotile. Facultatively anaerobic in most
instances, possessing a truncated flavin-terminated electron
transport chain devoid of quinones and cytochromes. Colonies
of Mycoplasma are usually less than l mm in diameter and colonies of Ureaplasma are much smaller than that. The typical colony has a fried-egg or “cauliflower head” appearance. Usually
catalase-negative. Chemo-organotrophic, usually using either
sugars or arginine, but sometimes both, or having an obligate
requirement for urea as the major energy source. Require cholesterol or related sterols for growth. Commensals or pathogens
of a wide range of vertebrate hosts. The genome size ranges
from about 580 to 1350 kbp, as measured by pulsed field gel
electrophoresis (PFGE) or complete DNA sequencing.
DNA G+C content (mol%): about 23–40 (Bd, Tm).
Type genus: Mycoplasma Nowak 1929, 1349 nom. cons. Jud.
Comm. Opin. 22, 1958, 166.
This family and its type genus Mycoplasma are polyphyletic. Two
genera, Mycoplasma and Ureaplasma, are currently accepted
within the family. The genus Mycoplasma is further divisible into
phylogenetic groups on the basis of 16S rRNA gene sequence
similarities (Johansson and Pettersson, 2002), including an ecologically, phenotypically, and genetically cohesive group called
the mycoides cluster, which includes the type species Mycoplasma
mycoides and other major pathogens of ruminant animals. The
taxonomic position of the mycoides cluster is an important
anomaly because molecular markers based upon rRNA and
other gene sequences indicate that it is closely related to other
genera usually associated with plant and insect hosts and currently classified within the order Entomoplasmatales. Members of
the genus Ureaplasma are distinguished by their tiny colony size
and capacity to hydrolyze urea.
Genus I. Mycoplasma Nowak 1929, 1349 nom. cons. Jud. Comm. Opin. 22, 1958, 166AL
Daniel R. Brown, Meghan May, Janet M. Bradbury, Mitchell F. Balish, Michael J. Calcutt, John I. Glass, Séverine Tasker,
Joanne B. Messick, Karl-Erik Johansson and Harold Neimark
My.co.plas¢ma. Gr. masc. n. myces a fungus; Gr. neut. n. plasma something formed or molded, a form; N.L.
neut. n. Mycoplasma fungus form.
Pleomorphic cells, 300–800 nm in diameter, varying in shape
from spherical, ovoid or flask-shaped, or twisted rods, to slender
branched filaments ranging in length from 50 to 500 nm. Cells
lack a cell wall and are bounded by a single plasma membrane.
Gram-stain-negative due to the absence of a cell wall. Some have
a complex internal cytoskeleton. Some have a specific tip structure that mediates attachment to host cells or other surfaces.
Usually nonmotile, but gliding motility has been demonstrated
in some species. Aerobic or facultatively anaerobic. Optimum
growth at 37°C is common, but permissive growth temperatures
range from 20 to 45°C. Chemo-organotrophic, usually using
either sugars or arginine as the major energy source. Require
cholesterol or related sterols for growth. Colonies are usually
less than l mm in diameter. The typical colony has a fried-egg
appearance. The genome size of species examined ranges from
580 kbp to about 1350 kbp. The codon UGA encodes tryptophan in all species examined. Commensals or pathogens in a
wide range of vertebrate hosts.
DNA G+C content (mol%): 23–40.
Type species: Mycoplasma mycoides (Borrel, DujardinBeaumetz, Jeantet and Jouan 1910) Freundt 1955, 73 (Asterococcus mycoides Borrel, Dujardin-Beaumetz, Jeantet and Jouan
1910, 179).
The shape of these organisms (trivial name, mycoplasmas)
can depend on the osmotic pressure, nutritional quality of the
culture medium, and the growth phase. Some mycoplasmas
are filamentous in their early and exponential growth phases
or when attached to surfaces or other cells. This form can
be transitory, and the filaments may branch or fragment into
chains of cocci or individual vegetative cells. Many species are
typically coccoid and never develop a filamentous phase. Some
species develop specialized attachment tip structures involved
in colonization and virulence (Figure 106). In Romanowskytype stained blood smears, hemotropic species (trivial name,
hemoplasmas) appear as round to oval cells on the surface of
erythrocytes (Figure 107). They may be found individually or,
during periods of high parasitemia, in pairs or chains giving the
appearance of pleomorphism. Their small size and the absence
of cell wall components provide considerable plasticity to
the organisms, so that cells of most species are readily filterable
through 450 nm pores, and many species have some cells in the
population that are able to pass through 220 nm or even 100
nm filters (Tully, 1983). Descriptions of the morphology, ultrastructure, and motility of mycoplasmas should be based on correlation of the appearance of young exponential-phase broth
cultures under phase-contrast or dark-field microscopy with
their appearance using negative-staining or electron microscopy (Biberfeld and Biberfeld, 1970; Boatman, 1979; Carson
et al., 1992; Cole, 1983). Special attention to the osmolarity of
the fixatives and buffers is required since these may alter the
size and shape of the organisms. The classical isolated colony
is umbonate with a fried-egg appearance, but others may have
576
Family I. Mycoplasmataceae
Figure 106. Diverse cellular morphology in the genus Mycoplasma. Scanning electron micrographs of cells
of (a) Mycoplasma penetrans, (b) Mycoplasma pneumoniae, (c) “Mycoplasma insons”, and (d) Mycoplasma genitalium.
Bar = 1 mm. Images provided by Dominika Jurkovic, Jennifer Hatchel, Ryan Relich and Mitchell Balish.
Figure 107. Hemotropic mycoplasmas. (a) Scanning electron micrograph of Mycoplasma ovis cells colonizing
the surface of an erythrocyte (Neimark et al., 2004); bar = 500 nm. (b) Transmission electron micrograph showing
fibrils bridging the space between a “Candidatus Mycoplasma kahaneii” cell and a depression in the surface of a
colonized erythrocyte (Neimark et al., 2002a); bar = 250 nm. Images used with permission.
either cauliflower-like or smooth colony surfaces (Figure 108),
with smooth, irregular or scalloped margins, depending on the
species, agar concentration, and other growth conditions.
A significant minority of species exhibit cell polarization.
This depends on Triton X-100-insoluble cytoskeletal structures
involved in morphogenesis, motility, cytadherence, and cell division (Balish and Krause, 2006). In the distantly related species
Mycoplasma pneumoniae and Mycoplasma mobile, the cytoskeleton
underlies a terminal organelle. This prominent extension of the
cytoplasm and cell membrane is the principal focus of adherence
Genus I. Mycoplasma
577
Figure 108. Diverse colonial morphology in the genus Mycoplasma. (a) Mycoplasma mycoides PG1T (diameter
0.50–0.75 mm), (b) Mycoplasma hyopneumoniae NCTC 10110T (diameter 0.15–0.20 mm), (c) Mycoplasma pneumoniae
NCTC 10119T (diameter 0.05–0.10 mm), and (d) Mycoplasma hyorhinis ATCC 29052 (diameter 0.25–0.30 mm) after
3, 7, 5, and 6 d growth, respectively, on Mycoplasma Experience Solid Medium at 36°C in 95% nitrogen/5% carbon
dioxide. Original magnification 25×. Images provided by Helena Windsor and David Windsor.
and is the leading end of cells engaged in gliding motility. In
Mycoplasma pneumoniae, adhesin proteins are located either all
over the surface of the organelle or at its distal tip; an unrelated
adhesin is concentrated at the base of the terminal organelle
in Mycoplasma mobile (Balish, 2006). Both the formation of this
attachment organelle of Mycoplasma pneumoniae and the localization of the adhesins depend upon cytoskeletal proteins that
form an electron-dense core within its cytoplasm, which is surrounded by an electron-lucent space (Krause and Balish, 2004).
The overall appearance of this core is that of two parallel, flat
rods of differing thickness, with a bend near the cell-proximal
end (Henderson and Jensen, 2006; Seybert et al., 2006). A bilobed button constitutes its distal end and its proximal base terminates in a bowl-like structure. Overall, both the core and the
attachment organelle are 270–300 nm in length (Hatchel and
Balish, 2008). Around the onset of DNA replication, a second
attachment organelle is constructed (Seto et al., 2001). The
motile force provided by the first organelle reorganizes the cell
such that the new organelle is moved to the opposite cell pole
before cell division (Hasselbring et al., 2006). These observations suggest that complex coordination exists between attachment organelle biogenesis, motor activity, the DNA replication
machinery, and the cytokinetic machinery. Similar structures are
present in other species of the Mycoplasma pneumoniae cluster, but
in most cases the attachment organelle is shorter, resulting in
much of the core protruding into the cell body (Hatchel and Balish, 2008). In Mycoplasma mobile, the terminal organelle is completely dissimilar, consisting of a cell-distal sphere with numerous
tentacle-like strands extending into the cytoplasm (Nakane and
Miyata, 2007). It is comprised of proteins unrelated to those
found in Mycoplasma pneumoniae, suggesting that it has evolved
independently. Further distinct cytoskeletal structures appear in
Mycoplasma penetrans (Jurkovic and Balish, unpublished), “Mycoplasma insons” (Relich et al., 2009), and several species of the
mycoides cluster (Peterson et al., 1973).
Attachment to eukaryotic host cells is important for the natural survival and transmission of mycoplasmas. The prominent
attachment organelle of species in the Mycoplasma pneumoniae
cluster is the most extensively characterized determinant of cytadherence. In other species, multiple adhesin proteins are involved
in cytadherence. When one adhesin is blocked, cytadherence is
reduced, but not completely lost. For this reason, the adhesins
appear to be functionally redundant rather than synergistic in
action. Numerous species possess multigene families of antigenically variable proteins, some of which have been implicated in
host cell attachment or hemagglutination. While this attachment may serve as a supplemental binding mechanism in species
such as Mycoplasma gallisepticum and Mycoplasma hominis, variable
surface proteins are currently the only known mechanism for
cytadherence and hemagglutination of Mycoplasma synoviae and
Mycoplasma pulmonis. The avidity of adherence may differ among
variants in Mycoplasma pulmonis and Mycoplasma hominis. Though
one or more attachment mechanisms have been described for
numerous species, there remains a greater number of species
with no documented system for cytadherence. Strains that lose
the capacity to cytadhere are almost invariably unable to survive
in their hosts, but highly invasive species such as Mycoplasma
alligatoris may not require host cell attachment for infection.
The mycoplasmas possess a typical prokaryotic plasma
membrane composed of amphipathic lipids and proteins
578
( McElhaney, 1992a, b, c; Smith, 1992; Wieslander et al., 1992).
At one time, demonstration of a single unit membrane was
mandatory for defining all novel species of mollicutes (Tully,
1995a). Now, when the 16S rRNA gene sequence of a novel species is determined and the candidate is placed in one of the phylogenetic clusters of mollicutes, in the majority of cases it can
be safely inferred that the organism lacks a cell wall, because
the majority of others in that cluster will have been shown to
be solely membrane-bound (Brown et al., 2007). The lack of
a cell wall explains the resistance of the organisms to lysis by
lysozyme and their susceptibility to lysis by osmotic shock and
various agents causing the lysis of bacterial protoplasts (Razin,
1979, 1983). In certain species, the extracellular surface is textured with capsular material or a nap, which can be stained with
ruthenium red in some cases (Rosenbusch and Minion, 1992).
These organisms represent some of the most nutritionally
fastidious prokaryotes, as expected from their greatly reduced
or minimalist genomes, close association with vertebrate hosts
as commensals and pathogens, and total dependence upon the
host to meet all nutritional requirements. They have very limited
capacity for intermediary metabolism, which restricts the utility of
conventional biochemical tests for identification. Detailed information on carbohydrate (Pollack, 1992, 1997, 2002; Pollack et al.,
1996), lipid (McElhaney, 1992a), and amino acid (Fischer et al.,
1992) metabolism is available. All species examined have truncated respiratory systems, lack a complete tricarboxylic acid cycle,
and lack quinones or cytochromes, which precludes their capacity
to carry out oxidative phosphorylation. Instead only low levels of
ATP may be generated through glycolysis or the arginine dihydrolase pathway (Miles, 1992a, b). Fermentative species catabolize glucose or other carbohydrates to produce ATP and acid and,
consequently, lower the pH of the medium. Non-fermentative species hydrolyze arginine to yield ammonia, some ATP, and carbon
dioxide, and consequently raise the pH of the medium. Species
such as Mycoplasma fermentans have both pathways. Species such as
Mycoplasma bovis evidently lack both pathways, but are capable oxidizing pyruvate or lactate to yield ATP (Miles, 1992a; Taylor et al.,
1994). Some species cause a pronounced “film and spots” reaction on media incorporating heat-inactivated horse serum or egg
yolk: a wrinkled film composed of cholesterol and phospholipids
forms on the surface of the medium and dark spots containing
salts of fatty acids appear around the colonies.
Most mycoplasmas are aerobes or facultative anaerobes, but
some species such as Mycoplasma muris prefer an anaerobic
environment. The optimum growth of species isolated from
homeothermic hosts is commonly at 37°C and the permissive
temperature range of species from poikilothermic fish and
reptiles is always above 20–25°C. Thus, growth of the mycoplasmas is restricted to mesophilic temperatures. Growth in
liquid cultures usually produces at most light turbidity and few
sedimented cells, except for the heavy turbidity and sediments
usually observed with members of the Mycoplasma mycoides cluster. Tully (1995b) described in detail the most commonly used
culture media formulations. Although colonies are occasionally
first detected on blood agar, complex undefined media such as
American Type Culture Collection (ATCC) medium 988 (SP-4)
are usually required for primary isolation and maintenance.
Cell-wall-targeting antibiotics are included to discourage
growth of other bacteria. Phenol red facilitates detection of
species that excrete acidic or alkaline metabolites. Growth of
a rginine-hydrolyzing species can be enhanced by supplementing
media with arginine. Commonly used alternatives such as Frey’s,
Hayflick’s and Friis’ media differ from SP-4 mainly in the proportions of inorganic salts, amino acids, serum sources, and types
of antibiotics. For species that utilize both sugars and arginine
as carbon sources, the pH of the medium may initially decrease
before rising later during the course of growth (Razin et al.,
1998). Defined mycoplasma culture media have been described
in detail (Rodwell, 1983), but provision of lipids and amino acids
in the appropriate ratios is difficult technically (Miles, 1992b).
Many mobile genetic elements occur in the genus. Four plasmids have been identified in members of the mycoides cluster (Bergemann and Finch, 1988; Djordjevic et al., 2001; King
and Dybvig, 1994). Each plasmid is apparently cryptic, with no
discernible determinants for virulence or antibiotic resistance.
DNA viruses have been isolated from Mycoplasma bovirhinis
(Howard et al., 1980), Mycoplasma hyorhinis (Gourlay et al.,
1983) Mycoplasma pulmonis (Tu et al., 2001), and Mycoplasma
arthritidis (Voelker and Dybvig, 1999). The Mycoplasma pulmonis
P1 virus and the lysogenic bacteriophage MAV1 of Mycoplasma
arthritidis do not share sequence similarity (Tu et al., 2001;
Voelker and Dybvig, 1999), whereas the Mycoplasma fermentans
MFV1 prophage is strikingly similar in genetic organization to
MAV1 (Röske et al., 2004). No role in pathobiology has been
demonstrated for any virus or prophage.
The most abundant mobile DNAs in Mycoplasma are insertion sequence (IS) elements. The first identified units (IS1138
of Mycoplasma pulmonis, IS1221 of Mycoplasma hyorhinis, IS1296
of Mycoplasma mycoides subsp. mycoides and ISMi1 of Mycoplasma
fermentans) are members of the IS3 family (Bhugra and Dybvig, 1993; Ferrell et al., 1989; Frey et al., 1995; Hu et al., 1990).
More recently, multiple IS elements of divergent subgroups have
been identified. Members of the IS4 family include IS1634 and
ISMmy1 of Mycoplasma mycoides subsp. mycoides (Vilei et al., 1999;
Westberg et al., 2002), ISMhp1 of Mycoplasma hyopneumoniae,
ISMhp1-like unit of Mycoplasma synoviae, and four distinct elements of Mycoplasma bovis (Lysnyansky et al., 2009). Among the
IS30 family members identified are IS1630 of Mycoplasma fermentans, ISMhom1 from Mycoplasma hominis, ISMag1 of Mycoplasma
agalactiae (Pilo et al., 2003), and two IS units of Mycoplasma bovis.
IS-like elements have also been identified in Mycoplasma leachii,
Mycoplasma penetrans (belonging to four different families), Mycoplasma hyopneumoniae, Mycoplasma flocculare, and Mycoplasma orale.
Transposases that reside within IS units are also discernable in
the genome of Mycoplasma gallisepticum (Papazisi et al., 2003).
In select instances, almost identical IS units have been found
in species from different phylogenetic clades, which strongly
suggests lateral gene transfer between species. Despite their
widespread distribution, IS elements are not ubiquitous in the
genus. Although the type strains of Mycoplasma bovis (54 IS units
of seven different types) and Mycoplasma mycoides subsp. mycoides
(97 elements of three different types) possess large numbers of
elements, the sequenced genomes of Mycoplasma arthritidis, Mycoplasma genitalium, Mycoplasma pneumoniae, and Mycoplasma mobile
lack detectable IS units.
Although IS units only encode genes related to transposition,
large integrating elements have also been identified in diverse
Mycoplasma species. The Integrative Conjugal Elements (ICE)
of Mycoplasma fermentans strain PG18T comprise >8% of the
genome and related units have been identified in Mycoplasma
Genus I. Mycoplasma
agalactiae, Mycoplasma bovis, Mycoplasma capricolum, Mycoplasma
hyopneumoniae, and Mycoplasma mycoides subsp. mycoides. In general, such units encode 18–30 genes, can be detected in extrachromosomal forms, and are strain-variable in distribution and
chromosomal insertion site. Two additional large mobile DNAs,
designated Tra Islands, were identified in Mycoplasma capricolum
California kidT. The presence of putative conjugation genes
and the variability in genomic location of Tra Islands and ICE
suggest that these are agents of lateral gene transfer.
The best-studied mycoplasmas are primary pathogens of
humans or domesticated animals (Baseman and Tully, 1997).
About half of the listed species occur in the absence of disease,
but are occasional opportunistic or secondary pathogens. The
principal human pathogens are Mycoplasma pneumoniae, Mycoplasma hominis, and Mycoplasma genitalium, with Mycoplasma penetrans added to this list due to its association with HIV infections
(Blanchard, 1997; Blanchard et al., 1997; Tully, 1993; Waites and
Talkington, 2005). Mycoplasma pneumoniae is one of the main
agents of community-acquired pneumonia, bronchitis, and
other respiratory complications (Atkinson et al., 2008). Mycoplasma pneumoniae infections can also involve extra-pulmonary
complications including central nervous system, cardiovascular,
and dermatological manifestations. Outbreaks cause considerable morbidity and require rapid and effective therapeutic intervention (Hyde et al., 2001; Meyer and Clough, 1993). Mycoplasma
hominis occurs more frequently in the urogenital tract of women
than men and is often found in the genital tract of women with
vaginitis, bacterial vaginosis, or localized intrauterine infections
(Keane et al., 2000). The organism can gain access to a fetus
from uterine sites and it is associated with perinatal morbidity
and mortality (Gonçalves et al., 2002; Waites et al., 1988). It is
also clearly associated with septicemias and respiratory infections
and with transplant or joint infections in immunosuppressed
persons (Brunner et al., 2000; Busch et al., 2000; Fernandez
Guerrero et al., 1999; Garcia-Porrua et al., 1997; Gass et al.,
1996; Hopkins et al., 2002; Mattila et al., 1999; Tully, 1993; Zheng
et al., 1997). Mycoplasma genitalium has been associated with nongonococcal urethritis in men (Gambini et al., 2000; Jensen, 2004;
Jensen et al., 2004; Taylor-Robinson et al., 2004; Taylor-Robinson
and Horner, 2001; Totten et al., 2001) and urogenital disease in
women (Baseman et al., 2004; Blaylock et al., 2004). Mycoplasma
genitalium occurs more frequently in the vagina than in the cervix or urethra, but it may be involved in cervicitis (Casin et al.,
2002; Manhart et al., 2001). Mycoplasmas are common agents
of chronic joint inflammation in a wide variety of hosts (Cole
et al., 1985). Species associated with arthritis in humans include
Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma genitalium,
Mycoplasma salivarium, and possibly Mycoplasma pneumoniae in
juvenile arthritis (Waites and Talkington, 2005). Humans are also
susceptible to opportunistic zoonotic mycoplasmosis; immunosuppressed persons are highly susceptible. The recent molecular
confirmation of a Mycoplasma haemofelis-like infection in an HIVpositive patient (dos Santos et al., 2008) highlights the zoonotic
potential of the hemoplasmas.
Mycoplasmas colonize fish, reptiles, birds, and terrestrial
and aquatic mammals. Some cause significant diseases of cattle and other ruminants, swine, poultry, or wildlife, and others
are opportunistic or secondary veterinary pathogens (Simecka
et al., 1992; Tully and Whitcomb, 1979). The principal bovine
pathogens include the serovars historically called “Small Colony”
579
types of Mycoplasma mycoides subsp. mycoides, and Mycoplasma
bovis. Mycoplasma mycoides subsp. mycoides has caused major losses
of livestock globally in the twentieth century due to contagious
bovine pleuropneumonia and currently remains a problem in
Asia and Africa (Lesnoff et al., 2004). Mycoplasma bovis is a widespread agent of otitis media, pneumonia, mastitis, polyarthritis, and urogenital disease in cattle and buffaloes. Mycoplasma
mycoides subsp. capri, Mycoplasma capricolum subsp. capricolum, and
Mycoplasma agalactiae are important causes of arthritis, mastitis,
and agalactia in goats and sheep. Mycoplasma mycoides subsp. capri
(type strain PG3T) properly includes all of the serovars historically called “Large Colony” types of subspecies mycoides (MansoSilván et al., 2009; Shahram et al., 2010). Mycoplasma capricolum
subsp. capripneumoniae (type strain F38T) causes severe contagious pleuropneumonia in goats (Leach et al., 1993; McMartin
et al., 1980). Contagious bovine and caprine pleuropneumonia,
and mycoplasmal agalactia of sheep or goats are subject to control through listing in the Terrestrial Animal Health Code of the
Office International des Epizooties (http://oie.int) as well as
strict notification and export regulations by individual countries.
Mycoplasma hyopneumoniae, one of the most difficult species to
cultivate, causes primary enzootic pneumonia in pigs and exacerbates other porcine respiratory diseases leading to substantial
economic burdens. Mycoplasma hyosynoviae is carried in the upper
respiratory tract, but causes nonsuppurative polyarthritis, usually
without other serositis, especially in growing pigs.
The most important poultry pathogens are Mycoplasma gallisepticum, Mycoplasma synoviae, and Mycoplasma meleagridis, but
more than 20 other species have been isolated from birds as
diverse as ostriches, raptors, and penguins (Bradbury and
Morrow, 2008). Mycoplasma gallisepticum can be transmitted vertically, venereally, by other direct contact, or by aerosol to cause
respiratory disease and its sequelae in chickens, turkeys, and
other birds. It also causes decreased egg production and egg
quality in chickens. Mycoplasma synoviae can cause a syndrome
of synovitis, tendonitis, and bursitis in addition to respiratory
disease in chickens and turkeys, whereas the developmental
abnormalities and airsacculitis associated with congenital or
acquired Mycoplasma meleagridis infection seem restricted to turkeys. Mycoplasma gallisepticum and Mycoplasma synoviae are also
listed in the OIE’s Terrestrial Animal Health Code.
Pathogenicity of specific mycoplasmas has also been reported
for companion animals (Chalker, 2005; Lemcke, 1979; Messick,
2003) and a number of wild animal hosts (Brown et al., 2005).
The respiratory, reproductive, and joint diseases caused in
rodents by Mycoplasma pulmonis and Mycoplasma arthritidis
(Schoeb, 2000) are important models of infection and immunity in humans and other animals.
Hemoplasmas infect a variety of wild and domesticated
animals and are relatively host-specific, although cross-infection
of related hosts has been reported. Transmission can be achieved
by ingestion of infected blood or by percutaneous inoculation.
Arthropod vector transmission of some species is also supported
by experimentation and by the clustered geographic distribution of hemoplasmosis in some studies (Sykes et al., 2007; Willi
et al., 2006a). The pathogenicity of different hemoplasma species is variable and strain virulence also likely plays a key role in
the development of disease. For example, Mycoplasma haemofelis
can induce acute clinical disease in non-splenectomized, immunocompetent cats, whereas Mycoplasma haemocanis appears able
580
to induce disease only in immunosuppressed or splenectomized
dogs. Clinical syndromes range from acute fatal hemolytic anemia to chronic insidious anemia and ill-thrift. Signs may include
anemia, pyrexia, anorexia, dehydration, weight loss, and infertility. The presence of erythrocyte-bound antibodies (including
cold agglutinins), indicated by positive Coombs’ testing, has
been demonstrated in some hemoplasma-infected animals and
may contribute to anemia. Animals can remain chronic asymptomatic carriers of hemoplasmas after acute infection. PCR is the
diagnostic test of choice for hemoplasma infection.
Contamination of eukaryotic cell cultures with mollicutes is still
a common and important yet often unrecognized problem (Tully
and Razin, 1996). More than 20 species have been isolated from
contaminated cell lines, but more than 90% of the contamination
is thought to be caused by just five species of mycoplasma: Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hominis, Mycoplasma hyorhinis, and Mycoplasma orale, plus Acholeplasma laidlawii.
Mycoplasma pirum and Mycoplasma salivarium account for most of
the remainder (Drexler and Uphoff, 2002). Culture medium components of animal origin, passage of contaminated cultures, and
laboratory personnel are likely to be the most significant sources
of cell culture contaminants. PCR-based approaches to detection
achieve sensitivity and specificity far superior to fluorescent staining methods (Masover and Becker, 1996). Another method of
detection is based on mycoplasma-specific ATP synthesis activity
present in contaminated culture medium (Robertson and Stemke,
1995; MycoAlert, Lonza Group). Eradication through treatment of
contaminated cultures with antibiotics (Del Giudice and Gardella,
1996) is rarely successful. Strategies for prevention and control of
mycoplasmal contamination of cell cultures have been described
in detail (Smith and Mowles, 1996).
Several categories of potential virulence determinants are
encoded in the metagenome of pathogenic mycoplasmas. Some
species possess multiple types of virulence factors. Determinants
such as adhesins and accessory proteins, extracellular polysaccharide structures, and pro-inflammatory or pro-apoptotic membrane
lipoproteins are produced by multiple species. Several species
excrete potentially toxic by-products of intermediary metabolism,
including hydrogen peroxide, superoxide radicals, or ammonia.
Other determinants such as extracellular endopeptidases, nucleases, and glycosidases seem irregularly distributed in the genus,
whereas the ADP-ribosylating and vacuolating cytotoxin (pertussis exotoxin S1 subunit analog) of Mycoplasma pneumoniae and the
T-lymphocyte mitogen (superantigen) of Mycoplasma arthritidis are
evidently unique to those species. Reports of a putative exotoxin
elaborated by Mycoplasma neurolyticum have not been substantiated
by later work (Tryon and Baseman, 1992).
Candidate virulence mechanisms, such as motility, biofilm
formation, or facultative intracellular invasion, are expressed
by a range of pathogenic species. Several species possess
systems of variable surface antigens that are thought to be
important in evasion of the hosts’ adaptive immune responses.
In addition, a large number of species can suppress or inappropriately stimulate host immune cells and their receptors and
cytokines through diverse, poorly characterized mycoplasmal
components. Although candidate virulence factor discovery has
accelerated significantly in recent years through whole genome
annotation, the molecular basis for pathogenicity and causal
relationships with disease still remain to be definitively established for most of these factors (Razin and Herrmann, 2002;
Razin et al., 1998).
Because they lack lipopolysaccharide and a cell wall, and do
not synthesize their own nucleotides, mycoplasmas are intrinsically resistant to polymixins, b-lactams, vancomycin, fosfomycin, sulfonamides, and trimethoprim. They are also resistant to
rifampin because their RNA polymerase is not affected by that
antibiotic (Bébéar and Kempf, 2005). Individual species exhibit
an even broader spectrum of antibiotic resistance, such as the
resistance to erythromycin and azithromycin exhibited by several species, which is apparently mediated by mutation in the
23S rRNA (Pereyre et al., 2002). Treatment of mycoplasmosis
often involves the use of antibiotics that inhibit protein synthesis or DNA replication. Certain macrolides or ketolides are
used when tetracyclines or fluoroquinolones are inappropriate. Fluoroquinolones, aminoglycosides, pleuromutilins, and
phenicols are not widely used to treat human mycoplasmosis
at present, with the exception of chloramphenicol for neonates
with mycoplasmosis of the central nervous system unresponsive
to other antibiotics (Waites et al., 1992), but their use in veterinary medicine is more common. The long-term antimicrobial
therapy often required may be due to mycoplasmal sequestration in privileged sites, potentially including inside host cells.
Mycoplasmosis in immunodeficient patients is very difficult to
control with antibiotic drugs (Baseman and Tully, 1997).
Enrichment and isolation procedures
Techniques for isolation of mycoplasmas from humans, various
species of animals, and from cell cultures have been described
(Neimark et al., 2001; Tully and Razin, 1983). Typical steps in the
isolation of mycoplasmas were outlined in the recently revised
minimal standards for descriptions of new species (Brown et al.,
2007). Initial isolates may contain a mixture of species, so cloning
by repeated filtration through membrane filters with a pore size
of 450 or 220 nm is essential. The initial filtrate and dilutions of
it are cultured on solid medium and an isolated colony is subsequently picked from a plate on which only a few colonies have
developed. This colony is used to found a new cultural line, which
is then expanded, filtered, plated, and picked two additional times.
Hemoplasmas have not yet been successfully grown in continuous
culture in vitro, although recent work (Li et al., 2008) suggests that
in vitro maintenance of Mycoplasma suis may be possible.
Maintenance procedures
Cultures of mycoplasma can be preserved by lyophilization or cryogenic storage (Leach, 1983). The serum in the culture medium
provides effective cryoprotection, but addition of sucrose may
enhance survival following lyophilization. Hemoplasmas can be
frozen in heparin- or EDTA-anticoagulated blood cryopreserved
with dimethylsulfoxide. Most species can be recovered with little
loss of viability even after storage for many years.
Taxonomic comments
This polyphyletic genus is divisible on the basis of 16S rRNA and
other gene sequence similarities into a large paraphyletic clade
of over 100 species in two groups called hominis and pneumoniae (Johansson and Pettersson, 2002; Figure 109), plus the
ecologically, phenotypically, and genetically cohesive “mycoides
cluster” of five species including the type species Mycoplasma
mycoides (Cottew et al., 1987; Manso-Silván et al., 2009; Shahram
et al., 2010). The priority of Mycoplasma mycoides as the type species of the genus Mycoplasma and, hence, the family Mycoplasmataceae and the order Mycoplasmatales is, in retrospect, unfortunate.
Genus I. Mycoplasma
The phylogenetic position of the mycoides cluster is eccentrically
situated to the remaining species of the order Mycoplasmatales,
amidst genera that are properly classified in the order Entomoplasmatales. When the order Entomoplasmatales was established, a century after the discovery of Mycoplasma mycoides, it was explicitly
accepted that the taxonomic anomaly created by the phylogenetic position of the mycoides cluster will remain impractical to
resolve (Tully et al., 1993). The few species in the mycoides cluster
cannot simply be renamed, because confusion and peril would
result, especially regarding “Small Colony” PG1T-like strains of
Mycoplasma mycoides subsp. mycoides and F38T-like strains of Mycoplasma capricolum subsp. capripneumoniae, which are highly virulent
pathogens and subject to strict international regulations.
Another controversy involves the nomenclature of uncultivated
hemotropic bacteria originally assigned to the genera Eperythrozoon or Haemobartonella. It is now undisputed that, on the bases of
their lack of a cell wall, small cell size, low G+C content, use of
the codon UGA to encode tryptophan, regular association with
vertebrate hosts, and 16S rRNA gene sequences that are most similar (80–84%) to species in the pneumoniae group of Mycoplasma,
these organisms are properly affiliated with the Mycoplasmatales.
However, the proposed transfers of Eperythrozoon and Haemobartonella species to the genus Mycoplasma (Neimark et al., 2001, 2005)
were opposed on the grounds that the degree of 16S rRNA gene
sequence similarity is insufficient (Uilenberg et al., 2004, 2006).
The principal objection to establishing the hemoplasmas in a third
genus in the Mycoplasmataceae (Uilenberg et al., 2006) is that this
would compound the polyphyly within the pneumoniae group
solely on the basis of a capacity to adhere to erythrocytes in vivo. In
addition, the transfer of the type species Eperythrozoon coccoides to
the genus Mycoplasma is complicated by priority because Eperythrozoon predates Mycoplasma. The alternative, to transfer all mycoplasmas to the genus Eperythrozoon, would be completely impractical
and perilous in part because the epithet Eperythrozoon does not
indicate an affiliation with Mycoplasmatales. The Judicial Commission of the International Committee on Systematics of Prokaryotes
(ICSP) declined to rule on a request for an opinion in this matter
(Neimark et al., 2005) during their 2008 meeting, but a provisional
placement in the genus Mycoplasma has otherwise been embraced
by specialists in the molecular biology and clinical pathogenicity of
these and similar hemotropic organisms. At present, the designation “Candidatus” must still be used for new types.
Mycoplasma feliminutum was first described during a time when
the only named genus of mollicutes was Mycoplasma. Its publication
coincided with the first proposal of the genus Acholeplasma (Edward
and Freundt, 1969, 1970), with which Mycoplasma feliminutum is
properly affiliated through established phenotypic (Heyward et al.,
1969) and 16S rRNA gene sequence (Brown et al., 1995) similarities. This explains the apparent inconsistencies with its assignment
to the genus Mycoplasma. The name Mycoplasma feliminutum should
therefore be revised to Acholeplasma feliminutum comb. nov. The
type strain is BenT (=ATCC 25749T; Heyward et al., 1969).
Recent work at the J. Craig Venter Institute, including complete chemical synthesis and cloning of an intact Mycoplasma
genitalium chromosome (Gibson et al., 2008) and other work
with Mycoplasma mycoides “Large Colony” (Lartigue et al., 2009),
suggests that de novo synthesis of two species of mollicute is imminent. Transplantation of isolated deproteinized tetR-selectable
chromosomes from donor Mycoplasma mycoides “Large Colony”
into recipient Mycoplasma capricolum cells displaced the recipient
genome and conferred the genotype and phenotype of the donor
581
(Lartigue et al., 2007). Cloning in yeast and subsequent resurrection of Mycoplasma mycoides “Large Colony” genomes as living
bacteria demonstrate that it is possible to enliven a prokaryotic
genome constructed in a eukaryotic cell (Lartigue et al., 2009).
The ICSP subcommittee on the taxonomy of Mollicutes may be
the first to accommodate a system of nomenclature and classification for species of novel prokaryotes that originate by entirely
artificial speciation events (Brown and Bradbury., 2008).
Differentiation of the genus Mycoplasma
from other genera
Properties that partially fulfill criteria for assignment to the
class Mollicutes (Brown et al., 2007) include absence of a cell
wall, filterability, and the presence of conserved 16S rRNA gene
sequences. Aerobic or facultatively anaerobic growth in artificial
medium and a growth requirement for sterols exclude assignment to the genera Anaeroplasma, Asteroleplasma, Acholeplasma,
or “Candidatus Phytoplasma”. Non-spiral cellular morphology
and regular association with a vertebrate host or fluids of vertebrate origin support exclusion from the genera Spiroplasma,
Entomoplasma, or Mesoplasma. The inability to hydrolyze urea
excludes assignment to the genus Ureaplasma.
Acknowledgements
The lifetime achievements in mycoplasmology and substantial
contributions to the preparation of this material by Joseph G.
Tully are gratefully acknowledged. Daniel R. Brown and Meghan
May were supported by NIH grant 5R01GM076584. Séverine
Tasker was supported by Wellcome Trust grant WT077718.
Further reading
Blanchard, A. and G. Browning (editors). 2005. Mycoplasmas:
Molecular Biology, Pathogenicity, and Strategies for Control.
Horizon Press, Norwich, UK.
Maniloff, J., R.N. McElhaney, L.R. Finch and J.B. Baseman (editors). 1992. Mycoplasmas: Molecular Biology and Pathogenesis. American Society for Microbiology, Washington, D.C.
Razin, S. and J.G. Tully (editors). 1995. Molecular and Diagnostic Procedures in Mycoplasmology, vol. 1, Molecular Characterization. Academic Press, San Diego.
Tully, J.G. and S. Razin (editors). 1996. Molecular and Diagnostic Procedures in Mycoplasmology, vol. 2, Diagnostic Procedures. Academic Press, San Diego.
Differentiation of the species of the genus Mycoplasma
Glucose fermentation and arginine hydrolysis are discriminating phenotypic markers (Table 137), but the pleomorphism and
metabolic simplicity of mycoplasmas has led to a current reliance
principally on the combination of 16S rRNA gene sequencing and
reciprocal serology for species differentiation. Failure to crossreact with antisera against previously recognized species provides
substantial evidence for species novelty. For this reason, deposition of antiserum against a novel type strain into a recognized
collection is still mandatory for novel species descriptions (Brown
et al., 2007). Preliminary differentiation can be by PCR and DNA
sequencing using primers specific for bacterial 16S rRNA genes
or the 16S–23S intergenic region. A similarity matrix relating the
candidate strain to its closest neighbors, usually species with >94%
16S rRNA gene sequence similarity, will suggest related species
that should be examined for serological cross-reactivities.
582
hominis group
*
*
*
*
*
*
Scale:
Figure 109. (Continued)
Mycoplasma equigenitalium
Mycoplasma elephantis
Mycoplasma bovis
Mycoplasma agalactiae
*
Mycoplasma primatum
Mycoplasma opalescens
Mycoplasma spermatophilum
Mycoplasma fermentans
*
Mycoplasma caviae
Mycoplasma adleri
**
Mycoplasma felifaucium
*
Mycoplasma leopharyngis
Mycoplasma maculosum
Mycoplasma lipofaciens
Mycoplasma bovigenitalium
Mycoplasma californicum
**
Mycoplasma simbae
Mycoplasma phocirhinis
Mycoplasma meleagridis
*
Mycoplasma gallinarum
Mycoplasma iners
Mycoplasma columbinasale
*
Mycoplasma columbinum
*
Mycoplasma lipophilum
Mycoplasma hyopharyngis
Mycoplasma sphenisci
Mycoplasma synoviae
Mycoplasma verecundum
Mycoplasma gallinaceum
Mycoplasma corogypsi
Mycoplasma glycophilum
Mycoplasma gallopavonis
Mycoplasma buteonis
*
Mycoplasma felis
*
Mycoplasma mustelae
*
Mycoplasma leonicaptivi
Mycoplasma bovirhinis
Mycoplasma cynos
*
Mycoplasma edwardii
Mycoplasma canis
Mycoplasma columborale
Mycoplasma oxoniensis
Mycoplasma citelli
Mycoplasma sturni
Mycoplasma pullorum
Mycoplasma anatis
Mycoplasma crocodyli
Mycoplasma alligatoris
Mycoplasma hominis
Mycoplasma equirhinis
Mycoplasma phocidae
*
Mycoplasma falconis
Mycoplasma spumans
Mycoplasma arthritidis
*
Mycoplasma phocicerebrale
*
Mycoplasma auris
*
Mycoplasma alkalescens
* * Mycoplasma canadense
Mycoplasma gateae
Mycoplasma arginini
Mycoplasma cloacale
Mycoplasma anseris
Mycoplasma buccale
Mycoplasma hyosynoviae
Mycoplasma orale
*
Mycoplasma indiense
Mycoplasma faucium
Mycoplasma subdolum
Mycoplasma gypis
Mycoplasma pulmonis strain UAB CTIP
Mycoplasma agassizii
Mycoplasma testudineum
Mycoplasma sualvi
Mycoplasma moatsii
Mycoplasma mobile
Mycoplasma neurolyticum
Mycoplasma cricetuli
Mycoplasma collis
Mycoplasma molare
Mycoplasma lagogenitalium
Mycoplasma iguanae
Mycoplasma hyopneumoniae
*
Mycoplasma flocculare
Mycoplasma ovipneumoniae
Mycoplasma dispar
Mycoplasma bovoculi
Mycoplasma conjunctivae
Mycoplasma hyorhinis
Mycoplasma vulturis
0.1 substitutions/site
equigenitalium cluster
bovis cluster
lipophilum cluster
synoviae cluster
hominis cluster
pulmonis cluster
sualvi cluster
neurolyticum cluster
583
Genus I. Mycoplasma
Mycoplasma insons
Mycoplasma cavipharyngis
Mycoplasma fastidiosum
pneumoniae group
*
hemotropic cluster
Mycoplasma genitalium
Mycoplasma amphoriforme
Mycoplasma testudinis
Mycoplasma alvi
Mycoplasma pirum
Mycoplasma gallisepticum strain R
Mycoplasma imitans
Ureaplasma parvum serovar 3
Ureaplasma gallorale
Ureaplasma diversum
Ureaplasma felinum
Ureaplasma canigenitalium
Mycoplasma penetrans strain HF-2
Mycoplasma iowae
Mycoplasma muris
Mycoplasma microti
*
hemotropic cluster
*
Scale:
fastidiosum cluster
pneumoniae cluster
Ureaplasma cluster
muris cluster
Mycoplasma coccoides
Mycoplasma haemofelis
Mycoplasma haemocanis
‘Candidatus Mycoplasma haemobos’
Mycoplasma haemomuris
Mycoplasma suis
Mycoplasma wenyonii
Mycoplasma ovis
0.1 substitutions /site
Figure 109. Phylogenetic relationships in the Mycoplasma hominis and Mycoplasma pneumoniae groups of the order Mycoplasmatales. The phylogram
was based on a Jukes–Cantor corrected distance matrix and weighted neighbor-joining analysis of the 16S rRNA gene sequences of the type strains,
except where noted. Acholeplasma (formerly Mycoplasma) feliminutum was the outgroup. The major groups and clusters are defined in terms of positions in 16S rRNA showing characteristic base composition and signature positions, plus higher-order structural synapomorphies (Johansson and
Pettersson, 2002; Weisburg et al., 1989). Bootstrap values (100 replicates) <50% are indicated (*); the branching order is considered to be equivocal.
List of species of the genus Mycoplasma
1. Mycoplasma mycoides (Borrel, Dujardin-Beaumetz, Jeantet
and Jouan 1910) Freundt 1955, 73AL (Asterococcus mycoides
Borrel, Dujardin-Beaumetz, Jeantet and Jouan 1910, 179)
my.co.i¢des. Gr. n. mukês -êtos mushroom or other fungus; L.
suff. -oides (from Gr. suff. -eides, from Gr. n. eidos that which
is seen, form, shape, figure) resembling, similar; N.L. neut.
adj. mycoides fungus-like.
This is the type species of the genus. Cells are pleomorphic and capable of forming long filaments. Nonmotile. An
extracellular capsule can be visualized by electron microscopy following staining with ruthenium red. Colonies on
solid agar have a characteristic fried-egg appearance. Grows
in modified Hayflick medium supplemented with glucose
at 37°C.
The species has subsequently been divided as follows.
1a. Mycoplasma mycoides subsp. capri Manso-Silván, Vilei,
Sachse, Djordjevic, Thiaucourt and Frey 2009, 1357VP (Asterococcus
mycoides var. capri Edward 1953, 874; Mycoplasma mycoides
subsp. mycoides var. large colony Cottew and Yeats 1978, 294)
ca¢pri. L. n. caper, -pri goat; L. gen. n. capri of the goat.
Cells are pleomorphic and capable of forming long filaments and long, helical rods known as rho forms. Nonmotile. An extracellular capsule can be visualized by electron
microscopy following staining with ruthenium red. Colonies
on solid agar have a characteristic fried-egg appearance and
are notably larger than those of Mycoplasma mycoides subsp.
mycoides. Grows in modified Hayflick medium supplemented
with glucose at 37°C. Formation of biofilms has been demonstrated (McAuliffe et al., 2006).
Pathogenic; causes polyarthritis, mastitis, conjunctivitis (a syndrome collectively termed contagious agalactia),
pneumonia, peritonitis, and septicemia in goats; and balanitis and vulvitis in sheep. Transmission occurs via direct
contact between animals or with fomites, or can be vectorborne by the common ear mite (Psoroptes cuniculi).
Tetracyclines are effective therapeutic agents. Eradication from herds is difficult due to the tendency of healthy
animals to harbor the organism in the ear canal without
seroconverting. Antigenic cross-reactivity with Mycoplasma
584
Table 137. Descriptive characteristics of species of Mycoplasma a
Species
M. mycoides subsp. mycoides
M. mycoides subsp. capri
M. adleri
M. agalactiae
M. agassizii
M. alkalescens
M. alligatoris
M. alvi
M. amphoriforme
M. anatis
M. anseris
M. arginini
M. arthritidis
M. auris
M. bovigenitalium
M. bovirhinis
M. bovis
M. bovoculi
M. buccale
M. buteonis
M. californicum
M. canadense
M. canis
M. capricolum subsp. capricolum
M. capricolum subsp.
capripneumoniae
M. caviae
M. cavipharyngis
M. citelli
M. cloacale
M. coccoides b
M. collis
M. columbinasale
M. columbinum
M. columborale
M. conjunctivae
M. corogypsi
M. cottewii
M. cricetuli
M. crocodyli
M. cynos
M. dispar
M. edwardii
M. elephantis
M. equigenitalium
M. equirhinis
M. falconis
M. fastidiosum
M. faucium
M. felifaucium
M. feliminutum
M. felis
M. fermentans
M. flocculare
M. gallinaceum
M. gallinarum
M. gallisepticum
M. gallopavonis
M. gateae
Morphology
DNA G+C
content
(mol%)
Energy
source
Medium pH
shift
Serum source
Pleomorphic
Pleomorphic
Coccoidal
Coccoidal
Pleomorphic
Coccobacillary
Coccoidal
Flask-shaped
Flask-shaped
Coccoidal
Spherical
Coccoidal
Filamentous
Pleomorphic
Pleomorphic
nr
Coccobacillary
Coccobacillary
Coccobacillary
Coccoidal
Pleomorphic
Coccobacillary
Pleomorphic
Coccobacillary
Coccobacillary
24
24
29.6
29.7
nr
25.9
nr
26.4
34
26.6
26
27.6
30.7
26.9
30.4
27.3
32.9
29
26.4
27
31.9
29
28.4
23
24.4
G
G
R
G
G
R
G
G, R
G
G
R
R
R
R
OH, OA
G
OH, OA
G
R
G
OH, OA
R
G
G
G
A
A
K
A
A
K
A
V
A
A
K
K
K
K
N
A
N
A
K
A
N
K
A
A
A
FB
FB
E
FB, E
FB
FB
FB
FB
FB
FB
E
E
FB
E
FB
FB
FB, E
E
E
P
E
FB
FB
FB, E
FB, E
Cattle
Goats
Goats
Goats
Tortoises
Cattle
Alligators
Cattle
Humans
Ducks
Goose
Mammals
Rats
Goats
Cattle
Cattle
Cattle
Cattle
Humans
Raptors
Cattle
Cattle
Dogs
Goats
Goats
Pathogen
Pathogen
Pathogen
Pathogen
Pathogen
Pathogen
Pathogen
Commensal
Opportunistic
Opportunistic
Opportunistic
Pathogen
Pathogen
Commensal
Pathogen
Opportunistic
Pathogen
Pathogen
Commensal
Commensal
Pathogen
Pathogen
Opportunistic
Pathogen
Pathogen
nr
Twisted rod
Pleomorphic
Spherical
Coccoidal
Coccoidal
Coccobacillary
Pleomorphic
Coccoidal
Coccobacillary
Pleomorphic
Coccoid
Pleomorphic
Coccoidal
Coccobacillary
Pleomorphic
Coccobacillary
Coccoidal
Pleomorphic
Coccobacillary
Coccoidal
Twisted rod
Coccoidal
Coccoidal
nr
Filamentous
Filamentous
Coccobacillary
Coccobacillary
Coccobacillary
Flask-shaped
Coccobacillary
nr
nr
30
27.4
26
nr
28
32
27.3
29.2
nr
28
27
nr
27.6
25.8
29.3
29.2
24
31.5
nr
27.5
32.3
nr
31
29.1
25.2
28.7
33
28
28
31
27
28.5
G
G
G
R
U
G
R
R
G
G
G
G
G
G
G
G
G
G
G
R
R
G
R
R
G
G
R, G
U
G
R
G
G
U
A
A
A
K
na
A
K
K
A
A
A
A
A
A
A
A
A
A
A
K
K
A
K
K
A
A
V
N
A
K
A
A
N
FB
E
FB
E
na
E
FB
P
P
FB
P
E
E
FB
FB
FB, P
FB
E
E
E
P
P
FB
FB, E
FB
FB
FB, E
P
P
P
FB, E
P
FB
Guinea pigs
Guinea pigs
Squirrels
Galliforms
Mice
Rodents
Pigeons
Pigeons
Pigeons
Goats
Vultures
Goats
Hamsters
Crocodiles
Dogs
Cattle
Dogs
Elephants
Horses
Horses
Falcons
Horses
Humans
Pumas
Cats
Cats
Humans
Pigs
Galliforms
Galliforms
Galliforms
Turkeys
Cats
Commensal
Commensal
Commensal
Commensal
Pathogen
Commensal
Commensal
Commensal
Commensal
Pathogen
Pathogen
Commensal
Commensal
Pathogen
Pathogen
Pathogen
Opportunistic
Commensal
Opportunistic
Opportunistic
Opportunistic
Commensal
Commensal
Commensal
Commensal
Pathogen
Unclear
Opportunistic
Pathogen
Commensal
Pathogen
Opportunistic
Opportunistic
Representative
host
Relation to host
(Continued)
585
Genus I. Mycoplasma
Table 137. (Continued)
Species
M. genitalium
M. glycophilum
M. gypis
M. haemocanisb
M. haemofelisb
M. haemomurisb
M. hominis
M. hyopharyngis
M. hyopneumoniae
M. hyorhinis
M. hyosynoviae
M. iguanae
M. imitans
M. indiense
M. iners
M. insonsc
M. iowae
M. lagogenitalium
M. leachii
M. leonicaptivi
M. leopharyngis
M. lipofaciens
M. lipophilum
M. maculosum
M. meleagridis
M. microti
M. moatsii
M. mobile
M. molare
M. mucosicanisc
M. muris
M. mustelae
M. neurolyticum
M. opalescens
M. orale
M. ovipneumoniae
M. ovisb
M. oxoniensis
M. penetrans
M. phocicerebrale
M. phocidae
M. phocirhinis
M. pirum
M. pneumoniae
M. primatum
M. pullorum
M. pulmonis
M. putrefaciens
M. salivarium
M. simbae
M. spermatophilum
M. spheniscic
M. spumans
M. sturni
M. sualvi
M. subdolum
M. suisb
M. synoviae
M. testudineum
Morphology
DNA G+C
content
(mol%)
Energy
source
Medium pH
shift
Serum source
Flask-shaped
Elliptical
Coccoidal
Coccoidal
Coccoidal
Coccoidal
Coccobacillary
Pleomorphic
Coccobacillary
Coccobacillary
Pleomorphic
Coccoidal
Flask-shaped
Pleomorphic
nr
Twisted rod
Pleomorphic
Coccoidal
Pleomorphic
Pleomorphic
Pleomorphic
Elliptical
Pleomorphic
Coccobacillary
Coccobacillary
Coccoidal
Spheroidal
Flask-shaped
Coccoidal
Coccoidal
Coccoidal
Pleomorphic
Filamentous
nr
Pleomorphic
nr
Coccoidal
Coccoidal
Flask-shaped
Dumbbell
Coccoidal
Coccoidal
Flask-shaped
Flask-shaped
Coccobacillary
Coccobacillary
Flask-shaped
Coccobacillary
Coccoidal
Pleomorphic
Coccoidal
Pleomorphic
Pleomorphic
Pleomorphic
Coccobacillary
Coccoidal
Coccoidal
Coccoidal
Coccoidal
31
27.5
27.1
nr
38.8
nr
33.7
24
28
27.8
28
nr
31.9
32
29.6
nr
25
23
nr
27
28
24.5
29.7
29.6
28.6
nr
25.7
25
26
nr
24.9
28
26.2
29.2
28.2
25.7
nr
29
25.7
25.9
27.8
26.5
25.5
40
28.6
29
26.6
28.9
27.3
37
32
28
28.4
31
23.7
28.8
31.1
34.2
nr
G
G
R
U
U
U
R
R
G
G
G
G
G
R
R
G
G, R
G
G
G
G
G, R
R
R
R
G
G, R
G, R
G
U
R
G
G
R
R
G
U
G
G, R
R
G, R
R
G
G
R
G
G
G
R
R
R
G
R
G
R, G
R
U
G
G
A
A
K
na
na
na
K
K
A
A
A
A
A
K
K
A
V
A
A
A
A
V
K
K
K
A
V
V
A
nr
K
A
A
K
K
A
na
A
V
K
V
K
A
A
K
A
A
A
K
K
K
A
K
A
V
K
na
A
A
FB
E
P
na
na
na
FB
P, E
P, FB
FB
FB
FB
FB
FB
P
FB
FB
FB
E
FB
FB
P
FB, E
FB, E
P
FB
FB, E
E
FB
E
FB
E
E
FB
FB, E
FB, P
na
FB
FB
FB
FB, E
E, P
FB
FB
FB, E
P, E
FB, E
FB, E
E
FB
FB
P
FB, E
FB
FB
FB, E, P
na
P
FB
Representative
host
Relation to host
Humans
Galliforms
Vultures
Dogs
Cats
Mice
Humans
Pigs
Pigs
Pigs
Pigs
Iguanas
Ducks, geese
Monkeys
Galliforms
Iguanas
Turkeys
Pikas
Cattle
Lions
Lions
Galliforms
Humans
Dogs
Turkeys
Voles
Monkeys
Tench
Dogs
Dogs
Mice
Minks
Mice
Dogs
Humans
Sheep
Sheep
Hamsters
Humans
Seals
Seals
Seals
Humans
Humans
Monkeys
Galliforms
Mice
Goats
Humans
Lions
Humans
Penguins
Dogs
Songbirds
Pigs
Horses
Pigs
Galliforms
Tortoises
Pathogen
Commensal
Opportunistic
Pathogen
Pathogen
Opportunistic
Pathogen
Commensal
Pathogen
Pathogen
Pathogen
Pathogen
Pathogen
Commensal
Commensal
Commensal
Pathogen
Commensal
Pathogen
Commensal
Commensal
Commensal
Unclear
Opportunistic
Pathogen
Commensal
Commensal
Pathogen
Opportunistic
Commensal
Commensal
Commensal
Unclear
Commensal
Commensal
Pathogen
Pathogen
Commensal
Opportunistic
Pathogen
Opportunistic
Pathogen
Commensal
Pathogen
Opportunistic
Pathogen
Pathogen
Pathogen
Opportunistic
Commensal
Pathogen
Pathogen
Opportunistic
Pathogen
Commensal
Opportunistic
Pathogen
Pathogen
Pathogen
(Continued)
586
Table 137. (Continued)
Species
Morphology
DNA G+C
content
(mol%)
M. testudinis
M. verecundum
“M. vulturis”b, c
M. wenyoniib
M. yeatsii
M. zalophic
“Candidatus M.
haematoparvum”b
“Candidatus M. haemobos”b
“Candidatus M.
haemodidelphidis”b
“Candidatus M. haemolamae”b
“Candidatus M.
haemominutum”b
“Candidatus M. kahaneii”b
“Candidatus M. ravipulmonis”b
“Candidatus M.
haemotarandirangiferis”b
“Candidatus M. turicensis”b
Flask-shaped
Pleomorphic
Coccoidal
Coccoidal
Coccoidal
nr
Coccoidal
35
27
nr
nr
26.6
nr
nr
G
OA
U
U
G
G
U
A
N
na
na
na
na
na
FB
FB, E
na
na
FB
FB
na
Tortoises
Cattle
Vultures
Cattle
Goats
Sea lions
Dogs
Commensal
Commensal
Unclear
Pathogen
Opportunistic
Pathogen
nr
Coccoidal
Coccoidal
nr
nr
U
U
na
na
na
na
Cattle
Opossum
nr
nr
Coccoidal
Coccoidal
nr
nr
U
U
na
na
na
na
Llamas
Cats
nr
nr
Coccoidal
Coccoidal
Coccoidal
nr
nr
nr
U
U
U
na
na
na
na
na
na
Monkeys
Mice
Reindeer
nr
Pathogen
nr
Coccoidal
nr
U
na
na
Cats
nr
Energy
source
Medium pH
shift
Serum source
Representative
host
Relation to host
nr, Not reported; na, not applicable; G, glucose; R, arginine; OH, alcohols; OA, organic acids; U, undefined; A, acidic pH shift; K, alkaline pH shift; N, pH remains
neutral; V, pH shift can be acidic or alkaline depending on the energy source provided; FB, fetal bovine serum; E, equine serum; P, porcine serum.
a
Not yet cultivated in cell-free artificial medium. The putative organism “Candidatus M. haemotarandirangiferis” remains to be definitively established and the name
has no standing in nomenclature.
b
Has been grown only in co-culture with eukaryotic cells.
c
mycoides subsp. mycoides precludes the exclusive reliance
on serological-based diagnostics. Experimental vaccines
using formalin-inactivated Mycoplasma mycoides subsp. capri
appear to protect goats from subsequent challenge (BarMoshe et al., 1984; de la Fe et al., 2007). This organism is
under certain quarantine regulations in most non-endemic
countries and is a List B pathogen in the World Organization for Animal Health (OIE) disease classification (http://
oie.int).
Source: isolated from the synovial fluid, synovial membranes, udders, expelled milk, conjunctivae, lungs, blood,
and ear canals of goats; and the urogenital tract of sheep
(Bergonier et al., 1997; Cottew, 1979; Kidanemariam et al.,
2005; Thiaucourt et al., 1996).
DNA G+C content (mol%): 24 (Tm).
Type strain: PG3, NCTC 10137, CIP 71.25.
Sequence accession nos (16S rRNA gene): U26037 (strain
PG3T), U26044 (strain Y-goat).
Further comment: Mycoplasma mycoides subsp. capri now
refers to strains once known as Mycoplasma mycoides subsp.
mycoides var. large colony as well as strains known as Mycoplasma mycoides subsp. capri (Manso-Silván et al., 2009;
Shahram et al., 2010).
1b.Mycoplasma mycoides subsp. mycoides Manso-Silván, Vilei,
Sachse, Djordjevic, Thiaucourt and Frey 2009, 1356VP (Mycoplasma mycoides subsp. mycoides var. small colony Cottew and
Yeats 1978, 294)
my.co.i¢des. Gr. n. mukês -êtos mushroom or other fungus; L.
suff. -oides (from Gr. suff. -eides from Gr. n. eidos that which
is seen, form, shape, figure) resembling, similar; N.L. neut.
adj. mycoides fungus-like.
Cells are pleomorphic and capable of forming long
filaments, but do not produce rho forms. Nonmotile. An
extracellular capsule can be visualized by electron microscopy following staining with ruthenium red. Colonies on
solid agar have a characteristic fried-egg appearance and
are notably smaller than those of Mycoplasma mycoides subsp.
capri. Grows in modified Hayflick medium supplemented
with glucose at 37°C. Formation of biofilms has been demonstrated (McAuliffe et al., 2008).
Pathogenic; causes a characteristic, highly lethal fibrinous interstitial pneumonia and pleurisy known as contagious bovine pleuropneumonia (CBPP) in adult cattle and
severe polyarthritis in calves. Transmission occurs primarily
via direct contact, but can occur by droplet aerosol as well.
Tetracyclines, chloramphenicol, and fluoroquinolones
are effective chemotherapeutic agents; however, treatment of endemic herds is often counterproductive as resistance can develop in carrier animals. Organisms are often
sequestered in areas of coagulative necrosis in subclinically
infected animals and can serve as a reservoir for reintroduction of resistant clones of Mycoplasma mycoides subsp. mycoides
into a herd. Culling of infecting herds and restricting the
movement of infected animals are more effective strategies
for controlling spread of the disease (Windsor and Masiga,
1977). Killed and live vaccines are available for the prevention of infection, but suffer from low antigenicity, poor
efficacy, and residual pathogenesis (Brown et al., 2005).
Antigenic cross-reactivity with Mycoplasma mycoides subsp.
capri and Mycoplasma leachii preclude the exclusive reliance
on serological-based diagnostics. Multiple molecular diagnostics have been described (Gorton et al., 2005; Lorenzon et al., 2008; Persson et al., 1999) and many additional
Genus I. Mycoplasma
molecular tools such as insertion sequence typing have led
to a greater understanding of the epidemiology of outbreaks
(Cheng et al., 1995; Frey et al., 1995; Vilei et al., 1999). This
organism is under certain quarantine regulations in most
non-endemic countries and is listed in the Terrestrial Animal Health Code of the Office International des Epizooties
(http://oie.int).
Source: isolated from the lungs, pleural fluid, lymph
nodes, sinuses, kidneys, urine, synovial fluid, and synovial membranes of cattle and water buffalo (Gourlay and
Howard, 1979; Scanziani et al., 1997; Scudamore, 1976);
the respiratory tract of bison; the respiratory tract of yak;
and the lungs, nasopharynx, and pleural fluid of sheep and
goats (Brandao, 1995; Kusiluka et al., 2000).
DNA G+C content (mol%): 26.1 (Tm), 24.0 (strain PG1T
complete genome sequence).
Type strain: PG1, NCTC 10114, CCUG 32753.
Sequence accession nos: U26039 (16S rRNA gene),
BX293980 (strain PG1T complete genome sequence).
Further comment: Mycoplasma mycoides subsp. mycoides now
refers exclusively to the agent of CBPP (Manso-Silván et al.,
2009).
2. Mycoplasma adleri Del Giudice, Rose and Tully 1995, 31VP
ad¢le.ri. N.L. masc. gen. n. adleri of Adler, referring to Henry
Adler, a Californian veterinarian whose studies contributed
much new information concerning the pathogenic role of
caprine and avian mycoplasmas.
Cells are primarily coccoid. Nonmotile. Colonies on solid
media have a typical fried-egg appearance. Grows well in
Hayflick medium supplemented with arginine at 35–37°C.
Pathogenic; associated with suppurative arthritis and
joint abscesses. Mode of transmission is unknown.
Source: isolated from an abscessed joint of a goat with suppurative arthritis (Del Giudice et al., 1995).
DNA G+C content (mol%): 29.6 (Bd).
Type strain: G145, ATCC 27948, CIP 105676.
Sequence accession no. (16S rRNA gene): U67943.
3. Mycoplasma agalactiae (Wróblewski 1931) Freundt 1955,
73AL (Anulomyces agalaxiae Wróblewski 1931, 111)
a.ga.lac.ti¢ae. Gr. n. agalactia want of milk, agalactia; N.L.
gen. n. agalactiae of agalactia.
Cells are primarily coccoid, but are occasionally branched
and filamentous. Nonmotile. Colonies on solid media have
a typical fried-egg appearance. Grows well in SP-4 or Hayflick medium supplemented with glucose at 37°C. Formation of biofilms has been demonstrated (McAuliffe et al.,
2006).
Pathogenic; causes polyarthritis, mastitis, conjunctivitis (a syndrome collectively termed contagious agalactia;
Bergonier et al., 1997), nonsuppurative arthritis, pneumonia, abortion, and granular vulvovaginitis (Cottew, 1983;
DaMassa, 1996) in goats and sheep. Transmission occurs via
direct contact, most commonly during feeding (kids and
lambs) or milking (dams and ewes).
Macrolides and fluoroquinolones are effective chemotherapeutic agents; however, antimicrobial therapy is not
often utilized in widespread outbreaks due to the potential
for infected animals to develop carrier states with resistant
587
strains and the tendency of antimicrobials to be excreted
in milk. Control measures such as disinfection of fomites
(endemic areas) and culling of infected animals (acute outbreaks) are more common practices. Mycoplasma agalactiae
reportedly shares surface antigens with Mycoplasma bovis
and Mycoplasma capricolum subsp. capricolum (Alberti et al.,
2008; Boothby et al., 1981), potentially complicating serology-based diagnosis of infection. Molecular diagnostics that
can distinguish Mycoplasma agalactiae from Mycoplasma bovis
have been described (Chávez Gonzalez et al., 1995). Commercially available vaccines are widely used, but exhibit
poor efficacy. Numerous experimental vaccines have been
described. This organism is under certain quarantine regulations in some countries and is listed in the Terrestrial Animal Health Code of the Office International des Epizooties
(http://oie.int).
Source: isolated from the joints, udders, milk, conjunctivae, lungs, vagina, liver, spleen, kidneys, and small intestine
of sheep and goats.
DNA G+C content (mol%): 30.5 (Tm), 29.7 (strain PG2T
Type strain: PG2, NCTC 10123, CIP 59.7.
Sequence accession nos: M24290 (16S rRNA gene),
NC_009497 (strain PG2T complete genome sequence).
4. Mycoplasma agassizii Brown, Brown, Klein, McLaughlin,
Schumacher, Jacobson, Adams and Tully 2001c, 417VP
a.gas.si¢zi.i. N.L. masc. gen. n. agassizii of Agassiz, referring
to Louis Agassiz, a naturalist whose name was assigned to a
species of desert tortoise (Gopherus agassizii) from which the
organism was isolated.
Cells are coccoid to pleomorphic, with some strains
appearing to possess a rudimentary terminal structure. Cells
exhibit gliding motility. Colony forms on solid medium vary
from those with a fried-egg appearance to some with mulberry characteristics. Grows well in SP-4 medium supplemented with glucose at 30°C.
Pathogenic; causes chronic upper respiratory tract disease characterized by severe rhinitis in desert tortoises,
gopher tortoises, Russian tortoises, and leopard tortoises.
Mode of transmission appears to be intranasal inhalation
(Brown et al., 1994).
Source: isolated from the nares and choanae of desert
tortoises, gopher tortoises, Russian tortoises, and leopard
tortoises (Brown et al., 2001c).
DNA G+C content (mol%): not determined.
Type strain: PS6, ATCC 700616.
5. Mycoplasma alkalescens Leach 1973, 149AL
al.ka.les¢cens. N.L. v. alkalesco to make alkaline, referring to
the reaction produced in arginine-containing media; N.L.
part. adj. alkalescens alkaline-making.
Coccoid to coccobacillary cells. Motility and colony morphology have not been described for this species. Grows
well in SP-4 medium supplemented with arginine at 37°C.
Pathogenic; causes febrile arthritis and sometimes mastitis, pneumonia, and otitis in cattle (Kokotovic et al., 2007;
Lamm et al., 2004; Leach, 1973). Mode of transmission has
not been established.
588
Tetracyclines and pleuromutilins are effective chemotherapeutic agents (Hirose et al., 2003). Mycoplasma alkalescens
reportedly shares surface antigens with many argininefermenting Mycoplasma species; however, this is only likely
to interfere with accurate diagnosis of infection in the case
of Mycoplasma arginini.
Source: isolated from the synovial fluid, expelled milk,
lungs, ears, prepuce, and semen of cattle.
DNA G+C content (mol%): 25.9 (Tm).
Type strain: D12, PG51, NCTC 10135, ATCC 29103.
Further comment: Bovine serogroup 8 of Leach (1967).
6. Mycoplasma alligatoris Brown, Farley, Zacher, Carlton, Clippinger, Tully and Brown 2001a, 423VP
al.li.ga.to¢ris. N.L. n. alligator, -oris an alligator; N.L. gen. n.
alligatoris of/from an alligator.
Cells are primarily coccoid. Nonmotile. Colonies on
solid medium exhibit typical fried-egg morphology. Growth
is very rapid in SP-4 medium supplemented with glucose
at 30°C.
Pathogenic; causes a multisystemic inflammatory illness
with a lethality unprecedented among mycoplasmas. Pathologic lesions in infected animals include meningitis, interstitial pneumonia, fibrinous pleuritis, polyserositis, fibrinous
pericarditis, myocarditis, endocarditis, synovitis, and splenic
and hepatic necrosis. A high level of mortality of naturally
and experimentally infected American alligators (Alligator
missisippiensis) and broad-nosed caimans (Caiman latirostris) occurs. In contrast, Mycoplasma alligatoris colonizes
the tonsils of experimentally infected Siamese crocodiles
(Crocodylus siamensis) without causing overt pathology. Such
findings led to the hypothesis that Mycoplasma alligatoris is
a natural commensal of a species more closely related to
crocodiles than alligators and that the extreme disease state
observed in alligators results from an enzoonotic infection
(Brown et al., 1996; Pye et al., 2001). The natural mode of
transmission has not been established definitively; however,
animals can be experimentally infected via inoculation of
the glottis.
Source: isolated from the blood, synovial fluid, cerebrospinal fluid, lungs, brain, heart, liver, and spleen of naturally
and experimentally infected American alligators, experimentally infected broad-nosed caimans, and from the tonsils of experimentally infected Siamese crocodiles.
Type strain: A21JP2, ATCC 700619.
Further comment: the name Mycoplasma alligatoris was
assigned for this organism in consideration of the initial
isolation from an alligator (Order: Crocodylia). The English word “alligator” is from the Spanish el lagarto (Latin
ille lacertu the lizard). However, the specific epithet lacerti,
originally proposed for this taxon, was ultimately rejected
because of the modern phylogenetic distinction between
lizards (Order: Lacertilia) and crocodilians.
7. Mycoplasma alvi Gourlay, Wyld and Leach 1977, 95AL
al¢vi. L. n. alvus bowel, womb, stomach; L. gen. n. alvi of
the bowel.
Cells are primarily coccoid; however, subsets of the
opulation display elongated, flask-shaped cells with wellp
defined terminal structures. Unlike most species that exhibit
polar structures, Mycoplasma alvi appears to be nonmotile
(Bredt, 1979; Hatchel and Balish, 2008). Colonies on solid
medium exhibit typical fried-egg morphology. Grows well in
SP-4 medium supplemented with either glucose or arginine
at 37°C.
No evidence of pathogenicity. Mode of transmission has
not been established definitively.
Source: isolated from the lower alimentary tract, feces,
bladder, and vagina of cows, and the intestinal tract of voles
(Gourlay and Howard, 1979).
Type strain: Ilsley, NCTC 10157, ATCC 29626.
8. Mycoplasma amphoriforme Pitcher, Windsor, Windsor,
Bradbury, Yavari, Jensen, Ling and Webster 2005, 2592VP
am.pho.ri.for¢me. L. n. amphora amphora; L. adj. suff.
-formis -e like, of the shape of; N.L. neut. adj. amphoriforme
amphora-shaped, having the form of an amphora.
Cells are flask-shaped with a distinct terminal structure
reminiscent of Mycoplasma gallisepticum. Cells exhibit lowspeed gliding motility and move in the direction of the terminal structure (Hatchel et al., 2006). Colony morphology
variable, from a typical fried-egg morphology to a “ground
glass” appearance. Grows well in SP-4 medium supplemented with glucose at 37°C.
Pathogenicity and mode of transmission have not been
established definitively.
Despite showing sensitivity to fluoroquinolones, tetracyclines, and macrolides in vitro, Mycoplasma amphoriforme
appears to be successfully evasive during treatment of
patients with these antibiotics. The veterinary antibiotic
valnemulin is successful at controlling infection (Webster
et al., 2003).
Source: isolated from the sputum of immunocompromised humans with bronchitis and related lower respiratory tract disease (Pitcher et al., 2005; Webster et al., 2003).
The prevalence of Mycoplasma amphoriforme in the general
human population is unsubstantiated.
DNA G+C content (mol%): 34.0 (fluorescent intensity).
Type strain: A39, NCTC 11740, ATCC BAA-992.
Sequence accession no. (16S rRNA gene): FJ226575.
9. Mycoplasma anatis Roberts 1964, 471AL
a.na¢tis. L. n. anas, -atis a duck; L. gen. n. anatis of a duck.
Cells have been described as coccoid with ring forms, but
cellular and colony morphology is generally poorly described.
Motility for this species has not been assessed. Grows well in
SP-4 medium supplemented with glucose 37°C.
Isolated from pathologic lesions, but attempts to reproduce disease following experimental infection have been
equivocal (Amin and Jordan, 1978; Roberts, 1964). The
mode of transmission has not been established definitively.
Source: isolated from pathologic lesions, the respiratory
tract, hock joint, pericardium, cloaca, and meninges of
ducks (Goldberg et al., 1995; Ivanics et al., 1988; Tiong,
1990).
Genus I. Mycoplasma
Type strain: 1340, ATCC 25524, NCTC 10156.
Sequence accession no. (16S rRNA gene): AF412970.
10. Mycoplasma anseris Bradbury, Jordan, Shimizu, Stipkovits
and Varga 1988, 76VP
an¢se.ris. L. gen. n. anseris of the goose.
Cells are primarily spherical. Nonmotile. Colonies on
solid medium have a typical fried-egg appearance. Grows
well in Hayflick medium supplemented with arginine at
37°C.
Opportunistic pathogen; associated with balanitis of
geese, but can be isolated from clinically normal animals.
Mode of transmission has not been established definitively.
Source: isolated from the phallus and cloaca of geese
(Hinz et al., 1994; Stipkovits et al., 1984a).
DNA G+C content (mol%): 24.7–26.0 (Bd/Tm).
Type strain: 1219, ATCC 49234.
11. Mycoplasma arginini Barile, Del Giudice, Carski, Gibbs and
Morris 1968, 490AL
ar.gi.ni¢ni. N.L. n. argininum arginine, an amino acid; N.L.
gen. n. arginini of arginine, referring to its hydrolysis.
Cells are primarily coccoid. Motility for this species has
not been assessed. Colonies have either the typical fried-egg
morphology or a granular, “berry-like” appearance. Grows
well in modified Hayflick medium supplemented with arginine at 37°C.
Pathogenic; associated with pneumonia, vesiculitis, keratoconjunctivitis, and mastitis in cattle; pneumonia and keratoconjunctivitis in sheep; possibly with arthritis in goats;
and septicemia in immunocompromised humans (Tully
and Whitcomb, 1979; Yechouron et al., 1992). Modes of
transmission have not been definitively assessed and likely
vary by anatomical site.
Mycoplasma arginini is commonly associated with contamination of eukaryotic cell culture and is frequently removed
by treatment of cells with antibiotics and/or maintenance of
cell lines in antibiotic-containing medium. The most effective classes of antibiotics for cell culture eradication are tetracyclines, macrolides, and fluoroquinolones. Additionally,
passage of eukaryotic cells in hyperimmune serum raised
against Mycoplasma arginini has been shown to be an effective method of eradication (Jeansson and Brorson, 1985).
Source: isolated from a wide array of mammalian hosts
including cattle, sheep, goats, pigs, horses, domestic dogs,
domestic cats, lions, lynxes, cheetahs, chamois, camels,
ibexes, humans, and mice.
Type strain: G230, ATCC 23838, NCTC 10129, CIP 71.23,
NBRC 14476.
12. Mycoplasma arthritidis (Sabin 1941) Freundt 1955, 73AL
(Murimyces arthritidis Sabin 1941, 57)
ar.thri¢ti.dis. Gr. n. arthritis -idos gout, arthritis; N.L. gen. n.
arthritidis of arthritis.
Cells are filamentous and vary in length. Motility for this
species has not been assessed. Colonies on solid medium
589
have a typical fried-egg appearance. Grows well in SP-4
medium supplemented with arginine.
Pathogenic; causes purulent polyarthritis, rhinitis, otitis
media, ocular lesions, and abscesses in rats. Mycoplasma
arthritidis is also known to superinfect lung lesions initiated
by Mycoplasma pulmonis. Experimental inoculations via various routes can result in septicemia, acute flaccid paralysis,
and pyelonephritis in rats, and chronic arthritis in mice and
rabbits. Mycoplasma arthritidis is unique among mycoplasmas
in harboring the lysogenized bacteriophage MAV1, whose
contribution to virulence is equivocal, and producing the
potent mitogen MAM, which appears to confer increased
toxicity and lethality but to be irrelevant to arthritogenicity
(Clapper et al., 2004; Luo et al., 2008; Voelker et al., 1995).
The mechanism of transmission is largely dependent on the
tissue infected.
Source: isolated from the synovial membranes, synovial
fluid, middle ear, eye, abscessed bone, abscessed ovary, and
oropharynx of wild and captive rats. Isolations have also
been reported from non-human primates including rhesus
monkeys and bush babies (Somerson and Cole, 1979); and
from joint fluid of wild boars (Binder et al., 1990). The true
origins of putative isolates from the human urethra, prostate, and cervix have been questioned (Cassell and Hill,
1979; Washburn et al., 1995).
DNA G+C content (mol%): 30.0 (Tm; strain PG6T), 30.7
(strain 158L3-1 complete genome sequence).
Type strain: PG6, ATCC 19611, NCTC 10162, CIP 104678,
NBRC 14860.
CP001047 (strain 158L3-1 complete genome sequence).
13. Mycoplasma auris DaMassa, Tully, Rose, Pitcher, Leach and
Cottew 1994, 483VP
au¢ris. L. gen. n. auris of the ear, referring to the provenance
of the organism, the ears of goats.
Cells are coccoid to pleomorphic. Nonmotile. Colonies on solid medium have a typical fried-egg appearance.
Grows well in Hayflick medium supplemented with arginine at 37°C.
No evidence of pathogenicity. Mechanism of transmission has not been established.
Source: isolated from the external ear canals of goats
(Damassa et al., 1994).
Type strain: UIA, ATCC 51348, NCTC 11731, CIP
105677.
14. Mycoplasma bovigenitalium Freundt 1955, 73AL
bo.vi.ge.ni.ta¢li.um. L. n. bos, bovis the ox, bull, cow; L. pl.
n. genitalia the genitals; N.L. pl. gen. n. bovigenitalium of
bovine genitalia.
Cells range from coccoid to filamentous. Motility for this
broth supplemented with glucose and/or arginine at 37°C
and produces a “film and spots” reaction.
Pathogenic; causes vulvovaginitis, vesiculitis, epididymitis, abortion, infertility, mastitis, pneumonia, conjunctivitis,
590
and arthritis in cattle; and pneumonia and conjunctivitis in
domestic dogs. Mode of transmission is via sexual contact
and/or droplet aerosol.
Control measures during outbreaks of Mycoplasma bovigenitalium infection include suspension of natural breeding
in favor of artificial insemination with disposable instruments and, in severe cases, culling of infected animals.
Source: isolated from the udders, seminal vesicles, prepuce, semen, vagina, cervix, lungs, conjunctivae, and joint
capsule of cattle; and from the lungs, prepuce, prostate,
vagina, cervix, and conjunctivae of domestic dogs (Chalker,
2005; Gourlay and Howard, 1979).
Type strain: PG11, ATCC 19852, NCTC 10122, NBRC
14862.
Sequence accession nos (16S rRNA gene): M24291,
AY121098.
Further comment: the collection of strains formerly referred
to as “Mycoplasma ovine/caprine serogroup 11” have been
reclassified as Mycoplasma bovigenitalium (Nicholas et al.,
2008).
15. Mycoplasma bovirhinis Leach 1967, 313AL
bo.vi.rhi¢nis. L. n. bos, bovis the ox; Gr. n. rhis, rhinos nose;
N.L. gen. n. bovirhinis of the nose of the ox.
Cell and colony morphology and motility for this species are poorly defined. Grows well in SP-4 medium supplemented with glucose at 37°C.
Pathogenic; causes pneumonia, otitis, conjunctivitis, and
mastitis in cattle. Mycoplasma bovirhinis is often found in coinfections with other pathogens, leading to speculation that
it often acts as a superinfecting agent. Mode of transmission
has not been established definitively.
Source: isolated from the lungs, nasopharynx, trachea,
udders, expelled milk, ears, conjunctivae, and rarely from
the urogenital tract of cattle (Gourlay and Howard, 1979).
Type strain: PG43, ATCC 27748, NCTC 10118, CIP 71.24,
NBRC 14857.
16. Mycoplasma bovis (Hale, Hemboldt, Plastridge and Stula
1962) Askaa and Ernø 1976, 325AL (Mycoplasma agalactiae
var. bovis Hale, Hemboldt, Plastridge and Stula 1962, 591;
Mycoplasma bovimastitidis Jain, Jasper and Dellinger 1967,
409)
bo¢vis. L. n. bos the ox; L. gen. n. bovis of the ox.
Cells range from coccoidal to short filaments. Nonmotile.
Formation of biofilms has been demonstrated (McAuliffe
et al., 2006). Colonies on solid medium have the typical
fried-egg appearance, with notably large centers. Grows
well in SP-4 medium supplemented with glucose at 37°C
Pathogenic; causes mastitis, polyarthritis, keratoconjunctivitis (Gourlay and Howard, 1979), pneumonia, and otitis
media (Caswell and Archambault, 2007; Maeda et al., 2003),
and is rarely associated with infertility, abortion, endometritis, salpingitis, and vesiculitis (Doig, 1981; Gourlay and Howard, 1979) in cattle; pneumonia and polyarthritis in bison
(Dyer et al., 2008); and is rarely associated with pneumonia,
mastitis, and arthritis in goats (Egwu et al., 2001; Gourlay
and Howard, 1979). Mode of transmission is via direct contact with infected animals or fomites, most commonly during feeding (suckling or trough), milking (cows), aerosol,
or sexual contact.
Macrolides and fluoroquinolones are effective chemotherapeutic agents in vitro; however, antimicrobial therapy
is not often utilized in animals with advanced disease due
to poor efficacy and the tendency of antimicrobials to be
excreted in milk. Control measures such as disinfection
of fomites, isolation of infected animals, and euthanasia
of animals showing clinical signs are more common practices. Mycoplasma bovis reportedly shares surface antigens
with Mycoplasma agalactiae (Boothby et al., 1981), potentially complicating serology-based diagnosis of infection.
Molecular diagnostics that can distinguish Mycoplasma agalactiae from Mycoplasma bovis have been described (Chávez
Gonzalez et al., 1995). Several vaccines are commercially
available, but exhibit poor efficacy in that they tend to allow
for the establishment of infection while only preventing
overt clinical signs.
Source: isolated from the udders, expelled milk, synovial
fluid, synovial membranes, conjunctivae, lungs, ear canals,
tympanic membranes, aborted calves, uterus, cervix, vagina,
and semen of cattle; from the lungs and synovial fluid of
bison; and from the lungs and udders of goats.
Type strain: Donetta, PG45, ATCC 25523, NCTC 10131.
Sequence accession no. (16S rRNA gene): AJ419905.
Further comment: Bovine serotype 5 of Leach (1967).
17. Mycoplasma bovoculi Langford and Leach 1973, 1443AL
bo.vo¢cu.li. L. n. bos, bovis ox, bull, cow; L. n. oculus the eye;
N.L. gen. n. bovoculi of the bovine eye.
Cells are coccoid to coccobacillary. Motility for this
have a typical fried-egg appearance. Grows well in Hayflick
medium supplemented with glucose at 37°C.
Pathogenic; causes conjunctivitis and keratoconjunctivitis in cattle. Face flies are the suggested mechanism of transmission, though this has yet to be established definitively.
Topical application of oxytetracycline is an effective treatment for infection.
Source: isolated from the conjunctivae and semen of cattle, and from aborted calves (Langford and Leach, 1973;
Singh et al., 2004).
Type strain: M165/69, NCTC 10141, ATCC 29104.
Further comment: Mycoplasma bovoculi was originally
described as Mycoplasma oculi by Leach in 1973, wherein
the defining publication referring to the species as Mycoplasma bovoculi (Langford and Leach, 1973) was cited as “in
press”.
18. Mycoplasma buccale Freundt, Taylor-Robinson, Purcell,
Chanock and Black 1974, 252AL
buc.ca¢le. L. n. bucca the mouth; L. neut. suff. -ale suffix
denoting pertaining to; N.L. neut. adj. buccale buccal, pertaining to the mouth.
Genus I. Mycoplasma
Cells range from coccoid to filamentous. Motility for this
medium supplemented with arginine and herring sperm
DNA at 37°C.
not been assessed definitively.
Source: isolated from the oropharynx of humans, rhesus
macaques, chimpanzees, orangutans, baboons, African
green monkeys, crab-eating macaques, and patas monkeys
(Somerson and Cole, 1979).
Type strain: CH20247, ATCC 23636, NCTC 10136, CIP
105530, NBRC 14851.
19. Mycoplasma buteonis Poveda, Giebel, Flossdorf, Meier and
Kirchhoff 1994, 97VP
bu.te.o¢nis. L. masc. n. buteo, -onis buzzard; L. masc. gen. n.
buteonis of the buzzard.
Cells are coccoid. Motility for this species has not been
assessed. Colonies on solid medium have typical fried-egg
appearance. Grows well in modified Frey’s medium supplemented with glucose at 37°C.
Mycoplasma buteonis may be pathogenic for saker falcons,
as it was found in the respiratory tract, nervous system, and
bone of a nestling with pneumonia, hepatitis, ataxia, and
dyschondroplasia. No evidence of pathogenicity for buzzards. Mode of transmission has not been assessed.
Source: isolated from the trachea of buzzards; from the
eggs of the lesser kestrel; and the trachea, lungs, brain, and
bone marrow of the saker falcon. Has been detected in the
common kestrel and the Western marsh harrier (Erdélyi
et al., 1999; Lierz et al., 2008a, 2008c).
Type strain: Bb/T2g, ATCC 51371.
20. Mycoplasma californicum Jasper, Ernø, Dellinger and
Christiansen 1981, 344VP
ca.li.for¢ni.cum. N.L. neut. adj. californicum pertaining to
California.
Cells are coccoid to filamentous. Motility for this species has not been assessed. Colonies are conical in shape
with distinct small centers. Grows well in modified Hayflick
broth at 37°C.
Pathogenic; causes purulent mastitis in cows and rarely
in sheep. Mode of transmission has not been established
definitively.
Source: isolated from the udders and expelled milk of
cows and ewes.
Type strain: ST-6, ATCC 33461, AMRC-C 1077, NCTC
10189.
Sequence accession no. (16S rRNA gene): M24582.
21. Mycoplasma canadense Langford, Ruhnke and Onoviran
1976, 218AL
ca.na.den¢se. N.L. neut. adj. canadense pertaining to
Canada.
591
s pecies has not been assessed. Colonies on solid agar have
a characteristic fried-egg appearance. Grows well in SP-4
medium supplemented with arginine at 37°C.
Pathogenic; causes mastitis and arthritis, and may be associated with infertility, abortion, and pneumonia of cattle.
Source: isolated from the udders, expelled milk, synovial
membranes, aborted calves, vagina, semen, and lungs
(Boughton et al., 1983; Friis and Blom, 1983; Gourlay and
Howard, 1979; Jackson et al., 1981).
Type strain: 275C, NCTC 10152, ATCC 29418.
22. Mycoplasma canis Edward 1955, 90AL
ca¢nis. L. n. canis, -is a dog; L. gen. n. canis of a dog.
Cells are pleomorphic, exhibiting branched and filamentous forms. Motility for this species has not been assessed.
Colonies on solid medium exhibit two stable forms: “smooth”
colonies with a nongranular appearance and round edges;
and “rough” colonies with a granular appearance and irregular or crenated edges. Each form maintains its characteristic appearance during repeated subculturing. Grows well in
SP-4 medium supplemented with glucose at 37°C.
Opportunistic pathogen; associated with infertility and
adverse pregnancy outcomes, endometritis, epididymitis, urethritis, cystitis, and pneumonia of domestic dogs
(Chalker, 2005); pneumonia of cattle (ter Laak et al.,
1992b); and pneumonia in immunocompromised humans
(Armstrong et al., 1971). Mode of transmission is via sexual contact or aerosol. Mycoplasma canis appears to have a
greater tendency toward upper respiratory tract commensalism and urogenital tract pathogenicity in dogs, while
exhibiting pathogenicity for the respiratory tract of cattle.
Source: isolated from the cervix, vagina, prepuce,
epididymis, prostate, semen, urine, bladder, oropharynx,
nares, lungs, trachea, conjunctivae, kidneys, spleen, pericardium, liver, and lymph nodes of domestic dogs; from the
lungs and oropharynx of cattle; from the lungs and pharynx of humans; and from the throat and rectum of baboons
and African green monkeys.
14846.
23. Mycoplasma capricolum Tully, Barile, Edward, Theodore
and Ernø 1974, 116AL
ca.pri.co¢lum. L. n. caper, -pri the male goat; N.L. -suff. colus,
-a, -um (from L. v. incolere to dwell) dwelling; N.L. neut. adj.
capricolum dwelling in a male goat.
Cells are coccobacillary. Nonmotile. Colonies on solid
agar have a characteristic fried-egg appearance. Grows in
SP-4 or modified Hayflick medium supplemented with glucose at 37°C.
Type strain: California kid, ATCC 27343, NCTC 10154,
CIP 104620.
The species has subsequently been divided as follows.
592
23a.Mycoplasma capricolum subsp. capricolum (Tully, Barile,
Edward, Theodore and Ernø 1974) Leach, Ernø and
MacOwan 1993, 604VP (Mycoplasma capricolum Tully, Barile,
Edward, Theodore and Ernø 1974, 116)
ca.pri.co¢lum. L. n. caper, -pri the male goat; N.L. -suff. colus,
-a, -um (from L. v. incolere to dwell) dwelling; N.L. neut. adj.
capricolum dwelling in a male goat.
Cells are coccobacillary and can produce long, helical
rods known as rho forms. Nonmotile. Colonies on solid
Pathogenic; causes fibrinopurulent polyarthritis, mastitis,
conjunctivitis (a syndrome collectively termed contagious
agalactia) and septicemia in goats. Transmission occurs via
direct contact between animals or with fomites.
Tetracyclines, macrolides, and tylosin are effective chemotherapeutic agents. Treatment of acutely infected
animals often leads to the eradication of the organism,
whereas treatment of chronically infected animals does not.
Early intervention with antibiotics and improved sanitation
are effective control measures (Thiaucourt et al., 1996).
Antigenic cross-reactivity with Mycoplasma capricolum subsp.
capripneumoniae and Mycoplasma leachii preclude the exclusive reliance on serological-based diagnostics. Multiple
molecular diagnostics have been described (Fitzmaurice
et al., 2008; Greco et al., 2001). This organism is under certain quarantine regulations in some countries and is listed
in the Terrestrial Animal Health Code of the Office International des Epizooties (http://oie.int).
Source: isolated from the synovial fluid, synovial membranes, udders, expelled milk, conjunctivae, spleen,
nasopharynx, oral cavity, and ear canal of goats; and the
nasopharynx of sheep (Cottew, 1979).
DNA G+C content (mol%): 24.1 (Tm), 23 (strain California
kidT complete genome).
Type strain: California kid, ATCC 27343, NCTC 10154,
CIP 104620.
Sequence accession nos: U26046 (16S rRNA gene),
NC_007633 (strain California kidT complete genome).
23b.Mycoplasma capricolum subsp. capripneumoniae Leach,
Ernø and MacOwan 1993, 604VP
ca.pri.pneu.mo.ni¢ae. L. n. capra, -ae a goat; Gr. n. pneumonia
disease of the lungs, pneumonia; N.L. gen. n. capripneumoniae of a pneumonia of a goat.
Cells are coccobacillary. Nonmotile. Colonies on solid
Pathogenic; causes characteristic, highly lethal fibrinous pleuropneumonia known as contagious caprine pleuropneumonia (CCPP) in goats (McMartin et al., 1980).
A respiratory tract disease of similar pathology found in
association with Mycoplasma capricolum subsp. capripneumoniae has been reported in sheep, mouflon, and ibex (Arif
et al., 2007; Shiferaw et al., 2006). Transmission occurs via
droplet aerosol.
Tetracyclines and tylosin are effective chemotherapeutic
agents; however, treatment of endemic herds is not often
undertaken. Culling of infected herds and restricting the
movement of infected animals are more common strategies for controlling spread of the disease (Thiaucourt et al.,
1996). Live and killed vaccines have been described; however, each appear to afford delayed or partial protection
from morbidity, and few have been completely successful
at preventing infection (Browning et al., 2005). Antigenic
cross-reactivity with Mycoplasma capricolum subsp. capricolum and Mycoplasma leachii preclude the exclusive reliance on serological-based diagnostics. Multiple molecular
diagnostics have been described (Lorenzon et al., 2008;
March et al., 2000; Woubit et al., 2004). This organism is
under certain quarantine regulations in some countries
and is listed in the Terrestrial Animal Health Code of the
Office International des Epizooties (http://www.oie.int).
Source: isolated from the lower respiratory tract of goats,
sheep, mouflon, and ibex.
Type strain: F38, NCTC 10192.
Further comment: previously known as the F38-type caprine
mycoplasmas.
24. Mycoplasma caviae Hill 1971, 112AL
ca.vi¢ae. N.L. n. cavia guinea pig (Cavia cobaya); N.L. gen. n.
caviae of a guinea pig.
Cell morphology for this species has not been described
and motility has not been assessed. Colonies on solid agar
have a characteristic fried-egg appearance. Grows in SP-4
Source: isolated from the nasopharynx and urogenital
tract of guinea pigs (Hill, 1971).
Type strain: G122, ATCC 27108, NCTC 10126.
25. Mycoplasma cavipharyngis Hill 1989, 371VP (Effective publication: Hill 1984, 3187)
ca.vi.pha.ryn¢gis. N.L. n. cavia the guinea pig (Cavia cobaya);
N.L. n. pharynx -yngis (from Gr. n. pharugx pharuggos throat)
throat; N.L. gen. n. cavipharyngis of the throat of a guinea
pig.
Cells are highly filamentous and filaments are twisted at
intervals along their length. Regular helical forms like those
of Spiroplasma species are not produced. Nonmotile. Growth
on solid medium shows small, granular colonies with poorly
defined centers. Grows in Hayflick medium supplemented
with glucose at 35°C.
Source: isolated from the nasopharynx of guinea pigs
(Hill, 1984).
Type strain: 117C, NCTC 11700, ATCC 43016.
26. Mycoplasma citelli Rose, Tully and Langford 1978, 571AL
ci.tel¢li. N.L. n. Citellus a genus of ground squirrel; N.L. gen.
n. citelli of Citellus.
Genus I. Mycoplasma
Cells are highly pleomorphic. Motility for this species
has not been assessed. Colonies on solid agar have a characteristic fried-egg appearance. Grows well in SP-4 medium
supplemented with glucose at 37°C.
Source: isolated from the trachea, lung, spleen, and liver
of ground squirrels (Rose et al., 1978).
Type strain: RG-2C, ATCC 29760, NCTC 10181.
27. Mycoplasma cloacale Bradbury and Forrest 1984, 392
VP
clo.a.ca¢le. L. neut. adj. cloacale pertaining to a cloaca.
Cells are primarily spherical. Nonmotile. Colonies on solid
medium exhibit typical fried-egg appearance. Grows well in
Hayflick medium supplemented with arginine at 37°C.
not been established definitively
Source: isolated from the cloaca of a turkey; from the lungs,
trachea, ovaries, and eggs of ducks; and from chickens,
pheasants, and geese (Benčina et al., 1987, 1988; Bradbury
et al., 1987; Goldberg et al., 1995; Hinz et al., 1994).
DNA G+C content (mol%): 26 (Bd).
28. Mycoplasma collis Hill 1983b, 849VP
col¢lis. L. gen. n. collis of a hill, alluding to the author who
described the species.
Cells are primarily coccoidal and are nonmotile. Growth
on a solid medium shows colonies with a typical fried-egg
appearance. Grows in Hayflick medium supplemented with
glucose at 35–37°C.
not been assessed.
Source: isolated from the conjunctivae of captive rats and
mice (Hill, 1983b). References to isolation of Mycoplasma
collis from domestic dogs appear to have been in error
(Chalker and Brownlie, 2004).
Type strain: 58B, NCTC 10197, ATCC 35278.
29. Mycoplasma columbinasale Jordan, Ernø, Cottew, Hinz and
Stipkovits 1982, 114VP
co.lum.bi.na.sa¢le. L. n. columbus a pigeon; L. n. nasus nose;
L. neut. suff. -ale suffix used with the sense of pertaining to;
N.L. neut. adj. nasale pertaining to the nose; N.L. neut. adj.
columbinasale pertaining to the nose of a pigeon.
exhibit typical fried-egg appearance. Grows well in SP-4
medium supplemented with arginine at 35–37°C. Produces
a “film and spots” reaction.
Source: isolated from the turbinates of rock pigeons,
racing pigeons, and fantail pigeons (Benčina et al., 1987;
Keymer et al., 1984; Nagatomo et al., 1997; Yoder and
Hofstad, 1964).
593
Further comment: previously known as avian serovar (serotype) L (Yoder and Hofstad, 1964).
30. Mycoplasma columbinum Shimizu, Ernø and Nagatomo
1978, 545AL
co.lum.bi¢num. L. neut. adj. columbinum pertaining to a
pigeon.
Cells are pleomorphic and vary from coccoid to ring
forms. Motility for this species has not been assessed. Colonies on solid medium have typical fried-egg morphology.
Grows in Frey’s medium supplemented with arginine at
37°C. Produces a “film and spots” reaction.
Source: isolated from the trachea and oropharynx of feral
pigeons and from the brain and lungs of racing pigeons
(Benčina et al., 1987; Jordan et al., 1981; Keymer et al.,
1984; Reece et al., 1986).
Type strain: MMP1, ATCC 29257, NCTC 10178.
31. Mycoplasma columborale Shimizu, Ernø and Nagatomo
1978, 545AL
co.lum.bo.ra¢le. L. n. columba pigeon; L. n. os, oris the mouth;
L. neut. suff. -ale suffix used with the sense of pertaining
to; N.L. neut. adj. orale of or pertaining to the mouth; N.L.
neut. adj. columborale of the pigeon mouth.
Cells are pleomorphic but predominantly coccoid or
exhibiting ring forms. Motility for this species has not been
assessed. Growth on solid medium yields medium to large
colonies with very small central zones. Grows in Frey’s
Pathogenicity for pigeons is unconfirmed, but one report
described airsacculitis in experimentally inoculated chickens. Mode of transmission has not been assessed definitively.
Source: isolated from the trachea and oropharynx of feral
pigeons and fantail pigeons; from the oropharynx and
sinuses of racing pigeons; and from corvids and house flies
(Benčina et al., 1987; Bradbury et al., 2000; Jordan et al.,
1981; Kempf et al., 2000; Keymer et al., 1984; MacOwan
et al., 1981; Nagatomo et al., 1997; Reece et al., 1986).
Type strain: MMP4, ATCC 29258, NCTC 10179.
32. Mycoplasma conjunctivae Barile, Del Giudice and Tully
1972, 74AL
con.junc.ti¢va.e. N.L. n. conjunctiva the membrane joining the
eyeball to the lids; N.L. gen. n. conjunctivae of conjunctiva.
Cells are coccoid to coccobacillary. Motility for this species has not been assessed. Colonies grown on solid medium
may have elevated centers and a greenish, brownish, or
olive color. Grows well in SP-4 medium supplemented with
glucose at 37°C.
Pathogenic; causes infectious keratoconjunctivitis that
can either resolve into a carrier state or result in complete
594
or near-complete blindness in goats, sheep, chamois, and
ibex (Cottew, 1979; Mayer et al., 1997). Mode of transmission is via direct contact.
Though tetracyclines are an effective antimicrobial therapy in vivo, treatment to eradicate Mycoplasma conjunctivae
from herds is not often attempted as the economic burden
of infection is low. Topical treatment of secondary infections is often necessary (Slatter, 2001). Several commercial
diagnostic assays have been described.
Source: isolated from the conjunctivae of goats, sheep,
chamois, and Alpine ibex.
Type strain: HRC581, ATCC 25834, NCTC 10147.
Sequence accession no. (16S rRNA gene): AY816349.
33. Mycoplasma corogypsi Panangala, Stringfellow, Dybvig,
Woodard, Sun, Rose and Gresham 1993, 589VP
co.ro.gyp¢si. Gr. n. korax -acos a raven (black); Gr. n. gyps gypos
a vulture; N.L. gen. n. corogypsi (sic) of a raven vulture.
Cells are highly pleomorphic and show small circular
budding processes abutting elongated cells. Motility for this
have a fried-egg appearance. Grows in Frey’s medium supplemented with glucose at 37°C.
Pathogenicity has not been established, although associated with abscess formation in a black vulture. Mycoplasma corogypsi has been isolated from clinically normal
captive falcons, and may represent a commensal of this
species. Mode of transmission has not been assessed
definitively.
Source: isolated from the abscessed footpad of a black vulture, and from captive falcons (Lierz et al., 2002; Panangala
et al., 1993).
Type strain: BV1, ATCC 51148.
Sequence accession no. (16S rRNA gene): L08054.
34. Mycoplasma cottewii DaMassa, Tully, Rose, Pitcher, Leach
and Cottew 1994, 483VP
cot.te¢wi.i. N.L. masc. gen. n. cottewii of Cottew, named for
Geoffrey S. Cottew, an Australian veterinarian who was a coisolator of the organism.
Cells are primarily coccoid. Nonmotile. Formation of
biofilms has been demonstrated (McAuliffe et al., 2006).
Growth on solid medium shows colonies with a typical friedegg appearance. Grows well in Hayflick medium supplemented with glucose at 37°C.
Source: isolated from the external ear canals and rarely
the sinuses of goats (Damassa et al., 1994).
Type strain: VIS, ATCC 51347, NCTC 11732, CIP 105678.
35. Mycoplasma cricetuli Hill 1983a, 117VP
cri.ce.tu¢li. N.L. n. Cricetulus generic name of the Chinese
hamster, Cricetulus griseus; N.L. gen. n. cricetuli of Cricetulus.
Cells are coccoid to pleomorphic. Nonmotile. Colony
growth on solid medium has a fried-egg appearance with
markedly small centers. Grows well in Hayflick broth supplemented with glucose at 37°C.
Source: isolated from the conjunctivae and nasopharynx
of Chinese hamsters (Hill, 1983a).
Type strain: CH, NCTC 10190, ATCC 35279.
36. Mycoplasma crocodyli Kirchhoff, Mohan, Schmidt, Runge,
Brown, Brown, Foggin, Muvavarirwa, Lehmann and Flossdorf 1997, 746VP
cro.co.dy¢li. N.L. n. Crocodylus (from L. n. crocodilus crocodile) generic name of the crocodile; N.L. gen. n. crocodyli
of Crocodylus.
Cells are coccoid. Nonmotile. Colonies on solid medium
show a typical fried-egg appearance. Grows very rapidly in
Pathogenic; causes exudative polyarthritis and rarely
pneumonia in crocodiles. The natural mode of transmission has not been assessed definitively; however, experimental infection resulting in the reproduction of disease was
achieved by intracoelomic and/or intrapulmonary inoculation.
Tetracyclines are effectively used to alleviate clinical signs
in farmed crocodiles. A bacterin vaccine effective at controlling infection and preventing disease has been described
(Mohan et al., 2001).
Source: isolated from the joints and lungs of Nile crocodiles (Mohan et al., 1997).
Type strain: MP145, ATCC 51981.
37. Mycoplasma cynos Røsendal 1973, 53AL
cy¢nos. Gr. n. cyon, cynos a dog; N.L. gen. n. cynos of a dog.
Cells are coccoid to coccobacillary. Motility for this species has not been assessed. Colonies on solid medium have
defined centers and scalloped perimeters. Grows well in
SP-4 medium supplemented with arginine at 37°C.
Pathogenic; causes pneumonia, bronchitis, and rarely cystitis in domestic dogs. The mode of transmission is via droplet aerosol, as demonstrated by studies housing infected
and sentinel dogs (Røsendal and Vinther, 1977).
Source: isolated from the lungs, trachea, nasopharynx,
urine, prepuce, prostate, cervix, vagina, and conjunctivae
of domestic dogs (Chalker, 2005).
Type strain: H 831, ATCC 27544, NCTC 10142.
38. Mycoplasma dispar Gourlay and Leach 1970, 121AL
dis¢par. L. neut. adj. dispar dissimilar, different.
Cells range from coccoid to short and filamentous. Motility for this species has not been assessed. An extracellular
capsule can be visualized by electron microscopy following
staining with ruthenium red. Colonies on solid medium
have a granular, lacy, or reticulated appearance with no or
a poorly defined central area. Grows in SP-4 medium or
Genus I. Mycoplasma
modified Friis medium supplemented with glucose and calf
thymus DNA at 37°C.
Pathogenic; causes pneumonia and rarely mastitis in cattle. Mode of transmission is by droplet aerosol.
Source: isolated from the lower respiratory tract and
udders of cattle (Gourlay and Howard, 1979; Hodges et al.,
1983).
DNA G+C content (mol%): 28.5–29.3 (Tm).
Type strain: 462/2, ATCC 27140, NCTC 10125.
39. Mycoplasma edwardii Tully, Barile, Del Giudice, Carski,
Armstrong and Razin 1970, 349AL
ed.war¢di.i. N.L. masc. gen. n. edwardii of Edward, named
after Derrick Graham ff. Edward (1910–1978), who first isolated this organism.
595
Cells are pleomorphic. Motility for this species has not
been assessed. Colonies on solid medium show a typical
fried-egg appearance. Grows in Hayflick medium supplemented with glucose at 37°C.
Opportunistic pathogen. Associated with endometritis, vulvitis, balanoposthitis, impaired fecundity, and abortion in horses; however, Mycoplasma equigenitalium is highly
prevalent in clinically normal horses (Spergser et al., 2002).
Mode of transmission is via sexual contact.
Source: isolated from the cervix, semen, and aborted foals,
and rarely from the trachea, of horses (Lemcke, 1979).
Type strain: T37, ATCC 29869, NCTC 10176.
42. Mycoplasma equirhinis Allam and Lemcke 1975, 405AL
Cells are coccobacillary to short and filamentous. Motility for this species has not been assessed. Colonies on solid
medium show a typical fried-egg appearance. Grows well in
Opportunistic pathogen; commonly found as a commensal of the oral and/or nasal cavities and urogenital tract of
domestic dogs. Mycoplasma edwardii is rarely associated with
pneumonia, arthritis, and septicemia of domestic dogs, often
as a secondary pathogen compounding an existing lesion.
Source: isolated from the oropharynx, nasopharynx,
trachea, lungs, prepuce, vagina, cervix, blood, and synovial fluid of domestic dogs (Chalker, 2005; Stenske et al.,
2005).
Type strain: PG-24, ATCC 23462, NCTC 10132.
e.qui.rhi¢nis. L. n. equus, equi a horse; Gr. n. rhis, rhinos nose;
N.L. gen. n. equirhinis of the nose of a horse.
40. Mycoplasma elephantis Kirchhoff, Schmidt, Lehmann,
Clark and Hill 1996, 440VP
43. Mycoplasma falconis Poveda, Giebel, Flossdorf, Meier and
e.le.phan¢tis. L. n. elephas, -antis elephant; L. gen. n. elephantis of the elephant.
fal.co¢nis. L. gen. n. falconis of the falcon, the host from
which the organism was first isolated.
Cells are coccoidal. Nonmotile. Colonies on solid
medium show a typical fried-egg appearance. Grows well in
Hayflick medium supplemented with glucose at 37°C.
Probable commensal. No pathology was observed at the
site of isolation (i.e., the vagina and urethra); however,
isolation was achieved almost exclusively from arthritic
animals with evidence of rheumatoid factor. The possibility thus exists that the clinical status of the animals was
due to sexually acquired reactive arthritis, which has been
observed with other Mycoplasma species known to parasitize
the urogenital tract (Blanchard and Bébéar, 2002). Mode of
transmission has not been established definitively.
Source: isolated from the vagina and urethra of captive
elephants (Clark et al., 1980, 1978).
Type strain: E42, ATCC 51980.
assessed. Colonies on solid medium have a fried-egg appearance. Grows well in modified Frey’s medium supplemented
with arginine at 37°C.
Pathogenicity has not been established. Associated with
respiratory tract infections of saker falcons, although can
also be isolated from clinically normal birds. Mode of transmission has not been established definitively.
Source: isolated from the trachea of falcons (Lierz et al.,
2002, 2008a, b).
Type strain: H/T1, ATCC 51372.
41. Mycoplasma equigenitalium Kirchhoff 1978, 500AL
e.qui.ge.ni.ta¢li.um. L. n. equus, equi the horse; L. pl. n. genitalia the genitals; N.L. pl. gen. n. equigenitalium of equine
genitalia.
Cells are coccoid to coccobacillary. Motility for this species has not been assessed. Colonies on solid medium show
a typical fried-egg appearance. Grows in SP-4 or Hayflick
Opportunistic pathogen; associated with rhinitis and
pneumonitis in horses, but can also be found in clinically
normal animals. Mode of transmission is via droplet aerosol.
Source: isolated from the nasopharynx, nasal turbinates,
trachea, tonsils, and semen of horses, and from the
nasopharynx of cattle (Lemcke, 1979; Spergser et al., 2002;
ter Laak et al., 1992a).
Type strain: M432/72, ATCC 29420, NCTC 10148.
44. Mycoplasma fastidiosum Lemcke and Poland 1980, 161VP
fas.ti.di.o¢sum. L. neut. adj. fastidiosum fastidious, referring
to the nutritionally fastidious nature of the organism on primary isolation.
Cells are highly filamentous and filaments are twisted at
regular intervals along their length. Helical forms like those
of Spiroplasma species are not produced. Nonmotile. Colonies on solid medium show a typical fried-egg appearance.
596
Grows in SP-4 or Frey’s medium supplemented with glucose
at 37°C.
Source: isolated from the nasopharynx of horses (Lemcke
and Poland, 1980).
Type strain: 4822, NCTC 10180, ATCC 33229.
45. Mycoplasma faucium Freundt, Taylor-Robinson, Purcell,
Chanock and Black 1974, 253AL
fau¢ci.um. L. pl. n. fauces, -ium the throat; L. gen. pl. n. faucium of the throat.
Cells are coccoidal. Motility for this species has not been
assessed. Colonies on solid medium show a typical friedegg appearance, but are more loosely attached to the agar
surface than are the colonies of most other mycoplasmas.
Grows well in SP-4 medium supplemented with arginine at
Probable commensal. Most commonly found as a commensal of the human oropharynx; however, recent isolations of Mycoplasma faucium have been made from brain
abscesses (Al Masalma et al., 2009). Mode of transmission
Source: isolated from the oropharynx and brain of
humans, and from the oral cavity of numerous species of
nonhuman primates (Freundt et al., 1974; Somerson and
Cole, 1979).
Type strain: DC-333, ATCC 25293, NCTC 10174.
46. Mycoplasma felifaucium Hill 1988, 449VP (Effective publication: Hill 1986, 1927)
fe.li.fau¢ci.um. L. n. felis cat; L. pl. n. fauces, -ium throat; N.L.
gen. pl. n. felifaucium of the feline throat.
Cells are primarily coccoidal. Nonmotile. Colonies on
solid medium show a typical fried-egg appearance. Grows
well in SP-4 or Hayflick medium supplemented with arginine at 37°C. Produces a “film and spots” reaction.
Source: isolated from the oropharynx of captive pumas
(Felis concolor; Hill, 1986).
Type strain: PU, NCTC 11703, ATCC 43428.
47. Mycoplasma feliminutum Heyward, Sabry and Dowdle
1969, 621AL
fe.li.mi.nu¢tum. L. n. felis a cat; L. neut. part. adj. minutum
small; N.L. neut. adj. feliminutum a small colony organism
isolated from cats.
Morphology is poorly defined. Motility for this species has
not been assessed. Colonies are relatively small and irregular in shape. Grows well in SP-4 medium supplemented with
glucose at 37°C.
Source: isolated from the oropharynx of domestic cats;
from the nasopharynx, lungs, and urogenital tract of domestic dogs; and from the respiratory tract of horses (Chalker,
2005; Heyward et al., 1969; Lemcke, 1979).
Type strain: Ben, ATCC 25749, NCTC 10159.
Further comment: this organism was first described during
a time when the only named genus of mollicutes was Mycoplasma. Its publication coincided with the first proposal of
the genus Acholeplasma (Edward and Freundt, 1969, 1970),
with which Mycoplasma feliminutum is properly affiliated
through established phenotypic (Heyward et al., 1969)
and 16S rRNA gene sequence (Brown et al., 1995) similarities. This explains the apparent inconsistencies with its
assignment to the genus Mycoplasma. The name Mycoplasma
feliminutum should therefore be revised to Acholeplasma
feliminutum comb. nov.
48. Mycoplasma felis Cole, Golightly and Ward 1967, 1456AL
fe¢lis. L. n. felis a cat, L. gen. n. felis of a cat.
Cells are coccobacillary to filamentous. Motility for this
species has not been assessed. Colonies on solid media display the typical fried-egg morphology. Grows well in SP-4
Pathogenic; associated with conjunctivitis, rhinitis, ulcerative keratitis, and polyarthritis in domestic cats, and upper
and lower respiratory tract infection in horses. Mycoplasma
felis can also be isolated from clinically normal domestic
cats, domestic dogs, and horses. The mode of transmission
Source: isolated from the conjunctivae, nasopharynx,
lungs, and urogenital tract of domestic cats; from the lungs,
tonsils, trachea nasopharynx of horses; from the oropharynx and trachea of domestic dogs; and from the synovial
fluid of an immunocompromised human (Lemcke, 1979)
Røsendal, 1979; (Bonilla et al., 1997; Gray et al., 2005;
Hooper et al., 1985).
Type strain: CO, ATCC 23391, NCTC 10160.
Further comment: the proposed species “Mycoplasma
equipharyngis” (Kirchoff, 1974) has been reported in horses.
Further characterization has demonstrated unequivocally
that these isolates are Mycoplasma felis and all mention of
“Mycoplasma equipharyngis” should be considered equivalent
to Mycoplasma felis (Lemcke, 1979).
49. Mycoplasma fermentans Edward 1955, 90AL
fer.men¢tans. L. part. adj. fermentans fermenting.
Cells are filamentous. Motility has not been established
for this species. Colonies on solid media display typical friedegg morphology. Grows well in SP-4 or Hayflick medium
supplemented with either arginine or glucose at 37°C.
Pathogenicity unclear; associated with balanitis, vulvovaginitis, salpingitis, respiratory distress syndrome, pneumonia, and development of rheumatoid arthritis. Mycoplasma
fermentans has also been tenuously linked with the progression of AIDS, chronic fatigue syndrome, Gulf War syndrome, Adamantiades-Behçet’s disease, and fibromyalgia.
Genus I. Mycoplasma
The connection of the preceding clinical syndromes with
Mycoplasma fermentans is highly equivocal, as different studies have reached markedly different conclusions. Mode of
Source: isolated from the urine, urethra, rectum, penis,
cervix, vagina, fallopian tube, amniotic fluid, blood, synovial
fluid, and throat of humans; from the cervix of an African
green monkey (Chlorocebus sp.); and from the vagina of a
sheep (Blanchard et al., 1993; Nicholas et al., 1998; TaylorRobinson and Furr, 1997; Waites and Talkington, 2005).
DNA G+C content (mol%): 28.7 (Tm)
14854.
50. Mycoplasma flocculare Meyling and Friis 1972, 289AL
floc.cu.la¢re. L. dim. n. flocculus a small flock or tuft of wool;
L. neut. suff. -are suffix denoting pertaining to; N.L. neut.
adj. flocculare resembling a small floc of wool, referring to
the tendency of the organism to form clumps of flocculent
material in broth culture.
Cells are coccoid to coccobacillary. Motility for this species
has not been assessed. Colonies on solid media are slightly
convex with a coarsely granular surface and lack a defined
center. Aggregates of cells may be produced during growth
in broth, appearing as small floccular elements upon gentle
shaking of the culture. Grows slowly in Friis medium at 37°C.
Opportunistic pathogen; normally regarded as a commensal of the nasopharynx that can cause pneumonia in
association with other pathogens, most notably Mycoplasma
hyopneumoniae. Mode of transmission is via droplet aerosol.
Mycoplasma flocculare reportedly shares surface antigens
with Mycoplasma hyopneumoniae, potentially complicating
serology-based diagnosis of infection (Whittlestone, 1979).
Source: isolated from the nasopharynx, lungs, pericardium, and conjunctivae of pigs.
Type strain: Ms42, ATCC 27399, NCTC 10143.
51. Mycoplasma gallinaceum Jordan, Ernø, Cottew, Hinz and
gal.li.na¢ce.um. L. neut. adj. gallinaceum pertaining to a
domestic fowl.
Cells are coccoid to coccobacillary. Motility for this species has not been assessed. Colonies on solid medium have
typical fried-egg morphology although some are devoid of
a central core. Grows well in Frey’s medium supplemented
Opportunistic pathogen associated with tracheitis, airsacculitis, or conjunctivitis in chickens, turkeys, ducks, and
pheasants. Mycoplasma gallinaceum has been reported to
complicate cases of infectious synovitis due to Mycoplasma
synoviae in chickens. The mode of transmission has not
been assessed definitively.
Source: isolated from upper and lower respiratory tract of
chickens, turkeys, pheasants, partridges, and ducks; from
the conjunctivae of pheasants; and from the synovial fluid
of chickens (Bradbury et al., 2001; Tiong, 1990; Welchman
et al., 2002; Yagihashi et al., 1983).
597
Type strain: DD, ATCC 33550, NCTC 10183.
Further comment: previously known as avian serotype D
(Kleckner, 1960).
52. Mycoplasma gallinarum Freundt 1955, 73AL
gal.li.na¢rum. L. n. gallina a hen; L. gen. pl. n. gallinarum
of hens.
Cells are coccoid to coccobacillary. Nonmotile. Colonies on solid medium have a typical fried-egg appearance.
Grows well in Frey’s medium supplemented with arginine
at 37°C. The organism shares some antigens in immunodiffusion tests with Mycoplasma iners, Mycoplasma columbinasale,
and Mycoplasma meleagridis.
Commensal of gallinaceous birds; little evidence exists
for the pathogenicity of isolates in such hosts. Mycoplasma
gallinarum may have a role in airsacculitis of geese and participate in complex infection of chickens. Mode of transmission has not been assessed definitively.
Source: isolated from the respiratory tract of chickens, turkeys, ducks, geese, red jungle fowl, bamboo partridge, sparrow, swan, and demoisella crane; and from sheep (Kisary
et al., 1976; Kleven et al., 1978; Shimizu et al., 1979; Singh
and Uppal, 1987).
DNA G+C content (mol%): 26.5–28.0 (Tm, Bd).
Type strain: PG16, ATCC 19708, NCTC 10120.
Further comment: previously known as avian serotype B
(Kleckner, 1960).
53. Mycoplasma gallisepticum Edward and Kanarek 1960,
699AL
gal.li.sep¢ti.cum. L. n. gallus rooster, chicken; L. adj. septicus -a -um producing a putrefaction, putrefying, septic; N.L.
neut. adj. gallisepticum hen-putrefying (infecting).
Cells are coccoid, ovoid, and elongated pear-shaped with
a highly structured polar body, called the bleb. Cells are
motile and glide in the direction of the terminal bleb. Gliding speed varies among strains. Colonies on solid medium
may be small and not necessarily of typical fried-egg appearance. Grows well in SP-4 or Hayflick medium supplemented
with glucose at 37°C (Balish and Krause, 2006; Hatchel
et al., 2006; Nakane and Miyata, 2009).
Pathogenic. Causes a characteristic combination of
pneumonia, tracheitis, and airsacculitis (collectively
termed chronic respiratory disease); salpingitis and atrophy of the ovaries, isthmus, and cloaca resulting in poor
egg quality and reduced hatchability; arthritis or synovitis;
and keratoconjunctivitis in chickens; infectious sinusitis,
coryza, airsacculitis, arthritis or synovitis, encephalitis,
meningitis, ataxia, and torticollis in turkeys; conjunctivitis and coryza featuring a high mortality rate in finches
and grosbeaks; and respiratory disease in additional game
birds including quail, partridges, pheasants, and peafowl. Lesions established by Mycoplasma gallisepticum are
often complicated by additional avian pathogens including Mycoplasma synoviae, avian strains of Escherichia coli,
Newcastle disease virus, and infectious bronchitis virus.
Established mechanisms of transmission include droplet
598
aerosols, direct contact with infected animals or fomites,
and vertical transmission.
Tetracyclines, macrolides, aminoglycosides, fluoroquinolones, and pleuromutilins are effective chemotherapeutic agents; however, treatment is typically only sought for
individual birds, as medicating a commercial flock is not
considered an effective control strategy. Vaccination and
management strategies (i.e., single age “all in/all out” systems and culling of endemic flocks) are more commonly
utilized. Multiple live and killed vaccines are commercially
available, but suffer from residual pathogenicity, the need
to develop a carrier state to provide protective immunity,
adverse reactions, or low efficacy. Experimental vaccines
have also been described. Mycoplasma gallisepticum shares
surface antigens with Mycoplasma imitans and Mycoplasma
synoviae, potentially complicating serology-based diagnosis
of infection. Numerous molecular diagnostics have been
described. This organism is listed in the Terrestrial Animal
Health Code of the Office International des Epizooties
(http://oie.int; Yogev et al., 1989; Kempf, 1998; Markham
et al., 1999; Gautier-Bouchardon et al., 2002; Ferguson et al.,
2004; Browning et al., 2005; Crespo and McMillan, 2008;
Gates et al., 2008; Gerchman et al., 2008; Kleven, 2008).
Source: isolated from the trachea, lungs, air sacs, ovaries,
oviducts, brain, arterial walls, synovial membranes, synovial
fluid, conjunctivae, and eggs of chickens; from the infraorbital sinuses, air sacs, brain, meninges, conjunctivae, synovial membranes, and synovial fluid of turkeys; from the
conjunctivae, infraorbital sinuses, and trachea of finches;
and from the respiratory tract of quail, partridges, pheasants, peafowl, ducks, grosbeak, crows, robins, and blue jays
(Benčina et al., 2003, 1988; Bradbury and Morrow, 2008;
Bradbury et al., 2001; Dhondt et al., 2005, 2007; Levisohn
and Kleven, 2000; Ley et al., 1996; Mikaelian et al., 2001;
Murakami et al., 2002; Nolan et al., 2004; Nunoya et al.,
1995; Welchman et al., 2002; Wellehan et al., 2001).
DNA G+C content (mol%): 31.8 (Tm), 31 (strain R complete genome sequence).
Type strain: PG31, X95, ATCC 19610, NCTC 10115.
Sequence accession nos: M22441 (16S rRNA gene of
strain A5969), NC_004829 (strain R complete genome
sequence).
Further comment: previously known as avian serotype A
(Kleckner, 1960).
54. Mycoplasma gallopavonis Jordan, Ernø, Cottew, Hinz and
gal.lo.pa.vo¢nis. N.L. n. gallopavo, -onis a turkey (Meleagris
gallopavo); N.L. gen. n. gallopavonis of a turkey.
Cells are coccoid to coccobacillary. Motility has not been
assessed for this species. Colonies on solid medium have
typical fried-egg morphology. Grows well in Frey’s medium
supplemented with glucose at 37°C.
Opportunistic pathogen; occasionally associated with
airsacculitis in turkeys, but is also isolated from clinically
normal turkeys. Mode of transmission has not been assessed
definitively.
Source: isolated from the choanae, trachea, and air sacs of
domestic and wild turkeys (Benčina et al., 1987; Cobb et al.,
1992; Hoffman et al., 1997; Luttrell et al., 1992).
Type strain: WR1, ATCC 33551, NCTC 10186.
Further comment: previously known as avian serotype F
(Kleckner, 1960).
55. Mycoplasma gateae Cole, Golightly and Ward 1967, 1456AL
ga.te¢ae. N.L. gen. n. gateae (probably from Spanish gato, a
cat) of a cat.
Morphology is poorly defined. Motility for this species
has not been assessed. Colonies on solid medium are vacuolated and lack a well-defined central spot. Grows well in
SP-4 medium at 37°C.
Opportunistic pathogen; can cause polyarthritis in
domestic cats (Moise et al., 1983), but appears to be primarily a commensal species of the oral cavity. Mycoplasma
gateae also appears to be a commensal species of domestic
dogs and cattle. Mode of transmission has not been assessed
definitively.
Source: isolated from the synovial membrane, oropharynx, saliva, and urogenital tract of domestic cats; from the
lungs, oropharynx, trachea, and urogenital tract of domestic dogs; and from the urogenital tract of cattle (Chalker,
2005; Gourlay and Howard, 1979; Røsendal, 1979).
Type strain: CS, ATCC 23392, NCTC 10161.
Further comment: the original specific epithet “gateae”,
which has been perpetuated in lists of bacterial names
approved by the International Committee on Systematics of
Prokaryotes, the American Type Culture Collection’s Catalog of Bacteria and Bacteriophages, and in GenBank, is illegitimate because the genitive of the medieval Latin word gata
(female cat) would have been gatae and there is no word for
which gateae would have been a legitimate genitive (Brown
et al., 1995).
56. Mycoplasma genitalium Tully, Taylor-Robinson, Rose, Cole
and Bové 1983, 395VP
ge.ni.ta¢li.um. L. pl. n. genitalia, -ium the genitals; L. gen. pl.
n. genitalium of the genitals.
Cells are predominantly flask-shaped with a terminal
organelle protruding from the cell pole that is narrower
than that of Mycoplasma gallisepticum and shorter than that
of Mycoplasma pneumoniae. The leading end of the terminal
structure is often curved. Cells exhibit gliding motility in
circular patterns and glide in the direction of the terminal
organelle’s curvature (Hatchel and Balish, 2008). Colonies
on solid media are round and possess a defined center that
is somewhat less distinct than most mycoplasma species.
Grows well in SP-4 medium supplemented with glucose at
37°C.
Pathogenic; causes urethritis, cervicitis, endometritis,
and pelvic inflammatory disease. Mycoplasma genitalium is
associated with infertility in humans. Mode of transmission is via sexual contact, congenitally, and possibly, in rare
instances, via droplet aerosol.
Macrolides and fluoroquinolones are effective chemotherapeutic agents; however, reports indicate that treatment should be extensive, as clinical signs and detection
Genus I. Mycoplasma
of Mycoplasma genitalium tend to recur following cessation.
This is potentially due to sequestration within host cells.
Mycoplasma genitalium shares numerous surface antigens
with Mycoplasma pneumoniae, complicating serology-based
diagnosis of infection. Numerous molecular diagnostics
have been described, but few have been developed commercially (Jensen, 2004; Waites and Talkington, 2005).
Source: isolated or detected in the urogenital tract, urine,
rectum, synovial fluid, conjunctiva, and nasopharynx of
humans (Baseman et al., 1988; de Barbeyrac et al., 1993;
Jensen, 2004; Waites and Talkington, 2005).
DNA G+C content (mol%): 32.4 (Bd), 31 (strain G-37T complete genome sequence).
Type strain: G-37, ATCC 33530, CIP 103767, NCTC
10195.
Sequence accession nos: X77334 (16S rRNA gene),
NC_000908 (strain G-37T complete genome sequence),
CP000925 (strain JCVI-1.0 complete genome sequence).
599
59. Mycoplasma haemocanis (Kikuth 1928) Messick, Walker,
Raphael, Berent and Shi 2002, 697VP [Bartonella canis Kikuth
1928, 1730; Haemobartonella (Bartonella) canis (Kikuth 1928)
Tyzzer and Weinman 1939, 151; Kreier and Ristic 1984, 726]
ha.e.mo.ca¢nis Gr. neut. n. haema blood; L. fem. gen. n. canis
of the dog; N.L. gen. n. haemocanis of dog blood.
57. Mycoplasma glycophilum Forrest and Bradbury 1984, 355VP
(Effective publication: Forrest and Bradbury 1984, 602)
Cells are coccoid to pleomorphic. Motility for this species
has not been assessed. The morphology of infected erythrocytes is altered, demonstrating a marked depression at the
site of Mycoplasma haemocanis attachment. This species has
not been grown on any artificial medium; therefore, notable biochemical parameters are not known.
Pathogenic; causes hemolytic anemia in domestic dogs.
Transmission is vector-borne and mediated by the brown
dog tick (Rhipicephalus sanguineus).
Source: observed in association with erythrocytes of
domestic dogs (Hoskins, 1991).
Type strain: not established.
gly.co.phi¢lum. Gr. adj. glykys sweet (this adjective was used
to coin the noun glucose); N.L. neut. adj. philum (from Gr.
neut. adj. philon) friend, loving; N.L. neut. adj. glycophilum
sweet-loving, intended to mean glucose-loving.
60. Mycoplasma haemofelis (Clark 1942) Neimark, Johansson,
Rikihisa and Tully 2002b, 683VP [Eperythrozoon felis Clark
1942, 16; Haemobartonella felis (Clark 1942) Flint and
McKelvie 1956, 240 and Kreier and Ristic 1984, 725]
Cells are spherical or elliptical with an extracellular
layer. Nonmotile. Growth on solid medium shows colonies
with typical fried-egg appearance. Grows well in Hayflick
Pathogenicity has not been established, although there
may be an association with a slight decrease in hatchability.
Mode of transmission has not been assessed definitively.
Source: isolated from the respiratory tract and cloaca of
chickens; and from the respiratory tract of turkeys, pheasants, partridges, ducks, and geese (Forrest and Bradbury,
1984).
58. Mycoplasma gypis Poveda, Giebel, Flossdorf, Meier and
gy¢pis. Gr. n. gyps, gypos vulture; N.L. gen. n. gypis of the vulture, the host from which the organism was first isolated.
Cells are coccoid or round. Motility for this species has
not been assessed. Colonies on solid medium have a friedegg appearance. Grows in Frey’s medium supplemented
with arginine at 37°C. Produces a “film and spots” reaction.
Pathogenicity has not been established. Associated with
respiratory tract disease of griffon vultures, but has also
been isolated from healthy birds of prey. Mode of transmission has not been assessed definitively.
Source: isolated from the trachea of griffon vultures (Griffon fulvus), and from the trachea and air sacs of Eurasian
buzzards, red kites, and Western marsh harriers (Lierz
et al., 2008a, 2000; Poveda et al., 1994).
Type strain: B1/T1, ATCC 51370.
ha.e.mo.fe¢lis. Gr. neut. n. haema blood; L. fem. gen. n. felis
of the cat; N.L. gen. n. haemofelis of cat blood.
assessed. This species has not been grown on artificial
medium; therefore, notable biochemical parameters are
not known.
Pathogenic; causes hemolytic anemia in cats. The mode
of transmission is percutaneous or oral; an insect vector has
not been identified, although fleas have been implicated
(Woods et al., 2005).
Tetracyclines and fluoroquinolones are effective therapeutic agents (Dowers et al., 2002; Tasker et al., 2006).
domestic cats.
DNA G+C content (mol%): 38.5–38.8 (genome sequence
survey of strain OH; Berent and Messick, 2003; J.B. Messick
et al., unpublished).
Type strain: not established
61. Mycoplasma haemomuris (Mayer 1921) Neimark, Johansson, Rikihisa and Tully 2002b, 683VP (Bartonella muris Mayer
1921, 151; Bartonella muris ratti Regendanz and Kikuth 1928,
1578; Haemobartonella muris Tyzzer and Weinman 1939,
143)
ha.e.mo.mu¢ris. Gr. neut. n. haema blood; L. masc. gen. n.
muris of the mouse; N.L. gen. n. haemomuris of mouse blood.
Cells are coccoid and some display dense inclusion particles. Motility for this species has not been assessed. The
morphology of infected erythrocytes is altered, demonstrating a marked depression at the site of Mycoplasma haemomuris attachment. This species has not been grown on any
artificial medium; therefore, notable biochemical para
meters are not known.
600
Opportunistic pathogen; causes anemia in splenectomized
or otherwise immunosuppressed mice. Transmission is vectorborne and mediated by the rat louse (Polypax spinulosa).
Source: observed in association with erythrocytes of wild
and captive mice, and hamsters.
62. Mycoplasma hominis (Freundt 1953) Edward 1955, 90AL
(Micromyces hominis Freundt 1953, 471)
ho¢mi.nis. L. n. homo, -inis man; L. gen. n. hominis of man.
Cells are coccoid to filamentous. Motility for this species has not been assessed. Colonies on solid media have
a typical fried-egg appearance. Grows well in SP-4 medium
supplemented with arginine at 37°C.
Pathogenic; causes pyelonephritis, pelvic inflammatory disease, chorioamnionitis, and postpartum fevers in
women; congenital pneumonia, meningitis, and abscesses
in newborns; and rarely extragenital pathologies including
bacteremia, arthritis, osteomyelitis, abscesses and wound
infections, mediastinitis, pneumonia, peritonitis, prosthetic- and catheter-associated infections, and infection
of hematomas. Extragenital manifestations of Mycoplasma
hominis infection are more commonly seen in immunosuppressed individuals, but can be seen in immunocompetent
patients as well. Synergism between Mycoplasma hominis and
Trichomonas vaginalis infections has been reported and a
recent report documents the intraprotozooal location and
transmission of Mycoplasma hominis with Trichomonas vaginalis (Dessi et al., 2006; Germain et al., 1994; Vancini and
Benchimol, 2008). Mode of transmission is via sexual contact, congenitally, or by artificial introduction on foreign
objects (e.g., catheters) or transplanted tissues.
Macrolides and fluoroquinolones are effective chemotherapeutic agents. Combination therapy with metronidazole is required for complex infections involving Trichomonas
vaginalis. Many commercial diagnostics are available for
routine clinical use.
Source: isolated from the urogenital tract, amniotic fluid,
placenta, umbilical cord blood, urine, semen, bloodstream,
cerebrospinal fluid, synovial fluid, bronchoalveolar lavage
fluid, peritoneal aspirates, conjunctivae, bone abscesses,
and hematoma aspirates of humans; and from several
species of nonhuman primates (Somerson and Cole,
1979; Taylor-Robinson and McCormack, 1979; Waites and
Talkington, 2005).
DNA G+C content (mol%): 33.7 (Tm)
Type strain: PG21, ATCC 23114, NCTC 10111, CIP 103715,
NBRC 14850.
63. Mycoplasma hyopharyngis Erickson, Ross, Rose, Tully and
Bové 1986, 58VP
hy.o.pha.ryn¢gis. Gr. n. hys, hyos a swine; N.L. n. pharynx
-yngis (from Gr. n. pharugx, pharuggos throat) throat; N.L.
gen. n. hyopharyngis of a hog’s throat.
Cells are pleomorphic. Motility for this species has not
been assessed. Colonies on solid medium have a typical
fried-egg appearance. Grows in medium D-TS or Hayflick
medium supplemented with arginine at 37°C and produces
a “film and spots” reaction.
Source: isolated from the nasopharynx of pigs (Erickson
et al., 1986).
DNA G+C content (mol%): 24 (Bd, Tm).
Type strain: H3-6B F, ATCC 51909, NCTC 11705.
64. Mycoplasma hyopneumoniae (Goodwin, Pomeroy and
Whittlestone 1965) Maré and Switzer 1965, 841AL (Mycoplasma suipneumoniae Goodwin, Pomeroy and Whittlestone
1965, 1249)
hy.o.pneu.mo¢ni.ae. Gr. n. hys, hyos a swine; Gr. n. pneumonia
pneumonia; N.L. gen. n. hyopneumoniae of swine pneumonia.
Cells are coccoid to coccobacillary. Nonmotile. Colonies on solid medium are very small, lack a defined central region, and are usually convex with a granular surface.
Grows very slowly in modified Friis medium, medium A26,
and modified SP-4 medium supplemented with glucose at
Pathogenic; causes a very characteristic chronic pneumonitis associated with ciliostasis and marked sloughing
of the epithelial lining in pigs. This collection of lesions in
conjunction with Mycoplasma hyopneumoniae is referred to as
enzootic pneumonia of pigs (EPP), and is associated with
high morbidity and poor feed conversion (with proportional economic loss). The mechanism of transmission is
via droplet aerosol.
Tetracyclines and tylosin do not typically eradicate
Mycoplasma hyopneumoniae from infected animals, but are
effective in limiting sequelae. Maintenance of pigs on antibiotics in conjunction with management practices involving adequate nutrition, air quality, and stress reduction
are commonly employed to control the effects of disease
in endemic herds. Many molecular diagnostics have been
described (Dubosson et al., 2004). Serological diagnostic
techniques utilizing monoclonal antibodies have proven
successful at distinguishing Mycoplasma hyopneumoniae from
Mycoplasma flocculare, though the two share surface antigens
(Armstrong et al., 1987). Numerous experimental vaccines
have been described and several commercial vaccines are
available. The latter appear to reduce or eliminate clinical
signs rather than prevent infection (Browning et al., 2005).
Source: isolated from the lungs, nasopharynx, tonsils, trachea, and bronchiolar lavage fluid of pigs (Marois et al.,
2007; Whittlestone, 1979).
DNA G+C content (mol%): 27.5 (Bd), 28 (strain JT and
strain 7448 complete genome sequence), 28.6 (strain 232
Type strain: J, ATCC 25934, NCTC 10110.
Sequence accession nos: AY737012 (16S rRNA gene),
AE017243 (strain JT complete genome sequence), AE017244
(strain 7448 complete genome sequence), AE017332 (strain
232 complete genome sequence).
65. Mycoplasma hyorhinis Switzer 1955, 544AL
hy.o.rhi¢nis. Gr. n. hys, hyos a swine; Gr. n. rhis, rhinos nose;
N.L. gen. n. hyorhinis of a hog’s nose.
Genus I. Mycoplasma
Cells are coccoid to coccobacillary. Motility for this species
has not been assessed. Colonies on solid media display typical fried-egg morphology. Grows well on S-4 supplemented
Mycoplasma hyorhinis is associated with contamination
of eukaryotic cell culture and can be removed by treatment of cells with antibiotics and/or maintenance of cell
lines in antibiotic-containing medium. The noted effects
of Mycoplasma hyorhinis on cell-cycle regulation make the
detection and elimination of this organism particularly pertinent (Goodison et al., 2007; Schmidhauser et al., 1990).
The most effective classes of antibiotics for cell culture
eradication are tetracyclines and fluoroquinolones (BorupChristensen et al., 1988; Schmitt et al., 1988).
Pathogenic; associated with arthritis, polyserositis, and
otitis media in pigs. Mycoplasma hyorhinis is also regarded
as a commensal of the nasopharynx that can occasionally
cause pneumonia, often in association with other pathogens (most notably Mycoplasma hyopneumoniae and Bordetella
bronchiseptica). Mode of transmission is via droplet aerosol.
Source: isolated from the nasopharynx, lungs, ear canal,
synovial fluid, serous cavity, and pericardium of pigs (Friis
and Szancer, 1994; Ross, 1992; Whittlestone, 1979).
Type strain: BTS-7, ATCC 17981, NCTC 10130, CIP
104968, NBRC 14858.
66. Mycoplasma hyosynoviae Ross and Karmon 1970, 710AL
hy.o.sy.no.vi¢ae. Gr. n. hys, hyos a swine; N.L. n. synovia fluid
in the joints; N.L. gen. n. hyosynoviae of joint fluid of swine.
Cells are coccoid to filamentous. Motility of this species
has not been assessed. Colonies display a typical fried-egg
appearance at 37°C. Grows well in SP-4 medium supplemented with glucose at 37°C. A granular deposit and a waxy
surface pellicle are produced during growth in broth.
Pathogenic; causes infectious synovitis, arthritis, and
rarely pericarditis in pigs. Transmission occurs from sows to
piglets or between adults via aerosol.
Lincosamides, fluoroquinolones, and macrolides have
been used effectively for treatment in conjunction with
improved disinfection and quarantine during husbandry.
Source: isolated from the synovial fluid, nasopharynx, tonsils, lymph nodes, and pericardium of pigs (Whittlestone,
1979).
Type strain: S16, ATCC 25591, NCTC 10167.
67. Mycoplasma iguanae Brown, Demcovitz, Plourdé, Potter,
Hunt, Jones and Rotstein 2006, 763VP
i.gua¢nae. N.L. gen. n. iguanae of the iguana lizard.
Cells are predominantly coccoid. Nonmotile. Colonies on
solid medium exhibit variable (convex to umbonate) forms;
mature colonies display sectored centers. Grows well in SP-4
medium supplemented with glucose between 25 and 42°C.
Associated with pathologic lesions, but unable to reproduce disease following experimental inoculation (Brown
et al., 2007). Mechanism of transmission has not been established.
601
Source: isolated from vertebral abscesses of green
iguanas.
Type strain: 2327, ATCC BAA-1050, NCTC 11745.
Sequence accession no. (partial 16S rRNA gene sequence):
AY714305.
68. Mycoplasma imitans Bradbury, Abdul-Wahab, Yavari,
Dupiellet and Bové 1993, 726VP
i¢mi.tans. L. part. adj. imitans imitating, mimicking, referring to the organism’s phenotypic resemblance to Mycoplasma gallisepticum.
Cells are oval and flask-shaped with a short, wide attachment organelle. Cells are motile and exhibit gliding motility in the direction of the attachment organelle (Hatchel
and Balish, 2008). Colonies have typical fried-egg morphology on solid medium. Grows well in SP-4 medium supplemented with glucose at 37°C.
Pathogenic; causes sinusitis in ducks, geese, and partridges. Disease has been reproduced experimentally. Mode
of transmission has not been assessed definitively.
Diagnosis of infection is potentially complicated by
numerous factors. Serological cross-reactions occur with
Mycoplasma gallisepticum due to known epitopes including
PvpA, the VlhA hemagglutinins, pyruvate dehydrogenase,
lactate dehydrogenase, and elongation factor Tu (Jan et al.,
2001; Lavrič et al., 2005; Markham et al., 1999; Rosengarten
et al., 1995). In addition, the 16S rRNA genes share 99.9%
identity, despite whole genome hybridization showing a
relatedness of only 40–46%, potentially complicating molecular diagnostics based on the 16S rRNA gene. Two molecular methods for distinguishing these two species have been
described (Harasawa et al., 2004; Marois et al., 2001).
Source: isolated from the nasal turbinates and sinuses of
ducks, geese, and partridges (Dupiellet, 1984; Ganapathy
and Bradbury, 1998; Kleven, 2003).
69. Mycoplasma indiense Hill 1993, 39VP
in.di.en¢se. N.L. neut. adj. indiense pertaining to India
(source of the infected primates).
Cells are pleomorphic. Nonmotile. Colonies on agar
have a characteristic fried-egg appearance. Grows well in
Source: isolated from the throats of a rhesus monkey and
a baboon (Hill, 1993).
Type strain: 3T, NCTC 11728, ATCC 51125.
70. Mycoplasma iners Edward and Kanarek 1960, 699AL
i¢ners. L. neut. adj. iners inactive, inert.
The cell morphology is poorly defined. Motility for this
species has not been assessed. Colonies on solid medium are
relatively small and of a typical fried-egg appearance. Grows
in Frey’s medium supplemented with arginine at 37°C.
602
Source: isolated from the respiratory tract of chickens, turkeys, geese pigeons, pheasants, and partridges; and from
tissues of swine (Benčina et al., 1987; Bradbury et al., 2001;
Taylor-Robinson and Dinter, 1968).
DNA G+C content (mol%): 29.1 (Tm), 29.6 (Bd).
Further comment: previously known as avian serotype E
(Kleckner, 1960).
71. Mycoplasma iowae Jordan, Ernø, Cottew, Hinz and Stipkovits 1982, 114VP
i.o¢wa.e. N.L. gen. n. iowae of Iowa.
Cells are pleomorphic and some display a terminal
rotrusion with possible attachment properties (Gallagher
p
and Rhoades, 1983; Mirsalimi et al., 1989). Motile. Colonies
on agar show typical fried-egg appearance. Grows well in
SP-4 medium supplemented with either glucose or arginine
at 41–43°C (Grau et al., 1991; Yoder and Hofstad, 1964).
Pathogenic; causes airsacculitis and embryo lethality
resulting in reduced hatchability in turkeys. Transmission
occurs vertically and by direct contact.
Tetracyclines, macrolides, and fluoroquinolones are
effective chemotherapeutic agents; however, medicating
a commercial flock is not considered an effective control
strategy. Management tactics are more commonly utilized. Molecular diagnostic methods have been described
(Ramírez et al., 2008; Raviv and Kleven, 2009). Mycoplasma
iowae strains show considerable intra-species antigenic
heterogeneity and a cross-reactive epitope with both Mycoplasma gallisepticum and Mycoplasma imitans potentially complicating serology-based diagnosis of infection (Al-Ankari
and Bradbury, 1996; Dierks et al., 1967; Rosengarten et al.,
1995).
Source: isolated from the air sacs, intestinal tract, and eggs
of turkeys, and from the seed of an apple tree with apple
proliferation disease (Bradbury and Kleven, 2008; Grau
et al., 1991; Mirsalimi et al., 1989).
Further comment: previously known as avian serotype I
(Yoder and Hofstad, 1964).
72. Mycoplasma lagogenitalium Kobayashi, Runge, Schmidt,
Kubo, Yamamoto and Kirchhoff 1997, 1211VP
la.go.ge.ni.ta¢li.um. Gr. masc. n. lagos hare; L. neut. pl. gen.
n. genitalium of genitals; N.L. gen. pl. n. lagogenitalium of
hare’s genitals.
Cells are primarily coccoid. Nonmotile. Colonies on agar
have a characteristic fried-egg appearance. Grows well in
Source: isolated from the preputial smegma of Afghan
pikas (Ochotona rufescens; Kobayashi et al., 1997).
Type strain: 12MS, ATCC 700289, CIP 105489, DSM
22062.
73. Mycoplasma leachii Manso-Silván, Vilei, Sachse, Djordjevic,
Thiaucourt and Frey 2009, 1356VP
le.a.chi¢i. N.L. masc. gen. n. leachii of Leach, named in
honor of Dr R.H. Leach, who first characterized this taxon.
Cells are pleomorphic. Nonmotile. Colonies on solid agar
have a characteristic fried-egg appearance. Grows in modified Hayflick medium supplemented with glucose at 37°C.
Pathogenic; causes polyarthritis, mastitis, abortion, and
pneumonia in cattle. Mode of transmission has not been
Tetracyclines appear to control infection during acute
outbreaks (Hum et al., 2000). Mycoplasma leachii shares
surface antigens with Mycoplasma mycoides subsp. mycoides,
Mycoplasma capricolum subsp. capripneumoniae, and Mycoplasma capricolum subsp. capricolum, potentially compounding serology-based diagnosis of infection.
Source: isolated from the synovial fluid, udders, expelled
milk, lungs, lymph nodes, pericardium, cervix, vagina, prepuce, semen, and aborted calves of cattle (Alexander et al.,
1985; Gourlay and Howard, 1979; Hum et al., 2000).
Type strain: PG50, NCTC 10133, DSM 21131.
Further comment: the assignment of strains formerly called
“Mycoplasma species bovine group 7 of Leach” to the species
Mycoplasma leachii came in response to a request from the Subcommittee on the Taxonomy of Mollicutes of the International
Committee on Systematics of Prokaryotes for a proposal for
an emended taxonomy for the members of the Mycoplasma
mycoides phylogenetic cluster (Manso-Silván et al., 2009).
74. Mycoplasma leonicaptivi corrig. Hill 1992, 521VP
le.o.ni.cap¢ti.vi. L. n. leo, -onis the lion; L. adj. captivus captive; N.L. gen. n. leonicaptivi of the captive lion.
Cells are pleomorphic (primarily coccoid). Nonmotile.
Colonies on solid medium have a typical fried-egg appearance. Growth in SP-4 broth supplemented with glucose
occurs between 35 and 37°C.
Source: isolated from the throat and respiratory tract of
captive lions and leopards (Hill, 1992).
Type strain: 3L2, NCTC 11726, ATCC 49890.
Further comment: the original spelling of the specific epithet, leocaptivus (sic), has been corrected by Trüper and
De’Clari (1998).
75. Mycoplasma leopharyngis Hill 1992, 521VP
le.o.pha.ryn¢gis. L. masc. n. leo, -onis lion; N.L. n. pharynx,
-yngis (from Gr. n. pharugx, pharuggos throat) throat; N.L.
gen. n. leopharyngis (sic) of the throat of a lion.
Cells are coccoid to pleomorphic. Nonmotile. Colonies
on solid medium have a typical fried-egg appearance under
Genus I. Mycoplasma
anaerobic conditions. Grows in SP-4 broth supplemented
with glucose at optimum temperatures of 35–37°C. Grows
well under both aerobic and anaerobic conditions.
Source: isolated from the throat of lions (Hill, 1992).
Type strain: LL2, NCTC 11725, ATCC 49889.
77. Mycoplasma lipofaciens Bradbury, Forrest and Williams
1983, 334VP
li.po.fa¢ci.ens. Gr. n. lipos animal fat, lard, tallow; L. v. facio
to make; N.L. part. adj. lipofaciens fat-making, intended to
refer to the production of a lipid film on solid media.
Cells are mainly spherical and elliptical. Nonmotile. Colonies on solid medium have typical fried-egg appearance. Grows
in Hayflick medium supplemented with glucose or arginine
at 37°C. Produces a strong “film and spots” reaction.
Commensal of birds; little evidence exists for naturally
occurring pathogenicity of the isolates, although experimental inoculation of chicken or turkey eggs can result in
embryo mortality. Inadvertent transmission to an investigator during experimental inoculation studies resulted in
clinical signs including rhinitis and pharyngitis. Aerosol
transmission has been documented in turkeys.
Source: isolated from the infraorbital sinuses of chickens;
from tissues or eggs of turkeys and ducks; and from eggs of
Northern goshawks (Benčina et al., 1987; Bradbury et al.,
1983; Lierz et al., 2007a, b, c, 2008b).
Type strain: R171, ATCC 35015, NCTC 10191.
78. Mycoplasma lipophilum Del Giudice, Purcell, Carski and
Chanock 1974, 152AL
li.po.phi¢lum. Gr. n. lipos animal fat; N.L. neut. adj. philum
(from Gr. neut. adj. philon) friend, loving; N.L. neut. adj.
lipophilum fat-loving.
Cells are pleomorphic and granular. Motility for this species has not been assessed. Colonies display typical fried-egg
morphology. Growth on solid medium is associated with
heavy production of film that spreads over the surface of
the agar, with the development of numerous internal particles in the colonies. A film similar to that produced on
agar medium develops on the surface of broth-grown cultures (Del Giudice et al., 1974). Grows in SP-4 or Hayflick
Pathogenicity for this species is unclear. This species was
first isolated from a human patient with primary atypical
pneumonia; however, subsequent isolations from similarly
symptomatic patients have not been achieved. Mode of
transmission has not been formally assessed.
Source: isolated from the upper and lower respiratory
tract of a human with primary atypical pneumonia and the
lower respiratory tract of rhesus monkeys (Hill, 1977).
Type strain: MaBy, ATCC 27104, NCTC 10173, NBRC
14895.
603
79. Mycoplasma maculosum Edward 1955, 90AL
ma.cu.lo¢sum. L. neut. adj. maculosum spotted, alluding to a
crinkled film covering the colonies and spreading between
them, and spots appearing in the medium beneath and
around the colonies.
Cells are short and filamentous, with occasional branching. Motility for this species has not been assessed. Colonies on solid medium have a typical fried-egg appearance.
Grows well in Hayflick or SP-4 medium supplemented with
arginine at 37°C.
Opportunistic pathogen. Cause of pneumonia in domestic dogs and rarely of meningitis in immunocompromised
humans. The route of transmission is via droplet aerosol.
Source: isolated from the nasopharynx, lungs, conjunctivae, and urogenital tract of domestic dogs and the cerebrospinal fluid of an immunocompromised human (Chalker,
2005).
14848.
80. Mycoplasma meleagridis Yamamoto, Bigland and Ortmayer
1965, 47AL
me.le.a¢gri.dis. L. n. meleagris, -idis a turkey; L. gen. n. meleagridis of a turkey.
Cells are coccoid to coccobacillary. Motility for this species has not been assessed. An extracellular capsule can be
visualized by electron microscopy following staining with
ruthenium red (Green III and Hanson, 1973). Colonies on
solid medium are not necessarily of typical fried-egg appearance. Grows in Frey’s medium supplemented with arginine
at 37–38°C (Yamamoto et al., 1965).
Pathogenic; causes airsacculitis, pneumonia, sinusitis,
perosis, chondrodystrophy, bursitis, synovitis, and reduced
hatchability due to embryo lethality in turkeys. Transmission is primarily vertical, but can also occur through droplet
aerosol or sexual contact.
Tetracyclines, macrolides, and fluoroquinolones are
effective chemotherapeutic agents; however, medicating
a commercial flock is not considered an effective control
strategy. The temporary use of in ovo antimicrobial therapy
can be used to eradicate Mycoplasma meleagridis from a flock.
Management tactics (e.g., single age “all in/all out” systems
and culling of endemic flocks) are more commonly utilized
(Kleven, 2008). Serological and molecular diagnostic methods have been described (Ben Abdelmoumen Mardassi
et al., 2007; Ramírez et al., 2008; Raviv and Kleven, 2009).
Source: isolated from the air sacs, trachea, infraorbital
sinuses, oviduct, cloaca, phallus, and eggs of turkeys, and
the air sacs of buzzards, kites, and kestrels (Chin et al., 2008;
Jordan, 1979; Lam et al., 2004; Lierz et al., 2000).
Further comment: previously known as avian serotype H
(Kleckner, 1960).
81. Mycoplasma microti (Dillehay, Sander, Talkington, Thacker
and Brown 1995) Brown, Talkington, Thacker, Brown,
604
Dillehay and Tully 2001b, 412VP (Mycoplasma volis Dillehay,
Sander, Talkington, Thacker and Brown 1995, 633)
mi.cro¢ti. N.L. n. Microtus a genus of field vole; N.L. gen. n.
microti of Microtus.
Cells are predominantly coccoid in shape. Nonmotile. Colonies on solid medium exhibit a typical fried-egg
appearance. Grows well in SP-4 supplemented with glucose
in temperatures ranging from 35 to 37°C.
Opportunistic pathogen. No evidence exists for pathogenicity in the natural host; however, pneumonitis was
experimentally induced in mice and rats (Evans-Davis et al.,
1998).
Source: isolated from the nasopharynx and lung of prairie
voles (Dillehay et al., 1995).
Type strain: IL371, ATCC 700935.
82. Mycoplasma moatsii Madden, Moats, London, Matthew
and Sever 1974, 464AL
mo.at¢si.i. N.L. gen. masc. n. moatsii of Moats, named after
Kenneth E. Moats, whose primary interest has been in the
mycoplasmas of nonhuman primates.
Cells are spheroidal and some exhibit protrusions from
the membrane. Motility for this species has not been
assessed. Colonies exhibit typical fried-egg morphology.
Grows readily in SP-4 or Hayflick broth supplemented with
either arginine or glucose at an optimum temperature of
37°C.
No evidence of pathogenicity.
Source: isolated from the respiratory and reproductive
tracts of grivet monkeys and from the cecum, jejunum, and
colon of wild Norway rats (Giebel et al., 1990).
Type strain: MK 405, ATCC 27625, NCTC 10158.
83. Mycoplasma mobile Kirchhoff, Beyene, Fischer, Flossdorf,
Heitmann, Khattab, Lopatta, Rosengarten, Seidel and
Yousef 1987, 197VP
mo¢bi.le. L. neut. adj. mobile motile.
Cells are conical or flask-shaped and have a distinct terminal protrusion referred to as the “head-like structure”.
Cells demonstrate rapid gliding motility when adhering to
charged surfaces and move in the directional of the headlike structure (Miyata et al., 2002, 2000). Colonies on solid
medium have a typically fried-egg appearance. Grows well
in Aluotto’s medium supplemented with glucose or arginine. The temperature range for growth is 17–30°C, with
optimum growth at 30°C.
Pathogenic; causes necrotic erythrodermatitis in tench.
The mode of transmission has not been established.
Source: isolated from the gills of a freshwater fish (Tinca
tinca) with “red disease” (Kirchhoff et al., 1987).
DNA G+C content (mol%): 23.5 (Bd), 24.9.
Type strain: 163K, ATCC 43663, NCTC 11711.
NC_006908 (complete genome sequence of strain
163K).
84. Mycoplasma molare Røsendal 1974, 130AL
mo.la¢re. L. neut. adj. molare of or belonging to a mill, here
millstone-like, referring to the heavy film reaction, which
resembles the pattern on the surface of a millstone.
Cells are coccoid to pleomorphic. Nonmotile. Colonies
medium supplemented with glucose at 37°C. A lipid film of
characteristic appearance develops on the surface and along
the circumference of colonies grown on egg-yolk agar.
Opportunistic pathogen. Associated with pharyngitis and
mild inflammatory lesions of the lower respiratory tract and
may be associated with infertility, vaginitis, and posthitis of
domestic dogs. No clear evidence for primary pathogenicity
of the species. Mode of transmission has not been established definitively.
Source: isolated from the oral cavity, pharynx, cervix,
vagina, and prepuce of domestic dogs (Røsendal, 1979;
(Chalker, 2005).
Type strain: H 542, ATCC 27746, NCTC 10144.
85. “Mycoplasma mucosicanis” Spergser, Langer,
Macher, Szostak, Rosengarten and Busse 2010
Muck,
mu.co.si.ca¢nis. N.L. n. mucosa mucous membrane; L. n.
canis a dog; N.L. gen. n. mucosicanis of mucous membranes
of a dog.
Cells are pleomorphic, but primarily coccoid. Nonmotile. Colonies on solid media have a typical fried-egg morphology. Grows wells in modified Hayflick medium at 37°C
Source: isolated from the prepuce, semen, vagina, cervix,
and oral cavity of domestic dogs (Spergser et al., 2010).
Type strain: 1642, ATCC BAA-1895, DSM 22457.
Sequence accession no. (16S rRNA gene): AM774638.
86. Mycoplasma muris McGarrity, Rose, Kwiatkowski, Dion,
Phillips and Tully 1983, 355VP
mu¢ris. L. n. mus, muris mouse; L. gen. n. muris of a mouse.
Cells are primarily coccoid or coccobacillary, but exhibit
a few other pleomorphic forms. Motility for this species has
not been assessed. Colonies usually have a granular appearance and few colonies demonstrate the typical fried-egg
appearance. Grow well in SP-4 broth supplemented with
arginine at 37°C and produces a “film and spots” reaction.
Source: isolated from the vagina of a pregnant mouse (laboratory strain RIII; McGarrity et al., 1983).
DNA G+C content (mol%): 24.9 (TLC).
Type strain: RIII-4, ATCC 33757, NCTC 10196.
87. Mycoplasma mustelae Salih, Friis, Arseculeratne, Freundt
and Christiansen 1983, 478VP
mu.ste¢lae. N.L. n. Mustela (from L. n. mustela a weasel) the
generic name of the mink Mustela vison; N.L. gen. n. mustelae of Mustela.
Genus I. Mycoplasma
Cells are highly pleomorphic; most common morphologies include pleomorphic rings, short filamentous forms,
and coccoid elements. Nonmotile. Colonies on solid
medium show a typical fried-egg appearance. Growth in
Hayflick medium supplemented with glucose occurs at
37°C.
Source: isolated from the trachea and lungs of juvenile
minks (Mustela vison; Salih et al., 1983).
Type strain: MX9, ATCC 35214, NCTC 10193, AMRC-C
1486.
88. Mycoplasma neurolyticum (Sabin 1941) Freundt 1955, 73AL
(Musculomyces neurolyticus Sabin 1941, 57)
neu.ro.ly¢ti.cum. Gr. n. neuron nerve; N.L. adj. lyticus -a -um
(from Gr. adj. lutikos -ê -on) able to loosen, able to dissolve;
N.L. neut. adj. neurolyticum nerve-destroying.
Cells are filamentous and highly variable length. Nonmotile (Nelson and Lyons, 1965). Colonies show a typical friedegg appearance after incubation at 37°C. Grows in Hayflick
medium supplemented with glucose at 37°C (Naot et al.,
1977).
Pathogenicity is currently uncertain. Potentially associated with spongiform encephalopathy and ischemic necrosis of the brain resulting in a clinical state referred to as
“rolling disease” in mice and rats. Pathology may be exacerbated in the presence of additional neurotropic organisms (i.e., Toxoplasma gondii, Chlamydia spp., Plasmodium
spp., and yellow fever virus) or during leukemic syndromes.
Transmission to suckling rodents occurs shortly after birth.
Treatment of Mycoplasma neurolyticum infections is uncommon, as pathology is typically not resolvable after the onset
of clinical signs.
Source: isolated from the brain, conjunctivae, nasopharynx, and middle ear of captive mice and rats.
DNA G+C content (mol%): 22.8 (Bd), 26.2 (Tm).
Type strain: Type A, ATCC 19988, NCTC 10166, CIP
103926, NBRC 14799.
Further comment: a putative exotoxin with neurological
effects on rodents was formerly thought to be produced
by most freshly isolated strains, although a few non-toxic
strains were described (Tully and Ruchman, 1964). The
findings were not substantiated by later work (Tryon and
Baseman, 1992).
89. Mycoplasma opalescens Røsendal 1975, 469AL
o.pa.les¢cens. L. n. opalus precious stone; N.L. neut. adj.
opalescens opalescent, referring to the opalescent film produced on solid medium.
Morphology by light microscopy or ultrastructural examination is not defined. Motility for this species has not been
assessed. Colonies on solid medium have a typical fried-egg
appearance and possess an iridescent quality. Grows well in
Source: isolated from the oral cavity, prepuce, and prostate gland of domestic dogs (Røsendal, 1975).
605
Type strain: MH5408, ATCC 27921, NCTC 10149.
90. Mycoplasma orale Taylor-Robinson, Canchola, Fox and
Chanock 1964, 141AL
o.ra¢le. L. n. os, oris the mouth; L. neut. suff. -ale suffix
denoting pertaining to; N.L. neut. adj. orale pertaining to
the mouth.
Cells can be either coccoid or filamentous. Motility for
this species has not been assessed. Colonies on solid medium
or SP-4 medium supplemented with arginine at 37°C.
Mycoplasma orale is most commonly associated with contamination of eukaryotic cell culture and is frequently
removed by treatment of cells with antibiotics and/or maintenance of cell lines in antibiotic-containing medium. The
most effective classes of antibiotics for cell culture eradication are tetracyclines, macrolides, and fluoroquinolones.
Additionally, passage of eukaryotic cells in hyperimmune
serum raised against Mycoplasma orale has been shown to be
an effective method of eradication (Vogelzang and Compeer-Dekker, 1969).
Commensal/opportunistic pathogen. Commonly found
as a commensal of the human oral cavity; can cause respiratory tract infections, osteomyelitis, infectious synovitis, and
abscesses in immunocompromised individuals (Paessler
et al., 2002; Roifman et al., 1986).
Source: isolated from the oral cavity of subclinical humans,
the sputum of an immunocompromised human with acute
respiratory illness, and from synovial fluid, bone, and splenic
abscesses of another immunocompromised individual.
Type strain: CH19299, ATCC 23714, NCTC 10112, CIP
104969, NBRC 14477.
91. Mycoplasma ovipneumoniae Carmichael, St George, Sullivan
and Horsfall 1972, 677AL
o.vi.pneu.mo.ni′ae. L. fem. n. ovis a sheep; Gr. n. pneumonia
pneumonia; N.L. gen. n. ovipneumoniae of sheep pneumonia.
Morphology and motility are poorly described. The
organism produces a polysaccharide capsule with variable
thickness that is dependent upon culture conditions and
strain (Niang et al., 1998). Colonies grown on standard
agar are convex and have a lacy or vacuolated appearance.
Grows well in Friis medium or SP-4 broth supplemented
Pathogenic; causes chronic proliferative interstitial pneumonia, pulmonary adenomatosis, conjunctivitis (Jones
et al., 1976), and mastitis under experimental conditions
(Jones, 1985) of sheep and goats. Transmission occurs via
droplet aerosol and can occur via intravenous inoculation
in experimental infection studies.
Source: isolated from the lungs, trachea, nose, and conjunctivae of sheep and goats.
Type strain: Y98, NCTC 10151, ATCC 29419.
606
92. Mycoplasma ovis (Neitz, Alexander and du Toit 1934)
Neimark, Hoff and Ganter 2004, 369VP (Eperythrozoon ovis
Neitz, Alexander and du Toit 1934, 267)
o¢vis. L. fem. n. ovis, -is a sheep; L. gen. n. ovis of a sheep.
Cells are coccoid and motility for this species has not
been assessed. The morphology of infected erythrocytes
is altered demonstrating a marked depression at the site
of Mycoplasma ovis attachment. This species has not been
grown on artificial medium; therefore, notable biochemical
parameters are not known.
Neoarsphenamine is an effective therapeutic agent. Mycoplasma ovis is reported to share antigens with Mycoplasma
wenyonii (Kreier and Ristic, 1963), potentially complicating
serology-based diagnosis of infection.
Pathogenic; causes mild to severe anemia in sheep and
goats that often results in poor feed conversion. Transmission occurs via blood-feeding arthropods, e.g., Haemophysalis
plumbeum, Rhipicephalus bursa, Aedes camptorhynchus, and
Culex annulirostris (Daddow, 1980; Howard, 1975; Nikol’skii
and Slipchenko, 1969), and likely via fomites such as reused
needles, shearing tools, and ear-tagging equipment (BrunHansen et al., 1997; Mason and Statham, 1991).
Source: observed in association with erythrocytes or unattached in suspension in the blood of sheep, goats, and
rarely in eland and splenectomized deer.
93. Mycoplasma oxoniensis Hill 1991b, 24VP
oxo.ni.en¢sis. N.L. adj. oxoniensis (sic) pertaining to Oxon,
an abbreviation of Oxfordshire, where the mycoplasma was
first isolated.
Cells are primarily coccoid. Nonmotile. Colonies on agar
have a typical fried-egg appearance. Growth in SP-4 broth
supplemented with glucose occurs at 35–37°C.
No evidence of pathogenicity. Mode of transmission is
unknown.
Source: isolated from the conjunctivae of the Chinese
hamster (Cricetulus griseus; Hill, 1991b).
94. Mycoplasma penetrans Lo, Hayes, Tully, Wang, Kotani,
Pierce, Rose and Shih 1992, 363VP
pe.ne¢trans. L. part. adj. penetrans penetrating, referring to
the ability of the organism to penetrate into mammalian
cells.
Cells are flask-shaped, with a distinct terminal structure reminiscent of the Mycoplasma pneumoniae attachment
organelle. Cells demonstrate gliding motility when adhering to charged surfaces and move in the direction of the
terminal structure. Colonies on agar plates display a typical fried-egg appearance. Grows well in SP-4 broth supplemented with either glucose or arginine at 37°C.
Opportunistic pathogen; found in the urogenital tract of
immunocompromised humans, most notably HIV-positive
individuals (serological detection in HIV-negative individuals is rare). Speculation regarding the ability of Mycoplasma
penetrans to act as a cofactor in the progression of AIDS by
modulation of the immune system remains intriguing, but
in need of further substantiation (Blanchard, 1997). Transmission is presumed to be via sexual contact.
Source: isolated from the urine of HIV-positive humans,
and from the blood, respiratory secretions, and trachea of
an HIV-negative patient with multiple autoimmune syndromes (Yanez et al., 1999).
DNA G+C content (mol%): 30.5 (Tm), 25.7 (HF-2 genome
sequence; Sasaki et al., 2002).
Type strain: GTU-54-6A1, ATCC 55252.
Sequence accession nos: L10839 (16S rRNA gene),
NC_004432 (HF-2 complete genome sequence).
95. Mycoplasma phocicerebrale corrig. Giebel, Meier, Binder,
Flossdorf, Poveda, Schmidt and Kirchhoff 1991, 43VP
pho.ci.ce.re.bra¢le. L. n. phoca seal; N.L. neut. adj. cerebrale
of or pertaining to the brain; N.L. neut. adj. phocicerebrale
pertaining to the brain of a seal.
Cells are coccoid or exhibit a dumbbell shape. Motility
for this species has not been assessed. Colonies on solid
medium typically show a fried-egg appearance. Grows well
in SP-4 medium supplemented with arginine at 37°C.
Pathogenic; associated with respiratory disease and conjunctivitis in harbor seals (Kirchhoff et al., 1989) and a distinctive ulcerative keratitis subsequent to seal bites (known
as “seal finger”) and secondary arthritis in humans (Baker
et al., 1998; Ståby, 2004). Mode of transmission between
harbor seals has not been established definitively; transmission to humans appears to be zoonotic following seal bites.
Source: isolated from the brains, noses, throats, lungs, and
hearts of seals (Phoca vitulina) during an outbreak of respiratory disease (Kirchhoff et al., 1989), and from cutaneous
lesions of humans with seal finger (Baker et al., 1998).
Further comment: the original spelling of the specific epithet, phocacerebrale (sic), has been corrected by Königsson
et al. (2001).
96. Mycoplasma phocidae Ruhnke and Madoff 1992, 213VP
pho.ci¢da.e. L. n. phoca seal; N.L. gen. n. phocidae (sic) of a
seal.
Cells are primarily coccoid. Motility for this species has
not been assessed. Colonies on solid medium have a typical fried-egg appearance. Grows well in SP-4 or Hayflick
medium supplemented with arginine at 37°C. Produces
“film and spots” reaction.
Opportunistic pathogen; associated with secondary
pneumonia of harbor seals subsequent to influenza infection. Attempts to produce disease in gray or harp seals with
Mycoplasma phocidae in pure culture were not successful
(Geraci et al., 1982). Mode of transmission has not been
Source: isolated from the lungs, tracheae, and heart of
harbor seals.
Genus I. Mycoplasma
Further comment: the species designation “Mycoplasma
p hocae” was suggested by Königsson et al. (2001), but the
original epithet “phocidae” should be retained. The suggested change is forbidden by Rule 61 (Note) of the Bacteriological Code because it would change the first syllable of
the original epithet without correcting any orthographic or
typographical error.
97. Mycoplasma phocirhinis corrig. Giebel, Meier, Binder,
Flossdorf, Poveda, Schmidt and Kirchhoff 1991, 43VP
pho.ci.rhi¢nis. L. n. phoca seal; Gr. n. rhis, rhinos nose; N.L.
gen. n. phocirhinis of the nose of a seal.
Cells are coccoid; motility for this species has not been
assessed. Colonies on solid medium usually have a friedegg appearance. Grows well in Friis or Hayflick medium at
37°C. Produces “film and spots” reaction.
Pathogenic; associated with respiratory disease and conjunctivitis of harbor seals (Kirchhoff et al., 1989). Mode of
Source: isolated from the nose, pharynx, trachea, lungs,
and heart of seals (Phoca vitulina; Kirchhoff et al., 1989).
Further comment: the original spelling of the specific
epithet, phocarhinis (sic), has been corrected by Königsson
et al. (2001)
98. Mycoplasma pirum Del Giudice, Tully, Rose and Cole 1985,
290VP
pi¢rum. L. neut. n. pirum (nominative in apposition) pear,
referring to the pear-shaped morphology of the cells.
Cells are predominantly flask or pear-shaped and possess
an organized terminal structure, with an outer, finely particulate nap covering the entire surface of the cell. Cells
exhibit low-speed gliding motility and move in the direction of the terminal organelle (Hatchel and Balish, 2008).
Colonies display a typical fried-egg appearance. Grows well
in SP-4 medium supplemented with glucose at 37°C.
Source: isolated from the rectum of immunocompetent
humans and whole blood and circulating lymphocytes of
HIV-positive humans (Montagnier et al., 1990). Originally
isolated from cultured eukaryotic cells that were of human
origin (Del Giudice et al., 1985).
Type strain: HRC 70-159, ATCC 25960, NCTC 11702.
99. Mycoplasma pneumoniae Somerson, Taylor-Robinson and
Chanock 1963, 122AL
pneu.mo.ni¢ae. Gr. n. pneumonia pneumonia; N.L. gen. n.
pneumoniae of pneumonia.
Cells are highly pleomorphic; the predominant shape
includes a long, thin terminal structure at one cell pole,
with or without a trailing filament at the opposite pole.
Cells are motile and glide in the direction of the terminal
organelle when attached to cell surfaces, plastic, or glass.
Colonies on solid medium usually lack the light peripheral
zone, appearing rather as circular dome-shaped, granular
607
structures. Growth is best achieved in SP-4 medium supplemented with glucose at 37°C.
Pathogenic; causes interstitial pneumonitis, tracheobronchitis, desquamative bronchitis, and pharyngitis [collectively
referred to as primary atypical pneumonia (PAP); Krause
and Taylor-Robinson (1992)]. Less commonly, Mycoplasma
pneumoniae causes meningoencephalitis, otitis media, bullous
myringitis, infectious synovitis, glomerulonephritis, pancreatitis, hepatitis, myocarditis, pericarditis, hemolytic anemia,
and rhabdomyolysis (Waites and Talkington, 2005). The
preceding can be primary lesions, but are often secondary
to respiratory disease. Dysfunction of the immune system by
inappropriate cytokine responses or possibly molecular mimicry following infection are associated with long-term sequelae including the development or exacerbation of asthma and
chronic obstructive pulmonary disease; Stevens-Johnson syndrome and other exanthemas; and Guillain-Barre syndrome,
Bell’s palsy, and demyelinating neuropathies (Atkinson et al.,
2008). Mode of transmission is via droplet aerosols (PAP) or
sexual contact (urogential colonization).
Clinical manifestations are successfully treated with tetracyclines, fluoroquinolones, macrolides, and lincosamides
(Waites and Talkington, 2005). Signs can be treated with
inhaled or injected steroids. Experimental vaccinations
aimed at preventing infection have been unsuccessful due
to failure to elicit immune responses, retention of virulence, or invocation of immune responses that exacerbated
clinical signs (Barile, 1984; Jacobs et al., 1988). Mycoplasma
pneumoniae is reported to share antigens with Mycoplasma
genitalium (Taylor-Robinson, 1983a), potentially complicating serology-based diagnosis of infection.
Source: isolated from the upper and lower respiratory
tract, cerebrospinal fluid, synovial fluid, and urogential
tract of humans.
DNA G+C content (mol%): 38.6 (Tm), 40.0 (strain M129
genome sequence).
Type strain: FH, ATCC 15531, NCTC 10119, CIP 103766,
NBRC 14401.
Sequence accession nos: M29061 (16S rRNA gene), U00089
(strain M129 genome sequence).
100.Mycoplasma primatum Del Giudice, Carski, Barile, Lemcke
and Tully 1971, 442AL
pri.ma¢tum. L. n. primas, primatis chief, from which primates, the highest order of mammals originates; L. pl. gen.
n. primatum of chiefs, of primates.
Cells are both spherical and coccobacillary. Motility for
this species has not been assessed. Colony morphology
has a fried-egg appearance. Grows well in SP-4 or Hayflick
Opportunistic pathogen; rarely associated with keratitis
in humans (Ruiter and Wentholt, 1955). Mode of transmission has not been established.
Source: isolated from the oral cavity and/or urogenital
tract of baboons, African green monkeys, rhesus macaques,
squirrel monkeys, and humans (Hill, 1977; Somerson and
Cole, 1979; Thomsen, 1974).
Type strain: HRC292, ATCC 25948, NCTC 10163.
608
101.Mycoplasma pullorum Jordan, Ernø, Cottew, Hinz and
pul.lo¢rum. L. n. pullus a young animal, especially chicken;
L. gen. pl. n. pullorum of young chickens.
display typical fried-egg appearance. Grows in Frey’s or
Hayflick medium supplemented with glucose at 37°C.
Pathogenicity not fully established, but has been associated with tracheitis and airsacculitis in chickens, and
embryo lethality resulting in reduced hatchability in
chickens and turkeys. Mode of transmission has not been
assessed definitively.
Source: isolated from the trachea, air sacs, and eggs of
chickens; from the eggs of turkeys; and from tissues of
pheasants, partridges, pigeons, and quail (Benčina et al.,
1987; Bradbury et al., 2001; Kempf et al., 1991; Kleven,
2003; Lobo et al., 2004; Moalic et al., 1997; Poveda et al.,
1990).
Type strain: CKK, ATCC 33553, NCTC 10187.
Further comment: previously known as avian serotype C
(Adler et al., 1958).
102.Mycoplasma pulmonis (Sabin 1941) Freundt 1955, 73AL
(Murimyces pulmonis Sabin 1941, 57)
pul.mo¢nis. L. n. pulmo, -onis the lung; L. gen. n. pulmonis
of the lung.
Cells are predominantly coccoid with a well-organized
terminal structure. Cells are motile and glide in the direction of the terminal structure. An extracellular capsular
matrix can be demonstrated by staining with ruthenium
red and formation of biofilms has been demonstrated.
Colonies on solid medium have a coarsely granulated and
vacuolated appearance, with a lesser tendency to grow
into the agar, and the central spot is consistently less well
defined than in most other Mycoplasma species. Grows in
SP-4 or modified Hayflick broth supplemented with glucose at an optimum temperature of 37°C.
Pathogenic; causes rhinitis, laryngotracheitis, bronchopneumonia (collectively described as murine respiratory
mycoplasmosis in mice), otitis media, conjunctivitis, acute
and chronic arthritis, oophoritis, salpingitis, epididymitis,
and urethritis of rodents (chiefly mice, rats, guinea pigs,
and hamsters). Transmission occurs via aerosol, fomites,
sexual contact, or vertically during gestation.
Macrolides, fluoroquinolones, and tetracyclines are
effective against Mycoplasma pulmonis in vitro; however, control measures such as decontamination of fomites, culling
of infected colonies, and treatment of clinical signs with
steroids are more commonly employed in clinical settings.
Several candidate vaccines have been described.
Source: isolated from the respiratory and urogenital tracts,
eyes, synovial fluid, and synovial membranes of (principally
captive) rodents, and rarely from the nasopharynx of rabbits and horses (Allam and Lemcke, 1975; Cassell and Hill,
1979; Deeb and Kenny, 1967; S
imecka et al., 1992).
DNA G+C content (mol%): 27.5–29.2 (Bd), 26.6 (strain
UAB CTIP genome sequence).
Type strain: Ash, PG34, ATCC 19612, NCTC 10139, CIP
75.26, NBRC 14896.
Sequence accession nos: M23941 (16S rRNA gene
sequence), NC_002771 (strain UAB CTIP genome sequence).
103.Mycoplasma putrefaciens Tully, Barile, Edward, Theodore
and Ernø 1974, 116AL
pu.tre.fa¢ci.ens. L. v. putrefacio to make rotten; L. part.
adj. putrefaciens making rotten or putrefying, connoting the production of a putrid odor in broth and agar
cultures.
Cells are predominantly coccobacillary to pleomorphic.
Nonmotile. Formation of biofilms has been demonstrated
(McAuliffe et al., 2006). Colony morphology has a typical
fried-egg appearance. Grows well in SP-4 medium supplemented with glucose at 37°C.
Pathogenic; causes polyarthritis, mastitis, conjunctivitis (a syndrome collectively termed contagious agalactia)
(Bergonier et al., 1997), abortion, salpingitis, metritis, and
testicular atrophy (Gil et al., 2003) in goats.
Macrolides, fluoroquinolones, lincosamides, and tetracyclines are effective against Mycoplasma putrefaciens; however, control measures such as decontamination of fomites
and culling of infected herds are typically recommended
to discourage the development of antimicrobial-resistant
strains in carrier animals (Antunes et al., 2007; Bergonier
et al., 1997).
Source: isolated from the synovial fluid, udders, expelled
milk, conjunctivae, ear canal, uterus, and testes of goats.
Type strain: KS1, ATCC 15718, NCTC 10155.
104.Mycoplasma salivarium Edward 1955, 90AL
sa.li.va¢ri.um. L. neut. adj. salivarium slimy, saliva-like,
intended to denote of saliva.
Cells are coccoid to coccobacillary. Nonmotile. Colonies
are large with a typical fried-egg appearance. Grows well in
Hayflick medium supplemented with arginine at 37°C and
produces a “film and spots” reaction.
Mycoplasma salivarium is most frequently associated
with contamination of eukaryotic cell culture and is frequently removed by treatment of cells with antibiotics
and/or maintenance of cell lines in antibiotic-containing
medium. The most effective classes of antibiotics for cell
culture eradication are tetracyclines, macrolides, and fluoroquinolones.
Opportunistic pathogen; primarily found as a commensal of the human oral cavity, and rarely associated with
arthritis, submasseteric abscesses, gingivitis, and periodontitis in immunocompromised patients (Grisold et al., 2008;
Lamster et al., 1997; So et al., 1983). Mode of transmission
is via direct contact with human saliva.
Source: isolated from the oral cavity, synovial fluid, dental plaque, and abscessed mandibles of humans, and the
nasopharynx of pigs (Erickson et al., 1988).
Type strain: PG20, H110, ATCC 23064, NCTC 10113,
NBRC 14478.
Genus I. Mycoplasma
105.Mycoplasma simbae Hill 1992, 520VP
sim¢bae. Swahili n. simba lion; N.L. gen. n. simbae of a lion.
Cells are pleomorphic and nonmotile. Colonies on solid
medium have a typical fried-egg appearance. Film is produced by cultivation on egg yolk agar. Grows well in SP-4
medium at 37°C.
Source: isolated from the throats of lions (Hill, 1992).
Type strain: LX, NCTC 11724, ATCC 49888.
106.Mycoplasma spermatophilum Hill 1991a, 232VP
sper.ma.to.phi¢lum. Gr. n. sperma, -atos sperm or seed; N.L.
neut. adj. philum (from Gr. neut. adj. philon) friend, loving;
N.L. neut. adj. spermatophilum sperm-loving.
Cells are primarily coccoid. Nonmotile. Colonies are
convex to fried-egg shaped and are of below-average size.
Grows well in SP-4 medium supplemented with added arginine under anaerobic conditions at 37°C.
Pathogenic; potentially associated with infertility, as
infected spermatozoa do not fertilize ova and infected
fertilized ova were unable to implant following in vitro
fertilization (Hill et al., 1987; Hill, 1991a). Mode of transmission is via sexual contact.
Source: isolated from the semen and cervix of humans
with impaired fertility.
Type strain: AH159, NCTC 11720, ATCC 49695, CIP
105549.
107.Mycoplasma spumans Edward 1955, 90AL
spu¢mans. L. part. adj. spumans foaming, presumably alluding to thick dark markings that suggest the presence of
globules inside the coarsely reticulated colonies.
Cells are coccoid to filamentous. Motility for this species
has not been assessed. Colonies in early subcultures have a
coarsely reticulated and vacuolated appearance. A typical
fried-egg appearance of the colonies develops on repeated
subculturing. Grows well in SP-4 or modified Hayflick
medium supplemented with arginine.
Opportunistic pathogen; primarily found as a commensal of the nasopharynx, but has also been associated with
pneumonia and arthritis of domestic dogs. Mode of transmission has not been established definitively.
Source: isolated from the lungs, nasopharynx, synovial fluid,
cerebrospinal fluid, trachea, prepuce, prostate, bladder, cervix, vagina, and urine of domestic dogs (Chalker, 2005).
14849.
108.Mycoplasma sturni Forsyth, Tully, Gorton, Hinckley,
Frasca, van Kruiningen and Geary 1996, 719VP
stur¢ni. N.L. n. Sturnus (from L. n. sturnus a starling or stare)
a genus of birds, N.L. gen. n. sturni of the genus Sturnus, the
genus of the bird from which the organism was isolated.
609
Cells are primarily coccoid with some irregular
ask-shaped and filamentous forms seen. Motility for this
fl
species has not been assessed. Colonies on agar usually have
a fried-egg appearance when grown at 37°C. Grows well in
SP-4 medium supplemented with glucose at 34–37°C.
Pathogenicity has not been fully established, but it is
associated with conjunctivitis in the European starling,
mockingbirds, blue jays, and American crows. The organism is also found in clinically normal birds. Mode of transmission has not been established definitively.
Source: isolated from the conjunctivae of European starlings, mockingbirds, blue jays, American crows, American
robins, blackbirds, rooks, carrion crows, and magpies
(Frasca et al., 1997; Ley et al., 1998; Pennycott et al., 2005;
Wellehan et al., 2001).
Type strain: UCMF, ATCC 51945.
109.Mycoplasma sualvi Gourlay, Wyld and Leach 1978, 292AL
su.al¢vi. L. n. sus, suis swine; L. n. alvus bowel, womb, stomach; N.L. gen. n. sualvi of the bowel of swine.
Cells are coccobacillary and many possess organized
terminal structures. Nonmotile. Colonies have a typical
fried-egg appearance. Grows well in SP-4 medium supplemented with either arginine or glucose at 37°C.
Source: isolated from the rectum, colon, small intestines,
and vagina of pigs (Gourlay et al., 1978).
Type strain: Mayfield B, NCTC 10170, ATCC 33004.
110.Mycoplasma subdolum Lemcke and Kirchhoff 1979, 49AL
sub.do¢lum. L. neut. adj. subdolum somewhat deceptive,
alludes to the deceptive color change that led to the
original erroneous description of the strains as ureahydrolyzing.
Cells are coccoid to coccobacilliary. Motility for this
species has not been assessed. Colony growth on solid
medium exhibits the typical fried-egg appearance. Grows
well in SP-4, Frey’s, or Hayflick medium supplemented
with arginine at 37°C.
Opportunistic pathogen; equivocal evidence for virulence may represent variation among strains. Associated
with impaired fecundity and abortion in horses; however,
is highly prevalent in clinically normal horses (Spergser
et al., 2002). Mode of transmission is via sexual contact.
Source: isolated from the cervix, semen, and aborted
foals of horses (Lemcke and Kirchhoff, 1979).
Type strain: TB, ATCC 29870, NCTC 10175.
111.Mycoplasma suis corrig. (Splitter 1950) Neimark,
Johansson, Rikihisa and Tully 2002b, 683VP (Eperythrozoon
suis Splitter 1950, 513)
su¢is. L. gen. n. suis of the pig.
assessed. This species has not been grown on any artificial
610
not known.
Neoarsphenamine and tetracyclines are effective therapeutic agents. An enzyme-linked immunosorbant assay
(ELISA) and PCR-based detection assays to enable diagnosis of infection have been described (Groebel et al., 2009;
Gwaltney and Oberst, 1994; Hoelzle, 2008; Hsu et al., 1992).
Pathogenic; causes febrile icteroanemia in pigs. Transmission occurs via insect vectors including Stomoxys calcitrans and Aedes aegypti (Prullage et al., 1993).
Source: observed in association with the erythrocytes of
pigs.
DNA G+C content (mol%): 31.1 (complete genome
sequence of strain Illinois; J.B. Messick et al., unpublished).
Further comment: the original spelling of the specific epithet, haemosuis (sic), has been corrected by the List Editor.
112.Mycoplasma synoviae Olson, Kerr and Campbell 1964,
209AL
sy.novi¢ae. N.L. n. synovia the joint fluid; N.L. gen. n. synoviae of joint fluid.
Cells are coccoid and pleomorphic. Nonmotile. An
amorphous extracellular layer is described. Colony appearance on solid medium is variable with some showing typical fried-egg type colonies. Grows well in Frey’s medium
supplemented with glucose, l-cysteine, and nicotinamide
adenine dinucleotide at 37°C. Produces a “film and spots”
reaction (Ajufo and Whithear, 1980; Frey et al., 1968).
Pathogenic; causes infectious synovitis, osteoarthritis,
and upper respiratory disease which is often subclinical in
chickens and turkeys. Also associated with a reduction in
egg quality in chickens. Mycoplasma synoviae is often found
in association with additional avian pathogens including
Mycoplasma gallisepticum, avian strains of Escherichia coli,
Newcastle disease virus, and infectious bronchitis virus.
Direct contact with fomites and droplet aerosols are the
primary mechanisms of transmission.
Tetracyclines and fluoroquinolones are effective chemotherapeutic agents; however, treatment is typically only
sought for individual birds, as medicating a commercial
flock is not considered an effective control strategy. Vaccination and management strategies (i.e., single age “all
in/all out” systems and culling of endemic flocks) are
more commonly utilized. A live vaccine is commercially
available. Mycoplasma synoviae shares surface antigens with
Mycoplasma gallisepticum, potentially complicating serologybased diagnosis of infection. Numerous molecular diagnostics have been described. This organism is listed in the
Terrestrial Animal Health Code of the Office International
des Epizooties (http://oie.int; Yogev et al., 1989; Browning
et al., 2005; Kleven, 2008; Hammond et al., 2009; Raviv and
Kleven, 2009)
Source: isolated from the synovial fluid, synovial membranes, and respiratory tract tissues of chickens and turkeys, and from ducks, geese, pigeons, Japanese quail,
pheasants, red-legged partridges, wild turkeys, and house
sparrows (Bradbury and Morrow, 2008; Feberwee et al.,
2009; Jordan, 1979; Kleven, 1998).
Type strain: WVU 1853, ATCC 25204, NCTC 10124.
Sequence accession nos: X52083 (16S rRNA gene),
NC_007294 (strain 53 complete genome sequence).
113.Mycoplasma testudineum Brown, Merritt, Jacobson, Klein,
Tully and Brown 2004, 1529VP
tes.tu.di¢ne.um. L. neut. adj. testudineum of or pertaining
to a tortoise.
Cells are predominantly coccoid in shape, though some
exhibit a terminal protrusion. Cells exhibit gliding motility.
Colonies on solid medium exhibit typical fried-egg forms.
Grows well in SP-4 medium supplemented with glucose at
22–30°C.
Pathogenic; causes rhinitis and conjunctivitis in desert
and gopher tortoises. Mode of transmission appears to be
intranasal inhalation (Brown et al., 2004).
Source: isolated from the nares of desert tortoises
(Gopherus agassizii) and gopher tortoises (Gopherus
polyphemus).
Type strain: BH29, ATCC 700618, MCCM 03231.
114.Mycoplasma testudinis Hill 1985, 491VP
tes.tu¢di.nis. L. n. testudo, -inis tortoise; L. gen. n. testudinis
of a tortoise.
Cells are pleomorphic, with many possessing a terminal
organelle similar to those of Mycoplasma gallisepticum and
Mycoplasma amphoriforme that periodically exhibits curvature (Hatchel et al., 2006). A subset of cells are motile and
glide at high speed in the direction of the terminal structure. Colonies on solid medium have a typical fried-egg
appearance. Grows well in SP-4 medium supplemented
with glucose at 25–37°C, with optimum growth at 30°C.
Source: isolated from the cloaca of a Greek tortoise (Hill,
1985).
115.Mycoplasma verecundum Gourlay, Leach and Howard
1974, 483AL
ve.re¢cun.dum. L. neut. adj. verecundum shy, unobtrusive, free from extravagance, alluding to the lack of obvious biochemical characteristics of the species.
Cells are highly pleomorphic, exhibiting coccoid bodies, ring forms, and branched filaments. Motility for this
species has not been assessed. Growth on solid medium
produces colonies with the typical fried-egg appearance.
Grows well in SP-4 or Hayflick medium at 37°C.
Probable commensal; attempts to produce disease experi
mentally have been unsuccessful (Gourlay and Howard,
1979). Mode of transmission has not been established.
Source: isolated from the eyes of calves with conjunctivitis, the prepuce of clinically normal bulls, and from
in-market kale (Gourlay and Howard, 1979; Gourlay et al.,
1974; Somerson et al., 1982).
Genus I. Mycoplasma
611
116.Mycoplasma wenyonii (Adler and Ellenbogen 1934)
Neimark, Johansson, Rikihisa and Tully 2002b, 683VP
(Eperythrozoon wenyonii Adler and Ellenbogen 1934, 220)
117.Mycoplasma yeatsii DaMassa, Tully, Rose, Pitcher, Leach
and Cottew 1994, 483VP
we.ny.o¢ni.i. N.L. masc. gen. n. wenyonii of Wenyon, named
after Charles Morley Wenyon (1878–1948), an investigator
of these organisms.
not known.
Pathogenic; causes anemia and subsequent lameness and/
or infertility in cattle. Transmission is primarily vector-mediated
by Dermacentor andersoni and reportedly can also occur vertically
during gestation. Oxytetracycline is an effective therapeutic
agent (Montes et al., 1994). Mycoplasma wenyonii is reported to
share antigens with Mycoplasma ovis (Kreier and Ristic, 1963),
potentially complicating serology-based diagnosis of infection.
cattle; Kreier and Ristic (1968) reported in addition to
erythrocytes an association with platelets.
ye.at¢si.i. N.L. masc. gen. n. yeatsii of Yeats, named after F.R.
Yeats, an Australian veterinarian who was a co-isolator of
the organism.
Cells are coccoid and nonmotile. Colonies on agar have
a fried-egg appearance. Grows well in SP-4 medium supplemented with glucose at 37°C. Formation of biofilms has
been demonstrated (McAuliffe et al., 2006).
Opportunistic pathogen; commensal of the ear canal of
goats that has rarely been found in association with mastitis and arthritis (DaMassa et al., 1991). The mode of transmission has not been established.
Source: isolated from the external ear canals, retropharyngeal lymph node, nasal cavity, udders, and milk of
goats.
Type strain: GIH, ATCC 51346, NCTC 11730, CIP
105675.
Species incertae sedis
1. Mycoplasma coccoides (Schilling 1928) Neimark, Peters,
Robinson and Stewart 2005, 1389VP (Eperythrozoon coccoides
Schilling 1928, 1854)
coc.co¢ides. N.L. masc. n. coccus (from Gr. masc. n. kokkos
grain, seed) coccus; L. suff. -oides (from Gr. suff. eides, from
Gr. n. eidos that which is seen, form, shape, figure), resembling, similar; N.L. neut. adj. coccoides coccus-shaped.
assessed. This species has not been grown on artificial medium;
therefore, notable biochemical parameters are not known.
Pathogenic; causes anemia in wild and captive mice, and
captive rats, hamsters, and rabbits. Transmission is believed
to be vector-borne and mediated by the rat louse Polyplex
spinulosa and the mouse louse Polyplex serrata.
Neoarsphenamine and oxophenarsine were thought to
be effective chemotherapeutic agents for treatment of Mycoplasma coccoides infection in captive rodents, whereas tetracyclines are effective only at keeping infection at subclinical
levels (Thurston, 1953).
wild and captive rodents.
Species Candidatus
1. “Candidatus Mycoplasma haematoparvum” Sykes, Ball,
Bailiff and Fry 2005, 29
2. “Candidatus Mycoplasma haemobos” Tagawa, Matsumoto
and Inokuma 2008, 179
ha.e.ma.to.par¢vum. Gr. neut. n. haema, -atos blood; L. neut.
adj. parvum small; N.L. neut. adj. haemoatoparvum small
(mycoplasma) from blood.
ha.e.mo¢bos. Gr. neut. n. haema blood; L. n. bos an ox, a bull, a
cow; N.L. n. haemobos (sic) intended to mean of cattle blood.
Source: blood of infected canines (Sykes et al., 2005).
Host habitat: circulation of infected canines.
Phylogeny: assignment to the hemoplasma cluster of
the pneumoniae group of Mollicutes (Foley and Pedersen,
2001).
Cell morphology: wall-less; coccoid in shape.
Optimum growth temperature: not applicable.
Cultivation status: non-culturable.
Source: blood of infected cattle.
Host habitat: blood of cattle.
Phylogeny: assignment to the hemoplasma cluster of the
pneumoniae group of the genus Mycoplasma.
Sequence accession no. (16S rRNA gene): EF460765.
Further comment: this organism is synonymous with “Candidatus
Mycoplasma haemobovis” (sequence accession no. EF616468).
612
3. “Candidatus Mycoplasma haemodidelphidis”
Walker, Raphael, Berent and Shi 2002, 697
Messick,
7. “Candidatus Mycoplasma ravipulmonis” Neimark, Mitchelmore and Leach 1998, 393
ha.e.mo.di.del¢phi.dis. Gr. neut n. haema blood; N.L. fem.
gen. n. didelphidis of the opossum; N.L. gen. n. haemodidelphidis of opossum blood.
ra.vi.pul.mo¢nis. L. adj. ravus grayish; L. n. pulmo, -onis the
lung; N.L. gen. n. ravipulmonis of a gray lung.
Source: blood of an infected opossum.
Host habitat: circulation of an infected opossum.
pneumoniae group of mollicutes (Messick et al., 2002).
4. “Candidatus Mycoplasma haemolamae” Messick, Walker,
Raphael, Berent and Shi 2002, 697
ha.e.mo.la¢ma.e. Gr. neut n. haema blood; N.L. gen. n. lamae
of the alpaca; N.L. fem. gen. n. haemolamae of alpaca blood.
Source: blood of infected llamas.
Host habitat: circulation of infected llamas (McLaughlin
et al., 1991).
pneumoniae group of mollicutes (Messick et al., 2002).
5. “Candidatus Mycoplasma
Pedersen 2001, 817
haemominutum”
Foley
and
ha.e.mo.mi¢nu.tum. Gr. neut n. haema blood; L. neut. part.
adj. minutum small in size; N.L. neut. adj. haemominutum
small (mycoplasma) from blood.
Source: blood of infected felines (George et al., 2002;
Tasker et al., 2003).
Host habitat: circulation of infected felines.
pneumoniae group of mollicutes (Foley and Pedersen,
2001).
Cell morphology: wall-less; coccoid in shape; 300–600 nm in
diameter.
6. “Candidatus Mycoplasma kahaneii” Neimark, Barnaud,
Gounon, Michel and Contamin 2002a, 697
ka.ha.ne¢i.i. N.L. masc. gen. n. kahaneii of Kahane, named
for I. Kahane.
Source: blood of infected monkeys (Saimiri sciureus)
(Michel et al., 2000).
Host habitat: circulation of infected monkeys.
Phylogeny: assignment to the hemoplasma cluster of
the pneumoniae group of mycoplasmas (Neimark et al.,
2002a).
Source: lung tissue of mouse with respiratory infection
(gray lung disease).
Host habitat: respiratory tissue of mice with pneumonia.
Phylogeny: forms a single species line in the hominis group
of mollicutes (Neimark et al., 1998; Pettersson et al., 2000).
Cell morphology: wall-less; coccoid in shape; 650 nm in
diameter.
8. “Candidatus Mycoplasma turicensis” Willi, Boretti, Baumgartner, Tasker, Wenger, Cattori, Meli, Reusch, Lutz and
Hofmann-Lehmann 2006, 4430
tu.ri.cen¢sis. L. masc. (sic) adj. turicensis pertaining to Turicum, the Latin name of Zurich, the site of the organism’s
initial detection.
Source: blood of infected domestic cats.
Host habitat: blood of domestic cats.
pneumoniae group of the genus Mycoplasma.
Sequence accession no. (16S rRNA gene): DQ157150.
The following proposed species has been incidentally
cited, but the putative organism remains to be established
definitively and the name has no standing in nomenclature.
1. “Candidatus Mycoplasma haemotarandirangiferis” Stoff
regen, Alt, Palmer, Olsen, Waters and Stasko 2006, 254
ha.e.mo.ta.ran.di.ran.gi¢fe.ris. Gr. neut n. haema blood;
Rangifer tarandus scientific name of the reindeer; N.L. gen. n.
haemotarandirangiferis epithet intended to indicate occurrence in blood of reindeer.
Source: blood of reindeer.
Host habitat: blood of reindeer.
Phylogeny: partial 16S rRNA gene sequences suggest possible relationships to Mycoplasma ovis and Mycoplasma wenyonii
(suis cluster), and/or to Mycoplasma haemofelis (haemofelis
cluster).
Cell morphology: single punctate, chaining punctate, clustering punctate, single bacillary, chaining bacillary, single
rings, chaining rings, and clustering rings.
Sequence accession nos (16S rRNA gene): DQ524812–
DQ524818.
Further comment : sequence accession no. DQ524819, representing clone 107LSIA (Stoffregen et al., 2006), does not
support assignment of that clone to the genus Mycoplasma
because it is most similar to 16S rRNA genes of Fusobacterium spp. Several different partial 16S rRNA gene sequences
obtained from other clones are too variable to establish their
coherence as a species.
Genus II. Ureaplasma
613
Other organisms
1. “Mycoplasma insons” May, Ortiz, Wendland, Rotstein,
Relich, Balish and Brown 2007, 298
in¢sons. L. neut. adj. insons guiltless, innocent.
Source: trachea and choanae of a healthy green iguana
(Iguana iguana).
Host habitat: respiratory tract and blood of green iguanas
(Iguana iguana).
Phylogeny: assignment to the Mycoplasma fastidiosum cluster
of the pneumoniae group of the genus Mycoplasma.
Cultivation status: cells are culturable in SP-4 medium supplemented with glucose.
Cell morphology: pleomorphic, but many have a highly atypical shape for a mycoplasma, often resembling a twisted rod.
Optimum growth temperature: 30°C.
2. “Mycoplasma sphenisci” Frasca, Weber, Urquhart, Liao,
Gladd, Cecchini, Hudson, May, Gast, Gorton and Geary
2005, 2979
sphe.nis¢ci. N.L. gen. n. sphenisci of Spheniscus, the genus of
penguin that includes the jackass penguin (Spheniscus demersus) from which this mycoplasma was isolated.
Source: choanae of a jackass penguin (Spheniscus demersus)
with choanal discharge and halitosis.
Host habitat: upper respiratory tract of the jackass penguin.
Phylogeny: assignment to the Mycoplasma lipophilum cluster
of the hominis group of the genus Mycoplasma.
Cultivation status: cells are culturable in Frey’s medium
supplemented with glucose.
Cell morphology: pleomorphic; some cells exhibit terminal
structures.
3. “Mycoplasma vulturis” corrig. Oaks, Donahoe, Rurangirwa,
Rideout, Gilbert and Virani 2004, 5911
vul.tu¢ri.i. L. gen. n. vulturis of a vultures, named for the host
animal (Oriental white-backed vulture).
Source: lung and spleen tissue of an Oriental white-backed
vulture.
Host habitat: upper and lower respiratory tract of Oriental
white-backed vulture, where it replicates intracellularly.
Phylogeny: assignment to the Mycoplasma neurolyticum cluster of the hominis group of the genus Mycoplasma.
Cultivation status: cells can be grown in co-culture with
chicken embryo fibroblasts, but have not been grown in
pure in vitro culture.
Cell morphology: coccoid; cells display intracellular vacuoles
and intracellular granules of electron-dense material.
4. “Mycoplasma zalophi” Haulena, Gulland, Lawrence, Fauquier,
Jang, Aldridge, Spraker, Thomas, Brown, Wendland and
Davidson 2006, 43
za.lo¢phi. N.L. gen. n. zalophi of Zalophus, the genus of sea
lion that includes the California sea lion (Zalophus californianus) from which this mycoplasma was isolated.
Source: subdermal abscesses of captive sea lions.
Host habitat: subdermal and intramuscular abscesses,
joints, lungs, and lymph nodes of captive sea lions.
Phylogeny: assignment to the Mycoplasma hominis cluster of
the hominis group of the genus Mycoplasma.
Cultivation status: cells are culturable in SP-4 medium supplemented with glucose.
Cell morphology: not yet described.
Genus II. Ureaplasma Shepard, Lunceford, Ford, Purcell, Taylor-Robinson, Razin and Black 1974, 167AL
Janet A. Robertson and David Taylor-Robinson
U.re.a.plas¢ma. N.L. fem. n. urea urea; Gr. neut. n. plasma anything formed or moulded, image, figure; N.L.
neut. n. Ureaplasma urea form.
Coccoid cells about 500 nm in diameter; may appear as coccobacillary forms in exponential growth phase; filaments are
rare. Nonmotile. Facultative anaerobes. Form exceptionally
small colonies on solid media that are described either as tiny
(T) “fried-egg” colonies or as “cauliflower head” colonies having a lobed periphery. Unusual pH required for growth (about
6.0–6.5). Optimal incubation temperature for examined species is 35–37°C. Chemo-organotrophic. Like Mycoplasma, species
of Ureaplasma lack oxygen-dependent, NADH oxidase activity.
Unlike Mycoplasma, species of Ureaplasma lack hexokinase or
arginine deiminase activities but have a unique and obligate
requirement for urea and produce potent ureases that hydrolyze urea to CO2 and NH3 for energy generation and growth.
Genome sizes range from 760 to 1170 kbp (PFGE). Commensals
or opportunistic pathogens in vertebrate hosts, primarily birds
and mammals (mainly primates, ungulates, and carnivores).
DNA G+C content (mol%): 25–32 (Bd, Tm).
Type species: Ureaplasma urealyticum Shepard, Lunceford,
Ford, Purcell, Taylor-Robinson, Razin and Black 1974, 167
emend. Robertson, Stemke, Davis, Harasawa, Thirkell, Kong,
Shepard and Ford 2002, 593.
Although cellular diameters as small as 100 nm and as large
as 1000 nm, and minimal reproductive units of about 330 nm
in diameter have been reported (Taylor-Robinson and Gourlay,
1984), published thin sections of these organisms (trivial name,
ureaplasmas) show diameters only as large as 450–500 nm. The
exceptions are feline ureaplasmas with thin section diameters
of up to 800 nm (Harasawa et al., 1990a). Morphometric analysis of cells of the type strain of Ureaplasma urealyticum fixed in
the exponential phase of growth showed coccoid cells with
614
diameters of about 500 nm (Robertson et al., 1983). Similar
cellular diameters have been seen in hemadsorption studies in
which cells were pre-fixed during incubation before usual fixation for electron microscopy. Reports of budding and filamentous forms probably reflect the effect of cultural and handling
conditions on these highly plastic cells. Although the sequence
of the Ureaplasma parvum genome lacks any recognizable FtsZ
genes (Glass et al., 2000), ureaplasmas appear to reproduce by
binary fission. Because of their minute size, ureaplasma cells
are rarely seen by light microscopy. Although they are of the
Gram-stain-positive lineage, the lack of cell wall results in the
organisms appearing Gram-stain-negative. They are more easily
detected if stained with crystal violet alone. Electron micrographs have indicated hair-like structures, possibly pili, 5–8 nm
long, radiating from the membrane (Whitescarver and Furness,
1975). An extramembranous capsule was expected from light
microscopic studies. In cytochemical studies, a carbohydratecontaining, capsular structure has been demonstrated in a
strain of Ureaplasma urealyticum (Robertson and Smook, 1976;
Figure 110). The structure is a lipoglycan that has also been
demonstrated on the type strain T960T of Ureaplasma urealyticum
and on a serovar 3 strain of Ureaplasma parvum; the lipoglycan
composition was strain-variable (Smith, 1986). No viruses have
been seen nor has viral or plasmid nucleic acid been reported
in any ureaplasma.
Ureaplasma colonies are significantly smaller in diameter
(£10–175 nm) than those of Mycoplasma species (300–800 nm;
Figure 110). For this reason, they were first described as “tiny
(T) form PPLO (pleuropneumonia-like organisms) colonies”
(Shepard, 1954) and later called T-mycoplasmas (Meloni et al.,
1980; Shepard et al., 1974). Cultures on solid media may grow
in air, but more numerous and larger colonies result in 5–15%
CO2 in N2 or H2 (Robertson, 1982). Colonies of many strains
are detectable after overnight incubation and reach maximum
dimensions within 2 d. Isolates from ungulates may require
longer incubation. Temperatures of 20–40°C are permissive for
growth of examined strains, but their optimal incubation temperatures are 35–37°C (Black, 1973). Ureaplasma cultures in
liquid media are incubated aerobically with growth occurring
in the bottom of the tube, as revealed by changes in the pH
indicator. Mean generation times of 10 isolates from humans
ranged from 50 to 105 min (Furness, 1975). Maximum titers
of £108 organisms per ml of culture produce insufficient cell
mass for detectable turbidity, precluding growth measurement
by turbidometric or spectrophotometric methods. Growth is
best measured by broth dilution methods (Ford, 1972; Rodwell
and Whitcomb, 1983; Stemke and Robertson, 1982). When
immediate estimation of populations is required, ATP luminometry (Stemler et al., 1987) may be useful. An indicator system enhances colony detection and ureaplasma identification.
Ammonia from urea degradation causes a rise in pH and certain cations to form a golden to deep brown precipitate on the
colorless colonies, making them visible when viewed by directly
transmitted light. Initially, the urease spot test used a solution
of urea and 1 mM Mn2+ (as MnCl2 or MnSO4) dropped onto
agar (Shepard and Howard, 1970); later, Mn2+ was incorporated into the agar itself to create a differential solid medium
(e.g., Shepard, 1983). However, Mn2+ is toxic for ureaplasmas
(Robertson and Chen, 1984). Equimolar CaCl2 (Shepard and
Robertson, 1986) gave a similar response, but allowed the recovery of live cells. Manganese susceptibility has taxonomic value
(Table 138); animal isolates show differing responses (Stemke
et al., 1984; Stemler et al., 1987).
Ureaplasma urealyticum has some elements of the glycolytic cycle and pentose shunt (Cocks et al., 1985), but cannot degrade glucose and lacks arginine deiminase (Woodson
et al., 1965). Instead, ureaplasmas have a unique and absolute
requirement for urea (0.4–1.0 mM) and a slightly acidic environment (pH 6.0–7.0); pH values outside this range can be
associated with growth inhibition. The essential cytoplasmic
Figure 110. Ureaplasma colonial size and cellular morphology. (a) Many isolated Ureaplasma urealyticum colonies,
accentuated by a urease spot test, surround a single large Mycoplasma hominis colony on a solid genital mycoplasma
(GM) agar surface. Colonies of Ureaplasma urealyticum commonly have diameters of 15–125 mm; the diameter of
the Mycoplasma hominis colony shown is approximately 0.9 mm. (Reproduced with permission from Robertson
et al., 1983. Sexually Transmitted Diseases 10 (October-December Suppl.): 232–239 © Lippincott Williams & Wilkins.)
(b) Transmission electron micrograph of Ureaplasma urealyticum, serovar 4, strain 381/74 cells, showing coccoid
morphology, extramembranous capsule stained with ruthenium red, lack of a cell wall, the single limiting membrane, and apparently simple cytoplasmic contents in which only ribosomes are clearly evident. Cell diameters
were 485–585 nm. (Reproduced with permission from Robertson and Smook, 1976. Journal of Bacteriology 128:
658–660.)
615
Table 138. Phenotypic characteristics that partition the human serovar-standard strains of ureaplasmas to Ureaplasma speciesa
Characteristic
Isoelectric focusing and SDS-PAGE at pH 5.3 polypeptide
patterns:
Absent
Present
1-D SDS-PAGE T960T biovar band:b
Absent
Present
2-D SDS-PAGE:b
Biovar 1 pattern
Biovar 2 pattern
Growth inhibition by 1 mM Mn2+:
Temporary
Permanent
Polypeptide recognized by immunoblots:
51 and 58 kDa
47 kDa
Biovar 1 pattern
Biovar 2 pattern
mAb UU8/39 recognition of membrane proteins:
17 kDa only
16/17 kDa
mAb UU8/17 recognition of 72 kDa urease subunit:d
No
Yes
mAb VB10 recognition of 72 kDa urease subunit:
Yes
No
Urease dimorphism:
Lower MW
Higher MW
Pyrophosphatase dimorphism:
Lower MW
Higher MW
Diaphorase bands dimorphism:
Inapparent
Apparent
U. urealyticum serovar
U. parvum serovar
Reference
Sayed and Kenny (1980)
1, 3T, 6
2, 4, 5, 7, 8T
Howard et al. (1981)
1, 3T, 6
2, 4, 5, 7, 8T
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9, 11, 12
Mouches et al. (1981)
Swensen et al. (1983)
Robertson and Chen (1984)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–12c
1, 3T, 6, 14
Horowitz et al. (1986)
1, 3T
Lee and Kenny (1987)
2, 4, 5, 7, 8T, 9–13
2, 4, 8T
Thirkell et al. (1989)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Thirkell et al. (1990)
1, 3T, 6, 14
8T
MacKenzie et al. (1996)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Davis et al. (1987)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Davis and Villanueva (1990)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Davis and Villanueva (1990)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
T, Type strain of species.
a
1-D, One-dimensional; 2-D, two-dimensional.
b
Serovar 13 gave an intermediate response and was excluded from the initial partition scheme.
c
Hyperimmune rabbit sera and acute and convalescent sera from women with postpartum fever.
d
urease, comprising three subunits (Blanchard, 1990), produces a transmembrane potential, which leads to ATP synthesis (Romano et al., 1980; Smith et al., 1993). Ureaplasma
dependence upon catalysis of urea for energy is the basis
of growth inhibition by the urease inhibitor hydroxamic
acid and its derivatives (Ford, 1972) and by fluorofamide
(Kenny, 1983). The proton pump inhibitor lansoprazole and
its metabolites interfere with ATP synthesis at micromolar
concentrations (Nagata et al., 1995). Their small genomes
make ureaplasmas dependent upon the host for amino acids,
amino acid precursors, lipids, and other growth components.
Ureaplasmas from humans exhibit minimal levels of acetate
kinase activity (Muhlrad et al., 1981). Like other Mycoplasmataceae, ureaplasmas make superoxide dismutase (O’Brien et al.,
1983), but, unlike the rest of this family, oxygen-dependent
NADH oxidase has not been detected (Masover et al., 1977).
The Ureaplasma parvum genome sequence includes genes for
six hemin and/or Fe3+ transporters which are believed to be
related to respiration (Glass et al., 2000). For information
on additional physiological traits, see: Black (1973); Shepard
et al., (1974); Shepard and Masover (1979); Taylor-Robinson
and G
ourlay (1984); and Pollack (1986).
Except for the obligatory requirements for supplementary
urea and lower pH, ureaplasma growth requirements are similar to those of members of the genus Mycoplasma. The sterol
requirement is met by horse, bovine, or fetal bovine serum.
Because heat-sensitive pantothenic acid is a growth factor supplied by serum (Shepard and Lunceford, cited by Shepard and
Masover, 1979), the serum supplement should not be “inactivated” by heating to reduce complement activity. The effect of
yeast extract is variable, perhaps depending upon the particular
strain requirements or the batch of extract. Other defined additives have been reported to enhance growth but, in the absence
of a defined medium, evaluation is difficult. Dependence on
sterols for the integrity of the cell membrane renders ureaplasmas susceptible to digitonin and certain antifungal agents, but
most strains tolerate the polyene nystatin (50 U/ml) that prevents overgrowth by yeasts.
616
Information about ureaplasma genetics is now abundant. The
genomes of 26 strains of Ureaplasma, including six of the seven
named species and six unnamed strains, range in size from 760
to 1170 kbp (Kakulphimp et al., 1991; Robertson et al., 1990).
The largest genome belongs to Ureaplasma felinum. The G+C
content of ureaplasmal DNA is in the range 25.5–31.6 mol%,
which is lower than that for other Mollicutes and for all other
prokaryotes, greatly limiting the degeneracy in the genetic code
of ureaplasmas. For the type strain of Ureaplasma urealyticum,
UGA is a codon for tryptophan (Blanchard, 1990). The entire
sequences of the genomes of two strains of Ureaplasma parvum
serovar 3 (formerly known as Ureaplasma urealyticum biovar 1;
Glass et al., 2000) have been determined. At 752 kbp, the organisms share with other obligate symbions, such as Mycoplasma
genitalium (580 kbp) and Buchnera aphidicola (641 kbp), greatly
reduced genomes. Of 641–653 total genes, 32–39 code for structural RNAs and about 610 code for proteins, about 47% of which
have been classified as hypothetical genes of unknown function.
Some (19%) resemble genes present in other genomes, but many
(28%) appear unique. Unexpectedly absent in the Ureaplasma
parvum genome are recognizable genes for FtsZ, for the GroEl
and GroES chaperones, and for ribonucleoside-diphosphate
reductase. Attempts have been made to reconcile the anomalies
of gene functions assigned to this serovar of Ureaplasma parvum
with the activities and pathways found in various ureaplasmas
of humans (Glass et al., 2000; Pollack, 2001). One limitation to
such analysis is that most of the physiological data that has been
accumulated pertain not to Ureaplasma parvum but instead to
the type species, Ureaplasma urealyticum strain T960T, possessing a 7% larger genome. The entire sequences of the genomes
of Ureaplasma urealyticum serovars 8 and 10 (Glass et al., 2000,
2008) have also been determined and the genomes of the type
strains of all remaining Ureaplasma urealyticum and Ureaplasma
parvum serovars are nearly completely annotated (Glass et al.,
2008). Preliminary comparisons found that in addition to “core”
and “dispensable” genomes for each species, Ureaplasma urealyticum had partly duplicated multiple-banded antigen (mba) genes
(Kong et al., 1999a, 2000), and up to twice the number of genes
that Ureaplasma parvum has for lipoproteins. Such comparisons
are expected to substantially improve understanding of ureaplasmal pathogenicity and differential strain virulence.
Clinical studies based entirely on qualitative assessments of
ureaplasmas are often difficult to interpret and only a few investigators have presented quantitative data (Bowie et al., 1977;
De Francesco et al., 2009; Heggie et al., 2001; Taylor-Robinson
et al., 1977). Factors such as hormonal levels, specific genetic
attributes, and even socio-economic conditions may encourage urogenital colonization and proliferation. Nevertheless,
it is clear that ureaplasmas are commensals that, on occasion,
contribute to disease in susceptible human hosts. Infections
attributed to ureaplasmas are often associated with an immunological component (Bowie et al., 1977), including the subset
of the human population with common variable hypogammaglobulinemia (Cordtz and Jensen, 2006; Furr et al., 1994;
Lehmer et al., 1991; Webster et al., 1978), in which ureaplasmainduced septic arthritis and occasionally persistent ureaplasmal
urethritis is seen (Taylor-Robinson, 1985). Sexually acquired,
reactive arthritis is usually linked to Chlamydia trachomatis, but
immunological evidence of ureaplasmal involvement exists
(Horowitz et al., 1994). Through urea metabolism, ureaplasmas
can induce crystallization of struvite and calcium phosphates
in urine in vitro and produce urinary calculi in animal models
(Reyes et al., 2009). They are found in patients with infection
stones more often than in those with metabolic stones (Grenabo
et al., 1988). A statistical association with infection stones has
been made (Kaya et al., 2003).
Evidence for the association of ureaplasmas with acute
nongonococcal urethritis in men has been controversial, but
a significant association of Ureaplasma urealyticum (but not Ureaplasma parvum) with this disease in two of three recent studies
suggests a way forward to resolving this issue. Evidence for a
role for ureaplasmas in acute epididymitis is, at the most, meager and it is very unlikely that they have a role in chronic prostatitis. Ureaplasmas have been associated with bacterial vaginosis
and pelvic inflammatory disease, but are unlikely to be causal in
either condition. It follows, therefore, that there is no convincing evidence to implicate ureaplasmas as an important cause
of infertility in men or in couples. The most convincing data
relating ureaplasmas to poor pregnancy outcomes have been
seen when the organisms have been detected in amniotic fluid
prior to membrane rupture, but there are conflicting opinions
about the role of Ureaplasma urealyticum vs Ureaplasma parvum.
Data since the 1980s have supported the association of neonatal
ureaplasma infection with chronic lung disease and sometimes
death in very low birth-weight infants. The ability of Ureaplasma
parvum to induce chorioamnionitis and to contribute to preterm labor and fetal lung injury is supported by experimental
studies in rhesus monkeys (Novy et al., 2009).
Information on the range of animal species infected with
ureaplasmas and their geographic distribution is patchy, possibly because ureaplasmas are largely avirulent or, at least, not an
economic threat. Isolations have been reported from squirrel,
talapoin, patas, macaque and green monkeys; as well as marmosets and chimpanzees (Taylor-Robison and Gourlay, 1984).
Also, there are reports of isolation from domestic dogs, raccoon
dogs, cats and mink; cattle, sheep, goats, and camels; chickens
and other fowl; and swine (the latter needing confirmation).
Baboons, rats and mice are susceptible to experimental infection with ureaplasmas. While Ureaplasma diversum can inhabit all
mucosal membranes of cattle, the two natural diseases it causes
are subclinical respiratory infections in young calves, which
occasionally develop into bronchopneumonia, and the economically important urogenital infections transmitted by bulls
or their semen. The latter present as vulvovaginitis or ascend
to cause infertility or abortion (Ruhnke et al., 1984; ter Laak
et al., 1993). A species-specific PCR assay (Vasconcellos Cardosa
et al., 2000) can circumvent culture insensitivity. As in humans,
ureaplasmal diseases in animals may be influenced by the particular ureaplasma strain or many other factors. Although ureaplasmas may be present initially in certain organ and primary
cell cultures, the lack of urea and higher pH in most eukaryotic
cell culture systems would discourage persistence.
Although the spectrum of diseases of primary ureaplasmal
etiology remains controversial, many potential virulence factors
have been identified. Structural elements include a capsule,
pilus-like fibrils, and the antigens of the outer membrane that
constitute the serovar determinants. Erythrocytes from several
animal species adhere firmly to colonies of certain strains of
Ureaplasma parvum (Shepard and Masover, 1979), but most
human isolates exhibit transient or no binding (Robertson
and Sherburne, 1991). Strains of both Ureaplasma urealyticum
and Ureaplasma parvum adhere to HeLa cells (Manchee and
Taylor-Robinson, 1969) and to spermatozoa (Knox et al.,
2003). Demonstration of beta-hemolysis of erythrocytes by ureaplasmal products depends upon several variables; hemolysis
of guinea pig erythrocytes has been most consistently observed
(Black, 1973; Manchee and Taylor-Robinson, 1970; Shepard,
1967). Manchee and Taylor-Robinson (1970) described hemolysis of homologous erythrocytes by a canine ureaplasma as peroxide-associated and blocked lysis by adding catalase, except in
the presence of a catalase inhibitor. Genome sequencing has
revealed genes resembling those for the hemolysins HlyA and
HlyC of enterohemorrhagic Escherichia coli (Glass et al., 2000).
Ammonia and ammonium ions generated by urea hydrolysis
and the alkaline environment that they create are inhibitory
to the ureaplasmas (Ford and MacDonald, 1967; Shepard and
Lunceford, 1967). They have well-established deleterious effects
on eukaryotic cell and mammalian tissue cultures and may be
the “toxin” described but never substantiated (Furness, 1973).
An IgA1 protease activity has been demonstrated by ureaplasmas from both humans and canines. Human IgA1-specific protease activity, apparently similar to the type 2 serine protease of
certain bacterial pathogens of humans (Spooner et al., 1992), is
produced by both Ureaplasma parvum and Ureaplasma urealyticum
(Kilian et al., 1984; Robertson et al., 1984), but a gene responsible for the activity has not yet been identified. Putative phopholipase A1, A2, and C activities (De Silva and Quinn, 1986) could
not be confirmed, nor could such gene sequences be identified
(J. Glass, unpublished). Biofilm production has been described
recently (García-Castillo et al., 2008). Ureaplasmas have been
seen intracellularly during studies in cell cultures (Mazzali
and Taylor-Robinson, 1971), but may be there transiently after
phagocytosis. Cells in culture (Li et al., 2000), in either experimental infection models (Moss et al., 2008) or epidemiological
study subjects (Buss et al., 2003; Dammann et al., 2003; Jacobsson et al., 2003; Shobokshi and Shaarawy, 2002), have exhibited
proinflammatory responses. While assessment of clinical studies
is sometimes difficult because of inherent reporting bias (Klassen et al., 2002; Schelonka et al., 2005), it is anticipated that
the application of bioarray technologies will lead to improved
understanding of ureaplasmal mechanisms of pathogenesis.
Reviews of in vitro methodologies used for antimicrobial
susceptibility testing for both human (Bébéar and Robertson,
1996; Waites et al., 2001) and animal (Hannan, 2000) isolates
are available. Antimicrobial susceptibility patterns of Ureaplasma
urealyticum and Ureaplasma parvum appear to be similar (Matlow
et al., 1998). The choice of therapeutic agents active against
them is limited. Tetracyclines or the macrolides (excluding lincomycin and clindamycin) are the bacteriostatic agents usually
employed. Ureaplasmas from human, simian, bovine, caprine,
feline, and avian sources (Koshimizu et al., 1983) withstand
relatively high levels of lincomycin (10 mg/ml), to which most
mycoplasmas are susceptible. Ureaplasmas also show in vitro
resistance to rifampin and sulfonamides. Clinical resistance of
ureaplasmas to tetracyclines has been long known (Ford and
Smith, 1974). This high-level resistance is determined by the
presence of the tetM determinant (Roberts and Kenny, 1986),
which is now readily identified by PCR (Blanchard et al., 1997).
Tetracycline-resistant strains exposed to a variety of antibiotics demonstrate a broad range of responses (Robertson et al.,
1988). Clinical resistance to macrolides has also been reported
(e.g., Taylor-Robinson and Furr, 1986). Certain aminoglyco-
617
sides, chloramphenicol, and newer fluoroquinolones inhibit
ureaplasmas, but are inappropriate for broad clinical use. Fluoroquinolone resistance is increasing (Xie and Zhang, 2006) and
has been found in a previously susceptible strain (Duffy et al.,
2006). Ureaplasma strains exhibiting resistance to multiple antibiotics have been found in immunocompromised hosts. In one
case, the isolates were of the same serovar, but exhibited different susceptibility patterns at different anatomical sites (Lehmer
et al., 1991). Because of ongoing changes in antimicrobial susceptibility patterns, the recent literature should be consulted
(e.g., Beeton et al., 2009). To treat natural Ureaplasma diversum
infections, tiamulin hydrogen fumarate, a diterpene agent in
common use in veterinary medicine, may be at least as effective
as the macrolide tylosin (Stipkovits et al., 1984). Others have
examined the efficacy of single or combinations of antibiotics
in eradicating ureaplasmas from various sites in cattle as well
as from semen used for artificial insemination (ter Laak et al.,
1993). The urease inhibitor, fluorofamide, has been used with
varying success in eliminating ureaplasmas from animals.
When establishing antibiograms for ureaplasmas, the requirements established for common bacterial pathogens do not suffice. First, medium components may affect antibiotic activity.
For instance, serum-binding reduces tetracycline activity, while
the initial acidic culture reduces macrolide activity against ureaplasmas (Robertson et al., 1981). Conventional bacteria of
established, low-level susceptibility can be used to measure the
effect of the ureaplasmal medium on a particular antimicrobial agent. Second, the relatively slow growth of ureaplasmas
as compared with many pathogenic bacteria requires that the
half-life of the antibiotic be considered when test inoculum and
incubation period are established. Lastly, the end points of the
sensitivity tests on agar are about four-fold lower than for tests
in broth (Waites et al., 1991). On consideration of the in vivo
environment in which these organisms naturally occur, interpretation of susceptibility test end points continues to present
a challenge. An international subcommittee under the aegis
of the National Committee for Clinical Laboratory Standards
Institute (USA) is currently finalizing a “Final Report for Development of Quality Control Reference Standards and Methods
for Antimicrobial Susceptibility Testing” for Ureaplasma and
Mycoplasma species infecting humans that have demonstrated
variability in response to antimicrobial agents.
Media formulations in current use for the cultivation of
ureaplasmas from human sources include, in order of decreasing
supplementation: the 10B broth of Shepard and Lunceford
(Shepard, 1983); Taylor-Robinson’s broth (Taylor-Robinson,
1983a), made without thallium acetate; and bromothymol blue
broth (Robertson, 1978). Ureaplasmas are exquisitely sensitive to
thallium acetate and it should not be used to inhibit other bacteria. The U4 formulation for ungulate isolates was developed by
Howard et al. (1978). Consult Waites et al. (1991) for additional
media formulations for isolation from humans, and Shepard
(1983), Livingston and Gauer (1974), or Hannan (2000) for isolation from animals. The quality of medium components may be
as important as the medium formulation. Serum supplements
should be tested for their ability to support growth of the ureaplasmas of interest. Liquid medium may be stored at −20°C until
required. Transport medium (e.g., 2SP) should be free of antibi-
618
otics inhibitory to ureaplasmas. Broth cultures should be diluted
to ³1:100 to reduce effects of any growth inhibitors present in the
specimen. In general, Ureaplasma parvum is less demanding nutritionally than Ureaplasma urealyticum and isolates from human and
other animal sources together are less demanding than those
from ungulate hosts. Agar surfaces are examined at ³40× magnification. The less fastidious strains (e.g., Ureaplasma parvum type
strain) are more likely to produce recognizable “fried-egg” colonies than are the more fastidious strains (e.g., Ureaplasma urealyticum type strain), which are more likely to produce “cauliflower
head” or core colonies. Cultures may be thrice cloned and the
resultant culture used to initiate stocks.
Broth cultures of ureaplasmas commonly become sterile within
12–24 h incubation at 35–37°C. “Red is dead” was the mantra of
Shepard in regard to his phenol red-containing broth medium.
One way to lessen this problem is to change to an indicator that
changes color at a lower pH (Robertson, 1978). To Shepard’s
mantra we might add, “the higher the urea concentration, the
steeper the death phase”, an effect not countered by buffer. Incubation of diluted cultures at 30–34°C slows growth and maintains viability for up to 1 week, reducing the frequency of culture
transfer and helping in transport. Short exposure of broth cultures to refrigeration reduces viability. Cells within colonies on
solid medium may be recovered for approximately 1 week if the
cultures are removed from incubation when growth is detectable and stored under cool, humidified conditions. For longterm storage, ultra-low temperatures (−80°C or liquid nitrogen)
with a cryoprotectant [e.g., 10–20% (v/v) sterile glycerol] can
maintain viability for well over a decade. For lyophilization, the
pellets from broth cultures centrifuged at high speed are resuspended in a minimal volume of liquid medium or the serum
used for its supplementation before processing; on reconstitution, urease activity occasionally is not immediately evident.
Differentiation of the genus Ureaplasma from other genera
wall, filterability, and the presence of conserved 16S rRNA
gene sequences. Aerobic or facultatively anaerobic growth in
artificial media and the necessity for sterols for growth exclude
assignment to the genera Anaeroplasma, Asteroleplasma, Acholeplasma, and “Candidatus Phytoplasma”. Non-spiral cellular morphology and regular association with a vertebrate host or fluids
of vertebrate origin support exclusion from the genera Spiroplasma, Entomoplasma, and Mesoplasma. The ability to hydrolyze
urea, with the inability to metabolize either glucose or arginine,
excludes assignment to the genus Mycoplasma. For routine purposes, the colonial morphology characteristic of ureaplasmas
and demonstration of the isolate’s ability to catabolize urea,
using the urease spot test or indicator agar, suffice for preliminary differentiation from other mollicutes. Other methods for
urease detection have been largely replaced by PCRs that detect
urease genes.
Taxonomic comments
The hypothetical evolutionary relationships of the Mollicutes
have been based primarily upon 16S rRNA gene sequences
(Weisburg et al., 1989). The nucleotide sequence for the
16S rRNA genes (Kong et al., 1999b; Robertson et al., 1993;
obertson et al., 1994) and the 16S–23S rRNA intergenic spacer
R
regions of all named species plus the 14 serovars associated with
humans (Ureaplasma urealyticum and Ureaplasma parvum) have
been determined (Harasawa and Kanamoto, 1999; Kong et al.,
1999b). The genus Ureaplasma comprises two subclusters within
the highly diverse pneumoniae group of the family Mycoplasmataceae (Johansson, 2002). One subcluster contains the human,
avian, and mink isolates and the other contains feline, canine,
and bovine strains (see genus Mycoplasma Figure 109, pneumonia group) Although the three serovars A, B, and C of Ureaplasma
diversum are antigenically heterogeneous, the strains examined
meet the 70% DNA–DNA hybridization benchmark used as an
arbitrary species criterion. However, the available DNA–DNA
hybridization values suggest that Ureaplasma gallorale and Ureaplasma canigenitalium might each represent more than a single
species. The 16S rRNA gene sequences of ureaplasmas isolated
from nonhuman primates and some less-studied vertebrates are
unknown. The range of G+C contents of ureaplasmal DNA is
too narrow to have much taxonomic utility. Nevertheless, the
values for isolates from cattle, sheep, and goats are between
28.7 and 31.6 mol% (Howard et al., 1978), at the higher end of
the range for the genus.
It is generally assumed that all ureaplasmas from avian
sources belong to the species Ureaplasma gallorale. The seven
avian isolates examined were similar to each other serologically
and in SDS-PAGE, immunoblot, and RFLP profiles, and they
were distinct from Ureaplasma urealyticum and Ureaplasma diversum, although resemblances between Ureaplasma gallorale and
Ureaplasma urealyticum based upon one- and two-dimensional
PAGE analyses have been reported (Mouches et al., 1981). However, the DNA–DNA hybridization values among Ureaplasma gallorale strains fall into two clusters. Homology was 70–100% for
the five strains within cluster A and 96–100% for the two strains
within cluster B that were studied. Between strains of cluster
A and B, homology was only 51–59% and, vice versa, 52–69%
(Harasawa et al., 1985). The taxonomic status of avian ureaplasmas, therefore, might benefit from examination of additional
strains and reconsideration. The clusters appear to be more
closely related than Ureaplasma urealyticum and Ureaplasma parvum (Table 139).
The taxonomy of ureaplasmas of canines is also problematic.
The first report indicated G+C contents of 27.2–27.8 mol%
(Bd; Howard et al., 1978). Later, the representatives of four
serogroups (SI to SIV for strains DIM-C, D29M, D11N-A, and
D6P-CT, respectively) were reported to have G+C contents of
28.3–29.4 mol% (HPLC). However, the published data regarding their DNA reassociation values are confusing. Initially,
Barile (1986) and Harasawa et al. (1990b) reported that the
serogroup SI representative, DIM-C, had 73% homology with
the serotype 2 SII representative, D29M, indicating that these
two strains likely belong to the same species. However, perhaps
because of the borderline value, further work was undertaken,
also using [3H]DNA–DNA hybridization procedures (Harasawa
et al., 1993). The values obtained ranged from 41 to 63% homology among the four serogroups, i.e., each serogroup seemed to
represent a distinct species. At present, the only named canine
ureaplasma species is Ureaplasma canigenitalium; the SIV representative D6P-CT is the type strain (Harasawa et al., 1993).
Its degree of distinctiveness from the other serogroups is not
emphasized in the literature.
Table 139. Genotypic characteristics that partition the serovar-standard strains of ureaplasmas isolated from humans to the level of speciesa
Characteristic
U. urealyticum serovars
DNA–DNA relatedness with strain 27 DNA:
91–102%
38–60%
DNA–DNA relatedness with strain T960T DNA:
49–52%
69–100%
DNA–DNA relatedness with strain 27T DNA:
75–100%
38–57%
DNA–DNA relatedness with strain T960T DNA:
48–59%
76–101%
Cleavage of DNA by Fnu4HI:
Yes
No
BamHI, HindIII, and PstI RFLP probed with RNA genes:
Biovar l pattern
Biovar 2 pattern
EcoRI and HindIII RFLP probed with RNA genes:
Biovar 1 pattern
Biovar 2 pattern
Genome sizes determined by PFGE:
~760 kbp
840–1140 kbp
Heterogeneity of alpha polypeptide-associated urease genes:
Yes
No
Heterogeneity of HindIII site in subunit ureC of urease gene:
Absent
Present
Heterogeneity of HindIII fragments probed with
serovar 8IC61 urease probe:
Biovar 1 pattern
Biovar 2 pattern
Heterogeneity of urease subunit-associated genes:d
Biovar 1 pattern
Biovar 2 pattern
Heterogeneity of biovar-specific 16S rRNA genes determined
by PCR:d
Strain 27T
Strain T960T
Biovar-specific 16S rRNA gene sequences:d
Strain 27T sequence
Strain T960T sequence
16S rRNA, spacer regions, and urease subunit sequences:d
Biovar 1 pattern
Biovar 2 pattern
RFLP of 5¢ region of mba genes:
Biovar 1 pattern
Biovar 2 pattern
5¢ end sequences of mba genes:d,e
Biovar 1 pattern
Biovar 2 pattern
16S–23S intergenic spacer region:d
U. parvum serovars
Biovar 1 pattern
Biovar 2 pattern
Arbitrarily primed PCR:d
Biovar 1 pattern
Biovar 2 pattern
Reference(s)
Christiansen et al. (1981)
T
1, 3T, 6
2, 4, 5, 7, 8T
Christiansen et al. (1981)
1, 3T, 6
2, 4, 5, 7, 8T
Harasawa et al. (1991)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Cocks and Finch (1987)
1, 3T, 6
2, 4, 5, 7, 8T, 9
Razin (1983)
1, 3T, 6
2, 4, 5, 7, 8T, 9
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13b
Robertson et al. (1990)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Blanchard (1990)
1, 3T, 6, 10c, 12c, 14
2, 4, 5, 7, 8T, 9, 13
Neyrolles et al. (1996)
1, 3T, 6
2, 8T
Neyrolles et al. (1996)
1, 3T, 6
2, 8T
Kong et al. (1999b)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
1, 3T, 6, 14
2, 5, 8T
Kong et al. (1999b)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Teng et al. (1994)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Kong et al. (1999b)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Harasawa and Kanamoto
(1999), Kong et al. (1999b)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
Grattard et al. (1995)
1, 3T, 6, 14
2, 4, 5, 7, 8T, 9–13
T, Type strain of species.
a
Serovar 13 response to EcoRI was anomalous.
b
This anomalous pattern resulted from cultures being misidentified as serovars 10 and 12; the error was corrected by Teng et al. (1994). Kong et al. (1999b) have confirmed the efficacy of the Blanchard (1990) primers.
c
Primers used for biovar-defining PCR(s).
d
The multiple band (MB) antigens seen by PAGE are putative virulence markers. See text for details.
e
620
While DNA–DNA hybridization has been key to species level
identification for the genus Ureaplasma, serology led to our knowledge of intra-species relationships (Robertson et al., 2002) and
still figures importantly. Much interest has been focused specifically on the multiple-banded antigens (MBA) of the ureaplasmal cell surface. Watson et al. (1990), studying the type strain
(serovar 3) of Ureaplasma parvum, identified the predominant
ureaplasmal antigens recognized by the host during human
infection. The MBA were first seen as unusual, laddered bands
in immunoblots. These lipoproteins exhibited epitopes for both
serovar specificity and cross-reactivity, and showed in vitro size
variation (Teng et al., 1994; Zheng et al., 1995). Monecke et al.
(2003) added evidence that the MBA are involved in a phaseswitching process similar to that identified in several Mycoplasma
species. The mba gene sequences have since been used to further
define ureaplasma phylogeny (Knox et al., 1998; Kong et al.,
1999b), as well as to characterize isolates (Knox et al., 1998;
Knox and Timms, 1998; Kong et al., 1999a; Pitcher et al., 2001).
The sequences of the 5¢ ends of the mba genes of all 14 serovars
have been defined (Kong et al., 1999a, 2000). Kong et al. (2000)
found that for Ureaplasma parvum, a specific site on the gene
determines serovar identity, whereas for Ureaplasma urealyticum
at least the sequence of the 3¢ end is required and, for certain
other serovars, the entire sequence is involved.
The genes for the three urease subunits, ureA, ureB, and ureC,
and adjoining regions from many isolates have been sequenced
(Blanchard, 1990; Kong et al., 1999b; Neyrolles et al., 1996;
Ruifu et al., 1997). Rocha and Blanchard (2002) applied bioinformatics to predict how certain gene products (e.g., the MBA,
restriction and modification systems, transcription anti-termination elements, and GTP-binding proteins) might exhibit
species specificity. PCRs based upon the 16S rRNA genes are
the most highly conserved, followed by the 16S–23S intergenic
region, the urease-associated genes, the mba genes, and the
region upstream of them.
Acknowledgements
We thank J. Glass and collaborators at the J. Craig Venter Institute and the University of Alabama - Birmingham for the gift
of the genome sequences of Ureaplasma parvum and Ureaplasma
urealyticum, and J.G. Tully, K.E. Johansson, and C. Williams for
their suggestions regarding the initial chapter.
Differentiation of the species of the genus Ureaplasma
The first paper on ureaplasma taxonomy stated that “An advantage of forming a new genus is that it confers freedom to classify
new species within the genus without adhering to the principles
formulated for the other genera. A numbered serovar of a Ureaplasma from humans is broadly equivalent to a named species
within the genera Mycoplasma or Acholeplasma” (Shepard et al.,
1974). The serological diversity within the type strain, Ureaplasma
urealyticum, was regarded as no more than antigenic heterogeneity, possibly reflecting minor differences within a single epitope.
To ensure that taxonomy would develop rationally, official recognition of Ureaplasma subspecies was avoided. Thirty-five years
later, this taxonomic restraint can be appreciated.
When the first genus and species, Ureaplasma urealyticum,
was named, it had eight known antigenic specificities (Shepard
et al., 1974); 6 years later, the number had reached 14 (Robertson and Stemke, 1982) where, surprisingly, it has remained. It
is surprising because the serotypes were isolated over a 20-year
period representing antigens in Vancouver, BC, Canada (Ford,
1967), Camp Lejeune, NC (Shepard, 1954), and Boston, MA,
USA (Lin et al., 1972), i.e., on the west and east coasts of North
America. While putative untypable strains are occasionally
encountered, after cloning, these have usually turned out to be
serovar 3, probably the most likely to dominate in a mixed culture. However, additional serovars/genovars can be expected to
emerge, especially from other parts of the world.
The named species were identified primarily by DNA–DNA
hybridization and by serological tests. For taxonomic studies,
the metabolism inhibition test (Purcell et al., 1966; Robertson
and Stemke, 1979; Taylor-Robinson, 1983b) and either a direct
or indirect immunofluorescence test (Black and KrogsgaardJensen, 1974; Piot, 1977; Stemke and Robertson, 1981) have
been most useful. However, serological tests sometimes gave
confusing cross-reactivity patterns and were not ideal for differentiating strains within a single species (Stemke and Robertson,
1985). In an attempt to circumvent problems of cross-reactions
obtained with polyclonal antisera, monoclonal antibodies
(mAbs) to all 14 serovars of human isolates were developed
(e.g., Echahidi et al., 2001). The use of mAbs coincided with
and has been largely overshadowed by the genomic revolution.
In the 1980s, the same pattern of partitioning of the serovars
of human isolates was demonstrated by other phenotypic traits,
traits primarily related to protein structures and functions
(Table 138). When phenotypy failed to deliver a clear and
convenient means of discrimination, genotypy did. The initial
DNA–DNA relatedness studies of ureaplasmas from humans
(Christiansen et al., 1981) confirmed the two, distinct clusters;
however, the large cell biomass, special equipment, and (then)
rare expertise required resulted in few strains being tested. New
techniques, especially PCR, became more easily performed and
less expensive so that more strains were examined and nonambiguous results were obtained (Table 139). Supported by
these strong data, the taxonomy of Ureaplasma urealyticum was
emended and extended (Robertson et al., 2002). The ten antigenic specificities of the larger cluster (known as group 2 or as
the T960 biovar) retained the Ureaplasma urealyticum designation with strain T960T as the type strain. The remaining four
antigenic specificities (known as group 1 or the parvo biovar)
were renamed Ureaplasma parvum in recognition of that cluster’s considerably smaller genome size; strain 27T, the serovar
3 standard, was designated the type strain. Many PCR primers
to identify species and strains of ureaplasmas have been published; commercial PCR-based kits are currently available for all
named Ureaplasma species.
In summary, current ureaplasma taxonomy is based upon
pragmatic, polyphasic criteria, i.e., a synthesis of phylogeny,
phenotypy, and genotypy. For the specific requirements for
taxonomic studies of Mollicutes, consult the most recent minimal standards document (Brown et al., 2007). Some serological
testing is mandatory. Rabbit antisera to the 14 serovars of ureaplasmas of humans and certain animal species are currently
available from Jerry K. Davis, Curator of the Mollicutes Collection, School of Veterinary Medicine, Purdue University, West
Lafayette, IN, USA. Expertise in determining phenotypic traits
specific to ureaplasmas may be accessible through collaboration
with the appropriate working team of the International Research
Programme for Comparative Mycoplasmology (IRPCM) of the
International Organization for Mycoplasmology (IOM) at www.
the-iom.org.
621
List of species of the genus Ureaplasma
1. Ureaplasma urealyticum Shepard, Lunceford, Ford, Purcell,
Taylor-Robinson, Razin and Black 1974, 167AL emend. Robertson, Stemke, Davis, Harasawa, Thirkell, Kong, Shepard
and Ford 2002, 593
u.re.a.ly¢ti.cum. N.L. fem. n. urea urea; N.L. adj. lyticus -a -um
(from Gr. adj. lutikos -ê -on) able to loosen, able to dissolve;
N.L. neut. adj. urealyticum urea-dissolving or urea-digesting.
Cells are coccoid and approximately 500 nm in diameter.
Coccobacillary forms are seen in exponential phase cultures.
One strain has been shown to have a carbohydrate-containing
capsule; it and others contain lipoglycans. Grows at temperatures between 20 and 40°C, grows better at 30–35°C and best
at 36–37°C. Colonies are £20–50 mm in diameter with complete or partial fried-egg morphology.
Serologically distinct from all other named species in the
genus, but serologically heterogeneous. Ten specific antigenic determinants are known: 2, 4, 5, and 7–13. Multiplebanded antigens are serovar-related and recognized by the
host. Like Ureaplasma parvum, it has human IgA-specific protease activity that specifically cleaves human IgA1, but not
human IgA2. Has distinctive PAGE and RFLP patterns. DNA
is not restricted by endonuclease Uur9601. Genome size of
the type strain is 890 kbp, whereas the sizes of the 10 known
serovar standard strains range from 840 to 1140 kbp (PFGE).
DNA reassociation values: within the species (serovars 2, 4,
5, and 7), 69–100%; with Ureaplasma parvum (serovars 1, 3,
and 6), 49–52%. Opportunistic pathogen of humans; causes
some cases of nongonococcal urethritis, infectious kidney
stones, systemic infection in immunologically compromised
hosts. Associated with a broad variety of urogenital infections
for which causality remains to be established.
Source: primarily found in the genitourinary tract of
female and male humans; occasionally in the oral cavity and
rectum.
DNA G+C content (mol%): 25.5–27.8 (Tm; type strain) and
27.7–28.5 (Bd; serovars 2, 4, 5, and 7).
Type strain: T960, (CX8), ATCC 27618, NCTC 10177.
Sequence accession nos: M23935 and AF073450 (type strain 16S
rRNA gene), AB028088 and AF059330 (type strain 16S–23S
rRNA intergenic region). Complete and near-complete (>99%)
genomes: serovar 2 strain ATCC 27814, NZ_ABFL00000000;
serovar 4 strain ATCC 27816, NZ_AAYO00000000; serovar
5 strain ATCC 27817, NZ_AAZR00000000; serovar 7 strain
ATCC 27819, NZ_AAYP00000000; serovar 8 strain ATCC
27618, NZ_AAYN00000000; serovar 9 strain ATCC 33175, NZ_
AAYQ00000000; serovar 10 strain ATCC 33699, NC_011374;
serovar 11 strain ATCC 33695, NZ_AAZS00000000; serovar
12 strain ATCC 33696, NZ_AAZT00000000; serovar 13 strain
ATCC 33698, NZ_ABEV00000000.
2. Ureaplasma canigenitalium Harasawa, Imada, Kotani, Koshimizu and Barile 1993, 644VP
ca.ni.ge.ni.ta¢li.um. L. n. canis dog; L. pl. n. genitalia the genitals; N.L. pl. gen. n. canigenitalium of canine genitals.
Cells are coccoid and about 500 nm in diameter; coccobacillary forms are seen. Colonies are £20–140 mm diameter
with fried-egg morphology. Serogroup I strains represented
by D6P-CT are serologically distinct from all other established
species in the genus and from the other three serogroups of
ureaplasmas isolated from dogs (represented by the strains
DIM-C, D29M, and D11N-A). The species designation refers
only to serogroup I strains, although strain D11N-A shows a
one-way, serological cross-reaction with D6P-CT. It produces
an IgA protease which specifically cleaves canine myeloma
IgA, but not human or murine IgA. Genome size is 860 kbp
(PFGE). DNA reassociation values: between D6P-CT and the
other three canine strains (DIM-C, D29M, and D11N-A)
are 41–63% versus 33% with Ureaplasma urealyticum (strain
T960T).
Source: habitat is the prepuce, vagina, and oral and nasal
cavities of canines.
DNA G+C content (mol%): 29.4 (HPLC).
Type strain: D6P-C, ATCC 51252, CIP 106087.
Sequence accession no. (16S rRNA gene): D78648 (type strain).
3. Ureaplasma cati Harasawa, Imada, Ito, Koshimizu, Cassell
and Barile 1990a, 50VP
ca¢ti. L. gen. n. cati of a cat.
Cells are coccoid and ³675 nm diameter, exceeding the
450–550 nm range of most named Ureaplasma species. Coccobacillary forms are seen and occasionally filaments. Colonies are £15–140 mm in diameter with diffuse, granular
appearance; some fried-egg colonies may appear after passaging. Distinct from other established species in the genus,
including Ureaplasma felinum, antigenically and in PAGE
(Harasawa et al., 1990a) and RFLP patterns (Harasawa et al.,
1984). Genome size has not been determined. DNA reassociation values: 83–100% within feline serogroup SII strains
(Ureaplasma cati) versus <10% with serogroup SI strains (Ureaplasma felinum).
Source: found in the oral cavity of healthy domestic cats
(Felis domestica).
DNA G+C content (mol%): 27.9 (Bd), 28.1 (HPLC) for
strain F2T.
Type strain: F2, ATCC 49228, NCTC 11710, CIP 106088.
Sequence accession nos: D78649 (type strain 16S rRNA gene),
D63685 (type strain 16S–23S rRNA intergenic spacer region).
4. Ureaplasma diversum Howard and Gourlay 1982, 450VP
di.ver¢sum. L. neut. part. adj. diversum different, distinct, heterogeneous, referring to the difference in polypeptides and
G+C content as compared to Ureaplasma urealyticum and to
the heterogeneous antigenic structure of the species.
Cells are coccoid or coccobacillary and appear to be
within the size range of other named Ureaplasma species
although no measurements have been published. Colonies
are £100–175 mm in diameter based on photomicrographs.
Serologically distinct from other named species but antigenically heterogeneous, comprising serogroups A, B, and C,
and represented by strains A417T, D48, and T44. These show
three distinctive PAGE patterns (Howard and Gourlay, 1982),
but only one RFLP pattern (Harasawa et al., 1984), and, based
upon the latter criterion, were considered homogeneous.
Genome size range is 1100–1160 kbp for strains 95 TX,
1763, and 2065-B202 (PFGE). No DNA reassociation values
are available.
Source: the type strain originated from a pneumonic calf
lung.
622
DNA G+C content (mol%): 29.0 and 28.7–30.2 (Bd) for the
type strain and 10 bovine isolates, respectively; thus, higher
than and not overlapping the values for Ureaplasma urealyticum and Ureaplasma parvum.
Type strain: A417, ATCC 43321, NCTC 10182, CIP 106089.
Sequence accession nos: D78650 (type strain 16S rRNA
gene), D63686 (type strain 16S–23S rRNA intergenic spacer
region).
5. Ureaplasma felinum Harasawa, Imada, Ito, Koshimizu, Cassell
and Barile 1990a, 50VP
fe.li¢num. L. neut. adj. felinum of or belonging to a cat.
Coccoid cells of ³800 nm diameter exceed the 450–500 nm
range of most Ureaplasma species. Coccobacillary forms and
occasional filaments are seen. Colonies are £15–140 mm diameter with diffuse, granular appearance; some fried-egg colonies may appear after passaging. Distinct antigenically and by
PAGE and RFLP patterns (Harasawa et al., 1984) from other
established species in the genus, including Ureaplasma cati. The
genome size is 1170 kbp (PFGE). It is the largest of any ureaplasma strain examined. DNA reassociation values: 89–100%
within feline serogroup SI strains (Ureaplasma felinum) versus
<10% with serogroup II strains (Ureaplasma cati).
Source: found in the oral cavity of healthy domestic cats
(Felis domestica).
Type strain: FT2-B, ATCC 49229, NCTC 11709, CIP
106090.
Sequence accession nos: D78651 (type strain 16S rRNA gene),
D63687 (type strain 16S–23S rRNA intergenic spacer region).
6. Ureaplasma gallorale Koshimizu, Harasawa, Pan, Kotani,
Ogata, Stephens and Barile 1987, 337VP
gal.lo.ra¢le. L. n. gallus a barnyard fowl; L. n. os, oris the
mouth; L. neut. suff. -ale suffix used with the sense of pertaining to; N.L. neut. adj. gallorale relating to the mouth of
barnyard fowl.
Cells are coccoid and about 500 nm in diameter; coccobacillary forms are seen. Colonies are £15–60 mm in diameter
with fried-egg morphology. Serologically distinct from all
other established species in the genus. Isolates have similar
SDS-PAGE, immunoblot, and RFLP patterns, but demonstrate some species heterogeneity based on reassociation values (cluster A strains D6-1T and T9-1; cluster B strain Y8-1).
Genome size is 760 kbp (PFGE). DNA reassociation values fall
into two clusters: within cluster A (strains D6-1T, D23, F2, F5,
and T9-1), values are 70–100%; within cluster B (strains Y8-1
and Y4-2), values are 96–100%; between these clusters, values
are 51–69%. Although these values are below expectations for
a single species status, they exceed the 19–27% reassociation
values with Ureaplasma urealyticum and Ureaplasma diversum.
References
Adler, H.E., J. Fabricant, R. Yamamoto and J. Berg. 1958. Symposium on
chronic respiratory diseases of poultry. I. Isolation and identification
of pleuropneumonia-like organisms of avian origin. Am. J. Vet. Res.
19 : 440–447.
Adler, S. and V. Ellenbogen. 1934. A note on two new blood parasites of
cattle: Eperythrozoon and Bartonella. J. Comp. Pathol. 47: 220–221.
Source: found only in oropharynx of healthy red jungle
fowl (Gallus gallus) and chickens (Gallus gallus var. domesticus)
kept as laboratory or zoo animals in Japan and in chickens
and turkeys with pneumonia or airsacculitis in Hungary.
Type strain: D6-1, ATCC 43346, NCTC 11707.
Sequence accession nos: U62937 (type strain 16S rRNA
gene), D63688 (type strain 16S–23S rRNA intergenic spacer
region).
7. Ureaplasma parvum Robertson, Stemke, Davis, Harasawa,
Thirkell, Kong, Shepard and Ford 2002, 593VP
par¢vum. L. neut. adj. parvum small, referring to its significantly
smaller genome sizes compared to Ureaplasma urealyticum, the
other species from humans.
Cells are coccoid and about 500 nm in diameter. Coccobacillary forms are present in exponential phase cultures.
Lipoglycans have been identified in a serovar 3 strain. Colonies are £20–140 mm in diameter with complete or partial
fried-egg morphology.
Serologically distinct from all other named species in the
genus, but serological heterogeneity is exhibited within the
species. Four specific antigenic determinants are known: 1,
3, 6, and 14. Serovar specificities are related to the multiplebanded antigens recognized by the host. Like Ureaplasma
urealyticum, Ureaplasma parvum has an IgA protease activity
that specifically cleaves human IgA1, but not human IgA2.
Distinctive PAGE and RFLP patterns. DNA restricted by
endonuclease Uur9601. Genome size: 751,719 kbp for the
type strain. Complete genomic sequence of the organism has
been reported (Glass et al., 2000). DNA reassociation values:
within the species (serovars 1, 3, and 6), 91–102%; with Urea
plasma urealyticum (serovars 2, 4, 5, 7–13), 38–60%.
Source: primary habitat is the genitourinary tract of female
and male humans; occasionally found in the oral cavity and
rectum. As yet, unclear whether an opportunistic pathogen
in non-gonococcal urethritis, but likely to be so in systemic
infection in immunologically compromised hosts. Associated with a broad variety of urogenital infections for which
causality remains to be established.
DNA G+C content (mol%): 25.5 (from genome sequence)
and 27.8–28.2 (Tm) for serovars 1 and 6.
Sequence accession nos: L08642 and AF073456 (type strain
16S rRNA gene), AB028083 and AF059323 (type strain
16S–23S rRNA intergenic spacer region). Complete and
near-complete (>99%) genomes: serovar 1 strain ATCC
27813, NZ_ABES00000000; serovar 3 strain ATCC 27815,
NC_010503; serovar 3 strain ATCC 700970, NC_002162;
serovar 6 strain ATCC 27818, NZ_AAZQ00000000; serovar
14 strain ATCC 33697, NZ_ABER00000000.
Ajufo, J. and K. Whithear. 1980. The surface layer of Mycoplasma synoviae
as demonstrated by the negative staining technique. Res. Vet. Sci. 29:
268–270.
Al-Ankari, A.R. and J.M. Bradbury. 1996. Mycoplasma iowae : a review.
Avian Pathol. 25: 205–229.
Al Masalma, M., F. Armougom, W. Scheld, H. Dufour, P. Roche,
M. Drancourt and D. Raoult. 2009. The expansion of the
icrobiological spectrum of brain abscesses with use of multiple 16S
m
ribosomal DNA sequencing. Clin. Infect. Dis. 48: 1169–1178.
Alberti, A., P. Robino, B. Chessa, S. Rosati, M. Addis, P. Mercier, A. Mannelli, T. Cubeddu, M. Profiti, E. Bandino, R. Thiery and M. Pittau.
2008. Characterisation of Mycoplasma capricolum P60 surface lipoprotein and its evaluation in a recombinant ELISA. Vet. Microbiol. 128:
81–89.
Alexander, P., K. Slee, S. McOrist, L. Ireland and P. Coloe. 1985. Mastitis
in cows and polyarthritis and pneumonia in calves caused by Mycoplasma species bovine group 7. Aust. Vet. J. 62: 135–136.
Allam, N.M. and R.M. Lemcke. 1975. Mycoplasmas isolated from the
respiratory tract of horses. J. Hyg. (Lond.) 74: 385–407.
Amin, M.M. and F.T. Jordan. 1978. Experimental infection of ducklings
with Mycoplasma gallisepticum and Mycoplasma anatis. Res. Vet. Sci. 25:
86–88.
Antunes, N., M. Tavío, P. Mercier, R. Ayling, W. Al-Momani, P. Assunção, R. Rosales and J. Poveda. 2007. In vitro susceptibilities of Mycoplasma putrefaciens field isolates. Antimicrob. Agents Chemother. 51:
3452–3454.
Arif, A., J. Schulz, F. Thiaucourt, A. Taha and S. Hammer. 2007. Contagious caprine pleuropneumonia outbreak in captive wild ungulates
at Al Wabra Wildlife Preservation, State of Qatar. J. Zoo Wildl. Med.
38: 93–96.
Armstrong, C.H., M.J. Freeman and L. Sands-Freeman. 1987. Crossreactions between Mycoplasma hyopneumoniae and Mycoplasma flocculare – practical implications for the serodiagnosis of mycoplasmal
pneumonia of swine. Isr. J. Med. Sci. 23: 654–656.
Armstrong, D., B.H. Yu, A. Yagoda and M.F. Kagnoff. 1971. Colonization of humans by Mycoplasma canis. J. Infect. Dis. 124: 607–609.
Askaa, G. and H. Erno. 1976. Elevation of Mycoplasma agalactiae subsp.
bovis to species rank, Mycoplasma bovis (Hale et al.) comb. nov. Int. J.
Syst. Bacteriol. 26: 323–325.
Atkinson, T.P., M.F. Balish and K.B. Waites. 2008. Epidemiology, clinical
manifestations, pathogenesis and laboratory detection of Mycoplasma
pneumoniae infections. FEMS Microbiol. Rev. 32: 956–973.
Baker, A.S., K.L. Ruoff and S. Madoff. 1998. Isolation of Mycoplasma
species from a patient with seal finger. Clin. Infect. Dis. 27: 1168–
1170.
Balish, M.F. 2006. Subcellular structures of mycoplasmas. Front Biosci.
11: 2017–2027.
Balish, M.F. and D.C. Krause. 2006. Mycoplasmas: a distinct cytoskeleton for wall-less bacteria. J. Mol. Microbiol. Biotechnol. 11: 244–255.
Bar-Moshe, B., E. Rapoport and J. Brenner. 1984. Vaccination trials
against Mycoplasma mycoides subsp. mycoides (large-colony-type) infection in goats. Isr. J. Med. Sci. 20: 972–974.
Barile, M. 1986. DNA homologies and serologic relationships among
ureaplasmas from various hosts. Pediatr. Infect. Dis. 5: S296–299.
Barile, M.F., R.A. Del Giudice, T.R. Carski, C.J. Gibbs and J.A. Morris.
1968. Isolation and characterization of Mycoplasma arginini: spec.
nov. Proc. Soc. Exp. Biol. Med. 129: 489–494.
Barile, M.F., R.A. Del Giudice and J.G. Tully. 1972. Isolation and characterization of Mycoplasma conjunctivae sp. n. from sheep and goats with
keratoconjunctivitis. Infect. Immun. 5: 70–76.
Barile, M.F. 1984. Immunization against Mycoplasma pneumoniae disease:
a review. Isr. J. Med. Sci. 20: 912–915.
Baseman, J., M. Cagle, J. Korte, C. Herrera, W. Rasmussen, J. Baseman, R. Shain and J. Piper. 2004. Diagnostic assessment of Mycoplasma genitalium in culture-positive women. J. Clin. Microbiol. 42:
203–211.
Baseman, J.B., S.F. Dallo, J.G. Tully and D.L. Rose. 1988. Isolation and
characterization of Mycoplasma genitalium strains from the human
respiratory tract. J. Clin. Microbiol. 26: 2266–2269.
Baseman, J.B. and J.G. Tully. 1997. Mycoplasmas: sophisticated, reemerging, and burdened by their notoriety. Emerg. Infect. Dis. 3:
21–32.
623
Bébéar, C. and J.A. Robertson. 1996. Determination of minimal inhibitory concentration. In Molecular and Diagnostic Procedures in
Mycoplasmology, vol. 2 (edited by Tully and Razin). Academic Press,
New York, pp. 189–197.
Bébéar, C.M. and I. Kempf. 2005. Antimicrobial therapy and antimicrobial resistance. In Mycoplasmas Molecular Biology Pathogenicity and
Strategies for Control (edited by Blanchard and Browning). Horizon
Bioscience, Norfolk, UK, pp. 535–568.
Beeton, M., V. Chalker, N. Maxwell, S. Kotecha and O. Spiller. 2009.
Concurrent titration and determination of antibiotic resistance in
Ureaplasma species with identification of novel point mutations in
genes associated with resistance. Antimicrob. Agents Chemother. 53:
2020–2027.
Ben Abdelmoumen Mardassi, B., A.O. Béjaoui Khiari, L., A. Landoulsi,
C. Brik, B. Mlik and F. Amouna. 2007. Molecular cloning of a
Mycoplasma meleagridis -specific antigenic domain endowed with a
serodiagnostic potential. Vet. Microbiol. 119: 31–41.
Benčina, D., D. Dorrer and T. Tadina. 1987. Mycoplasma species isolated
from six avian species. Avian Pathol. 16: 653–664.
Benčina, D., T. Tadina and D. Dorrer. 1988. Natural infection of ducks
with Mycoplasma synoviae and Mycoplasma gallisepticum and Mycoplasma
egg transmission. Avian Pathol. 17: 441–449.
Benčina, D., I. Mrzel, O. Zorman Rojs, A. Bidovec and A. Dovc. 2003.
Characterisation of Mycoplasma gallisepticum strains involved in
respiratory disease in pheasants and peafowl. Vet. Rec. 152: 230–
234.
Berent, L.M. and J.B. Messick. 2003. Physical map and genome sequencing survey of Mycoplasma haemofelis (Haemobartonella felis). Infect.
Immun. 71: 3657–3662.
Bergemann, A.D. and L.R. Finch. 1988. Isolation and restriction
endonuclease analysis of a Mycoplasma plasmid. Plasmid 19 :
68–70.
Bergonier, D., X. Berthelot and F. Poumarat. 1997. Contagious agalactia of small ruminants: current knowledge concerning epidemiology,
diagnosis and control. Rev. Sci. Tech. 16: 848–873.
Bhugra, B. and K. Dybvig. 1993. Identification and characterization
of IS1138, a transposable element from Mycoplasma pulmonis that
belongs to the IS3 family. Mol. Microbiol. 7: 577–584.
Biberfeld, G. and P. Biberfeld. 1970. Ultrastructural features of Mycoplasma pneumoniae. J. Bacteriol. 102: 855–861.
Binder, A., R. Aumuller, B. Likitdecharote and H. Kirchhoff. 1990. Isolation of Mycoplasma arthritidis from the joint fluid of boars. Zentralbl.
Veterinarmed. B 37: 611–614.
Black, F.T. 1973. Biological and physical properties of human
T-mycoplasmas. Ann. N. Y. Acad. Sci. 225: 131–143.
Black, F.T. and A. Krogsgaard-Jensen. 1974. Application of indirect
immunofluorescence, indirect haemagglutination and polyacrylamide-gel electrophoresis to human T-mycoplasmas. Acta Pathol. Microbiol. Scand. [B] Microbiol. Immunol. 82: 345–353.
Blanchard, A. 1990. Ureaplasma urealyticum urease genes: use of a UGA
tryptophan codon. Mol. Microbiol. 4: 669–676.
Blanchard, A., W. Hamrick, L. Duffy, K. Baldus and G.H. Cassell. 1993.
Use of the polymerase chain reaction for detection of Mycoplasma fermentans and Mycoplasma genitalium in the urogenital tract and amniotic fluid. Clin. Infect. Dis. 17 Suppl 1: S272–279.
Blanchard, A. 1997. Mycoplasmas and HIV infection, a possible interaction through immune activation. Wien Klin. Wochenschr. 109:
590–593.
Blanchard, A., L. Montagnier and M.L. Gougeon. 1997. Influence
of microbial infections on the progression of HIV disease. Trends
Microbiol. 5: 326–331.
Blanchard, A. and C.M. Bébéar. 2002. Mycoplasmas of Humans. In
Molecular Biology and Pathogenicity of Mycoplasmas (edited by
Razin and Herrmann). Kluwer Academic/Plenum Publishers, New
York, pp. 45–72.
624
Blaylock, M., O. Musatovova, J. Baseman and J. Baseman. 2004. Determination of infectious load of Mycoplasma genitalium in clinical samples of human vaginal cells. J. Clin. Microbiol. 42: 746–752.
Boatman, E.S. 1979. Morphology and ultrastructure of the Mycoplasmatales. In The Mycoplasmas, vol. 1 (edited by Barile and Razin).
Academic Press, New York, pp. 63–102.
Bonilla, H.F., C.E. Chenoweth, J.G. Tully, L.K. Blythe, J.A. Robertson,
V.M. Ognenovski and C.A. Kauffman. 1997. Mycoplasma felis septic
arthritis in a patient with hypogammaglobulinemia. Clin. Infect. Dis.
24: 222–225.
Boothby, J.T., D.E. Jasper, M.H. Rollins and C.B. Thomas. 1981. Detection of Mycoplasma bovis specific IgG in bovine serum by enzymelinked immunosorbent assay. Am. J. Vet. Res. 42: 1242–1247.
Borrel, A., E. Dujardin-Beaumetz, Jeantet and C. Jouan. 1910.
Le microbe de la péripneumonie. Ann. Inst. Pasteur (Paris) 24:
168–179.
Borup-Christensen, P., K. Erb and J.C. Jensenius. 1988. Curing human
hybridomas infected with Mycoplasma hyorhinis. J. Immunol. Methods
110: 237–240.
Boughton, E., S.A. Hopper and P.J.R. Gayford. 1983. Mycoplasma
canadense from bovine fetuses. Vet. Rec. 112: 87.
Bowie, W., S. Wang, E. Alexander, J. Floyd, P. Forsyth, H. Pollock, J. Lin,
T. Buchanan and K. Holmes. 1977. Etiology of nongonococcal urethritis. Evidence for Chlamydia trachomatis and Ureaplasma urealyticum.
J. Clin. Invest. 59: 735–742.
Bradbury, J.M., F. M. and A. Williams. 1983. Mycoplasma lipofaciens, a
new species of avian origin. International Journal of Systematic and
Evolutionary Microbiology 33: 329–335.
Bradbury, J.M. and M. Forrest. 1984. Mycoplasma cloacale, a new species
isolated from a turkey. Int. J. Syst. Bacteriol. 34: 389–392.
Bradbury, J.M., A. Vuillaume, J.P. Dupiellet, M. Forrest, J.L. Bind and
G. Gaillardperrin. 1987. Isolation of Mycoplasma cloacale from a
number of different avian hosts in Great-Britain and France. Avian
Pathol. 16: 183–186.
Bradbury, J.M., F.T.W. Jordan, T. Shimizu, L. Stipkovits and Z. Varga.
1988. Mycoplasma anseris sp. nov. found in geese. Int. J. Syst. Bacteriol.
38: 74–76.
Bradbury, J.M., O.M.S. Abdulwahab, C.A. Yavari, J.P. Dupiellet and J.M.
Bove. 1993. Mycoplasma imitans sp. nov. is related to Mycoplasma gallisepticum and found in birds. Int. J. Syst. Bacteriol. 43: 721–728.
Bradbury, J.M., C.M. Dare and C.A. Yavari. 2000. Evidence of Mycoplasma
gallisepticum in British wild birds. Proceedings of the 13th Congress of
the International Organization for Mycoplasmology Fukouka, Japan,
p. 253.
Bradbury, J.M., C.A. Yavari and C.M. Dare. 2001. Mycoplasmas and
respiratory disease in pheasants and partridges. Avian Pathol. 30:
391–396.
Bradbury, J.M. and C.J. Morrow. 2008. Mycoplasma infections. In Poultry Diseases, 6th edn (edited by Pattison, Bradbury and Alexander).
Elsevier, Edinburgh, pp. 220–234.
Bradbury, J.M. and S.H. Kleven. 2008. Mycoplasma synoviae infection.
In Diseases of Poultry, 12th edn. (edited by Saif, Fadly, Glisson,
McDougald, Nolan and Swayne). Blackwell Publishing, Ames, pp.
856–864.
Brandao, E. 1995. Isolation and identification of Mycoplasma mycoides
subspecies mycoides SC strains in sheep and goats. Vet. Rec. 136:
98–99.
Bredt, W. 1979. Motility. In The Mycoplasmas, vol. 1 (edited by Barile
Brown, D.R., G. McLaughlin and M. Brown. 1995. Taxonomy of the
feline mycoplasmas Mycoplasma felifaucium, Mycoplasma feliminutum,
Mycoplasma felis, Mycoplasma gateae, Mycoplasma leocaptivus, Mycoplasma
leopharyngis, and Mycoplasma simbae by 16S rRNA gene sequence comparisons. Int. J. Syst. Bacteriol. 45: 560–564.
Brown, D.R., R. Whitcomb and J. Bradbury. 2007. Revised minimal standards for description of new species of the class Mollicutes (division
Brown, D.R., Clippinger T.L., Helmick K.E., Schumacher I.M., Bennett R.A., Johnson C.M., Vliet K.A., Jacobson E.R. and B. M.B. 1996.
Mycoplasma isolation during a fatal epizootic of captive alligators
(Alligator mississippiensis) in Florida. Int. Org. Mycoplasmol. Lett
4: 2.
Brown, D.R., J.M. Farley, L.A. Zacher, J.M.R. Carlton, T.L. Clippinger,
J.G. Tully and M.B. Brown. 2001a. Mycoplasma alligatoris sp. nov., from
American alligators. Int. J. Syst. Evol. Microbiol. 51: 419–424.
Brown, D.R., D.F. Talkington, W.L. Thacker, M.B. Brown, D.L. Dillehay
and J.G. Tully. 2001b. Mycoplasma microti sp. nov., isolated from the
respiratory tract of prairie voles (Microtus ochrogaster). Int. J. Syst.
Evol. Microbiol. 51: 409–412.
Brown, D.R., J.L. Merritt, E.R. Jacobson, P.A. Klein, J.G. Tully and M.B.
Brown. 2004. Mycoplasma testudineum sp. nov., from a desert tortoise
(Gopherus agassizii) with upper respiratory tract disease. Int. J. Syst.
Brown, D.R., L.A. Zacher, L.D. Wendland and M.B. Brown. 2005. Emerging Mycoplasmoses in Wildlife. In Mycoplasmas Molecular Biology
Pathogenicity and Strategies for Control (edited by Blanchard and
Browning). Horizon Bioscience, Norfolk, England, pp. 383–414.
Brown, D.R., D.L. Demcovitz, D.R. Plourde, S.M. Potter, M.E. Hunt,
R.D. Jones and D.S. Rotstein. 2006. Mycoplasma iguanae sp. nov., from
a green iguana (Iguana iguana) with vertebral disease. Int. J. Syst.
Brown, D.R. and J.M. Bradbury. 2008. International Committee on Systematics of Prokaryotes of the International Union of Microbiological Societies, Subcommittee on the taxonomy of mollicutes, minutes
of the meeting, 6 and 11 July 2008, Tianjin, PR China. International
Journal of Systematic and Evolutionary Microbiology 58: 2987–2990.
Brown, M.B., I.M. Schumacher, P.A. Klein, K. Harris, T. Correll and E.R.
Jacobson. 1994. Mycoplasma agassizii causes upper respiratory tract
disease in the desert tortoise. Infect. Immun. 62: 4580–4586.
Brown, M.B., D.R. Brown, P.A. Klein, G.S. McLaughlin, I.M. Schumacher,
E.R. Jacobson, H.P. Adams and J.G. Tully. 2001c. Mycoplasma agassizii
sp. nov., isolated from the upper respiratory tract of the desert tortoise (Gopherus agassizii) and the gopher tortoise (Gopherus polyphemus). Int. J. Syst. Evol. Microbiol. 51: 413–418.
Browning, G.F., K.G. Whithear and S.J. Geary. 2005. Vaccines to Control Mycoplasmosis. In Mycoplasmas: Molecular Biology, Pathogenicity, and Strategies for Control (edited by Blanchard and Browning).
Horizon Bioscience, Norfolk, UK, pp. 569–599.
Brun-Hansen, H., H. Gronstol, H. Waldeland and B. Hoff. 1997.
Eperythrozoon ovis infection in a commercial flock of sheep. Zentralbl.
Veterinarmed. B 44: 295–299.
Brunner, S., P. Frey-Rindova, M. Altwegg and R. Zbinden. 2000. Retroperitoneal abscess and bacteremia due to Mycoplasma hominis in a
polytraumatized man. Infection 28: 46–48.
Busch, U., H. Nitschko, F. Pfaff, B. Henrich, J. Heesemann and
M. Abele-Horn. 2000. Molecular comparison of Mycoplasma hominis
strains isolated from colonized women and women with various urogenital infections. Zentralbl. Bakteriol. 289: 879–888.
Buss, I., R. Senthilmohan, B. Darlow, N. Mogridge, A. Kettle and
C. Winterbourn. 2003. 3-Chlorotyrosine as a marker of protein
damage by myeloperoxidase in tracheal aspirates from preterm
infants: association with adverse respiratory outcome. Pediatr. Res.
53: 455–462.
Carmichael, L.E., T.D. St George, N.D. Sullivan and N. Horsfall. 1972.
Isolation, propagation, and characterization studies of an ovine Mycoplasma responsible for proliferative interstitial pneumonia. Cornell
Vet. 62: 654–679.
Carson, J.L., P.C. Hu and A.M. Collier. 1992. Cell structure and functional elements. In Mycoplasmas: Molecular Biology and Pathogen-
esis (edited by Maniloff, McElhaney, Finch, and Baseman). American
Society for Microbiology, Washington, DC, pp. 63–72.
Casin, I., D. Vexiau-Robert, P. De La Salmoniere, A. Eche, B. Grandry
and M. Janier. 2002. High prevalence of Mycoplasma genitalium
in the lower genitourinary tract of women attending a sexually
transmitted disease clinic in Paris, France. Sex. Transm. Dis. 29:
353–359.
Cassell, G.H. and A. Hill. 1979. Murine and other small animal mycoplasmas. In The Mycoplasmas, vol. 1 (edited by Tully and Whitcomb).
Caswell, J.L. and M. Archambault. 2007. Mycoplasma bovis pneumonia in
cattle. Anim. Health Res. Rev. 8: 161–186.
Chalker, V.J. and J. Brownlie. 2004. Taxonomy of the canine Mollicutes
by 16S rRNA gene and 16S/23S rRNA intergenic spacer region
sequence comparison. Int. J. Syst. Evol. Microbiol. 54: 537–542.
Chalker, V.J. 2005. Canine mycoplasmas. Res. Vet. Sci. 79: 1–8.
Chávez Gonzalez, Y.R., C. Ros Bascunana, G. Bolske, J.G. Mattsson,
C. Fernandez Molina and K.-E. Johansson. 1995. In vitro amplification of the 16S rRNA genes from Mycoplasma bovis and Mycoplasma
agalactiae by PCR. Vet. Microbiol. 47: 183–190.
Cheng, X., J. Nicolet, F. Poumarat, J. Regalla, F. Thiaucourt and J. Frey.
1995. Insertion element IS1296 in Mycoplasma mycoides subsp. mycoides
small colony identifies a European clonal line distinct from African
and Australian strains. Microbiology 141: 3221–3228.
Chin, R.P., G.Y. Ghazikhanian and I. Kempf. 2008. Mycoplasma meleagridis infection. In Diseases of Poultry, 12th edn. (edited by Saif,
Fadly, Glisson, McDougald, Nolan and Swayne). Blackwell Publishing, Ames, pp. 834–845.
Christiansen, C., F.T. Black and E.A. Freundt. 1981. Hybridization
experiments with DNA from Ureaplasma urealyticum, serovars I to VIII.
Clapper, B., A.H. Tu, W.L. Simmons and K. Dybvig. 2004. Bacteriophage
MAV1 is not associated with virulence of Mycoplasma arthritidis. Infect.
Immun. 72: 7322–7325.
Clark, H.W., J. S. Bailey, D. C. Laughlin and T.M. Brown. 1978. Isolation
of Mycoplasma from the genital tracts of elephants. Zentralbl. Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt. 1 Orig. 241:262.
Clark, H.W., D. C. Laughlin, J. S. Bailey and T.M. Brown. 1980. Mycoplasma Species and Arthritis in Captive Elephants. Journal of Zoo
Animal Medicine 11: 3–15.
Clark, R. 1942. Eperythrozoon felis (sp. nov.) in a cat. J. Afr. Vet. Med.
Assoc. 13: 15–16.
Cobb, D.T., D.H. Ley and P.D. Doerr. 1992. Isolation of Mycoplasma
gallopavonis from free-ranging wild turkeys in coastal North Carolina seropositive and culture-negative for Mycoplasma gallisepticum.
J. Wildl. Dis. 28: 105–109.
Cocks, B., F. Brake, A. Mitchell and L. Finch. 1985. Enzymes of intermediary carbohydrate metabolism in Ureaplasma urealyticum and
Mycoplasma mycoides subsp. mycoides. J. Gen. Microbiol. 131: 2129–
2135.
Cocks, B.G. and L.R. Finch. 1987. Characterization of a restriction
endonuclease from Ureaplasma urealyticum 960 and differences in
deoxyribonucleic acid modification of human ureaplasmas. Int. J.
Cole, B.C., L. Golightly and J.R. Ward. 1967. Characterization of Mycoplasma strains from cats. J. Bacteriol. 94: 1451–1458.
Cole, B.C., L.R. Washburn and D. Taylor-Robinson. 1985. Mycoplasmainduced arthritis. In The Mycoplasmas, vol. 4 (edited by Razin and
Barile). Academic Press, New York, pp. 108–160.
Cole, R.M. 1983. Transmission electron microscopy: Basic techniques.
In Methods in Mycoplasmology (edited by Razin and Tully). Academic Press, New York, p. 4350.
Cordtz, J. and J. Jensen. 2006. Disseminated Ureaplasma urealyticum
infection in a hypo-gammaglobulinaemic renal transplant patient.
Scand. J. Infect. Dis. 38: 1114–1117.
625
Cottew, G.S. and F.R. Yeats. 1978. Subdivision of Mycoplasma mycoides
subsp. mycoides from cattle and goats into two types. Aust. Vet. J. 54:
293–296.
Cottew, G.S. 1979. Caprine-ovine mycoplasmas. In The Mycoplasmas,
vol. 2 (edited by Tully and Whitcomb). Academic Press, New York,
pp. 103–132.
Cottew, G.S. 1983. Recovery and identification of caprine and ovine
mycoplasmas. In Methods in Mycoplasmology, vol. 2 (edited by Tully
Cottew, G.S., A. Breard, A.J. DaMassa, H. Erno, R.H. Leach, P.C. Lefevre, A.W. Rodwell and G.R. Smith. 1987. Taxonomy of the Mycoplasma
mycoides cluster. Isr. J. Med. Sci. 23: 632–635.
Crespo, R. and R. McMillan. 2008. Facial cellulitis induced in chickens
by Mycoplasma gallisepticum bacterin and its treatment. Avian Dis. 52:
698–701.
Daddow, K.N. 1980. Culex annulirostris as a vector of Eperythrozoon ovis
infection in sheep. Vet. Parasitol. 7: 313–317.
DaMassa, A.J., E.R. Nascimento, M.I. Khan, R. Yamamoto and D.L.
Brooks. 1991. Characteristics of an unusual Mycoplasma isolated from
a case of caprine mastitis and arthritis with possible systemic manifestations. J. Vet. Diagn. Invest. 3: 55–59.
Dammann, O., E. Allred, D. Genest, R. Kundsin and A. Leviton. 2003.
Antenatal Mycoplasma infection, the fetal inflammatory response
and cerebral white matter damage in very-low-birthweight infants.
Paediatr. Perinat. Epidemiol. 17: 49–57.
Damassa, A.J., J.G. Tully, D.L. Rose, D. Pitcher, R.H. Leach and G.S.
Cottew. 1994. Mycoplasma auris sp. nov., Mycoplasma cottewii sp. nov.,
and Mycoplasma yeatsii sp. nov., new sterol-requiring mollicutes from
the external ear canals of goats. Int. J. Syst. Bacteriol. 44: 479–484.
DaMassa, A.J. 1996. Mycoplasma infections of goat and sheep. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 2 (edited by
Tully and Razin). Academic Press, San Diego, pp. 265–273.
Davis, J.W., Jr., I.S. Moses, C. Ndubuka and R. Ortiz. 1987. Inorganic
pyrophosphatase activity in cell-free extracts of Ureaplasma urealyticum. J. Gen. Microbiol. 133: 1453–1459.
Davis, J.W., Jr. and I. Villanueva. 1990. Enzyme differences in serovar
clusters of Ureaplasma urealyticum. In Recent Advances in Mycoplasmology (edited by Stanek, Cassell, Tully and Whitcomb). Gustav
Fischer Verlag, New York, pp. 665–666.
de Barbeyrac, B., C. Bernet-Poggi, F. Fébrer, H. Renaudin, M. Dupon
and C. Bébéar. 1993. Detection of Mycoplasma pneumoniae and Mycoplasma genitalium in clinical samples by polymerase chain reaction.
Clin. Infect. Dis. 17 Suppl 1: S83–89.
De Francesco, M., R. Negrini, G. Pinsi, L. Peroni and N. Manca. 2009.
Detection of Ureaplasma biovars and polymerase chain reactionbased subtyping of Ureaplasma parvum in women with or without
symptoms of genital infections. Eur. J. Clin. Microbiol. Infect. Dis.
28: 641–646.
de la Fe, C., P. Assunção, P. Saavedra, S. Tola, C. Poveda and J. Poveda.
2007. Vaccine. Field trial of two dual vaccines against Mycoplasma agalactiae and Mycoplasma mycoides subsp. mycoides (large colony type) in
goats 25: 2340–2345.
De Silva, N. and P. Quinn. 1986. Endogenous activity of phospholipases A and C in Ureaplasma urealyticum. J. Clin. Microbiol. 23:
354–359.
Deeb, B. and G. Kenny. 1967. Characterization of Mycoplasma pulmonis
variants isolated from rabbits. I. Identification and properties of isolates. J. Bacteriol. 93: 1416–1424.
Del Giudice, R.A., T.R. Carski, M.F. Barile, R.M. Lemcke and J.G. Tully.
1971. Proposal for classifying human strain Navel and related simian
mycoplasmas as Mycoplasma primatum sp. n. J. Bacteriol. 108: 439–445.
Del Giudice, R.A., R.H. Purcell, T.R. Carski and R.M. Chanock. 1974.
Mycoplasma lipophilum sp. nov. Int. J. Syst. Bacteriol. 24: 147–153.
626
Del Giudice, R.A., J.G. Tully, D.L. Rose and R.M. Cole. 1985. Mycoplasma
pirum sp. nov., a terminal structured mollicute from cell cultures. Int.
Del Giudice, R.A., D.L. Rose and J.G. Tully. 1995. Mycoplasma adleri sp.
nov., an isolate from a goat. Int. J. Syst. Bacteriol. 45: 29–31.
Del Giudice, R.A. and R.S. Gardella. 1996. Antibiotic treatment of
mycoplasma-infected cell cultures. In Molecular and Diagnostic
Procedures in Mycoplasmology, vol. 2 (edited by Tully and Razin).
Dessì, D., P. Rappelli, N. Diaz, P. Cappuccinelli and P.L. Fiori. 2006.
Mycoplasma hominis and Trichomonas vaginalis: a unique case of symbiotic relationship between two obligate human parasites. Front.
Biosci. 11: 2028–2034.
Dhondt, A.A., S. Altizer, E.G. Cooch, A.K. Davis, A. Dobson, M.J. Driscoll,
B.K. Hartup, D.M. Hawley, W.M. Hochachka, P.R. Hosseini, C.S.
Jennelle, G.V. Kollias, D.H. Ley, E.C. Swarthout and K.V. Sydenstricker. 2005. Dynamics of a novel pathogen in an avian host: Mycoplasmal conjunctivitis in house finches. Acta Trop. 94: 77–93.
Dhondt, A.A., K.V. Dhondt, D.M. Hawley and C.S. Jennelle. 2007.
Experimental evidence for transmission of Mycoplasma gallisepticum
in house finches by fomites. Avian Pathol. 36: 205–208.
Dierks, R.E., J.A. Newman and B.S. Pomeroy. 1967. Characterization of
avian Mycoplasma. Ann. N. Y. Acad. Sci. 143: 170–189.
Dillehay, D.L., M. Sander, D.F. Talkington, W.L. Thacker and
D.R. Brown. 1995. Isolation of mycoplasmas from prairie voles
(Microtus ochrogaster). Lab. Anim. Sci. 45: 631–634.
Djordjevic, S.R., W.A. Forbes, J. Forbes-Faulkner, P. Kuhnert, S. Hum,
M.A. Hornitzky, E.M. Vilei and J. Frey. 2001. Genetic diversity among
Mycoplasma species bovine group 7: clonal isolates from an outbreak
of polyarthritis, mastitis, and abortion in dairy cattle. Electrophoresis
22: 3551–3561.
Doig, P.A. 1981. Bovine genital mycoplasmosis. Can. Vet. J. 22: 339–343.
dos Santos, A.P., R.P. dos Santos, A.W. Biondo, J.M. Dora, L.Z.
Goldani, S.T. de Oliveira, A.M. de Sa Guimaraes, J. Timenetsky,
H.A. de Morais, F.H. Gonzalez and J.B. Messick. 2008. Hemoplasma
infection in HIV-positive patient, Brazil. Emerg Infect Dis 14: 1922–
1924.
Dowers, K.L., C. Olver, S.V. Radecki and M.R. Lappin. 2002. Use
of enrofloxacin for treatment of large-form Haemobartonella felis
in experimentally infected cats. J. Am. Vet. Med. Assoc. 221:
250–253.
Drexler, H.G. and C.C. Uphoff. 2002. Mycoplasma contamination of cell
cultures: Incidence, sources, effects, detection, elimination, prevention. Cytotechnology 39: 75–90.
Dubosson, C.R., C. Conzelmann, R. Miserez, P. Boerlin, J. Frey, W.
Zimmermann, H. Hani and P. Kuhnert. 2004. Development of two
real-time PCR assays for the detection of Mycoplasma hyopneumoniae
in clinical samples. Vet. Microbiol. 102: 55–65.
Duffy, L., J. Glass, G. Hall, R. Avery, R. Rackley, S. Peterson and
K. Waites. 2006. Fluoroquinolone resistance in Ureaplasma parvum in
the United States. J. Clin. Microbiol. 44: 1590–1591.
Dupiellet, J.-P. 1984. Mycoplasmes et acholeplasmes des palmipedes a
foie gras: isolement, caracterisation, etude du role dans la pathologie. Universite de Bordeaux II.
Dyer, N., L. Hansen-Lardy, D. Krogh, L. Schaan and E. Schamber. 2008.
An outbreak of chronic pneumonia and polyarthritis syndrome
caused by Mycoplasma bovis in feedlot bison (Bison bison). J. Vet.
Diagn. Invest. 20: 369–371.
Echahidi, F., G. Muyldermans, S. Lauwers and A. Naessens. 2001. Development of an enzyme-linked immunosorbent assay for serotyping
Ureaplasma urealyticum strains using monoclonal antibodies. Clin.
Diagn. Lab. Immunol. 8: 52–57.
Edward, D.G. and A.D. Kanarek. 1960. Organisms of the pleuropneumonia group of avian origin: their classification into species. Ann. N.
Y. Acad. Sci. 79: 696–702.
Edward, D.G. and E.A. Freundt. 1969. Proposal for classifying organisms related to Mycoplasma laidlawii in a family Sapromycetaceae, genus
Sapromyces, within the Mycoplasmatales. J. Gen. Microbiol. 57: 391–
395.
Edward, D.G. and E.A. Freundt. 1970. Amended nomenclature for
strains related to Mycoplasma laidlawii. J. Gen. Microbiol. 62: 1–2.
Edward, D.G.f. 1953. Organisms of the pleuropneumonia group causing disease in goats. Vet. Rec. 63: 873–874.
Edward, D.G.f. 1955. A suggested classification and nomenclature
for organisms of the pleuropneumonia group. Int. Bull. Bacteriol.
Nomencl. Taxon. 5: 85–93.
Egwu, G., J. Ameh, M. Aliyu and F. Mohammed. 2001. Caprine Mycoplasmal mastitis in Nigeria. Small Rumin. Res. 39: 87–91.
Erdélyi, K., M. Tenk and A. Dan. 1999. Mycoplasmosis associated perosis type skeletal deformity in a saker falcon nestling in Hungary. J.
Wildl. Dis. 35: 586–590.
Erickson, B.Z., R.F. Ross, D.L. Rose, J.G. Tully and J.M. Bové. 1986.
Mycoplasma hyopharyngis, a new species from swine. Int. J. Syst. Bacteriol. 36: 55–59.
Erickson, B.Z., R.F. Ross and J.M. Bove. 1988. Isolation of Mycoplasma
salivarium from swine. Vet. Microbiol. 16: 385–390.
Evans-Davis, K.D., D.L. Dillehay, D.N. Wargo, S.K. Webb, D.F. Talkington, W.L. Thacker, L.S. Small and M.B. Brown. 1998. Pathogenicity of Mycoplasma volis in mice and rats. Lab. Anim. Sci. 48:
38–44.
Feberwee, A., J. de Wit and W. Landman. 2009. Induction of eggshell
apex abnormalities by Mycoplasma synoviae : field and experimental
studies. Avian Pathol. 38: 77–85.
Ferguson, N.M., V.A. Leiting and S.H. Klevena. 2004. Safety and efficacy
of the avirulent Mycoplasma gallisepticum strain K5054 as a live vaccine
in poultry. Avian Dis. 48: 91–99.
Fernandez Guerrero, M.L., J. Manuel Ramos and F. Soriano. 1999. Mycoplasma hominis bacteraemia not associated with genital infections. J.
Infect. 39: 91–94.
Ferrell, R., M. Heidari, K. Wise and M. McIntosh. 1989. A Mycoplasma
genetic element resembling prokaryotic insertion sequences. Mol.
Fischer, R.S., B.E. Fischer and R.A. Jensen. 1992. Sources of amino
acids. In Mycoplasmas: Molecular Biology and Pathogenesis (edited
by Maniloff, McElhaney, Finch and Baseman). American Society for
Microbiology, Washington, D.C., pp. 201–209.
Fitzmaurice, J., M. Sewell, L. Manso-Silvan, F. Thiaucourt, W.L.
McDonald and J.S. O’Keefe. 2008. Real-time polymerase chain reaction assays for the detection of members of the Mycoplasma mycoides
cluster. N Z Vet. J. 56: 40–47.
Flint, J.C. and McKelvie D.H. 1956. Feline infectious anemia-diagnosis
and treatment. Presented at the Proc. 92nd Ann. Meet. Amer. Vet.
Med. Assoc. 1955, 240–242.
Foley, J.E. and N.C. Pedersen. 2001. “Candidatus Mycoplasma haemominutum”, a low-virulence epierythrocytic parasite of cats. Int. J. Syst.
Ford, D. 1967. Relationships between Mycoplasma and the etiology of
nongonococcal urethritis and Reiter’s syndrome. Ann. N. Y. Acad.
Sci. 143: 501–504.
Ford, D. and J. MacDonald. 1967. Influence of urea on the growth of
T-strain mycoplasmas. J. Bacteriol. 93: 1509–1512.
Ford, D. 1972. Inhibition of growth of T-strain mycoplasmas by
hydroxamic acids and by aurothiomalate. Antimicrob Agents
Chemother 2: 340–343.
Ford, D. and J. Smith. 1974. Non-specific urethritis associated with a
tetracycline-resistant T-mycoplasma. Br. J. Vener. Dis. 50: 373–374.
Forrest, M. and J.M. Bradbury. 1984. Mycoplasma glycophilum, a new species of avian origin. J. Gen. Microbiol. 130: 597–603.
Forrest, M. and J. M. Bradbury. 1984. Mycoplasma glycophilum sp. nov.
Validation List No. 15. Int. J. Syst. Bacteriol. 34: 355–357.
Forsyth, M.H., J.G. Tully, T.S. Gorton, L. Hinckley, S. Frasca, Jr., H.J.
van Kruiningen and S.J. Geary. 1996. Mycoplasma sturni sp. nov., from
the conjunctiva of a European starling (Sturnus vulgaris). Int. J. Syst.
Bacteriol. 46: 716–719.
Frasca, S., Jr, L. Hinckley, M. Forsyth, T. Gorton, S. Geary and H. Van
Kruiningen. 1997. Mycoplasmal conjunctivitis in a European starling. J. Wildl. Dis. 33: 336–339.
Frasca, S., Jr, E. Weber, H. Urquhart, X. Liao, M. Gladd, K. Cecchini, P.
Hudson, M. May, R. Gast, T. Gorton and S. Geary. 2005. Isolation and
characterization of Mycoplasma sphenisci sp. nov. from the choana of
an aquarium-reared jackass penguin (Spheniscus demersus). J. Clin.
Microbiol. 43: 2976–2979.
Freundt, E.A. 1953. The occurrence of Micromyces (pleuropneumonialike organisms) in the female genito-urinary tract. Acta Pathol.
Microbiol. Scand. 32: 468–480.
of organisms (Borrelomycetales). Int. Bull. Bacteriol. Nomencl.
Taxon. 5: 67–78.
Freundt, E.A., Taylorro.D, R.H. Purcell, R.M. Chanock and F.T. Black.
1974. Proposal of Mycoplasma buccale nom. nov. and Mycoplasma faucium nom. nov. for Mycoplasma orale types 2 and 3, respectively. Int. J.
Frey, J., X. Cheng, P. Kuhnert and J. Nicolet. 1995. Identification and
characterization of IS1296 in Mycoplasma mycoides subsp. mycoides SC
and presence in related mycoplasmas. Gene 160: 95–100.
Frey, M.L., R.P. Hanson and D.P. Anderson. 1968. A medium for the
isolation of avian mycoplasmas. American Journal of Veterinary
Research 29: 2163–2171.
Friis, N.F. and E. Blom. 1983. Isolation of Mycoplasma canadense from
bull semen. Acta Vet. Scand. 24: 315–317.
Friis, N.F. and J. Szancer. 1994. Sensitivity of certain porcine and bovine
mycoplasmas to antimicrobial agents in a liquid medium test compared to a disc assay. Acta Vet. Scand. 35: 389–394.
Furness, G. 1973. T-mycoplasmas: their growth and production of a
toxic substance in broth. J. Infect. Dis. 127: 9–16.
Furness, G. 1975. T-mycoplasmas: Growth patterns and physical characteristics of some human strains. J. Infect. Dis. 132: 592–596.
Furr, P.M., D. Taylor-Robinson and A.D. Webster. 1994. Mycoplasmas
and ureaplasmas in patients with hypogammaglobulinaemia and
their role in arthritis: microbiological observations over twenty years.
Ann. Rheum. Dis. 53: 183–187.
Gallagher, J. and K. Rhoades. 1983. Scanning electron and light microscopy
of selected avian strains of Mycoplasma iowae. Avian Dis. 27: 211–217.
Gambini, D., I. Decleva, L. Lupica, M. Ghislanzoni, M. Cusini and
E. Alessi. 2000. Mycoplasma genitalium in males with nongonococcal urethritis: prevalence and clinical efficacy of eradication. Sex.
Transm. Dis. 27: 226–229.
Ganapathy, K. and J.M. Bradbury. 1998. Pathogenicity of Mycoplasma
gallisepticum and Mycoplasma imitans in red-legged partridges (Alectoris rufa). Avian Pathol. 27: 455–463.
García-Castillo, M., M. Morosini, M. Gálvez, F. Baquero, R. del Campo
and M. Meseguer. 2008. Differences in biofilm development and
antibiotic susceptibility among clinical Ureaplasma urealyticum and Ureaplasma parvum isolates. J. Antimicrob. Chemother. 62: 1027–1030.
Garcia-Porrua, C., F.J. Blanco, A. Hernandez, A. Atanes, F. Galdo, R.
Moure and A. Alonso. 1997. Septic arthritis by Mycoplasma hominis: a
case report and review of the medical literature. Ann. Rheum. Dis.
56: 699–700.
Gass, R., J. Fisher, D. Badesch, M. Zamora, A. Weinberg, H. Melsness,
F. Grover, J.G. Tully and F.C. Fang. 1996. Donor-to-host transmission
of Mycoplasma hominis in lung allograft recipients. Clin. Infect. Dis.
22: 567–568.
Gates, A.E., S. Frasca, A. Nyaoke, T.S. Gorton, L.K. Silbart and
S.J. Geary. 2008. Comparative assessment of a metabolically
attenuated Mycoplasma gallisepticum mutant as a live vaccine for
627
the prevention of avian respiratory mycoplasmosis. Vaccine 26:
2010–2019.
Gautier-Bouchardon, A.V., A.K. Reinhardt, M. Kobisch and I. Kempf.
2002. In vitro development of resistance to enrofloxacin, erythromycin, tylosin, tiamulin and oxytetracycline in Mycoplasma gallisepticum, Mycoplasma iowae and Mycoplasma synoviae. Vet. Microbiol. 88:
47–58.
George, J.W., B.A. Rideout, S.M. Griffey and N.C. Pedersen. 2002. Effect
of preexisting FeLV infection or FeLV and feline immunodeficiency
virus coinfection on pathogenicity of the small variant of Haemobartonella felis in cats. Am. J. Vet. Res. 63: 1172–1178.
Geraci, J.R., D.J. Staubin, I.K. Barker, R.G. Webster, V.S. Hinshaw, W.J.
Bean, H.L. Ruhnke, J.H. Prescott, G. Early, A.S. Baker, S. Madoff and
R.T. Schooley. 1982. Mass mortality of harbor seals: pneumonia associated with influenza A virus. Science 215: 1129–1131.
Gerchman, I., I. Lysnyansky, S. Perk and S. Levisohn. 2008. In vitro susceptibilities to fluoroquinolones in current and archived Mycoplasma
gallisepticum and Mycoplasma synoviae isolates from meat-type turkeys.
Vet. Microbiol. 131: 266–276.
Germain, M., M.A. Krohn, S.L. Hillier and D.A. Eschenbach. 1994. Genital flora in pregnancy and its association with intrauterine growth
retardation. J. Clin. Microbiol. 32: 2162–2168.
Gibson, D.G., G.A. Benders, C. Andrews-Pfannkoch, E.A. Denisova, H.
Baden-Tillson, J. Zaveri, T.B. Stockwell, A. Brownley, D.W. Thomas,
M.A. Algire, C. Merryman, L. Young, V.N. Noskov, J.I. Glass, J.C. Venter, C.A. Hutchison, 3rd and H.O. Smith. 2008. Complete chemical
synthesis, assembly, and cloning of a Mycoplasma genitalium genome.
Science 319: 1215–1220.
Giebel, J., A. Binder and H. Kirchhoff. 1990. Isolation of Mycoplasma
moatsii from the intestine of wild Norway rats (Rattus norvegicus).
Giebel, J., J. Meier, A. Binder, J. Flossdorf, J.B. Poveda, R. Schmidt and
H. Kirchhoff. 1991. Mycoplasma phocarhinis sp. nov. and Mycoplasma
phocacerebrale sp. nov., two new species from harbor seals (Phoca vitulina L). Int. J. Syst. Bacteriol. 41: 39–44.
Gil, M.C., F.J. Pena, J. Hermoso De Mendoza and L. Gomez. 2003. Genital lesions in an outbreak of caprine contagious agalactia caused
by Mycoplasma agalactiae and Mycoplasma putrefaciens. J. Vet. Med. B
Infect. Dis. Vet. Public Health 50: 484–487.
Glass, J., E. Lefkowitz, J. Glass, C. Heiner, E. Chen and G. Cassell. 2000.
The complete sequence of the mucosal pathogen Ureaplasma urealyticum. Nature 407: 757–762.
Glass, J.I., B.A. Methe, V. Paralanov, L.B. Duffy and K.B. Waites. 2008.
Comparative genome analysis of all 14 Ureaplasma parvum and
Ureaplasma urealyticum serovars. Presented at the International Organization for Mycoplasmology, Tianjin, China.
Goldberg, D.R., M.D. Samuel, C.B. Thomas, P. Sharp, G.L. Krapu, J.R.
Robb, K.P. Kenow, C.E. Korschgen, W.H. Chipley, M.J. Conroy and
et al. 1995. The occurrence of mycoplasmas in selected wild North
American waterfowl. J. Wildl. Dis. 31: 364–371.
Gonçalves, L.F., T. Chaiworapongsa and R. Romero. 2002. Intrauterine
infection and prematurity. Ment. Retard. Dev. Disabil. Res. Rev. 8: 3–13.
Goodison, S., K. Nakamura, K.A. Iczkowski, S. Anai, S.K. Boehlein and
C.J. Rosser. 2007. Exogenous mycoplasmal p37 protein alters gene
expression, growth and morphology of prostate cancer cells. Cytogenet Genome Res. 118: 204–213.
Goodwin, R.F.W., A.P. Pomeroy and P. Whittlestone. 1965. Production
of enzootic pneumonia in pigs with a Mycoplasma. Vet. Rec. 77: 1247–
1249.
Gorton, T.S., M.M. Barnett, T. Gull, R.A. French, Z. Lu, G.F. Kutish,
L.G. Adams and S.J. Geary. 2005. Development of real-time diagnostic assays specific for Mycoplasma mycoides subspecies mycoides Small
Colony. Vet. Microbiol. 111: 51–58.
628
Gourlay, R.N. and R.H. Leach. 1970. A new Mycoplasma species isolated
from pneumonic lungs of calves (Mycoplasma dispar sp. nov.). J. Med.
Gourlay, R.N., R.H. Leach and C.J. Howard. 1974. Mycoplasma verecundum, a new species isolated from bovine eyes. J. Gen. Microbiol. 81:
475–484.
Gourlay, R.N., S.G. Wyld and R.H. Leach. 1977. Mycoplasma alvi, a new
species from bovine intestinal and urogenital tracts. Int. J. Syst. Bacteriol. 27: 86–96.
Gourlay, R.N., S.G. Wyld and R.H. Leach. 1978. Mycoplasma sualvi, a
new species from intestinal and urogenital tracts of pigs. Int. J. Syst.
Bacteriol. 28: 289–292.
Gourlay, R.N. and C.J. Howard. 1979. Bovine mycoplasmas. In The
Mycoplasmas, vol. 2 (edited by Tully and Whitcomb). Academic
Press, New York, pp. 49–102.
Gourlay, R.N., S.G. Wyld and M.E. Poulton. 1983. Some characteristics
of Mycoplasma virus Hr 1, isolated from and infecting Mycoplasma
hyorhinis. Brief report. Arch. Virol. 77: 81–85.
Grattard, F., B. Pozzetto, B. de Barbeyrac, H. Renaudin, M. Clerc, O.
Gaudin and C. Bébéar. 1995. Arbitrarily-primed PCR confirms the
differentiation of strains of Ureaplasma urealyticum into two biovars.
Mol. Cell Probes 9: 383–389.
Grau, O., F. Laigret, P. Carle, J. Tully, D. Rose and J. Bové. 1991. Identification of a plant-derived mollicute as a strain of an avian pathogen,
Mycoplasma iowae, and its implications for mollicute taxonomy. Int. J.
Gray, L.D., K.L. Ketring and Y.W. Tang. 2005. Clinical use of 16S
rRNA gene sequencing to identify Mycoplasma felis and M. gateae
associated with feline ulcerative keratitis. J. Clin. Microbiol. 43:
3431–3434.
Greco, G., M. Corrente, V. Martella, A. Pratelli and D. Buonavoglia.
2001. A multiplex-PCR for the diagnosis of contagious agalactia of
sheep and goats. Mol. Cell. Probes 15: 21–25.
Green III, F. and R.P. Hanson. 1973. Ultrastructure and capsule of Mycoplasma meleagridis. Journal of Bacteriology 116: 1011–1018.
Grenabo, L., H. Hedelin and S. Pettersson. 1988. Urinary infection
stones caused by Ureaplasma urealyticum: a review. Scand. J. Infect.
Dis. Suppl. 53: 46–49.
Grisold, A., M. Hoenigl, E. Leitner, K. Jakse, G. Feierl, R. Raggam and
E. Marth. 2008. Submasseteric abscess caused by Mycoplasma salivarium infection. J. Clin. Microbiol. 46: 3860–3862.
Groebel, K., K. Hoelzle, M.M. Wittenbrink, U. Ziegler and L.E. Hoelzle.
2009. Mycoplasma suis invades porcine erythrocytes. Infect. Immun.
77: 576–584.
Gwaltney, S.M. and R.D. Oberst. 1994. Comparison of an improved
polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection. J. Vet.
Diagn. Invest. 6: 321–325.
Hale, H.H., C.F. Helmboldt, W.N. Plastridge and E.F. Stula. 1962.
Bovine mastitis caused by a Mycoplasma species. Cornell Vet. 52:
582–591.
Hammond, P., A. Ramírez, C. Morrow and J. Bradbury. 2009. Development and evaluation of an improved diagnostic PCR for Mycoplasma synoviae using primers located in the haemagglutinin
encoding gene vlhA and its value for strain typing. Vet. Microbiol.
136: 61–68.
Hannan, P. 2000. Guidelines and recommendations for antimicrobial
minimum inhibitory concentration (MIC) testing against veterinary
Mycoplasma species. International Research Programme on Comparative Mycoplasmology. Vet. Res. 31: 373–395.
Harasawa, R., K. Koshimizu, I. Pan, E. Stephens and M. Barile. 1984.
Genomic analysis of avian and feline Ureaplasmas by restriction endonucleases. Isr. J. Med. Sci. 20: 942–945.
Harasawa, R., K. Koshimizu, I. Pan and M. Barile. 1985. Genomic and
phenotypic analyses of avian Ureaplasma strains. Nippon Juigaku
Zasshi 47: 901–909.
Harasawa, R., Y. Imada, M. Ito, K. Koshimizu, G.H. Cassell and
M.F. Barile. 1990a. Ureaplasma felinum sp. nov. and Ureaplasma cati sp. nov.
isolated from the oral cavities of cats. Int. J. Syst. Bacteriol. 40: 45–51.
Harasawa, R., E.B. Stephens, K. Koshimizu, I.J. Pan and M.F. Barile.
1990b. DNA relatedness among established Ureaplasma species and
unidentified feline and canine serogroups. Int. J. Syst. Bacteriol. 40:
52–55.
Harasawa, R., K. Dybvig, H.L. Watson and G.H. Cassell. 1991. Two
genomic clusters among 14 serovars of Ureaplasma urealyticum. Syst.
Appl. Microbiol. 14: 393–396.
Harasawa, R., Y. Imada, H. Kotani, K. Koshimizu and M.F. Barile. 1993.
Ureaplasma canigenitalium sp. nov., isolated from dogs. Int. J. Syst. Bacteriol. 43: 640–644.
Harasawa, R. and Y. Kanamoto. 1999. Differentiation of two biovars of
Ureaplasma urealyticum based on the 16S-23S rRNA intergenic spacer
region. J. Clin. Microbiol. 37: 4135–4138.
Harasawa, R., D.G. Pitcher, A.S. Ramirez and J.M. Bradbury. 2004. A
putative transposase gene in the 16S-23S rRNA intergenic spacer
region of Mycoplasma imitans. Microbiology 150: 1023–1029.
Hasselbring, B.M., J.L. Jordan, R.W. Krause and D.C. Krause. 2006.
Terminal organelle development in the cell wall-less bacterium
Mycoplasma pneumoniae. Proc. Natl. Acad. Sci. USA 103: 16478–
16483.
Hatchel, J., R. Balish, M. Duley and M. Balish. 2006. Ultrastructure and
gliding motility of Mycoplasma amphoriforme, a possible human respiratory pathogen. Microbiology 152: 2181–2189.
Hatchel, J.M. and M.F. Balish. 2008. Attachment organelle ultrastructure correlates with phylogeny, not gliding motility properties, in
Mycoplasma pneumoniae relatives. Microbiology 154: 286–295.
Haulena, M., F. Gulland, J. Lawrence, D. Fauquier, S. Jang, B. Aldridge,
T. Spraker, L. Thomas, D. Brown, L. Wendland and M. Davidson.
2006. Lesions associated with a novel Mycoplasma sp. in California sea
lions (Zalophus californianus) undergoing rehabilitation. J. Wildl.
Dis. 42: 40–45.
Heggie, A.D., D. Bar-Shain, B. Boxerbaum, A.A. Fanaroff, M.A.
O’Riordan and J.A. Robertson. 2001. Identification and quantification of ureaplasmas colonizing the respiratory tract and assessment
of their role in the development of chronic lung disease in preterm
infants. Pediatr. Infect. Dis. J. 20: 854–859.
Henderson, G. and G. Jensen. 2006. Three-dimensional structure of
Mycoplasma pneumoniae’s attachment organelle and a model for its
role in gliding motility. Mol. Microbiol. 60: 376–385.
Heyward, J.T., M.Z. Sabry and W.R. Dowdle. 1969. Characterization of
Mycoplasma species of feline origin. Am. J. Vet. Res. 30: 615–622.
Hill, A. 1971. Mycoplasma caviae, a new species. J. Gen. Microbiol. 65:
109–113.
Hill, A. 1977. The isolation of mycoplasmas from non-human primates.
Vet. Rec. 101: 117.
Hill, A.C. 1983a. Mycoplasma cricetuli, a new species from the conjunctivas of chinese hamsters. Int. J. Syst. Bacteriol. 33: 113–117.
Hill, A.C. 1983b. Mycoplasma collis, a new species isolated from rats and
mice. Int. J. Syst. Bacteriol. 33: 847–851.
Hill, A.C. 1984. Mycoplasma cavipharyngis, a new species isolated from
the nasopharynx of guinea pigs. J. Gen. Microbiol. 130: 3183–3188.
Hill, A.C. 1985. Mycoplasma testudinis, a new species isolated from a tortoise. Int. J. Syst. Bacteriol. 35: 489–492.
Hill, A.C. 1986. Mycoplasma felifaucium, a new species isolated from the
respiratory tract of pumas. J. Gen. Microbiol. 132: 1923–1928.
Hill, A.C. 1988. In Validation of the publication of new names and new
combinations previously effectively published outside the IJSB. List
no. 27. Int. J. Syst. Bacteriol. 38: 449.
Hill, A.C. 1989. In Validation of the publication of new names and new
combinations previously effectively published outside the IJSB. List
no. 30. Int. J. Syst. Bacteriol. 39: 371.
Hill, A.C. 1991a. Mycoplasma spermatophilum, a new species isolated
from human spermatozoa and cervix. Int. J. Syst. Bacteriol. 41:
229–233.
Hill, A.C. 1991b. Mycoplasma oxoniensis, a new species isolated from chinese hamster conjunctivas. Int. J. Syst. Bacteriol. 41: 21–25.
Hill, A.C. 1992. Mycoplasma simbae sp. nov., Mycoplasma leopharyngis sp.
nov., and Mycoplasma leocaptivus sp. nov., isolated from lions. Int. J.
Hill, A.C. 1993. Mycoplasma indiense sp. nov., isolated from the throats of
nonhuman primates. Int. J. Syst. Bacteriol. 43: 36–40.
Hill, A., M. Tucker, D. Whittingham and I. Craft. 1987. Mycoplasmas
and in vitro fertilization. Fertil. Steril. 47: 652–655.
Hinz, K.H., H. Pfutzner and K.P. Behr. 1994. Isolation of mycoplasmas
from clinically healthy adult breeding geese in Germany. Zentralbl
Veterinarmed B 41: 145–147.
Hirose, K., H. Kobayashi, N. Ito, Y. Kawasaki, M. Zako, K. Kotani, H.
Ogawa and H. Sato. 2003. Isolation of Mycoplasmas from nasal swabs
of calves affected with respiratory diseases and antimicrobial susceptibility of their isolates. J. Vet. Med. B Infect. Dis. Vet. Public Health
50: 347–351.
Hodges, R.T., M. MacPherson, R.H. Leach and S. Moller. 1983. Isolation of Mycoplasma dispar from mastitis in dry cows. NZ Vet. J. 31:
60–61.
Hoelzle, L. 2008. Haemotrophic mycoplasmas: recent advances in Mycoplasma suis. Vet. Microbiol. 130: 215–226.
Hoffman, R.W., M.P. Luttrell, W.R. Davidson and D.H. Ley. 1997. Mycoplasmas in wild turkeys living in association with domestic fowl. J.
Wildl. Dis. 33: 526–535.
Hooper, P.T., L.A. Ireland and A. Carter. 1985. Mycoplasma polyarthritis in
a cat with probable severe immune deficiency. Aust. Vet. J. 62: 352.
Hopkins, P.M., D.S. Winlaw, P.N. Chhajed, J.L. Harkness, M.D. Horton,
A.M. Keogh, M.A. Malouf and A.R. Glanville. 2002. Mycoplasma hominis infection in heart and lung transplantation. J. Heart Lung Transplant 21: 1225–1229.
Horowitz, S., L. Duffy, B. Garrett, J. Stephens, J. Davis and G. Cassell.
1986. Can group- and serovar-specific proteins be detected in Ureaplasma urealyticum? Pediatr. Infect. Dis. 5: S325–331.
Horowitz, S., J. Horowitz, D. Taylor-Robinson, S. Sukenik, R.N. Apte, J.
Bar-David, B. Thomas and C. Gilroy. 1994. Ureaplasma urealyticum in
Reiter’s syndrome. J. Rheumatol. 21: 877–882.
Hoskins, J.D. 1991. Canine haemobartonellosis, canine hepatozoonosis,
and feline cytauxzoonosis. Vet. Clin. North Am. Small Anim. Pract.
21: 129–140.
Howard, C.J., D.H. Pocock and R.N. Gourlay. 1978. Base composition
of deoxyribonucleic acid from ureaplasmas isolated from various animal species. Int. J. Syst. Bacteriol. 28: 599–601.
Howard, C.J., R.N. Gourlay and S.G. Wyld. 1980. Isolation of a Virus,
Mvbr1, from Mycoplasma-Bovirhinis. FEMS Microbiol. Lett. 7: 163–
165.
Howard, C.J., D.H. Pocock and R.N. Gourlay. 1981. Polyacrylamide gel
electrophoretic comparison or the polypeptides from Ureaplasma
species isolated from cattle and humans. Int. J. Syst. Bacteriol. 31:
128–130.
Howard, C.J. and R.N. Gourlay. 1982. Proposal for a second species
within the genus Ureaplasma, Ureaplasma diversum sp. nov. Int. J. Syst.
Bacteriol. 32: 446–452.
Howard, G.W. 1975. The experimental transmission of Eperythrozoon ovis
by mosquitoes. Parasitology 71: xxxiii.
Hsu, F.S., M.C. Liu, S.M. Chou, J.F. Zachary and A.R. Smith. 1992.
Evaluation of an enzyme-linked immunosorbent assay for detection of Eperythrozoon suis antibodies in swine. Am. J. Vet. Res. 53:
352–354.
Hu, W.S., R.Y. Wang, R.S. Liou, J.W. Shih and S.C. Lo. 1990. Identification of an insertion-sequence-like genetic element in the
629
newly recognized human pathogen Mycoplasma incognitus. Gene
93: 67–72.
Hum, S., A. Kessell, S. Djordjevic, R. Rheinberger, M. Hornitzky, W.
Forbes and J. Gonsalves. 2000. Mastitis, polyarthritis and abortion
caused by Mycoplasma species bovine group 7 in dairy cattle. Aust.
Vet. J. 78: 744–750.
Hyde, T., M. Gilbert, S. Schwartz, E. Zell, J. Watt, W. Thacker, D.
Talkington and R. Besser. 2001. Azithromycin prophylaxis during a
hospital outbreak of Mycoplasma pneumoniae pneumonia. J. Infect.
Dis. 183: 907–912.
Ivanics, E., R. Glávitis, G. Takacs, E. Molnár, Z. Bitay and M. Meder.
1988. An outbreak of Mycoplasma anatis infection associated with nervous symptoms in large-scale duck flocks. Zentralbl. Veterinarmed. B
35: 368–378.
Jackson, G., E. Boughton and S.G. Hamer. 1981. An outbreak of
bovine mastitis associated with Mycoplasma canadense. Vet. Rec. 108:
31–32.
Jacobs, E., A. Stuhlert, M. Drews, K. Pumpe, H. Schaefer, M. Kist and
W. Bredt. 1988. Host reactions to Mycoplasma pneumoniae infections
in guinea-pigs preimmunized systemically with the adhesin of this
pathogen. Microb. Pathog. 5: 259–265.
Jacobsson, B., I. Mattsby-Baltzer, B. Andersch, H. Bokstrom, R.M. Holst,
N. Nikolaitchouk, U.B. Wennerholm and H. Hagberg. 2003. Microbial invasion and cytokine response in amniotic fluid in a Swedish
population of women with preterm prelabor rupture of membranes.
Acta Obstet. Gynecol. Scand. 82: 423–431.
Jain, N.C., D.E. Jasper and J.D. Dellinger. 1967. Cultural characteristics and serological relationships of some mycoplasmas isolated from
bovine sources. J. Gen. Microbiol. 49: 401–410.
Jan, G., M. Le Hénaff, C. Fontenelle and H. Wróblewski. 2001. Biochemical and antigenic characterisation of Mycoplasma gallisepticum
membrane proteins P52 and P67 (pMGA). Arch. Microbiol. 177:
81–90.
Jasper, D.E., H. Erno, J.D. Dellinger and C. Christiansen. 1981. Mycoplasma californicum, a new species from cows. Int. J. Syst. Bacteriol.
31: 339–345.
Jeansson, S. and J.E. Brorson. 1985. Elimination of mycoplasmas from
cell cultures utilizing hyperimmune sera. Exp. Cell. Res. 161: 181–
188.
Jensen, J.S. 2004. Mycoplasma genitalium: the aetiological agent of urethritis and other sexually transmitted diseases. J. Eur. Acad. Dermatol. Venereol. 18: 1–11.
Jensen, J.S., E. Bjornelius, B. Dohn and P. Lidbrink. 2004. Comparison
of first void urine and urogenital swab specimens for detection of
Mycoplasma genitalium and Chlamydia trachomatis by polymerase chain
reaction in patients attending a sexually transmitted disease clinic.
Sex. Transm. Dis. 31: 499–507.
Johansson, K.E. and Pettersson B. 2002. Taxonomy of Mollicutes. In
Molecular biology and pathogenicity of mycoplasmas (edited by
Razin and Herrmann). Kluwer Academic, New York, pp. 1–30.
Jones, G.E., A. Foggie, A. Sutherland and D.B. Harker. 1976. Mycoplasmas and ovine keratoconjunctivitis. Vet. Rec. 99: 137–141.
Jones, G.E. 1985. The pathogenicity of some ovine or caprine mycoplasmas in the lactating mammary gland of sheep and goats. J. Comp.
Pathol. 95: 305–318.
Jordan, F.T., J.N. Howse, M.P. Adams and O.O. Fatunmbi. 1981. The
isolation of Mycoplasma columbinum and M columborale from feral
pigeons. Vet. Rec. 109: 450.
Jordan, F.T.W. 1979. Avian mycoplasmas. In The Mycoplasmas, vol. 2
(edited by Tully Maniloff Whitcomb). Academic Press, New York,
pp. 1–48.
Jordan, F.T.W., H. Ernø, G.S. Cottew, K.H. Kunz and L. Stipkovits. 1982.
Characterization and taxonomic description of five Mycoplasma serovars (serotypes) of avian origin and their elevation to species rank
630
and further evaluation of the taxonomic status of Mycoplasma synoviae. Int. J. Syst. Bacteriol. 32: 108–115.
Judicial Commission. 1958. Opinion 22. Status of the generic name
Asterococcus and conservation of the generic name Mycoplasma. Int.
Bull. Bacteriol. Nomencl. Taxon. 8: 166–168.
Kakulphimp, J., L.R. Finch and J.A. Robertson. 1991. Genome sizes of
mammalian and avian Ureaplasmas. Int. J. Syst. Bacteriol. 41: 326–
327.
Kaya, S., O. Poyraz, G. Gokce, H. Kilicarslan, K. Kaya and S. Ayan. 2003.
Role of genital mycoplasmata and other bacteria in urolithiasis.
Scand. J. Infect. Dis. 35: 315–317.
Keane, F.E., B.J. Thomas, C.B. Gilroy, A. Renton and D. Taylor-Robinson.
2000. The association of Mycoplasma hominis, Ureaplasma urealyticum
and Mycoplasma genitalium with bacterial vaginosis: observations on
heterosexual women and their male partners. Int. J. STD AIDS 11:
356–360.
Kempf, I., F. Gesbert, E. Guinebert, G. M. and G. Bennejean. 1991.
Isolement et caractérisation d’une souche mycoplasmique chez
des faisans d’élevage. Recuil de Medecine Veterinaire 167: 1133–
1139.
Kempf, I. 1998. DNA amplification methods for diagnosis and epidemiological investigations of avian mycoplasmosis. Avian Pathol. 27:
7–14.
Kempf, I., C. Chastel, S. Ferris, F. Dufour-Gesbert, K.E. Johansson, B.
Pettersson and A. Blanchard. 2000. Isolation of Mycoplasma columborale from a fly (Musca domestica). Vet. Rec. 147: 304–305.
Kenny, G. 1983. Inhibition of the growth of Ureaplasma urealyticum by a
new urease inhibitor, flurofamide. Yale J. Biol. Med. 56: 717–722.
Keymer, I.F., R.H. Leach, R.A. Clarke, M.E. Bardsley and R.R. McIntyre.
1984. Isolation of Mycoplasma spp. from racing pigeons (Columba
livia). Avian Pathol. 13: 65–74.
Kidanemariam, A., J. Gouws, M. van Vuuren and B. Gummow. 2005.
Ulcerative balanitis and vulvitis of Dorper sheep in South Africa:
a study on its aetiology and clinical features. JS Afr. Vet. Assoc. 76:
197–203.
Kikuth, W. 1928. Über Einen neuen Anämeerreger; Bartonella canis nov.
spec. Klin. Wochenschr. 7: 1729–1730.
Kilian, M., M.B. Brown, T.A. Brown, E.A. Freundt and G.H. Cassell.
1984. Immunoglobulin A1 protease activity in strains of Ureaplasma
urealyticum. Acta Pathol. Microbiol. Immunol. Scand. [B]. 92: 61–64.
King, K.W. and K. Dybvig. 1994. Mycoplasmal cloning vectors derived
from plasmid pKMK1. Plasmid 31: 49–59.
Kirchhoff, H. 1978. Mycoplasma equigenitalium, a new species from cervix
region of mares. Int. J. Syst. Bacteriol. 28: 496–502.
Kirchhoff, H., P. Beyene, M. Fischer, J. Flossdorf, J. Heitmann, B.
Khattab, D. Lopatta, R. Rosengarten, G. Seidel and C. Yousef. 1987.
Mycoplasma mobile sp. nov., a new species from fish. Int. J. Syst. Bacteriol. 37: 192–197.
Kirchhoff, H., A. Binder, B. Liess, K.T. Friedhoff, J. Pohlenz, M. Stede
and T. Willhaus. 1989. Isolation of mycoplasmas from diseased seals.
Vet. Rec. 124: 513–514.
Kirchhoff, H., R. Schmidt, H. Lehmann, H.W. Clark and A.C. Hill.
1996. Mycoplasma elephantis sp. nov., a new species from elephants.
Kirchhoff, H., K. Mohan, R. Schmidt, M. Runge, D.R. Brown, M.B.
Brown, C.M. Foggin, P. Muvavarirwa, H. Lehmann and J. Flossdorf.
1997. Mycoplasma crocodyli sp. nov., a new species from crocodiles. Int.
Kirchoff, H. 1974. Neue spezies der Fam. Acholeplasmataceae und der
Fam. Mykoplasmataceae bei Pferden. Zentralbl. Veterinarmed. B 21:
207–210.
Kisary, J., A. El-Ebeedy and L. Stipkovits. 1976. Mycoplasma infection of
geese. II. Studies on pathogenicity of mycoplasmas in goslings and
goose and chicken embryos. Avian Pathol. 5: 15–20.
Klassen, T.P., N. Wiebe, K. Russell, K. Stevens, L. Hartling, W.R.
Craig and D. Moher. 2002. Abstracts of randomized controlled
trials presented at the society for pediatric research meeting: an
example of publication bias. Arch. Pediatr. Adolesc. Med. 156:
474–479.
Kleckner, A.L. 1960. Serotypes of avian pleuropneumonia-like organisms. Am. J. Vet. Res. 21: 274–280.
Kleven, S.H., C.S. Eidson and O.J. Fletcher. 1978. Airsacculitis induced
in broilers with a combination of Mycoplasma gallinarum and respiratory viruses. Avian Dis. 22: 707–716.
Kleven, S.H. 1998. Mycoplasmas in the etiology of multifactorial respiratory disease. Poult. Sci. 77: 1146–1149.
Kleven, S.H. 2003. Mycoplasmosis: other mycoplasmal infections. In
Diseases of Poultry, 11th edn (edited by Saif and Barnes). WileyBlackwell, Hoboken, NJ.
Kleven, S.H. 2008. Control of avian Mycoplasma infections in commercial poultry. Avian Dis. 52: 367–374.
Knox, C., P. Giffard and P. Timms. 1998. The phylogeny of Ureaplasma
urealyticum based on the mba gene fragment. Int. J. Syst. Bacteriol.
48 Pt 4: 1323–1331.
Knox, C. and P. Timms. 1998. Comparison of PCR, nested PCR, and
random amplified polymorphic DNA PCR for detection and typing
of Ureaplasma urealyticum in specimens from pregnant women. J. Clin.
Microbiol. 36: 3032–3039.
Knox, C., J. Allan, J. Allan, W. Edirisinghe, D. Stenzel, F. Lawrence, D.
Purdie and P. Timms. 2003. Ureaplasma parvum and Ureaplasma urealyticum are detected in semen after washing before assisted reproductive technology procedures. Fertil. Steril. 80: 921–929.
Kobayashi, H., M. Runge, R. Schmidt, M. Kubo, K. Yamamoto and
H. Kirchhoff. 1997. Mycoplasma lagogenitalium sp. nov., from the
preputial smegma of Afghan pikas (Ochotona rufescens rufescens). Int.
Kokotovic, B., N.F. Friis and P. Ahrens. 2007. Mycoplasma alkalescens
demonstrated in bronchoalveolar lavage of cattle in Denmark. Acta
Vet. Scand. 49: 2.
Kong, F., G. James, Z. Ma, S. Gordon, W. Bin and G. Gilbert. 1999a.
Phylogenetic analysis of Ureaplasma urealyticum–support for the establishment of a new species, Ureaplasma parvum. Int. J. Syst. Bacteriol.
49 Pt 4: 1879–1889.
Kong, F., X. Zhu, W. Wang, X. Zhou, S. Gordon and G.L. Gilbert.
1999b. Comparative analysis and serovar-specific identification of
multiple-banded antigen genes of Ureaplasma urealyticum biovar 1. J.
Clin. Microbiol. 37: 538–543.
Kong, F., Z. Ma, G. James, S. Gordon and G. Gilbert. 2000. Molecular
genotyping of human Ureaplasma species based on multiple-banded
antigen (MBA) gene sequences. Int. J. Syst. Evol. Microbiol. 50 Pt 5:
1921–1929.
Königsson, M.H., B. Pettersson and K.E. Johansson. 2001. Phylogeny of the seal mycoplasmas Mycoplasma phocae corrig., Mycoplasma
phocicerebrale corrig. and Mycoplasma phocirhinis corrig. based on
sequence analysis of 16S rDNA. Int. J. Syst. Evol. Microbiol. 51:
1389–1393.
Koshimizu, K., M. Ito, T. Magaribuchi and H. Kotani. 1983. Selective
medium for isolation of ureaplasmas from animals. Nippon Juigaku
Zasshi 45: 263–268.
Koshimizu, K., R. Harasawa, I.J. Pan, H. Kotani, M. Ogata, E.B. Stephens
and M.F. Barile. 1987. Ureaplasma gallorale sp. nov. from the oropharynx of chickens. Int. J. Syst. Bacteriol. 37: 333–338.
Krause, D.C. and D. Taylor-Robinson. 1992. Mycoplasmas which infect
humans. In Mycoplasmas: Molecular Biology and Pathogenesis
(edited by Maniloff, McElhaney, Finch and Baseman). American
Society for Microbiology, Washington, D.C., pp. 417–444.
Krause, D.C. and M.F. Balish. 2004. Cellular engineering in a minimal
microbe: structure and assembly of the terminal organelle of Mycoplasma pneumoniae. Mol. Microbiol. 51: 917–924.
Kreier, J.P. and M. Ristic. 1963. Morphologic, antigenic, and pathogenic
characteristics of Eperythrozoon ovis and Eperythrozoon wenyoni. Am. J.
Vet. Res. 24: 488–500.
Kreier, J.P. and M. Ristic. 1968. Haemobartonellosis, eperythrozoonosis,
grahamellosis and ehrlichiosis. In Infectious Blood Diseases of Man
and Animals (edited by Weinman and Ristic). Academic Press, New
York, pp. 387–472.
Kreier, J.P. and M. Ristic. 1984. Genus III. Haemobartonella; Genus IV. Eperythrozoon. In Bergey’s Manual of Systematic Bacteriology, vol. 1 (edited by
Krieg and Holt). Williams & Wilkins, Baltimore, pp. 724–729.
Kusiluka, L.J., B. Ojeniyi, N.F. Friis, R.R. Kazwala and B. Kokotovic.
2000. Mycoplasmas isolated from the respiratory tract of cattle and
goats in Tanzania. Acta Vet. Scand. 41: 299–309.
Lam, K., A. DaMassa and G. Ghazikhanian. 2004. Mycoplasma meleagridisinduced lesions in the tarsometatarsal joints of turkey embryos. Avian
Dis. 48: 505–511.
Lamm, C.G., L. Munson, M.C. Thurmond, B.C. Barr and L.W. George.
2004. Mycoplasma otitis in California calves. J. Vet. Diagn. Invest.
16: 397–402.
Lamster, I., J. Grbic, R. Bucklan, D. Mitchell-Lewis, H. Reynolds and
J. Zambon. 1997. Epidemiology and diagnosis of HIV-associated periodontal diseases. Oral Dis. 3 Suppl 1: S141–148.
Langford, E.V. and R.H. Leach. 1973. Characterization of a Mycoplasma
isolated from infectious bovine keratoconjunctivitis: M. bovoculi sp.
nov. Can. J. Microbiol. 19: 1435–1444.
Langford, E.V., H.L. Ruhnke and O. Onoviran. 1976. Mycoplasma
canadense, a new bovine species. Int. J. Syst. Bacteriol. 26: 212–
219.
Lartigue, C., J.I. Glass, N. Alperovich, R. Pieper, P.P. Parmar, C.A.
Hutchison, 3rd, H.O. Smith and J.C. Venter. 2007. Genome transplantation in bacteria: changing one species to another. Science 317:
632–638.
Lartigue, C., S. Vashee, M.A. Algire, R.Y. Chuang, G.A. Benders, L.
Ma, V.N. Noskov, E.A. Denisova, D.G. Gibson, N. Assad-Garcia, N.
Alperovich, D.W. Thomas, C. Merryman, C.A. Hutchison, 3rd, H.O.
Smith, J.C. Venter and J.I. Glass. 2009. Creating bacterial strains from
genomes that have been cloned and engineered in yeast. Science
325: 1693–1696.
Lavrič, M., D. Benčina and M. Narat. 2005. Mycoplasma gallisepticum
hemagglutinin V1hA, pyruvate dehydrogenase PdhA, lactate dehydrogenase, and elongation factor Tu share epitopes with Mycoplasma
imitans homologues. Avian Dis. 49: 507–513.
Leach, R.H. 1967. Comparative studies of Mycoplasma of bovine origin.
Ann. N. Y. Acad. Sci. 143: 305–316.
Leach, R.H. 1973. Further studies on classification of bovine strains of
Mycoplasmatales, with proposals for new species, Acholeplasma modicum
and Mycoplasma alkalescens. J. Gen. Microbiol. 75: 135–153.
Leach, R.H. 1983. Preservation of Mycoplasma cultures and culture collections. In Methods in Mycoplasmology, vol. 1 (edited by Razin and
Tully). Academic Press, New York, pp. 197–204.
Leach, R.H., H. Ernø and K.J. MacOwan. 1993. Proposal for designation of F38-type caprine mycoplasmas as Mycoplasma capricolum subsp.
capripneumoniae and consequent obligatory relegation of strains currently classified as Mycoplasma capricolum (Tully, Barile, Edward, Theo
dore, and Ernø 1974) to an additional new subspecies, M. capricolum
subsp. capricolum subsp. nov. Int. J. Syst. Bacteriol. 43: 603–605.
Lee, G.Y. and G.E. Kenny. 1987. Humoral immune response to polypeptides of Ureaplasma urealyticum in women with postpartum fever.
J. Clin. Microbiol. 25: 1841–1844.
Lehmer, R.R., B.S. Andrews, J.A. Robertson, E.E. Stanbridge, L. de la
Maza and G.J. Friow. 1991. Polyarthiritis due to Ureaplasma urealyticum infection in a patient with common variable immunodeficiency
(CVID): Similarities to rheumatoid arthritis. Ann. Rheum. Dis. 50:
574–576.
631
Lemcke, R.M. 1979. Equine Mycoplasmas. In The Mycoplasmas Volume
2: Human and Animal Mycoplasmas, vol. 2 (edited by Tully and Whitcomb). Academic Press, New York, pp. 177–189.
Lemcke, R.M. and H. Kirchhoff. 1979. Mycoplasma subdolum, a new species isolated from horses. Int. J. Syst. Bacteriol. 29: 42–50.
Lemcke, R.M. and J. Poland. 1980. Mycoplasma fastidiosum: new species
from horses. Int. J. Syst. Bacteriol. 30: 151–162.
Lesnoff, M., G. Laval, P. Bonnet, S. Abdicho, A. Workalemahu, D. Kifle,
A. Peyraud, R. Lancelot and F. Thiaucourt. 2004. Within-herd spread
of contagious bovine pleuropneumonia in Ethiopian highlands. Prev
Vet Med 64: 27–40.
Levisohn, S. and S.H. Kleven. 2000. Avian mycoplasmosis (Mycoplasma
gallisepticum). Rev. Sci. Tech. 19: 425–442.
Ley, D.H., J.E. Berkhoff and J.M. McLaren. 1996. Mycoplasma gallisepticum isolated from house finches (Carpodacus mexicanus) with conjunctivitis. Avian Dis. 40: 480–483.
Ley, D.H., S.J. Geary, J.E. Berkhoff, J.M. McLaren and S. Levisohn.
1998. Mycoplasma sturni from blue jays and northern mockingbirds
with conjunctivitis in Florida. J. Wildl. Dis. 34: 403–406.
Li, X., X. Jia, D. Shi, Y. Xiao, S. Hu, M. Liu, Z. Yuan and D. Bi. 2008.
Continuous in vitro Cultivation of Mycoplasma suis. Acta Veterinaria et
Zootechnica Sinica 38: 1142–1146.
Li, Y., A. Brauner, B. Jonsson, I. van der Ploeg, O. Söder, M. Holst, J.
Jensen, H. Lagercrantz and K. Tullus. 2000. Ureaplasma urealyticuminduced production of proinflammatory cytokines by macrophages.
Pediatr. Res. 48: 114–119.
Lierz, M., R. Schmidt, L. Brunnberg and M. Runge. 2000. Isolation of
Mycoplasma meleagridis from free-ranging birds of prey in Germany. J.
Vet. Med. B Infect. Dis. Vet. Public Health 47: 63–67.
Lierz, M., R. Schmidt and M. Runge. 2002. Mycoplasma species isolated
from falcons in the Middle East. Vet. Rec. 151: 92–93.
Lierz, M., S. Deppenmeier, A. Gruber, S. Brokat and H. Hafez. 2007a.
Pathogenicity of Mycoplasma lipofaciens strain ML64 for turkey
embryos. Avian Pathol. 36: 389–393.
Lierz, M., N. Hagen, N. Harcourt-Brown, S.J. Hernandez-Divers, D.
Luschow and H.M. Hafez. 2007b. Prevalence of mycoplasmas in eggs
from birds of prey using culture and a genus-specific Mycoplasma
polymerase chain reaction. Avian Pathol. 36: 145–150.
Lierz, M., R. Stark, S. Brokat and H. Hafez. 2007c. Pathogenicity of
Mycoplasma lipofaciens strain ML64, isolated from an egg of a Northern Goshawk (Accipiter gentilis), for chicken embryos. Avian Pathol.
36: 151–153.
Lierz, M., N. Hagen, S.J. Hernadez-Divers and H.M. Hafez. 2008a.
Occurrence of mycoplasmas in free-ranging birds of prey in Germany. J. Wildl. Dis. 44: 845–850.
Lierz, M., A. Jansen and H. Hafez. 2008b. Avian Mycoplasma lipofaciens
transmission to veterinarian. Emerg. Infect. Dis. 14: 1161–1163.
Lierz, M., E. Obon, B. Schink, F. Carbonell and H.M. Hafez. 2008c. The
role of mycoplasmas in a conservation project of the lesser kestrel
(Falco naumanni). Avian Dis. 52: 641–645.
Lin, J., M. Kendrick and E. Kass. 1972. Serologic typing of human genital T-mycoplasmas by a complement-dependent mycoplasmacidal
test. J. Infect. Dis. 126: 658–663.
Livingston, C.J. and B. Gauer. 1974. Serologic typing of T-strain Mycoplasma isolated from the respiratory and reproductive tracts of cattle
in the United States. Am. J. Vet. Res. 35: 1469–1471.
Lo, S.C., M.M. Hayes, J.G. Tully, R.Y. Wang, H. Kotani, P.F. Pierce,
D.L. Rose and J.W. Shih. 1992. Mycoplasma penetrans sp. nov., from
the urogenital tract of patients with AIDS. Int. J. Syst. Bacteriol. 42:
357–364.
Lobo, E., M.C. García, H. Moscoso, S. Martínez and S.H. Kleven. 2004.
Strain heterogeneity in Mycoplasma pullorum isolates identified by
random amplified polymorphic DNA techniques. Spanish Journal of
Agricultural Research 2: 500–503.
632
Lorenzon, S., L. Manso-Silvan and F. Thiaucourt. 2008. Specific realtime PCR assays for the detection and quantification of Mycoplasma
mycoides subsp. mycoides SC and Mycoplasma capricolum subsp. capripneumoniae. Mol. Cell. Probes. 22: 324–328.
Luo, W., H. Yu, Z. Cao, T.R. Schoeb, M. Marron and K. Dybvig. 2008.
Association of Mycoplasma arthritidis mitogen with lethal toxicity but
not with arthritis in mice. Infect. Immun. 76: 4989–4998.
Luttrell, M.P., T.H. Eleazer and S.H. Kleven. 1992. Mycoplasma gallopavonis in eastern wild turkeys. J. Wildl. Dis. 28: 288–291.
Lysnyansky, I., M. Calcutt, I. Ben-Barak, Y. Ron, S. Levisohn, B. Methé
and D. Yogev. 2009. Molecular characterization of newly identified
IS3, IS4 and IS30 insertion sequence-like elements in Mycoplasma
bovis and their possible roles in genome plasticity. FEMS Microbiol.
Lett. 294: 172–182.
MacKenzie, C., B. Henrich and U. Hadding. 1996. Biovar-specific
epitopes of the urease enzyme of Ureaplasma urealyticum. J. Med.
Microbiol. 45: 366–371.
MacOwan, K.J., H.G. Jones, C.J. Randall and F.T. Jordan. 1981. Mycoplasma columborale in a respiratory condition of pigeons and experimental air sacculitis of chickens. Vet. Rec. 109: 562.
Madden, D.L., K.E. Moats, W.T. London, E.B. Matthew and J.L. Sever.
1974. Mycoplasma moatsii, a new species isolated from recently
imported Grivit monkeys (Cercopithecus aethiops). Int. J. Syst. Bacteriol. 24: 459–464.
Maeda, T., T. Shibahara, K. Kimura, Y. Wada, K. Sato, Y. Imada,
Y. Ishikawa and K. Kadota. 2003. Mycoplasma bovis-associated suppurative otitis media and pneumonia in bull calves. J. Comp. Pathol. 129:
100–110.
Manchee, R. and D. Taylor-Robinson. 1969. Enhanced growth of T-strain
mycoplasmas with N-2-hydroxyethylpiperazone-N¢-2-ethanesulfonic
acid buffer. J. Bacteriol. 100: 78–85.
Manchee, R. and D. Taylor-Robinson. 1970. Lysis and protection of
erythrocytes by T-mycoplasmas. J. Med. Microbiol. 3: 539–546.
Manhart, L.E., S.M. Dutro, K.K. Holmes, C.E. Stevens, C.W. Critchlow,
D.A. Eschenbach and P.A. Totten. 2001. Mycoplasma genitalium is
associated with mucopurulent cervicitis. Int. J. STD AIDS 12
(suppl. 2): 69.
Manso-Silván, L., E.M. Vilei, K. Sachse, S.P. Djordjevic, F. Thiaucourt
and J. Frey. 2009. Mycoplasma leachii sp. nov. as a new species designation for Mycoplasma sp. bovine group 7 of Leach, and reclassification
of Mycoplasma mycoides subsp. mycoides LC as a serovar of Mycoplasma
mycoides subsp. capri. Int. J. Syst. Evol. Microbiol. 59: 1353–1358.
March, J.B., C. Gammack and R. Nicholas. 2000. Rapid detection of
contagious caprine pleuropneumonia using a Mycoplasma capricolum subsp. capripneumoniae capsular polysaccharide-specific
antigen detection latex agglutination test. J. Clin. Microbiol. 38:
4152–4159.
Maré, C.J. and W.P. Switzer. 1965. Mycoplasm hyopneumoniae, a causative
agent of virus pig pneumonia. Vet. Med. 60: 841–845.
Markham, P.F., M.F. Duffy, M.D. Glew and G.F. Browning. 1999. A gene
family of Mycoplasma imitans closely related to the pMGA family of
Mycoplasma gallisepticum. Microbiology 145: 2095–2103.
Marois, C., F. Dufour-Gesbert and I. Kempf. 2001. Comparison of
pulsed-field gel electrophoresis with random amplified polymorphic DNA for typing of Mycoplasma synoviae. Vet. Microbiol. 79: 1–9.
Marois, C., J. Le Carrou, M. Kobisch and A.V. Gautier-Bouchardon.
2007. Isolation of Mycoplasma hyopneumoniae from different sampling
sites in experimentally infected and contact SPF piglets. Vet. Microbiol. 120: 96–104.
Mason, R.W. and P. Statham. 1991. The determination of the level of
Eperythrozoon ovis parasitaemia in chronically infected sheep and its
significance to the spread of infection. Aust. Vet. J. 68: 115–116.
Masover, G., S. Razin and L. Hayflick. 1977. Localization of enzymes
in Ureaplasma urealyticum (T-strain mycoplasma). J. Bacteriol. 130:
297–302.
Masover, G.K. and F.A. Becker. 1996. Detection of mycoplasmas by DNA
staining and fluorescent antibody methodology. In Molecular and
Diagnostic Procedures in Mycoplasmology, vol. 2 (edited by Tully
and Razin). Academic Press, San Diego, pp. 419–429.
Matlow, A., C. Th’ng, D. Kovach, P. Quinn, M. Dunn and E. Wang.
1998. Susceptibilities of neonatal respiratory isolates of Ureaplasma
urealyticum to antimicrobial agents. Antimicrob. Agents Chemother.
42: 1290–1292.
Mattila, P.S., P. Carlson, A. Sivonen, J. Savola, R. Luosto, J. Salo and M.
Valtonen. 1999. Life-threatening Mycoplasma hominis mediastinitis.
Clin. Infect. Dis. 29: 1529–1537.
Mayer, D., M.P. Degiorgis, W. Meier, J. Nicolet and M. Giacometti. 1997.
Lesions associated with infectious keratoconjunctivitis in alpine ibex.
J. Wildl. Dis. 33: 413–419.
May, M., G.J. Ortiz, L.D. Wendland, D.S. Rotstein, R.F. Relich, M.F. Balish and D.R. Brown. 2007. Mycoplasma insons sp. nov., a twisted Mycoplasma from green iguanas (Iguana iguana). FEMS Microbiol. Lett.
274: 298–303.
Mayer, D., M.P. Degiorgis, W. Meier, J. Nocolet and M. Giacometti. 1997.
Lesions associated with infectious keratoconjuctivitis in alpine ibex.
J. Widl. Dis. 33: 413–419.
Mayer, M. 1921. Über einige bakterienähnliche Parasiten der Erythrozyten
bei Menschen und Tieren. Arch. Schiffs Trop. Hyg. 25: 150–152.
Mazzali, R. and D. Taylor-Robinson. 1971. The behaviour of T-mycoplasmas
in tissue culture. J. Med. Microbiol. 4: 125–138.
McAuliffe, L., R. Ellis, K. Miles, R. Ayling and R. Nicholas. 2006. Biofilm
formation by Mycoplasma species and its role in environmental persistence and survival. Microbiology 152: 913–922.
McAuliffe, L., R.D. Ayling, R.J. Ellis and R.A. Nicholas. 2008. Biofilmgrown Mycoplasma mycoides subsp. mycoides SC exhibit both phenotypic and genotypic variation compared with planktonic cells. Vet.
Microbiol. 129: 315–324.
McElhaney, R.N. 1992a. Lipid composition, biosynthesis, and metabolism. In Mycoplasmas: Molecular Biology and Pathogenesis (edited
McElhaney, R.N. 1992b. Membrance function. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney,
Finch and Baseman). American Society for Microbiology, Washington, D.C., pp. 259–287.
McElhaney, R.N. 1992c. Membrane structure. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch
and Baseman). American Society for Microbiology, Washington,
D.C., pp. 113–155.
McGarrity, G.J., D.L. Rose, V. Kwiatkowski, A.S. Dion, D.M. Phillips and
J.G. Tully. 1983. Mycoplasma muris, a new species from laboratory
mice. Int. J. Syst. Bacteriol. 33: 350–355.
McLaughlin, B.G., P.S. McLaughlin and C.N. Evans. 1991. An Eperythrozoon-like parasite of llamas: attempted transmission to swine, sheep,
and cats. J. Vet. Diagn. Invest. 3: 352–353.
McMartin, D.A., K.J. MacOwan and L.L. Swift. 1980. A century of classical contagious caprine pleuropneumonia: from original description
to aetiology. Br. Vet. J. 136: 507–515.
Meloni, G.A., G. Bertoloni, F. Busolo and L. Conventi. 1980. Colony
morphology, ultrastructure and morphogenesis in Mycoplasma hominis, Acholeplasma laidlawii and Ureaplasma urealyticum. J. Gen. Microbiol. 116: 435–443.
Messick, J.B., P.G. Walker, W. Raphael, L. Berent and X. Shi. 2002.
‘Candidatus Mycoplasma haemodidelphidis’ sp. nov., ‘Candidatus
Mycoplasma haemolamae’ sp. nov. and Mycoplasma haemocanis comb.
nov., haemotrophic parasites from a naturally infected opossum
(Didelphis virginiana), alpaca (Lama pacos) and dog (Canis familiaris): phylogenetic and secondary structural relatedness of their
16S rRNA genes to other mycoplasmas. Int. J. Syst. Evol. Microbiol.
52: 693–698.
Messick, J.B. 2003. New perspectives about Hemotrophic Mycoplasma
(formerly, Haemobartonella and Eperythrozoon species) infections in
dogs and cats. Vet. Clin. N. Am. Small Anim. Pract. 33: 1453–1465.
Meyer, R. and W. Clough. 1993. Extragenital Mycoplasma hominis infections in adults: emphasis on immunosuppression. Clin. Infect. Dis.
17 Suppl 1: S243–249.
Meyling, A. and N.F. Friis. 1972. Serological identification of a new porcine Mycoplasma species, M. flocculare. Acta Vet. Scand. 13: 287–289.
Michel, J.C., B. de Thoisy and H. Contamin. 2000. Chemotherapy of
haemobartonellosis in squirrel monkeys (Saimiri sciureus). J. Med.
Primatol. 29: 85–87.
Mikaelian, I., D.H. Ley, R. Claveau, M. Lemieux and J.P. Berube. 2001.
Mycoplasmosis in evening and pine grosbeaks with conjunctivitis in
Quebec. J. Wildl. Dis. 37: 826–830.
Miles, R.J. 1992a. Catabolism in Mollicutes. J. Gen. Microbiol. 138: 1773–
1783.
Miles, R.J. 1992b. Cell nutrition and growth. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch
D.C., pp. 23–40.
Mirsalimi, S., S. Rosendal and R. Julian. 1989. Colonization of the intestine of turkey embryos exposed to Mycoplasma iowae. Avian Dis. 33:
310–315.
Miyata, M., H. Yamamoto, T. Shimizu, A. Uenoyama, C. Citti and
R. Rosengarten. 2000. Gliding mutants of Mycoplasma mobile: relationships between motility and cell morphology, cell adhesion and
microcolony formation. Microbiology 146: 1311–1320.
Miyata, M., W.S. Ryu and H.C. Berg. 2002. Force and velocity of Mycoplasma mobile gliding. J. Bacteriol. 184: 1827–1831.
Moalic, P.Y., I. Kempf, F. Gesbert and F. Laigret. 1997. Identification of
two pathogenic avian mycoplasmas as strains of Mycoplasma pullorum.
Mohan, K., C.M. Foggin, P. Muvavarirwa and J. Honywill. 1997. Vaccination of farmed crocodiles (Crocodylus niloticus) against Mycoplasma
crocodyli infection. Vet. Rec. 141: 476.
Mohan, K., C.M. Foggin, F. Dziva and P. Muvavarirwa. 2001. Vaccination
to control an outbreak of Mycoplasma crocodyli infection. Onderstepoort J. Vet. Res. 68: 149–150.
Moise, N.S., J.W. Crissman, J.F. Fairbrother and C. Baldwin. 1983.
Mycoplasma gateae arthritis and tenosynovitis in cats: case report
and experimental reproduction of the disease. Am. J. Vet. Res. 44:
16–21.
Monecke, S., J. Helbig and E. Jacobs. 2003. Phase variation of the multiple banded protein in Ureaplasma urealyticum and Ureaplasma parvum.
Int. J. Med. Microbiol. 293: 203–211.
Montagnier, L., A. Blanchard, D. Guetard, M. Lemaitre, A.M. Dirienzo,
S. Chamaret, Y. Henin, E. Bahraoui, C. Dauguet, C. Axler, M.
Kirstetter, R. Roue, G. pialoux and B. Dupont. 1990. A possible role
of mycoplasmas as cofactors in AIDS. In Retroviruses of Human AIDS
and Related Animal Diseases (edited by Girard and Valette). Colloque de Cent Gardes, Foundation Merieux, Lyon, pp. 9–17.
Montes, A., D. Wolfe, E. Welles, J. Tyler and E. Tepe. 1994. Infertility
associated with Eperythrozoon wenyonii infection in a bull. J. Am. Vet.
Med. Assoc. 204: 261–263.
Moss, T., C. Knox, S. Kallapur, I. Nitsos, C. Theodoropoulos, J. Newnham, M. Ikegami and A. Jobe. 2008. Experimental amniotic fluid
infection in sheep: effects of Ureaplasma parvum serovars 3 and 6
on preterm or term fetal sheep. Am. J. Obstet. Gynecol. 198: 122.
e121–128.
Mouches, C., D. Taylor-Robinson, L. Stipkovits and J. Bove. 1981. Comparison of human and animal Ureaplasmas by one- and two-dimensional protein analysis on polyacrylamide slab gel. Ann. Microbiol.
(Paris) 132B: 171–196.
633
Muhlrad, A., I. Peleg, J. Robertson, I. Robinson and I. Kahane. 1981.
Acetate kinase activity in mycoplasmas. J. Bacteriol. 147: 271–273.
Murakami, S., M. Miyama, A. Ogawa, J. Shimada and T. Nakane. 2002.
Occurrence of conjunctivitis, sinusitis and upper region tracheitis
in Japanese quail (Coturnix coturnix japonica), possibly caused by
Mycoplasma gallisepticum accompanied by Cryptosporidium sp. infection. Avian Pathol. 31: 363–370.
Nagata, K., E. Takagi, H. Satoh, H. Okamura and T. Tamura. 1995.
Growth inhibition of Ureaplasma urealyticum by the proton pump
inhibitor lansoprazole: direct attribution to inhibition by lansoprazole of urease activity and urea-induced ATP synthesis in U. urealyticum. Antimicrob. Agents Chemother. 39: 2187–2192.
Nagatomo, H., H. Kato, T. Shimizu and B. Katayama. 1997. Isolation of
mycoplasmas from fantail pigeons. J. Vet. Med. Sci. 59: 461–462.
Nakane, D. and M. Miyata. 2007. Cytoskeletal “jellyfish” structure of
Mycoplasma mobile. Proc. Natl. Acad. Sci. USA 104: 19518–19523.
Nakane, D. and M. Miyata. 2009. Cytoskeletal asymmetrical dumbbell
structure of a gliding mycoplasma, Mycoplasma gallisepticum, revealed
by negative-staining electron microscopy. J. Bacteriol. 191: 3256–
3264.
Naot, Y., J.G. Tully and H. Ginsburg. 1977. Lymphocyte activation by
various Mycoplasma strains and species. Infect. Immun. 18: 310–
317.
Neimark, H., D. Mitchelmore and R.H. Leach. 1998. An approach
to characterizing uncultivated prokaryotes the Grey Lung agent
and proposal of a Candidatus taxon for the organism, ‘Candidatus
Mycoplasma ravipulmonis’. Int. J. Syst. Bacteriol. 48: 389–394.
Neimark, H., K.E. Johansson, Y. Rikihisa and J.G. Tully. 2001. Proposal
to transfer some members of the genera Haemobartonella and Eperythrozoon to the genus Mycoplasma with descriptions of ‘Candidatus Mycoplasma haemofelis’, ‘Candidatus Mycoplasma haemomuris’, ‘Candidatus
Mycoplasma haemosuis’ and ‘Candidatus Mycoplasma wenyonii’. Int. J.
Syst. Evol. Microbiol. 51: 891–899.
Neimark, H., A. Barnaud, P. Gounon, J.-C. Michel and H. Contamin.
2002a. The putative haemobartonella that influences Plasmodium
falciparum parasitaemia in squirrel monkeys is a haemotrophic mycoplasma. Microbes Infect. 4: 693–698.
Neimark, H., B. Hoff and M. Ganter. 2004. Mycoplasma ovis comb. nov.
(formerly Eperythrozoon ovis), an epierythrocytic agent of haemolytic
anaemia in sheep and goats. Int. J. Syst. Evol. Microbiol. 54: 365–
371.
Neimark, H., W. Peters, B.L. Robinson and L.B. Stewart. 2005. Phylogenetic analysis and description of Eperythrozoon coccoides, proposal
to transfer to the genus Mycoplasma as Mycoplasma coccoides comb.
nov. and Request for an Opinion. Int. J. Syst. Evol. Microbiol. 55:
1385–1391.
Neimark, H.C., K.E. Johansson, Y. Rikihisa and J.G. Tully. 2002b. Revision of haemotrophic Mycoplasma species names. Int. J. Syst. Evol.
Microbiol. 52: 683.
Neitz, W.O., R.A. Alexander and P.J. de Toit. 1934. Eperythrozoon ovis (sp.
nov.) infection in sheep. Onderstepoort J. Vet. Sci. 3: 263–274.
Nelson, J.B. and M.J. Lyons. 1965. Phase-contrast and electron microscopy of murine strains of Mycoplasma. J. Bacteriol. 90: 1750–1763.
Neyrolles, O., S. Ferris, N. Behbahani, L. Montagnier and A. Blanchard.
1996. Organization of Ureaplasma urealyticum urease gene cluster and
expression in a suppressor strain of Escherichia coli. J. Bacteriol. 178:
2725.
Niang, M., R.F. Rosenbusch, J.J. Andrews and M.L. Kaeberle. 1998.
Demonstration of a capsule on Mycoplasma ovipneumoniae. Am. J. Vet.
Res. 59: 557–562.
Nicholas, R.A., A. Greig, S.E. Baker, R.D. Ayling, M. Heldtander, K.-E.
Johansson, B.M. Houshaymi and R.J. Miles. 1998. Isolation of Mycoplasma fermentans from a sheep. Vet. Rec. 142: 220–221.
634
Nicholas, R.A., Y.C. Lin, K. Sachse, H. Hotzel, K. Parham, L. McAuliffe,
R.J. Miles, D.P. Kelly and A.P. Wood. 2008. Proposal that the strains of
the Mycoplasma ovine/caprine serogroup 11 be reclassified as Mycoplasma bovigenitalium. Int. J. Syst. Evol. Microbiol. 58: 308–312.
Nikol’skii, S.N. and S.N. Slipchenko. 1969. Experiments in the transmission of Eperythrozoon ovis by the ticks H. plumbeum and Rh. bursa.
Veterinariia (Russian) 5: 46.
Nolan, P.M., S.R. Roberts and G.E. Hill. 2004. Effects of Mycoplasma gallisepticum on reproductive success in house finches. Avian Dis. 48:
879–885.
Novy, M.J., L. L. Duffy, M.K. Axhelm, D.W. Sadowsky, S.S. Witkin, M.G.
Gravett, Cassell, G.H. and K.B. Waites. 2009. Congenital and opportunistic infections: Ureaplasma species and Mycoplasma hominis. Reproductive Science 16: 56–70.
Nowak, J. 1929. Morphologie, nature et cycle évolutif du microbe de
la péripneumonie des bovidés. Ann. Inst. Pasteur (Paris) 43: 1330–
1352.
Nunoya, T., T. Yagihashi, M. Tajima and Y. Nagasawa. 1995. Occurrence
of keratoconjunctivitis apparently caused by Mycoplasma gallisepticum
in layer chickens. Vet. Pathol. 32: 11–18.
O’Brien, S.J., J. Simonson, S. Razin and M.F. Barile. 1983. On the distribution and characteristics of isozyme expression in Mycoplasma. The
Yale Journal of Biology and Medicine 56: 701–708.
Oaks, J.L., S.L. Donahoe, F.R. Rurangirwa, B.A. Rideout, M. Gilbert and
M.Z. Virani. 2004. Identification of a novel Mycoplasma species from
an Oriental white-backed vulture (Gyps bengalensis). J. Clin. Microbiol. 42: 5909–5912.
Olson, N.O., K.M. Kerr and A. Campbell. 1964. Control of infectious synovitis. 13. The antigen study of three strains. Avian Dis. 8: 209–214.
Paessler, M., A. Levinson, J.B. Patel, M. Schuster, M. Minda and
I. Nachamkin. 2002. Disseminated Mycoplasma orale infection in a
patient with common variable immunodeficiency syndrome. Diagn.
Microbiol. Infect. Dis. 44: 201–204.
Panangala, V.S., J.S. Stringfellow, K. Dybvig, A. Woodard, F. Sun, D.L.
Rose and M.M. Gresham. 1993. Mycoplasma corogypsi sp. nov., a new
species from the footpad abscess of a black vulture, Coragyps atratus.
Papazisi, L., T. Gorton, G. Kutish, P. Markham, G. Browning, D. Nguyen,
S. Swartzell, A. Madan, G. Mahairas and S. Geary. 2003. The complete genome sequence of the avian pathogen Mycoplasma gallisepticum strain R(low). Microbiology 149: 2307–2316.
Pennycott, T., C. Dare, C. Yavari and J. Bradbury. 2005. Mycoplasma
sturni and Mycoplasma gallisepticum in wild birds in Scotland. Vet. Rec.
156: 513–515.
Pereyre, S., P. Gonzalez, B. de Barbeyrac, A. Darnige, H. Renaudin, A.
Charron, S. Raherison, C. Bébéar and C.M. Bébéar. 2002. Mutations
in 23S rRNA account for intrinsic resistance to macrolides in Mycoplasma hominis and Mycoplasma fermentans and for acquired resistance
to macrolides in M. hominis. Antimicrob. Agents Chemother. 46:
3142–3150.
Persson, A., B. Pettersson, G. Bölske and K.-E. Johansson. 1999. Diagnosis of contagious bovine pleuropneumonia by PCR-laser- induced
fluorescence and PCR-restriction endonuclease analysis based on the
16S rRNA genes of Mycoplasma mycoides subsp. mycoides SC. J. Clin.
Microbiol. 37: 3815–3821.
Peterson, J.E., A.W. Rodwell and E.S. Rodwell. 1973. Occurrence and
ultrastructure of a variant (rho) form of Mycoplasma. J. Bacteriol. 115:
411–425.
Pilo, P., B. Fleury, M. Marenda, J. Frey and E. Vilei. 2003. Prevalence
and distribution of the insertion element ISMag1 in Mycoplasma agalactiae. Vet. Microbiol. 92: 37–48.
Piot, P. 1977. Comparison of growth inhibition and immunofluorescence tests in serotyping clinical isolates of Ureaplasma urealyticum. Br.
J. Vener. Dis. 53: 186–189.
Pitcher, D., M. Sillis and J.A. Robertson. 2001. Simple method for determining biovar and serovar types of Ureaplasma urealyticum clinical isolates using PCR-single-strand conformation polymorphism analysis.
Pitcher, D., D. Windsor, H. Windsor, J. Brabbury, C. Yavari, J.S. Jensen,
C. Ling and D. Webster. 2005. Mycoplasma amphoriforme sp. nov., a new
species isolated from a patient with chronic brochopneumonia. Int.
J. Syst. Evol. Microbiol. 55: 2589–2594.
Pollack, J. 1986. Metabolic distinctiveness of ureaplasmas. Pediatr.
Infect. Dis. 5: S305–307.
Pollack, J.D. 1992. Carbohydrate metabolism and energy conservation. In Mycoplasmas: Molecular Biology and Pathogenesis (edited
Pollack, J.D., M.V. Williams, J. Banzon, M.A. Jones, L. Harvey and
J.G. Tully. 1996. Comparative metabolism of Mesoplasma, Entomoplasma, Mycoplasma, and Acholeplasma. Int. J. Syst. Bacteriol. 46:
885–890.
Pollack, J.D. 1997. Mycoplasma genes: a case for reflective annotation.
Trends Microbiol. 5: 413–419.
Pollack, J.D. 2001. Ureaplasma urealyticum: an opportunity for combinatorial genomics. Trends Microbiol. 9: 169–175.
Pollack, J.D. 2002. The necessity of combining genomic and enzymatic
data to infer metabolic function and pathways in the smallest bacteria: amino acid, purine and pyrimidine metabolism in mollicutes.
Front. Biosci. 7: d1762–1781.
Poveda, J., J. Carranza, A. Miranda, A. Garrido, M. Hermoso, A. Fernandez and J. Domenech. 1990. An epizootiological study of avian
mycoplasmas in southern Spain. Avian Pathol. 19: 627–633.
Poveda, J.B., J. Giebel, J. Flossdorf, J. Meier and H. Kirchhoff. 1994.
Mycoplasma buteonis sp. nov. Mycoplasma falconis sp. nov. and Mycoplasma gypis sp. nov. three species from birds of prey. Int. J. Syst. Bacteriol. 44: 94–98.
Prullage, J.B., R.E. Williams and S.M. Gaafar. 1993. On the transmissibility of Eperythrozoon suis by Stomoxys calcitrans and Aedes aegypti. Vet.
Parasitol. 50: 125–135.
Purcell, R., D. Taylor-Robinson, D. Wong and R. Chanock. 1966. Color
test for the measurement of antibody to T-strain mycoplasmas. J.
Bacteriol. 92: 6–12.
Pye, G.W., D.R. Brown, M.F. Nogueira, K.A. Vliet, T.R. Schoeb, E.R.
Jacobson and R.A. Bennett. 2001. Experimental inoculation of
broad-nosed caimans (Caiman latirostris) and Siamese crocodiles
(Crocodylus siamensis) with Mycoplasma alligatoris. J. Zoo Wildl. Med.
32: 196–201.
Ramírez, A., C. Naylor, D. Pitcher and J. Bradbury. 2008. High interspecies and low intra-species variation in 16S-23S rDNA spacer
sequences of pathogenic avian mycoplasmas offers potential use as a
diagnostic tool. Vet. Microbiol. 128: 279–287.
Raviv, Z. and S. Kleven. 2009. The development of diagnostic real-time
TaqMan PCrs for the four pathogenic avian mycoplasmas. Avian Dis.
53: 103–107.
Razin, S. 1979. Isolation and characterization of Mycoplasma membranes. In The Mycoplasmas, vol. 1 (edited by Barile and Tully). Academic Press, New York, pp. 213–229.
Razin, S. 1983. Cell lysis and isolation of membranes. In Methods in
Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press,
Razin, S., D. Yogev and Y. Naot. 1998. Molecular biology and pathogenicity of mycoplasmas. Microbiol. Mol. Biol. Rev. 62: 1094–1156.
Razin, S. and R. Herrmann (editors). 2002. Molecular Biology and
Pathogenicity of Mycoplasmas. Academic/Plenum Press, L
ondon.
Reece, R.L., L. Ireland and P.C. Scott. 1986. Mycoplasmosis in racing
pigeons. Aust. Vet. J. 63: 166–167.
Regendanz, P. and W. Kikuth. 1928. Über Aktivierung labiler Infektionen duch Entmilzung (Piroplasma canis, Nuttalia brasiliensis, Bartonella opossum, Spirochaeta didelphydis). Arch. f. Schiffs. U. Tropenhyg.
32: 587–593.
Relich, R.F., A.J. Friedberg and M.F. Balish. 2009. Novel cellular organization in a gliding Mycoplasma, Mycoplasma insons. J. Bacteriol. 191:
5312–5314.
Reyes, L., M. Reinhard and M. Brown. 2009. Different inflammatory
responses are associated with Ureaplasma parvum-induced UTI and
urolith formation. BMC Infect. Dis. 9: 9.
Roberts, D.H. 1964. The isolation of an influenza A virus and a Mycoplasma associated with duck sinusitis. Vet. Rec. 76: 470–473.
Roberts, M. and G. Kenny. 1986. Dissemination of the tetM tetracycline
resistance determinant to Ureaplasma urealyticum. Antimicrob. Agents
Chemother. 29: 350–352.
Robertson, J. and E. Smook. 1976. Cytochemical evidence of extramembranous carbohydrates on Ureaplasma urealyticum (T-strain Mycoplasma). J. Bacteriol. 128: 658–660.
Robertson, J. 1978. Bromothymol blue broth: improved medium for
detection of Ureaplasma urealyticum (T-strain mycoplasma). J. Clin.
Robertson, J. and G. Stemke. 1979. Modified metabolic inhibition test
for serotyping strains of Ureaplasma urealyticum (T-strain Mycoplasma).
Robertson, J., J. Coppola and O. Heisler. 1981. Standardized method
for determining antimicrobial susceptibility of strains of Ureaplasma
urealyticum and their response to tetracycline, erythromycin, and
rosaramicin. Antimicrob. Agents Chemother. 20: 53–58.
Robertson, J. 1982. Effect of gaseous conditions on isolation and growth
of Ureaplasma urealyticum on agar. J. Clin. Microbiol. 15: 200–203.
Robertson, J. and G. Stemke. 1982. Expanded serotyping scheme for
Ureaplasma urealyticum strains isolated from humans. J. Clin. Microbiol. 15: 873–878.
Robertson, J., M. Alfa and E. Boatman. 1983. Morphology of the cells
and colonies of Mycoplasma hominis. Sex Transm. Dis. 10: 232–239.
Robertson, J. and M. Chen. 1984. Effects of manganese on the growth
and morphology of Ureaplasma urealyticum. J. Clin. Microbiol. 19:
857–864.
Robertson, J., M. Stemler and G. Stemke. 1984. Immunoglobulin A
protease activity of Ureaplasma urealyticum. J. Clin. Microbiol. 19:
255–258.
Robertson, J., G. Stemke, S. Maclellan and D. Taylor. 1988. Characterization of tetracycline-resistant strains of Ureaplasma urealyticum. J.
Antimicrob. Chemother. 21: 319–332.
Robertson, J., A. Vekris, C. Bebear and G. Stemke. 1993. Polymerase
chain reaction using 16S rRNA gene sequences distinguishes the two
biovars of Ureaplasma urealyticum. J. Clin. Microbiol. 31: 824–830.
Robertson, J.A., G. Stemke, J.J. Davis, R. Harasawa, D. Thirkell, F. Kong,
M. Shepard and D. Ford. 2002. Proposal of Ureaplasma parvum sp.
nov. and emended description of Ureaplasma urealyticum (Shepard
et al. 1974) Robertson et al. 2001. Int. J. Syst. Evol. Microbiol. 52:
587–597.
Robertson, J.A., L.E. Pyle, G.W. Stemke and L.R. Finch. 1990. Human
ureaplasmas show diverse genome sizes by pulsed-field electrophoresis. Nucleic Acids Res. 18: 1451–1455.
Robertson, J.A. and R. Sherburne. 1991. Hemadsorption by colonies of
Ureaplasma urealyticum. Infect. Immun. 59: 2203–2206.
Robertson, J.A., L.A. Howard, C.L. Zinner and G.W. Stemke. 1994.
Comparison of 16S rRNA genes within the T960 and parvo biovars
of ureaplasmas isolated from humans. Int. J. Syst. Bacteriol. 44:
836–838.
635
Robertson, J.A. and G.W. Stemke. 1995. Measurement of Mollicute
Growth by ATP-Dependent Luminometry. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 1 (edited by Razin and
Tully). Academic Press, San Diego, CA, pp. 65–71.
Rocha, E. and A. Blanchard. 2002. Genomic repeats, genome plasticity
and the dynamics of Mycoplasma evolution. Nucleic Acids Res. 30:
2031–2042.
Rodwell, A. and R.F. Whitcomb. 1983. Methods for direct and indirect
measurement of Mycoplasma growth. In Methods in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, New York,
pp. 185–196.
Rodwell, A.W. 1983. Defined and partly defined media. In Methods in
Roifman, C., C. Rao, H. Lederman, S. Lavi, P. Quinn and E. Gelfand.
1986. Increased susceptibility to Mycoplasma infection in patients with
hypogammaglobulinemia. Am. J. Med. 80: 590–594.
Romano, N., G. Tolone, F. Ajello and R. La Licata. 1980. Adenosine
5′-triphosphate synthesis induced by urea hydrolysis in Ureaplasma
urealyticum. J. Bacteriol. 144: 830–832.
Rose, D.L., J.G. Tully and E.V. Langford. 1978. Mycoplasma citelli, a
new species from ground squirrels. Int. J. Syst. Bacteriol. 28: 567–
572.
Rosenbusch, R.F. and F.C. Minion. 1992. Cell envelope: morphology
and biochemistry. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch and Baseman). American
Røsendal, S. 1974. Mycoplasma molare, a new canine Mycoplasma species.
Røsendal, S. 1975. Canine mycoplasmas: serological studies of type and
reference strains, with a proposal for the new species, Mycoplasma
opalescens. Acta Pathol. Microbiol. Scand. Sect. B 83: 463–470.
Røsendal, S. and O. Vinther. 1977. Experimental mycoplasmal pneumonia in dogs: electron microscopy of infected tissue. Acta Pathol.
Microbiol. Scand. (B) 85B: 462–465.
Røsendal, S. 1979. Canine and feline mycoplasmas. In The Mycoplasmas (edited by Tully and Whitcomb). Academic Press, New York,
pp. 217–234.
Rosengarten, R., S. Levisohn and D. Yogev. 1995. A 41-kDa variable surface protein of Mycoplasma gallisepticum has a counterpart in Mycoplasma imitans and Mycoplasma iowae. FEMS Microbiology Letters 132:
115–123.
Röske, K., M. Calcutt and K. Wise. 2004. The Mycoplasma fermentans
prophage phiMFV1: genome organization, mobility and variable
expression of an encoded surface protein. Mol. Microbiol. 52: 1703–
1720.
Ross, R.F. and J.A. Karmon. 1970. Heterogeneity among strains of Mycoplasma granularum and identification of Mycoplasma hyosynoviae, sp. n.
J. Bacteriol. 103: 707–713.
Ross, R.F. 1992. Mycoplasmal diseases. In Diseases of Swine, 7th edn
(edited by Lemanske, Jr, Straw, Mengeling, D’Allaire and Taylor).
Iowa State University Press, Ames, IA, pp. 537–551.
Ruhnke, H., N. Palmer, P. Doig and R. Miller. 1984. Bovine abortion
and neonatal death associated with Ureaplasma diversum. Theriogenology 21: 295–301.
Ruhnke, H.L. and S. Madoff. 1992. Mycoplasma phocidae sp. nov., isolated from harbor seals (Phoca vitulina L.). Int. J. Syst. Bacteriol. 42:
211–214.
Ruifu, Y., Z. Minli, Z. Guo and X. Wang. 1997. Biovar diversity is
reflected by variations of genes encoding urease of Ureaplasma urealyticum. Microbiol. Immunol. 41: 625–627.
Ruiter, M. and H.M. Wentholt. 1955. Isolation of a pleuropneumonialike organism from a skin lesion associated with a fusospirochetal
flora. J. Invest. Dermatol. 24: 31–34.
636
Sabin, A.B. 1941. The filterable microorganisms of the pleuropneumonia group. Bacteriol. Rev. 5: 1–66.
Salih, M.M., N.F. Friis, S.N. Aarseculeratne, E.A. Freundt and C. Christiansen. 1983. Mycoplasma mustelae, a new species from mink. Int. Syst.
Bacteriol. 33: 476–479.
Sasaki, Y., J. Ishikawa, A. Yamashita, K. Oshima, T. Kenri, K. Furuya, C.
Yoshino, A. Horino, T. Shiba, T. Sasaki and M. Hattori. 2002. The
complete genomic sequence of Mycoplasma penetrans, an intracellular bacterial pathogen in humans. Nucleic Acids Res. 30: 5293–
5300.
Sayed, I. and G.E. Kenny. 1980. Comparison of the proteins and polypeptides of the eight serotypes of Ureaplasma urealyticum by isoelectric
focussing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Int. J. Syst. Bacteriol. 30: 33–41.
Scanziani, E., S. Paltrinieri, M. Boldini, V. Grieco, C. Monaci, A.M.
Giusti and G. Mandelli. 1997. Histological and immunohistochemical findings in thoracic lymph nodes of cattle with contagious bovine
pleuropneumonia. J. Comp. Pathol. 117: 127–136.
Schelonka, R., B. Katz, K. Waites and D.J. Benjamin. 2005. Critical
appraisal of the role of Ureaplasma in the development of bronchopulmonary dysplasia with metaanalytic techniques. Pediatr. Infect. Dis.
J. 24: 1033–1039.
Schilling, V. 1928. Eperythrozoon coccoides, eine neue durch Splenektomie
aktivierbare Dauerinfektion der weissen Maus. Klin. Wochenschr. 7:
1854–1855.
Schmidhauser, C., R. Dudler, T. Schmidt and R.W. Parish. 1990. A mycoplasmal protein influences tumour cell invasiveness and contact inhibition in vitro. J. Cell Sci. 95: 499–506.
Schmitt, K., W. Daubener, D. Bitter-Suermann and U. Hadding. 1988.
A safe and efficient method for elimination of cell culture mycoplasmas using ciprofloxacin. J. Immunol. Methods 109: 17–25.
Schoeb, T.R. 2000. Respiratory diseases of rodents. Vet. Clin. North Am.
Exot. Anim. Pract. 3: 481–496, vii.
Scudamore, J.M. 1976. Observations on the epidemiology of contagious
bovine pleuropneumonia: Mycoplasma mycoides in urine. Res. Vet. Sci.
20: 330–333.
Seto, S., G. Layh-Schmitt, T. Kenri and M. Miyata. 2001. Visualization of
the attachment organelle and cytadherence proteins of Mycoplasma
pneumoniae by immunofluorescence microscopy. J. Bacteriol. 183:
1621–1630.
Seybert, A., R. Herrmann and A.S. Frangakis. 2006. Structural analysis of Mycoplasma pneumoniae by cryo-electron tomography. J. Struct.
Biol. 156: 342–354.
Shahram, M., R.A. Nicholas, A.P. Wood and D.P. Kelly. 2010. Further
evidence to justify reassignment of Mycoplasma mycoides subspecies
mycoides Large Colony type to Mycoplasma mycoides subspecies capri.
Syst. Appl. Microbiol. 33: 20–24.
Shepard, M. 1954. The recovery of pleuropneumonia-like organisms
from Negro men with and without nongonococcal urethritis. Am. J.
Syph. Gonorrhea Vener. Dis. 38: 113–124.
Shepard, M. 1967. Cultivation and properties of T-strains of Mycoplasma
associated with nongonococcal urethritis. Ann. N. Y. Acad. Sci. 143:
505–514.
Shepard, M. and C. Lunceford. 1967. Occurrence of urease in T strains
of Mycoplasma. J. Bacteriol. 93: 1513–1520.
Shepard, M. and D. Howard. 1970. Identification of “T” mycoplasmas
in primary agar cultures by means of a direct test for urease. Ann. N.
Y. Acad. Sci. 174: 809–819.
Shepard, M.C., C.D. Lunceford, D.K. Ford, R.H. Purcell, D. TaylorRobinson, S. Razin and F.T. Black. 1974. Ureaplasma urealyticum gen.
nov., sp. nov.: proposed nomenclature for Human-T (T-strain) mycoplasmas. Int. J. Syst. Bacteriol. 24: 160–171.
Shepard, M.C. and G.K. Masover. 1979. Special features of ureaplasmas.
In The Mycoplamas, vol. 1 (edited by Barile and Razin). Academic
Shepard, M.C. 1983. Culture media for ureaplasmas. In Methods in
Shepard, M.C. and J.A. Robertson. 1986. Calcium chloride as an indicator for colonies of Ureaplasma urealyticum. The Pediatric Infectious
Diseases Journal 5: S349.
Shiferaw, G., S. Tariku, G. Ayelet and Z. Abebe. 2006. Contagious
caprine pleuropneumonia and Mannheimia haemolytica-associated
acute respiratory disease of goats and sheep in Afar Region, Ethiopia.
Rev. Sci. Tech. 25: 1153–1163.
Shimizu, T., H. Erno and H. Nagatomo. 1978. Isolation and characterization of Mycoplasma columbinum and Mycoplasma columborale, two new species from pigeons. Int. J. Syst. Bacteriol. 28:
538–546.
Shimizu, T., K. Numano and K. Uchida. 1979. Isolation and identification of mycoplasmas from various birds: an ecological study. Jpn. J.
Vet. Sci. 41: 273–282.
Shobokshi, A. and M. Shaarawy. 2002. Maternal serum and amniotic
fluid cytokines in patients with preterm premature rupture of membranes with and without intrauterine infection. Int. J. Gynaecol.
Obstet. 79: 209–215.
Simecka, J.W., J.K. Davis, M.K. Davidson, S.R. Ross, C.T.K.-H. Städtlander
and G.H. Cassell. 1992. Mycoplasma diseases of animals. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch, and Baseman). American Society for Microbiology,
Washington, D.C., pp. 391–415.
Singh, K.C. and P.K. Uppal. 1987. Isolation of Mycoplasma gallinarum
from sheep. Vet. Rec. 120: 464.
Singh, Y., D.N. Garg, P.K. Kapoor and S.K. Mahajan. 2004. Isolation of
Mycoplasma bovoculi from genitally diseased bovines and its experimental pathogenicity in pregnant guinea pigs. Indian J. Exp. Biol.
42: 933–936.
Slatter, D.H. 2001. Fundamentals of Veterinary Ophthamology, 3rd
edn. Elsevier Health Sciences, Saint Louis, MO.
Smith, A. and J. Mowles. 1996. Prevention and control of Mycoplasma
infection of cell cultures. In Molecular and Diagnostic Procedures in
San Diego, pp. 445–451.
Smith, D., W. Russell, W. Ingledew and D. Thirkell. 1993. Hydrolysis of
urea by Ureaplasma urealyticum generates a transmembrane potential
with resultant ATP synthesis. J. Bacteriol. 175: 3253–3258.
Smith, P. 1986. Mass cultivation of ureaplasmas and some applications.
Pediatr. Infect. Dis. 5: S313–315.
Smith, P.F. 1992. Membrane lipid and lipopolysaccharide structures.
In Mycoplasmas: Molecular Biology and Pathogenesis (edited by
Maniloff, McElhaney, Finch and Baseman). American Society for
So, A.K., P.M. Furr, D. Taylor-Robinson and A.D. Webster. 1983. Arthritis caused by Mycoplasma salivarium in hypogammaglobulinaemia. Br.
Med. J. (Clin. Res. Ed.) 286: 762–763.
Somerson, N.L., D. Taylor-Robinson and R.M. Chanock. 1963.
Hemolyin production as an aid in the identification and quantitation of Eaton agent (Mycoplasma pneumoniae). Am. J. Hyg. 77:
122–128.
Somerson, N.L. and B.C. Cole. 1979. The Mycoplasma flora of human
and non-human primates. In The Mycoplasmas, vol. 1 (edited by
Tully and Whitcomb). Academic Press, New York, pp. 191–216.
Somerson, N.L., J.P. Kocka, D. Rose and R.A. Del Giudice. 1982.
Isolation of acholeplasmas and a Mycoplasma from vegetables. Appl.
Environ. Microbiol. 43: 412–417.
Spergser, J., C. Aurich, J.E. Aurich and R. Rosengarten. 2002. High
prevalence of mycoplasmas in the genital tract of asymptomatic stallions in Austria. Vet. Microbiol. 87: 119–129.
Spergser, J., S. Langer, S. Muck, K. Macher, M. Szostak, R. Rosengarten
and H.-J. Busse. 2010. Mycoplasma mucosicanis sp. nov., isolated from
the mucosa of dogs. Int. J. Syst. Evol. Microbiol.: published 23 April
2010 as doi:10.1099/ijs.0.015750-0.
Splitter, E.J. 1950. Eperythrozoon suis, the etiologic agent of icteroanemia–an anaplasmosis-like disease in swine. Am. J. Vet. Res. 11: 324–
329.
Spooner, R., W. Russell and D. Thirkell. 1992. Characterization of the
immunoglobulin A protease of Ureaplasma urealyticum. Infect. Immun.
60: 2544–2546.
Ståby, M. 2004. Seal finger-a problem among hunters once again.
Lakartidningen 101: 1910–1911.
Stemke, G. and J. Robertson. 1981. Modified colony indirect epifluorescence test for serotyping Ureaplasma urealyticum and an adaptation to detect common antigenic specificity. J. Clin. Microbiol. 14:
582–584.
Stemke, G. and J. Robertson. 1982. Comparison of two methods for
enumeration of mycoplasmas. J. Clin. Microbiol. 16: 959–961.
Stemke, G.W. and J.A. Robertson. 1985. Problems associated with serotyping strains of Ureaplasma urealyticum. Diagn. Microbiol. Infect. Dis.
3: 311–320.
Stemke, G., M. Stemler and J. Robertson. 1984. Growth characteristics
of ureaplasmas from animal and human sources. Isr. J. Med. Sci. 20:
935–937.
Stemler, M.E., G.W. Stemke and J.A. Robertson. 1987. ATP measurements obtained by luminometry provide rapid estimation of
Ureaplasma urealyticum growth. J. Clin. Microbiol. 25: 427–429.
Stenske, K.A., D.A. Bemis, K. Hill and D.J. Krahwinkel. 2005. Acute
polyarthritis and septicemia from Mycoplasma edwardii after surgical
removal of bilateral adrenal tumors in a dog. J. Vet. Intern. Med. 19:
768–771.
Stipkovits, L., Z. Varga, M. Dobos-Kovacs and M. Santha. 1984a. Biochemical and serological examination of some Mycoplasma strains of
goose origin. Acta Vet. Hung. 32: 117–125.
Stipkovits, L., Z. Varga, G. Laber and J. Bockmann. 1984b. A comparison of the effect of tiamulin hydrogen fumarate and tylosin tartrate
on mycoplasmas of ruminants and some animal ureaplasmas. Vet.
Stoffregen, W.C., D.P. Alt, M.V. Palmer, S.C. Olsen, W.R. Waters and J.A.
Stasko. 2006. Identification of a Haemomycoplasma species in anemic
reindeer (Rangifer tarandus). J. Wildl. Dis. 42: 249–258.
Swensen, C., J. VanHamont and B.S. Dunbar. 1983. Specific protein differences among strains of Ureaplasma urealyticum as determined by
two-dimensional gel electrophoresis and a sensitive silver stain. Int. J.
Switzer, W.P. 1955. Studies on infectious atrophic rhinitis. IV. Characterization of a pleuropneumonia-like organism isolated from the nasal
cavities of swine. Am. J. Vet. Res. 16: 540–554.
Sykes, J.E., L.M. Ball, N.L. Bailiff and M.M. Fry. 2005. ‘Candidatus Mycoplasma haematoparvum’, a novel small haemotropic mycoplasma
from a dog. Int. J. Syst. Evol. Microbiol. 55: 27–30.
Sykes, J.E., N.L. Drazenovich, L.M. Ball and C.M. Leutenegger. 2007.
Use of conventional and real-time polymerase chain reaction to
determine the epidemiology of hemoplasma infections in anemic
and nonanemic cats. J. Vet. Intern. Med. 21: 685–693.
Tagawa, M., K. Matsumoto and H. Inokuma. 2008. Molecular detection
of Mycoplasma wenyonii and ‘Candidatus Mycoplasma haemobos’ in
cattle in Hokkaido, Japan. Vet. Microbiol. 132: 177–180.
Tasker, S., S.H. Binns, M.J. Day, T.J. Gruffydd-Jones, D.A. Harbour, C.R.
Helps, W.A. Jensen, C.S. Olver and M.R. Lappin. 2003. Use of a PCR
assay to assess the prevalence and risk factors for Mycoplasma haemofelis and ‘Candidatus Mycoplasma haemominutum’ in cats in the United
Kingdom. Vet. Rec. 152: 193–198.
Tasker, S., S.M. Caney, M.J. Day, R.S. Dean, C.R. Helps, T.G.
Knowles, P.J. Lait, M.D. Pinches and T.J. Gruffydd-Jones. 2006.
Effect of chronic FIV infection, and efficacy of marbofloxacin
637
treatment, on Mycoplasma haemofelis infection. Vet. Microbiol.
117: 169–179.
Taylor-Robinson, D., J. Canchola, H. Fox and R.M. Chanock. 1964. A
newly identified oral Mycoplasma (M. orale) and its relationship to
other human mycoplasmas. Am. J. Hyg. 80: 135–148.
Taylor-Robinson, D. and Z. Dinter. 1968. Unexpected serotypes of
mycoplasmas isolated from pigs. J. Gen. Microbiol. 53: 221–229.
Taylor-Robinson, D., G.W. Csonka and M.J. Prentice. 1977.
Human intra-urethral inoculation of ureplasmas. Q. J. Med. 46:
309–326.
Taylor-Robinson, D. and W.M. McCormack. 1979. Mycoplasmas in
human genitourinary infections. In The Mycoplasmas, Volume 2,
Human and Animal Mycoplasmas (edited by Tully and Whitcomb).
Taylor-Robinson, D. 1983a. Recovery of mycoplasmas from the genitourinary tract. In Methods in Mycoplasmology, vol. 2 (edited by Razin
and Tully). Academic Press, New York, pp. 19–26.
Taylor-Robinson, D. 1983b. Metabolism inhibition tests. In Methods in
Taylor-Robinson, D. and R.N. Gourlay. 1984. Genus II. Ureaplasma
Shepard, Lunceford, Ford, Purcell, Taylor-Robinson, Razin and
Black 1974. In Bergey’s Manual of Systematic Bacteriology, 8th edn,
vol. 1 (edited by Kreig and Holt). Williams & Wilkins, Baltimore, pp.
770–775.
Taylor-Robinson, D. 1985. Mycoplasmal and mixed infections of the
human male urogenital tract and their possible complications. In
The Mycoplasmas, vol. 4 (edited by Razin and Barile). Academic
Taylor-Robinson, D. and P. Furr. 1986. Clinical antibiotic resistance of
Ureaplasma urealyticum. Pediatr. Infect. Dis. 5: S335–337.
Taylor-Robinson, D. and P.M. Furr. 1997. Genital Mycoplasma infections.
Wien. Klin. Wochenschr. 109: 578–583.
Taylor-Robinson, D. and P. Horner. 2001. The role of Mycoplasma genitalium in non-gonococcal urethritis. Sex. Transm. Infect. 77: 229–
231.
Taylor-Robinson, D., C. Gilroy, B. Thomas and P. Hay. 2004. Mycoplasma
genitalium in chronic non-gonococcal urethritis. Int. J. STD AIDS 15:
21–25.
Taylor, R.R., H. Varsani and R.J. Miles. 1994. Alternatives to arginine
as energy sources for the non-fermentative Mycoplasma gallinarum.
FEMS Microbiol. Lett. 115: 163–167.
Teng, L., X. Zheng, J. Glass, H. Watson, J. Tsai and G. Cassell. 1994.
Ureaplasma urealyticum biovar specificity and diversity are encoded
in multiple-banded antigen gene. J. Clin. Microbiol. 32: 1464–
1469.
ter Laak, E.A., J.H. Noordergraaf and M.H. Verschure. 1993. Susceptibilities of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma
diversum strains to antimicrobial agents in vitro. Antimicrob. Agents
Chemother. 37: 317–321.
ter Laak EA, Noordergraaf JH and Boomsluiter E. 1992a. The nasal
mycoplasmal flora of healthy calves and cows. Zentralbl Veterinarmed B 39: 610–616.
ter Laak EA, Noordergraaf JH and Dieltjes RP. 1992b. Prevalence of
mycoplasmas in the respiratory tracts of pneumonic calves. Zentralbl
Veterinarmed B 39: 553–562.
Thiaucourt, F., G. Bolske, B. Leneguersh, D. Smith and H. Wesonga.
1996. Diagnosis and control of contagious caprine pleuropneumonia. Rev. Sci. Tech. 15: 1415–1429.
Thirkell, D., A. Myles and W. Russell. 1989. Serotype 8- and seroclusterspecific surface-expressed antigens of Ureaplasma urealyticum. Infect.
Immun. 57: 1697–1701.
638
Thirkell, D., A. Myles and D. Taylor-Robinson. 1990. A comparison of
four major antigens in five human and several animal strains of urea
plasmas. J. Med. Microbiol. 32: 163–168.
Thomsen, A.C. 1974. The isolation of Mycoplasma primatum during an
autopsy study of the Mycoplasma flora of the human urinary tract. Acta
Pathol. Microbiol. Scand. B Microbiol. Immunol. 82B: 653–656.
Thurston, J.P. 1953. The chemotherapy of Eperythrozoon coccoides (Schilling 1928). Parasitology 43: 170–174.
Tiong, S.K. 1990. Mycoplasmas and acholeplasmas isolated from
ducks and their possible association with pasteurellas. Vet. Rec. 127:
64–66.
Totten, P.A., M.A. Schwartz, K.E. Sjostrom, G.E. Kenny, H.H. Handsfield, J.B. Weiss and W.L. Whittington. 2001. Association of Mycoplasma genitalium with nongonococcal urethritis in heterosexual
men. J. Infect. Dis. 183: 269–276.
Trüper, H.G. and L. de’Clari. 1998. Taxonomic note: erratum and correction of further specific epithets formed as substantives (nouns)
“in apposition”. Int. J. Syst. Bacteriol. 48: 615.
Tryon, V.V. and J.B. Baseman. 1992. Pathogenic determinants and
mechanisms. In Mycoplasmas: Molecular Biology and Pathogenesis
Tu, A.H., L.L. Voelker, X. Shen and K. Dybvig. 2001. Complete nucleotide sequence of the Mycoplasma virus P1 genome. Plasmid 45:
122–126.
Tully, J.G. and I. Ruchman. 1964. Recovery, identification, and neurotoxicity of Sabin’s Type A and C mouse Mycoplasma (PPLO)
from lyophilized cultures. Proc. Soc. Exp. Biol. Med. 115: 554–
558.
Tully, J.G., M.F. Barile, R.A. Del Giudice, T.R. Carski, D. Armstrong
and S. Razin. 1970. Proposal for classifying strain PG-24 and related
canine mycoplasmas as Mycoplasma edwardii sp. n. J. Bacteriol. 101:
346–349.
Tully, J.G., M.F. Barile, D.G.F. Edward, T.S. Theodore and H. Erno.
1974. Characterization of some caprine mycoplasmas, with proposals
for new species, Mycoplasma capricolum and Mycoplasma putrefaciens. J.
Gen. Microbiol. 85: 102–120.
Tully, J.G. and R.F. Whitcomb. 1979. The Mycoplasmas Volume II:
Human and Animal Mycoplasmas. Academic Press, New York.
Tully, J.G. 1983. Cloning and filtration techniques for mycoplasmas. In
Methods in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, New York, pp. 173–177.
Tully, J.G. and S.E. Razin. 1983. Methods in Mycoplasmology, vol. 2.
Academic Press, New York.
Tully, J.G., D. Taylor-Robinson, D.L. Rose, R.M. Cole and J.M. Bové.
1983. Mycoplasma genitalium, a new species from the human urogenital tract. Int. J. Syst. Bacteriol. 33: 387–396.
Tully, J.G. 1993. Current status of the mollicute flora of humans. Clin.
Infect. Dis. 17 Suppl 1: S2–S9.
Tully, J.G., J.M. Bove, F. Laigret and R.F. Whitcomb. 1993. Revised taxonomy of the class Mollicutes - proposed elevation of a monophyletic
cluster of arthropod-associated Mollicutes to ordinal rank (Entomoplasmatales ord. nov.), with provision for familial rank to separate species
with nonhelical morphology (Entomoplasmataceae fam. nov.) from
helical species (Spiroplasmataceae), and emended descriptions of the
order Mycoplasmatales, family Mycoplasmataceae. Int. J. Syst. Bacteriol.
43: 378–385.
Tully, J.G. 1995a. International Committee on Systematic Bacteriology
Subcommittee on the Taxonomy of Mollicutes. Revised minimal standards for description of new species of the class Mollicutes. Int. J. Syst.
Bacteriol. 45: 605–612.
Tully, J.G. 1995b. Culture medium formulation for primary isolation
and maintenance of mollicutes. In Molecular and Diagnostic Pro-
cedures in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, San Diego, pp. 33–39.
Tully, J.G. and S. Razin (editors). 1996. Molecular and Diagnostic
Procedures in Mycoplasmology, vol. 2. Academic Press, San Diego,
CA.
Tyzzer, E.E. and D. Weinman. 1939. Haemobartonella n.g. (Bartonella olim
pro parte) H. microti n. sp. of the field vole, Microtus pennsylvanicus.
Am. J. Hyg. 30: 141–157.
Uilenberg, G., F. Thiaucourt and F. Jongejan. 2004. On molecular taxonomy: what is in a name? Exp. Appl. Acarol. 32: 301–312.
Uilenberg, G., F. Thiaucourt and F. Jongejan. 2006. Mycoplasma and
Eperythrozoon (Mycoplasmataceae). Comments on a recent paper. Int.
Vancini, R.G. and M. Benchimol. 2008. Entry and intracellular location
of Mycoplasma hominis in Trichomonas vaginalis. Arch. Microbiol. 189:
7–18.
Vasconcellos Cardosa, M., A. Blanchard, S. Ferris, R. Verlengia, J.
Timenetsky and R.A. Florio Da Cunha. 2000. Detection of Ureaplasma
diversum in cattle using a newly developed PCR-based detection assay.
Veterinary Microbiology 72: 241–250.
Vilei, E.M., J. Nicolet and J. Frey. 1999. IS1634, a novel insertion element creating long, variable-length direct repeats which is specific
for Mycoplasma mycoides subsp. mycoides small-colony type. J. Bacteriol.
181: 1319–1323.
Voelker, L.L., K.E. Weaver, L.J. Ehle and L.R. Washburn. 1995. Association of lysogenic bacteriophage MAV1 with virulence of Mycoplasma
arthritidis. Infect. Immun. 63: 4016–4023.
Voelker, L.L. and K. Dybvig. 1999. Sequence analysis of the Mycoplasma
arthritidis bacteriophage MAV1 genome identifies the putative virulence factor. Gene 233: 101–107.
Vogelzang, A. and G. Compeer-Dekker. 1969. Elimination of Mycoplasma from various cell cultures. Antonie Van Leeuwenhoek 35:
393–408.
Waites, K., D. Crouse and G. Cassell. 1992. Antibiotic susceptibilities
and therapeutic options for Ureaplasma urealyticum infections in neonates. Pediatr. Infect. Dis. J. 11: 23–29.
Waites, K., C.M. Bébéar, J.A. Robertson, D.F. Talkington and G.E. Kenny.
2001. Laboratory Diagnosis of Mycoplasmal Infections. Cumitech 34.
ASM Press, Washington, D.C.
Waites, K. and D. Talkington. 2005. New developments in human
disease due to mycoplasmas. In Mycoplasmas: Molecular Biology, Pathogenicity, and Strategies for Control (edited by
Blanchard and Browning). Horizon Bioscience, Norfolk, UK,
pp. 289–354.
Waites, K.B., P.T. Rudd, D.T. Crouse, K.C. Canupp, K.G. Nelson, C.
Ramsey and G.H. Cassell. 1988. Chronic Ureaplasma urealyticum and
Mycoplasma hominis infections of central nervous system in preterm
infants. Lancet 1: 17–21.
Waites, K.B., T.A. Figarola, T. Schmid, D.M. Crabb, L.B. Duffy and J.W.
Simecka. 1991. Comparison of agar versus broth dilution techniques
for determining antibiotic susceptibilities of Ureaplasma urealyticum.
Diagn. Microbiol. Infect. Dis. 14: 265–271.
Washburn, L.R., L.L. Voelker, L.J. Ehle, S. Hirsch, C. Dutenhofer,
K. Olson and B. Beck. 1995. Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis. J. Clin. Microbiol. 33:
2271–2279.
Watson, H.L., D.K. Blalock and G.H. Cassell. 1990. Variable antigens
of Ureaplasma urealyticum containing both serovar-specific and
serovar-cross-reactive epitopes. Infect. Immun. 58: 3679–3688.
Webster, A.D., D. Taylor-Robinson, P.M. Furr and G.L. Asherson. 1978.
Mycoplasmal (Ureaplasma) septic arthritis in hypogammaglobulinaemia. Br. Med. J. 1: 478–479.
Family II. Incertae sedis
Webster, D., H. Windsor, C. Ling, D. Windsor and D. Pitcher. 2003. Chronic
bronchitis in immunocompromised patients: association with a novel
Mycoplasma species. Eur. J. Clin. Microbiol. Infect. Dis. 22: 530–534.
Weisburg, W., J. Tully, D. Rose, J. Petzel, H. Oyaizu, D. Yang, L.
Mandelco, J. Sechrest, T. Lawrence and J. Van Etten. 1989. A phylogenetic analysis of the mycoplasmas: basis for their classification. J.
Bacteriol. 171: 6455–6467.
Welchman, D.B., J. Bradbury, D. Cavanagh and N. Aebischer. 2002.
Infectious agents associated with respiratory disease in pheasants.
Vet. Rec. 150: 658–664.
Wellehan, J.F., M. Calsamiglia, D.H. Ley, M.S. Zens, A. Amonsin and V.
Kapur. 2001. Mycoplasmosis in captive crows and robins from Minnesota. J. Wildl. Dis. 37: 547–555.
Westberg, J., A. Persson, B. Pettersson, M. Uhlen and K.E. Johansson.
2002. ISMmy1, a novel insertion sequence of Mycoplasma mycoides subsp.
mycoides small colony type. FEMS Microbiol. Lett. 208: 207–213.
Whitescarver, J. and G. Furness. 1975. T-mycoplasmas: a study of the
morphology, ultrastructure and mode of division of some human
strains. J. Med. Microbiol. 8: 349–355.
Whittlestone, P. 1979. Porcine mycoplasmas. In The Mycoplasmas,
vol. 2 (edited by Tully and Whitcomb). Academic Press, New York,
pp. 133–176.
Wieslander, A., M.J. Boyer and H. Wróblewski. 1992. Membrane protein structure. In Mycoplasmas: Molecular Biology and Pathogenesis
Willi, B., F.S. Boretti, C. Baumgartner, S. Tasker, B. Wenger, V. Cattori,
M.L. Meli, C.E. Reusch, H. Lutz and R. Hofmann-Lehmann. 2006a.
Prevalence, risk factor analysis, and follow-up of infections caused by
three feline hemoplasma species in cats in Switzerland. J. Clin. Microbiol. 44: 961–969.
Willi, B., S. Tasker, F. S. Boretti, M. G. Doherr, V. Cattori, M. L. Meli, R.
G. Lobetti, R. Malik, C. E. Reusch, H. Lutz, R. Hofmann-Lehmann.
2006b. Phylogenetic analysis of “Candidatus Mycoplasma turicensis”
isolates from pet cats in the United Kingdom, Australia, and South
Africa, with analysis of risk factors for infection. J. Clin. Microbiol.
44: 4430–4435.
Windsor, R.S. and W.N. Masiga. 1977. Investigations into the role of carrier animals in the spread of contagious bovine pleuropneumonia.
Res. Vet. Sci. 23: 224–229.
Woods, J.E., M.M. Brewer, J.R. Hawley, N. Wisnewski and M.R. Lappin.
2005. Evaluation of experimental transmission of Candidatus Mycoplasma haemominutum and Mycoplasma haemofelis by Ctenocephalides
felis to cats. Am. J. Vet. Res. 66: 1008–1012.
639
Woodson, B.A., K.S. McCarty and M.C. Shepard. 1965. Arginine metabolism in Mycoplasma and infected L929 fibroblasts. Arch. Biochem.
109: 364–371.
Woubit, S., S. Lorenzon, A. Peyraud, L. Manso-Silván and F. Thiaucourt. 2004. A specific PCR for the identification of Mycoplasma
capricolum subsp. capripneumoniae, the causative agent of contagious caprine pleuropneumonia (CCPP). Vet. Microbiol. 104:
125–132.
Wroblewski, W. 1931. Morphologie et cycle évolutif des microbnes de la
péripneumonie das bovides et de l’agalaxie contagieuse des chêvres
et des moutons. Ann. Inst. Pasteur (Paris) 47: 94–115.
Xie, X. and J. Zhang. 2006. Trends in the rates of resistance of Ureaplasma urealyticum to antibiotics and identification of the mutation
site in the quinolone resistance-determining region in Chinese
patients. FEMS Microbiol. Lett. 259: 181–186.
Yagihashi, T., T. Nunoya and Y. Otaki. 1983. Effects of dual infection
of chickens with Mycoplasma synoviae and Mycoplasma gallinaceum or
infectious bursal disease virus on infectious synovitis. Nippon Juigaku
Zasshi 45: 529–532.
Yamamoto, R., C.H. Bigland and H.B. Ortmayer. 1965. Characteristics
of Mycoplasma meleagridis sp. n. isolated from turkeys. J. Bacteriol. 90:
47–49.
Yanez, A., L. Cedillo, O. Neyrolles, E. Alonso, M.C. Prevost, J. Rojas,
H.L. Watson, A. Blanchard and G.H. Cassell. 1999. Mycoplasma penetrans bacteremia and primary antiphospholipid syndrome. Emerg.
Infect. Dis. 5: 164–167.
Yechouron, A., J. Lefebvre, H.G. Robson, D.L. Rose and J.G. Tully. 1992.
Fatal septicemia due to Mycoplasma arginini: a new human zoonosis.
Clin. Infect. Dis. 15: 434–438.
Yoder, H.W. and M.S. Hofstad. 1964. Characterization of avian mycoplasmas. Avian Dis. 8: 481–512.
Yogev, D., S. Levisohn and S. Razin. 1989. Genetic and antigenic relatedness between Mycoplasma gallisepticum and Mycoplasma synoviae.
Zheng, X., L. Teng, H. Watson, J. Glass, A. Blanchard and G. Cassell.
1995. Small repeating units within the Ureaplasma urealyticum MB
antigen gene encode serovar specificity and are associated with antigen size variation. Infect. Immun. 63: 891–898.
Zheng, X., D.A. Olson, J.G. Tully, H.L. Watson, G.H. Cassell, D.R.
Gustafson, K.A. Svien and T.F. Smith. 1997. Isolation of Myco
plasma hominis from a brain abscess. J. Clin. Microbiol. 35:
992–994.
Daniel R. Brown, Séverine Tasker, Joanne B. Messick and Harold Neimark
This family accommodates the genera Eperythrozoon and Haemobartonella. These wall-less hemotropic bacteria were once placed
in the family Anaplasmataceae, order Rickettsiales, because they are
obligate blood parasites. None have been cultivated on artificial
media, so no type strains have been established. Motility and biochemical parameters have not been definitively established for
any species. These organisms are now known to be unambiguously affiliated with the order Mycoplasmatales on the basis of 16S
rRNA similarities, plus morphology, DNA G+C contents, and evidence that they use the codon UGA to encode tryptophan (Berent and Messick, 2003), but their nomenclature remains a matter
of controversy (Neimark et al., 2005; Uilenberg et al., 2006).
640
Genus I. Eperythrozoon Schilling 1928, 293AL
E.pe.ry.thro.zo¢on. Gr. pref. epi on; Gr. adj. erythros red; Gr. neut. n. zoon living being, animal; N.L. neut. n.
Eperythrozoon (presumably intended to mean) animals on red (blood cells).
Cells adherent to host erythrocyte surfaces are coccoid and about
350 nm in diameter, but may arrange to appear as chains or
deform to appear rod- or ring-shaped in stained blood smears.
Type species: Eperythrozoon coccoides Schilling 1928, 1854.
Hemotropic mollicutes such as the species formerly called
Eperythrozoon coccoides (trivial name, hemoplasmas; Neimark
et al., 2005) infect a variety of mammals occasionally including humans. Transmission can be through ingestion of infected
blood, percutaneous inoculation, or by arthropod vectors
(Sykes et al., 2007; Willi et al., 2006). The pathogenicity of different hemoplasma species is variable, and strain virulence and
host immunocompetence likely play roles in the development
of disease. Clinical syndromes range from acute fatal hemolytic
anemia to chronic insidious anemia. Signs may include anemia, pyrexia, anorexia, dehydration, weight loss, and infertility. The presence of erythrocyte-bound antibodies has been
demonstrated in some hemoplasma-infected animals and may
contribute to anemia. Animals can remain chronic asymptomatic carriers of hemoplasmas after acute infection. PCR is the
diagnostic test of choice for hemoplasmosis. Tetracycline treatment reduces the number of organisms in peripheral blood,
but probably does not eradicate the organisms from infected
animals.
Hemoplasmas have not yet been successfully grown in continuous culture in vitro, although recent work (Li et al., 2008) suggests that in vitro maintenance of the species Mycoplasma suis
may be possible.
Hemoplasmas can be frozen in heparin- or EDTA-anticoagulated blood cryopreserved with dimethylsulfoxide.
Differentiation of the genus Eperythrozoon
from other genera
A distinctive characteristic of these organisms is that they are
found only in the blood of vertebrate hosts or transiently in
arthropod vectors of transmission. The tenuous distinction
between species of Eperythrozoon and those of Haemobartonella
was based on the relatively more common visualization of
eperythrozoa as ring forms (now known to be artifactual) in
stained blood smears and the perception that eperythrozoa were
observed with about equal frequency on erythrocytes and free in
plasma, while haemobartonellae were thought to occur less
often free in plasma. Properties that partially fulfill criteria for
assignment of this genus to the class Mollicutes (Brown et al.,
2007) include absence of a cell wall, filterability, and the presence
of conserved 16S rRNA gene sequences. Presumptive use of the
codon UGA to encode tryptophan (Berent and Messick, 2003)
supports exclusion from the genera Anaeroplasma, Asteroleplasma,
Acholeplasma, and “Candidatus Phytoplasma”. Non-spiral cellular
morphology and regular association with vertebrate hosts
support exclusion from the genera Spiroplasma, Entomoplasma,
and Mesoplasma, but sterol requirement, the degree of aerobiosis, and the capacity to hydrolyze arginine, characteristics that
would help to confirm their provisional 16S rRNA-based placement in the genus Mycoplasma, remain unknown.
Taxonomic comments
The taxonomy and nomenclature of the uncultivated hemotropic bacteria originally assigned to the genus Eperythrozoon remain
matters of current controversy. It is now undisputed that, on
the basis of their lack of a cell wall, small cell size, low G+C
content, use of the codon UGA to encode tryptophan, regular
association with vertebrate hosts, and 16S rRNA gene sequences
that are most similar (80–84%) to species in the pneumoniae
group of genus Mycoplasma, these organisms are properly affiliated with the Mycoplasmatales. However, the proposed transfers of Eperythrozoon and Haemobartonella species to the genus
Mycoplasma (Neimark et al., 2001, 2005) were opposed on the
grounds that the degree of 16S rRNA gene sequence similarity
is insufficient (Uilenberg et al., 2004, 2006). The alternative of
situating the hemoplasmas in a new genus in the Mycoplasmataceae (Uilenberg et al., 2006) would regrettably compound the
16S rRNA gene-based polyphyly within Mycoplasma on no other
basis than a capacity to adhere to the surface of erythrocytes
in vivo.
The proposed transfer of the type species Eperythrozoon
coccoides to the genus Mycoplasma (Neimark et al., 2005) is
complicated by priority because Eperythrozoon predates Mycoplasma. However, the alternative of uniting the genera by transferring all mycoplasmas to the genus Eperythrozoon is completely
unjustifiable considering the biological characteristics of the
non-hemotropic majority of Mycoplasma species. The Judicial
Commission of the International Committee on Systematics of
Prokaryotes declined to rule on a request for an opinion in this
matter during their 2008 meeting, but a provisional placement
of the former Eperythrozoon species in the genus Mycoplasma has
otherwise been embraced by specialists in the molecular biology
and clinical pathogenicity of these and similar hemotropic
organisms. At present, the designation “Candidatus Mycoplasma”
must still be used for new types.
Further reading
Kreier, J.P. and M. Ristic. 1974. Genus IV. Haemobartonella Tyzzer
and Weinman 1939, 143AL; Genus V. Eperythrozoon Schilling
1928, 1854AL. In Bergey’s Manual of Determinative Bacteriology, 8th edn (edited by Buchanan and Gibbons). Williams &
Wilkins, Baltimore, pp. 910–914.
Differentiation of the species of the genus
Eperythrozoon
Species differentiation relies principally on 16S rRNA gene
sequencing. Some species exhibit a degree of host specificity,
although cross-infection of related hosts has been reported.
Genus I. Eperythrozoon
641
List of species of the genus Eperythrozoon
1. Mycoplasma coccoides (Schilling 1928) Neimark, Peters,
Robinson and Stewart 2005, 1389VP (Eperythrozoon coccoides
Schilling 1928, 1854)
coc.co′ides. N.L. masc. n. coccus (from Gr. masc. n. kokkos
grain, seed) coccus; L. suff. -oides (from Gr. suff. eides from
Gr. n. eidos that which is seen, form, shape, figure), resembling, similar; N.L. neut. adj. coccoides coccus-shaped.
Pathogenic; causes anemia in wild and captive mice, and
captive rats, hamsters, and rabbits. Transmission is believed to
be vector-borne and mediated by the rat louse Polyplex spinulosa and the mouse louse Polyplex serrata. Neoarsphenamine
and oxophenarsine were thought to be effective chemotherapeutic agents for treatment of Mycoplasma coccoides infection
in captive rodents, whereas tetracyclines are effective only at
keeping infection at subclinical levels (Thurston, 1953).
wild and captive rodents.
2. Eperythrozoon parvum Splitter 1950, 513AL
par¢vum. L. neut. adj. parvum small.
A nonpathogenic epierythrocytic parasite of pigs. Organic
arsenicals are effective; tetracyclines suppress infection.
Transmissible by parenteral inoculation and sometimes by
massive oral inoculation. This is the only remaining species
of Eperythrozoon whose name has standing in nomenclature
that has not yet been examined by molecular genetic methods. It seems likely that, if a specimen of this organism can
be found, it will prove to be a mycoplasma.
3. Mycoplasma ovis (Neitz, Alexander and de Toit 1934) Neimark, Hoff and Ganter 2004, 369VP (Eperythrozoon ovis Neitz,
Alexander and de Toit 1934, 267)
o¢vis. L. fem. gen. n. ovis of a sheep.
Cells are coccoid and motility for this species has not
been assessed. The morphology of infected erythrocytes
is altered demonstrating a marked depression at the site
of Myplasma ovis attachment. This species has not been
grown on artificial medium; therefore, notable biochemical parameters are not known.
Neoarsphenamine is an effective therapeutic agent. Mycoplasma ovis is reported to share antigens with Mycoplasma
wenyonii (Kreier and Ristic, 1963), potentially complicating
serology-based diagnosis of infection.
Pathogenic; causes mild to severe anemia in sheep and
goats that often results in poor feed conversion. Transmission occurs via blood-feeding arthropods, e.g., Haemophysalis plumbeum, Rhipicephalus bursa, Aedes camptorhynchus, and
Culex annulirostris (Daddow, 1980; Howard, 1975; Nikol’skii
and Slipchenko, 1969), and likely via fomites such as reused
needles, shearing tools, and ear-tagging equipment (BrunHansen et al., 1997; Mason and Statham, 1991).
Source: observed in association with erythrocytes or unattached in suspension in the blood of sheep, goats, and
rarely in eland and splenectomized deer.
4. Mycoplasma suis corrig. (Splitter 1950) Neimark, Johansson,
Rikihisa and Tully 2002, 683VP (Eperythrozoon suis Splitter
1950, 513)
su¢is. L. gen. n. suis of the pig.
not known.
Neoarsphenamine and tetracyclines are effective therapeutic agents. An enzyme-linked immunosorbant assay
(ELISA) and PCR-based detection assays to enable diagnosis
of infection have been described (Groebel et al., 2009; Gwaltney and Oberst, 1994; Hoelzle, 2008; Hsu et al., 1992).
Pathogenic; causes febrile icteroanemia in pigs. Transmission occurs via insect vectors including Stomoxys calcitrans and Aedes aegypti (Prullage et al., 1993).
Source: observed in association with the erythrocytes of pigs.
Further comment: the original spelling of the specific
epithet, haemosuis (sic), has been corrected by the List
Editor.
5. Mycoplasma wenyonii (Adler and Ellenbogen 1934) Neimark, Johansson, Rikihisa and Tully 2002, 683VP (Eperythrozoon wenyonii Adler and Ellenbogen 1934, 220)
we.ny.o¢ni.i. N.L. masc. gen. n. wenyonii of Wenyon, named
after Charles Morley Wenyon (1878–1948), an investigator
of these organisms.
not known.
Pathogenic; causes anemia and subsequent lameness
and/or infertility in cattle. Transmission is primarily vectormediated by Dermacentor andersoni and reportedly can also
occur vertically during gestation. Oxytetracycline is an effective therapeutic agent (Montes et al., 1994). Mycoplasma
wenyonii is reported to share antigens with Mycoplasma ovis
(Kreier and Ristic, 1963), potentially complicating serologybased diagnosis of infection.
Source: observed in association with the erythrocytes and
platelets of cattle (Kreier and Ristic, 1968).
Species of unknown phylogenetic affiliation
The phylogenetic affiliations of the following proposed organisms are unknown and their names do not have standing in
nomenclature. They are listed here merely because they have
been incidentally cited as species of Eperythrozoon.
642
1. “Eperythrozoon mariboi” Ewers 1971
The name given to uncultivated polymorphic structures
observed on or in erythrocytes from flying foxes (Pteropus
macrotis) following splenectomy (Ewers, 1971). The structures,
described as fine lines, lines with rings, and rows of rings that
span the diameter of the erythrocytes, differ from those of
hemotropic mycoplasmas.
2. “Eperythrozoon teganodes” Hoyte 1962
The name given to uncultivated serially transmissible bodies observed in Giemsa-stained blood smears from cattle.
The bodies only occur free in the blood plasma and do not
attach to erythrocytes (Hoyte, 1962). The bodies differ from
Mycoplasma wenyonii in morphology and include “frying-pan”
shaped structures.
3. “Eperythrozoon tuomii” Tuomi and Von Bonsdorff 1967
Uncultivated transmissible cell wall-less bodies observed in
Giemsa-stained blood smears and electron micrographs of
blood from splenectomized calves. The bodies appeared in
blood smears predominantly as delicate rings that did not
attach to erythrocytes but were associated exclusively with
thrombocytes (Tuomi and Von Bonsdorff, 1967; Uilenberg,
1967; Zwart et al., 1969).
Genus II. Haemobartonella Tyzzer and Weinman 1939, 305AL
Ha.e.mo.bar.to.nel′la. Gr. n. haima (L. transliteration haema) blood; N.L. fem. n. Bartonella a bacterial genus;
N.L. fem. n. Haemobartonella the blood (-inhabiting) Bartonella.
Cells adherent to host erythrocyte surfaces are coccoid and
about 350 nm in diameter, but may occur as chains or deform
to appear rod- or ring-shaped in stained blood smears.
Type species: Haemobartonella muris (Mayer 1921) Tyzzer and
Weinman 1939AL (Bartonella muris Mayer 1921, 151; Bartonella
muris ratti Regendanz and Kikuth 1928, 1578; Haemobartonella
muris Tyzzer and Weinman 1939, 143).
Those organisms originally assigned to the genus Haemobartonella
are properly affiliated with the Mycoplasmatales, but their transfer
to the order has not yet been formalized. Any distinction between
Haemobartonella and Eperythrozoon is tenuous and possibly arbitrary
(Kreier and Ristic, 1974; Uilenberg et al., 2004). Enrichment, isolation and maintenance procedures, and methods of differentiation are essentially the same as those for genus Eperythrozoon.
List of species of the genus Haemobartonella
1. Mycoplasma haemomuris (Mayer 1921) Neimark, Johansson,
Rikihisa and Tully 2002, 683VP (Bartonella muris Mayer 1921,
151; Bartonella muris ratti Regendanz and Kikuth 1928, 1578;
Haemobartonella muris Tyzzer and Weinman 1939, 143)
ha.e.mo.mu¢ris. Gr. neut. n. haema blood; L. masc. gen. n.
muris of the mouse; N.L. gen. n. haemomuris of mouse blood.
Cells are coccoid and some display dense inclusion
articles. Motility for this species has not been assessed.
p
The morphology of infected erythrocytes is altered,
demonstrating a marked depression at the site of Mycoplasma
haemomuris attachment. This species has not been grown
on any artificial medium; therefore, notable biochemical
parameters are not known.
Opportunistic pathogen; causes anemia in splenectomized or otherwise immunosuppressed mice. Transmission is vector-borne and mediated by the rat louse (Polypax
spinulosa).
Source: observed in association with erythrocytes of wild
and captive mice, and hamsters.
2. Mycoplasma haemocanis (Kikuth 1928) Messick, Walker,
Raphael, Berent and Shi 2002, 697VP [Bartonella canis Kikuth
1928, 1730; Haemobartonella (Bartonella) canis (Kikuth 1928)
Tyzzer and Weinman 1939, 151; Kreier and Ristic 1984,
726]
ha.e.mo.ca¢nis Gr. neut. n. haema blood; L. fem. gen. n. canis
of the dog; N.L. gen. n. haemocanis of dog blood.
Cells are coccoid to pleomorphic. Motility for this species
has not been assessed. The morphology of infected erythrocytes is altered, demonstrating a marked depression at the
site of Mycoplasma haemocanis attachment. This species has
not been grown on any artificial medium; therefore, notable biochemical parameters are not known.
Pathogenic; causes hemolytic anemia in domestic dogs.
Transmission is vector-borne and mediated by the brown
dog tick (Rhipicephalus sanguineus).
domestic dogs (Hoskins, 1991).
DNA G+C content (mol%): Not determined.
3. Mycoplasma haemofelis (Clark 1942) Neimark, Johansson,
Rikihisa and Tully 2002, 683VP [Eperythrozoon felis Clark 1942,
16; Haemobartonella felis (Clark 1942) Flint and McKelvie
1956, 240 and Kreier and Ristic 1984, 725]
ha.e.mo.fe¢lis. Gr. neut. n. haema blood; L. fem. gen. n. felis
of the cat; N.L. gen. n. haemofelis of cat blood.
assessed. This species has not been grown on artificial medium;
therefore, notable biochemical parameters are not known.
Pathogenic; causes hemolytic anemia in cats. The mode
of transmission is percutaneous or oral; an insect vector has
not been identified although fleas have been implicated
(Woods et al., 2005).
Tetracyclines and fluoroquinolones are effective therapeutic agents (Dowers et al., 2002; Tasker et al., 2006).
Genus II. Haemobartonella
643
domestic cats.
DNA G+C content (mol%): 38.5 (genome sequence survey
of strain OH; Berent and Messick, 2003).
The phylogenetic affiliations of the following proposed
organism are unknown and its name does not have standing
in nomenclature. It is listed here merely because it has been
incidentally cited as a species of Haemobartonella.
References
Man and Animals (edited by Weinman and Ristic). Academic Press,
Kreier, J.P. and M. Ristic. 1974. Genus IV. Haemobartonella Tyzzer and
Weinman 1939, 143AL; Genus V. Eperythrozoon Schilling 1928, 1854AL.
In Bergey’s Manual of Determinative Bacteriology, 8th edn (edited
by Buchanan and Gibbons). Williams & Wilkins, Baltimore, pp.
910–914.
Kreier, J.P. and M. Ristic. 1984. Genus III. Haemobartonella; Genus IV.
Eperythrozoon. In Bergey’s Manual of Systematic Bacteriology, vol.
1 (edited by Krieg and Holt). Williams & Wilkins, Baltimore, pp.
724–729.
Li, X., X. Jia, D. Shi, Y. Xiao, S. Hu, M. Liu, Z. Yuan and D. Bi. 2008.
Continuous in vitro Cultivation of Mycoplasma suis. Acta Vet. Zootech.
Sinica 38: 1142–1146.
Mason, R.W. and P. Statham. 1991. The determination of the level of
Eperythrozoon ovis parasitaemia in chronically infected sheep and its
significance to the spread of infection. Aust. Vet. J. 68: 115–116.
Mayer, M. 1921. Über einige bakterienähnliche Parasiten der Erythrozyten
bei Menschen und Tieren. Arch. Schiffs Trop. Hyg. 25: 150–152.
Messick, J.B., P.G. Walker, W. Raphael, L. Berent and X. Shi. 2002. ‘Candidatus Mycoplasma haemodidelphidis’ sp. nov., ‘Candidatus Mycoplasma haemolamae’ sp. nov. and Mycoplasma haemocanis comb. nov.,
haemotrophic parasites from a naturally infected opossum (Didelphis
virginiana), alpaca (Lama pacos) and dog (Canis familiaris): phylogenetic and secondary structural relatedness of their 16S rRNA genes
to other mycoplasmas. Int. J. Syst. Evol. Microbiol. 52: 693–698.
Montes, A., D. Wolfe, E. Welles, J. Tyler and E. Tepe. 1994. Infertility
associated with Eperythrozoon wenyonii infection in a bull. J. Am. Vet.
Med. Assoc. 204: 261–263.
Neimark, H., K.E. Johansson, Y. Rikihisa and J.G. Tully. 2001. Proposal to transfer some members of the genera Haemobartonella and
Eperythrozoon to the genus Mycoplasma with descriptions of ‘Candidatus Mycoplasma haemofelis’, ‘Candidatus Mycoplasma haemomuris’,
‘Candidatus Mycoplasma haemosuis’ and ‘Candidatus Mycoplasma
wenyonii’. Int. J. Syst. Evol. Microbiol. 51: 891–899.
Neimark, H., B. Hoff and M. Ganter. 2004. Mycoplasma ovis comb. nov.
(formerly Eperythrozoon ovis), an epierythrocytic agent of haemolytic
anaemia in sheep and goats. Int. J. Syst. Evol. Microbiol. 54: 365–371.
Neimark, H., W. Peters, B.L. Robinson and L.B. Stewart. 2005. Phylogenetic analysis and description of Eperythrozoon coccoides, proposal
to transfer to the genus Mycoplasma as Mycoplasma coccoides comb.
nov. and Request for an Opinion. Int. J. Syst. Evol. Microbiol. 55:
1385–1391.
Neimark, H.C., K.E. Johansson, Y. Rikihisa and J.G. Tully. 2002. Revision of haemotrophic Mycoplasma species names. Int. J. Syst. Evol.
Microbiol. 52: 683.
Neitz, W.O., R.A. Alexander and P.J. de Toit. 1934. Eperythrozoon ovis (sp.
nov.) infection in sheep. Onderstepoort J. Vet. Sci. 3: 263–274.
Nikol’skii, S.N. and S.N. Slipchenko. 1969. Experiments in the transmission of Eperythrozoon ovis by the ticks H. plumbeum and Rh. bursa.
Veterinariia (Russian) 5: 46.
Prullage, J.B., R.E. Williams and S.M. Gaafar. 1993. On the transmissibility of Eperythrozoon suis by Stomoxys calcitrans and Aedes aegypti. Vet.
Parasitol. 50: 125–135.
Adler, S. and V. Ellenbogen. 1934. A note on two new blood parasites of
cattle: Eperythrozoon and Bartonella. J. Comp. Pathol. 47: 220–221.
Berent, L.M. and J.B. Messick. 2003. Physical map and genome sequencing survey of Mycoplasma haemofelis (Haemobartonella felis). Infect.
Immun. 71: 3657–3662.
Brown, D., R. Whitcomb and J. Bradbury. 2007. Revised minimal
standards for description of new species of the class Mollicutes
(division Tenericutes). Int. J. Syst. Evol. Microbiol. 57: 2703–2719.
Brun-Hansen, H., H. Gronstol, H. Waldeland and B. Hoff. 1997. Eperythrozoon ovis infection in a commercial flock of sheep. Zentralbl. Veterinarmed. B 44: 295–299.
Clark, R. 1942. Eperythrozoon felis (sp. nov.) in a cat. J. Afr. Vet. Med.
Assoc. 13: 15–16.
Daddow, K.N. 1980. Culex annulirostris as a vector of Eperythrozoon ovis
infection in sheep. Vet. Parasitol. 7: 313–317.
Dowers, K.L., C. Olver, S.V. Radecki and M.R. Lappin. 2002. Use of
enrofloxacin for treatment of large-form Haemobartonella felis in
experimentally infected cats. J. Am. Vet. Med. Assoc. 221: 250–253.
Ewers, W.H. 1971. Eperythrozoon mariboi sp. nov. (Protophyta: order
Richettsiales) a parasite of red blood cells of the flying fox Pteropus
macrotis epularius in New Guinea. Parasitology 63: 261–269.
Flint, J.C. and McKelvie D.H.. 1956. Feline infectious anemia-diagnosis
and treatment. Proc. 92nd Ann. Meet. Am. Vet. Med. Assoc. 1955
240–242.
Frerichs, W.M. and A.A. Holbrook. 1971. Haemobartonella procyoni sp. n.
in the raccoon, Procyon lotor. J. Parasitol. 57: 1309–1310.
Groebel, K., K. Hoelzle, M.M. Wittenbrink, U. Ziegler and L.E. Hoelzle.
2009. Mycoplasma suis invades porcine erythrocytes. Infect. Immun.
77: 576–584.
Gwaltney, S.M. and R.D. Oberst. 1994. Comparison of an improved
polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection. J. Vet.
Diagn. Invest. 6 : 321–325.
Hoelzle, L. 2008. Haemotrophic mycoplasmas: recent advances in Mycoplasma suis. Vet. Microbiol. 130 : 215–226.
Hoskins, J.D. 1991. Canine haemobartonellosis, canine hepatozoonosis,
and feline cytauxzoonosis. Vet. Clin. North Am. Small Anim. Pract.
21: 129–140.
Howard, G.W. 1975. The experimental transmission of Eperythrozoon ovis
by mosquitoes. Parasitology 71: xxxiii.
Hoyte, H.M.D. 1962. Eperythrozoon teganodes sp. nov. (Rickettsiales), parasitic in cattle. Parasitology 52: 527–532.
Hsu, F.S., M.C. Liu, S.M. Chou, J.F. Zachary and A.R. Smith. 1992. Evaluation of an enzyme-linked immunosorbent assay for detection of
Eperythrozoon suis antibodies in swine. Am. J. Vet. Res. 53: 352–354.
Kikuth, W. 1928. Über Einen neuen Anämeerreger; Bartonella canis nov.
spec. Klin. Wochenschr. 7: 1729–1730.
Kreier, J.P. and M. Ristic. 1963. Morphologic, antigenic, and pathogenic
characteristics of Eperythrozoon ovis and Eperythrozoon wenyoni. Am.
J. Vet. Res. 24: 488–500.
Kreier, J.P. and M. Ristic. 1968. Haemobartonellosis, eperythrozoonosis, grahamellosis and ehrlichiosis. In Infectious Blood Diseases of
1. “Haemobartonella procyoni” Frerichs and Holbrook 1971
Electron microscopy shows this epierythrocytic organism
from a raccoon (Procyon lotor) is wall-less and its description
indicates it probably will prove to be a hemotropic mycoplasma (Frerichs and Holbrook, 1971).
644
Regendanz, P. and W. Kikuth. 1928. Über Aktivierung labiler Infektionen
duch Entmilzung (Piroplasma canis, Nuttalia brasiliensis, Bartonella
opossum, Spirochaeta didelphydis). Arch. f. Schiffs. U. Tropenhyg. 32:
587–593.
Schilling, V. 1928. Eperythrozoon coccoides, eine neue durch Splenektomie
aktivierbare Dauerinfektion der weissen Maus. Klin. Wochenschr. 7:
1854–1855.
Splitter, E.J. 1950. Eperythrozoon suis, the etiologic agent of icteroanemia–an anaplasmosis-like disease in swine. Am. J. Vet. Res. 11:
324–329.
Sykes, J.E., N.L. Drazenovich, L.M. Ball and C.M. Leutenegger. 2007.
Use of conventional and real-time polymerase chain reaction to
determine the epidemiology of hemoplasma infections in anemic
and nonanemic cats. J. Vet. Intern. Med. 21: 685–693.
Tasker, S., S.M. Caney, M.J. Day, R.S. Dean, C.R. Helps, T.G.
Knowles, P.J. Lait, M.D. Pinches and T.J. Gruffydd-Jones. 2006.
Effect of chronic FIV infection, and efficacy of marbofloxacin
treatment, on Mycoplasma haemofelis infection. Vet. Microbiol.
117: 169–179.
Thurston, J.P. 1953. The chemotherapy of Eperythrozoon coccoides
(Schilling 1928). Parasitology 43: 170–174.
Tuomi, J. and C.H. Von Bonsdorff. 1967. Ultrastructure of a microorganism associated with bovine platelets. Experientia 23: 111–112.
Tyzzer, E.E. and D. Weinman. 1939. Haemobartonella n.g. (Bartonella olim
pro parte) H. microti n. sp. of the field vole, Microtus pennsylvanicus.
Am. J. Hyg. 30 : 141–157.
Uilenberg, G. 1967. [Eperythrozoon tuomii, n.sp. (Rickettsiales), the 3rd
species of Eperythrozoon of cattle in Madagascar]. Rev. Elev. Med. Vet.
Pays. Trop. 20 : 563–569.
Uilenberg, G., F. Thiaucourt and F. Jongejan. 2004. On molecular taxonomy: what is in a name? Exp. Appl. Acarol. 32: 301–312.
Uilenberg, G., F. Thiaucourt and F. Jongejan. 2006. Mycoplasma and
Eperythrozoon (Mycoplasmataceae). Comments on a recent paper. Int.
Willi, B., F.S. Boretti, C. Baumgartner, S. Tasker, B. Wenger, V. Cattori,
M.L. Meli, C.E. Reusch, H. Lutz and R. Hofmann-Lehmann. 2006.
Prevalence, risk factor analysis, and follow-up of infections caused
by three feline hemoplasma species in cats in Switzerland. J. Clin.
Microbiol. 44: 961–969.
Woods, J.E., M.M. Brewer, J.R. Hawley, N. Wisnewski and M.R. Lappin.
2005. Evaluation of experimental transmission of Candidatus Mycoplasma haemominutum and Mycoplasma haemofelis by Ctenocephalides
felis to cats. Am. J. Vet. Res. 66: 1008–1012.
Zwart, D., P. Leeflang and C.J. van Vorstenbosch. 1969. Studies on
an Eperythrozoon associated with bovine thrombocytes. Zentralbl.
Bakteriol. [Orig.] 210: 82–105.
Order II. Entomoplasmatales Tully, Bové, Laigret and Whitcomb 1993, 381VP
Daniel R. Brown, Janet M. Bradbury and Robert F. Whitcomb*
En.to.mo.plas.ma.ta¢les. N.L. neut. n. Entomoplasma type genus of the order; -ales ending to
denote an order: N.L. fem. pl. n. Entomoplasmatales the Entomoplasma order.
This order in the class Mollicutes has been assigned to a group
of nonhelical and helical mollicutes that are regularly associated
with arthropod or plant hosts. The description of organisms in
the order is essentially the same as for the class. Two families are
designated, Entomoplasmataceae for nonhelical mollicutes and
Spiroplasmataceae for helical ones. The order consists of four major
phylogenetic clades: the paraphyletic entomoplasmataceae clade,
which consists of the genera Entomoplasma and Mesoplasma; and
the Apis, Citri–Chrysopicola–Mirum, and Ixodetis clades of the
genus Spiroplasma. All cells are chemo-organotrophic, usually fermenting glucose through the phosphoenolpyruvate-dependent
sugar transferase system. Arginine may be hydrolyzed, but urea is
not. Cells may require sterol for growth. Nonhelical strains that
grow in serum-free media supplemented with polyoxyethylene
sorbitan (PES) are currently assigned to the genus Mesoplasma.
Temperature optimum for growth is usually 30–32°C, with a few
species able to grow at 37°C. Genome sizes range from 780 to
2220 kbp by pulsed-field gel electrophoresis (PFGE), with DNA
G+C contents ranging from 25 to 34 mol%. Like members of the
Mycoplasmatales, all organisms in this order are thought to utilize
the UGA codon to encode tryptophan.
Type genus: Entomoplasma Tully, Bové, Laigret and Whitcomb
1993, 379VP.
Taxonomic comments
*Deceased 21 December 2007.
The genera Entomoplasma and Mesoplasma constitute a polyphyletic sister lineage of the mycoides cluster of mycoplasmas
that are eccentrically situated in the paraphyletic family Entomoplasmataceae (Gasparich et al., 2004). There is no current
The basis for the proposal for the order Entomoplasmatales (Tully
et al., 1993) was the distinctive phylogenetic and phenotypic
characteristics of culturable mollicutes regularly associated
with arthropods or plants. Members of the family Entomoplasmataceae are nonhelical mollicutes that differ in their cholesterol or serum requirements for growth. Nonhelical organisms
with a strict requirement for cholesterol were placed in the
genus Entomoplasma (trivial name, entomoplasmas), whereas
nonhelical strains able to grow in a sterol-free medium supplemented with PES were assigned to the genus Mesoplasma (trivial
name, mesoplasmas). The proposal also included the transfer
of the family Spiroplasmataceae from the family Mycoplasmatales
to the family Entomoplasmatales. The helical organisms assigned
to the genus Spiroplasma were within the Spiroplasmataceae, and
genus and family descriptions of these organisms remained
as proposed previously (Skripal, 1983; Whitcomb and Tully,
1984). The order Entomoplasmatales is a phylogenetic sister to
the order Mycoplasmatales. These two orders together form a
lineage with several unique properties, including the use of
UGA as a tryptophan codon rather than a stop codon.
Family I. Entomoplasmataceae
645
hylogenetic support for separation of Entomoplasma and
p
Mesoplasma species based on neighbor-joining or maximumparsimony methods of 16S rRNA gene sequence similarity
analysis because they do not form coherent clusters, but are
instead intermixed in one paraphyletic group (Johansson
and Pettersson, 2002; Tully et al., 1998). No DNA–DNA reassociation experiments have been performed nor is there any
other polyphasic taxonomic basis to support the separation.
In particular, the growth requirement for sterols is not as profound a character as was initially believed and fails to justify
these two species (Gasparich et al., 2004; Rose et al., 1993).
For these reasons, and because Entomoplasma has priority
(Tully et al., 1993), the species currently assigned to the genus
Mesoplasma should most likely be transferred to the genus Entomoplasma. Because the transfer would include its type species,
the genus Mesoplasma would then become illegitimate. Moreover, Knight (2004) showed that the species formerly called
Mesoplasma pleciae (Tully et al., 1994) is properly affiliated with
the genus Acholeplasma on undisputed grounds of 16S rRNA
gene sequence similarity and preferred use of UGG rather
than UGA as the codon for tryptophan. Therefore, transfer of
the currently remaining members of genus Mesoplasma cannot
be endorsed until similar analyses have been completed for all
of those organisms (D.V. Volokhov, unpublished).
References
phyletic cluster of arthropod-associated mollicutes to ordinal rank
(Entomoplasmatales ord. nov.), with provision for familial rank to
separate species with nonhelical morphology (Entomoplasmataceae
fam. nov.) from helical species (Spiroplasmataceae), and emended
descriptions of the order Mycoplasmatales, family Mycoplasmataceae.
Tully, J.G., R.F. Whitcomb, K.J. Hackett, D.L. Rose, R.B. Henegar, J.M.
Bove, P. Carle, D.L. Williamson and T.B. Clark. 1994. Taxonomic
descriptions of eight new non-sterol-requiring Mollicutes assigned to
the genus Mesoplasma. Int. J. Syst. Bacteriol. 44: 685–693.
Tully, J.G., R.F. Whitcomb, K.J. Hackett, D.L. Williamson, F. Laigret, P.
Carle, J.M. Bove, R.B. Henegar, N.M. Ellis, D.E. Dodge and J. Adams.
1998. Entomoplasma freundtii sp. nov., a new species from a green tiger
beetle (Coleoptera: Cicindelidae). Int. J. Syst. Bacteriol. 48: 1197–
1204.
Whitcomb, R.F. and J.G. Tully. 1984. Family III. Spiroplasmataceae
Skripal 1983, 408VP. Genus I. Spiroplasma Saglio, L’Hospital, Laflèche,
Dupont, Bové, Tully and Freundt. In Bergey’s Manual of Systematic
Bacteriology, vol. 1 (edited by Krieg and Holt). Williams & Wilkins,
Baltimore, pp. 781–787.
Gasparich, G.E., R.F. Whitcomb, D. Dodge, F.E. French, J. Glass and
D.L. Williamson. 2004. The genus Spiroplasma and its non-helical
descendants: phylogenetic classification, correlation with phenotype
and roots of the Mycoplasma mycoides clade. Int. J. Syst. Evol. Microbiol. 54: 893–918.
Johansson, K.E. and B. Pettersson. 2002. Taxonomy of Mollicutes. In Molecular Biology and Pathogenicity of Mycoplasmas (edited by Razin and
Hermann). Kluwer Academic/Plenum Publishers, London, pp. 1–31.
Skripal, I.G. 1983. Revival of the name Spiroplasmataceae fam. nov., nom.
rev., omitted from the 1980 Approved Lists of Bacterial Names. Int.
J. Syst. Bacteriol. 33: 408.
Tully, J.G., J.M. Bové, F. Laigret and R.F. Whitcomb. 1993. Revised
taxonomy of the class Mollicutes–proposed elevation of a mono-
Family I. Entomoplasmataceae Tully, Bové, Laigret and Whitcomb 1993, 380VP
Daniel
R. Brown, Janet M. Bradbury and Robert F. Whitcomb*
En.to.mo.plas.ma.ta.ce¢ae. N.L. neut. n. Entomoplasma, -atos type genus of the family; -aceae ending
to denote a family; N.L. fem. pl. n. Entomoplasmataceae the Entomoplasma family.
Cells are usually coccoid or occur as short, branched or
unbranched, pleomorphic, nonhelical filaments. Filterable
through membranes with a mean pore diameter of 220–450
nm. Cells lack a cell wall and are bounded only by a plasma
membrane. Nonmotile. Facultatively anaerobic. The temperature range for growth varies from 10 to 37°C, with the optimum
usually at 30°C. The typical colony has a “fried-egg” appearance. Chemo-organotrophic; acid is produced from glucose,
with evidence of a phosphoenolpyruvate-dependent sugar
transport system(s) in some members. Arginine and urea are
not hydrolyzed. The organisms may require serum or cholesterol for growth or may grow in serum-free media plus 0.04%
PES. The genome sizes range from 790 to 1140 kbp.
Type genus: Entomoplasma Tully, Bové, Laigret and Whitcomb
1993, 379VP.
All members of this paraphyletic family are nonhelical and
are regularly associated with arthropod or plant hosts. They
may require cholesterol or serum for growth, and most have
an optimal growth temperature near 30°C. Separation of
members of the genera Entomoplasma and Mesoplasma within
the Entomoplasmataceae is based on the capacity of the Mesoplasma species to grow in a serum-free or cholesterol-free
medium supplemented with PES (Rose et al., 1993; Tully
et al., 1995), whereas Entomoplasma species have a growth
requirement for cholesterol. The family is derived from the
Spiroplasma lineage and is most closely related to the Apis
cluster of that group. The mycoides cluster of species in the
genus Mycoplasma is related to this family and seems to have
evolved from it.
646
Genus I. Entomoplasma Tully, Bové, Laigret and Whitcomb 1993, 379VP
En.to.mo.plas¢ma. Gr. n. entomon insect; Gr. neut. n. plasma something formed or molded, a form; N.L. neut.
n. Entomoplasma name intended to show association with insects.
Cells are nonhelical and nonmotile, frequently pleomorphic
and range in size from 200 to 1200 nm in diameter. Some cells
exhibit short filamentous forms. Most species ferment glucose.
Species possess the phosphoenolpyruvate-dependent sugarphosphotransferase system. Organisms require serum or cholesterol for growth. The temperature range for growth ranges
from 10 to 32°C, with the optimum usually at 30–32°C. The
genome sizes range from 870 to 900 kbp (PFGE). All currently
assigned species were isolated from insects or from plant surfaces where they were presumably deposited by insects.
Type species: Entomoplasma ellychniae Tully, Rose, Hackett,
Whitcomb, Carle, Bové, Colflesh and Williamson (Tully et al.,
1989) Tully, Bové, Laigret and Whitcomb 1993, 380VP (Mycoplasma ellychniae Tully, Rose, Hackett, Whitcomb, Carle, Bové,
Colflesh and Williamson 1989, 288).
Cells of these organisms vary from coccoid to pleomorphic forms
exhibiting short, branching, nonhelical filaments. Round cells
are usually in the size range of 200–300 nm, but may be larger.
Most strains were initially isolated in either M1D or SP-4 medium
and all entomoplasmas grow well in SP-4 broth containing a supplement of 17% fetal bovine serum. Some strains are able to grow
on media with reduced serum content. Most established species
have an optimal growth temperature of 30°C, but some species
grow better in broth medium maintained at 23–25°C or at 32°C.
Colony growth on solid medium is best obtained on SP-4 medium
incubated under anaerobic conditions at about 30°C. Under
these conditions, most species produce colonies with a classic
fried-egg appearance, although Entomoplasma freundtii is notable
for its granular colony m
orphology.
All species show strong fermentation of glucose with production of acid and a reduction in medium pH (Table 140). Actively
growing cultures in broth medium containing glucose may rapidly acidify the medium, causing partial or complete loss of
Table 140. Differential characteristics of species of the genus
E. ellychniae
E. freundtii
E. lucivorax
E. luminosum
E. melaleucae
E. somnilux
Entomoplasma a
Glucose fermentation
+
Arginine hydrolysis
−
“Film and spots”
−
Hemadsorption of guinea
−
pig red blood cells
DNA G+C content (mol%) 27.7
+
+
nd
nd
+
−
+
−
+
−
+
+
+
−
−
−
+
−
−
−
34
27.4
28.8
27
27
Characteristic
Symbols: +, >85% positive; −, 0–15% positive; nd, not determined.
a
v iability after 7–10 d. Arginine hydrolysis and “film and spot”
lipase reactions are rare among species described to date. Entomoplasmas were shown to lack some key metabolic activities
found in other mollicutes, especially PPi-dependent phosphofructokinase and dUTPase, and to possess uracil DNA glycosylase activity. Although the latter pyrimidine enzymic activity
distinguished Entomoplasma from Mesoplasma species, only two
Entomoplasma species and three Mesoplasma species have been
tested so far for these activities (Pollack et al., 1996).
Antisera to whole cell antigens of entomoplasmas have been
used extensively to provide specific identification to the species
level with a variety of serologic techniques, including growth
inhibition, metabolism inhibition, and agar plate immunofluorescence (Tully et al., 1989, 1990, 1998). There is no evidence
for the pathogenicity of entomoplasmas to either plant or insect
hosts. Like other mollicutes, the entomoplasmas are resistant to
500 U/ml penicillin G.
Flowers and other plant material should be cut in the field and
placed in plastic bags without touching by hand. In the laboratory, plant materials are rinsed briefly in either SP-4 or M1D
media (May et al., 2008). In both of these media, fetal bovine
serum is a critical component for successful growth of these
organisms (Hackett and Whitcomb, 1995; Tully, 1995). The rinse
medium is immediately decanted and passed through a sterile
membrane filter, usually of 450 nm porosity. The filtrate is then
passed through at least several tenfold dilutions in the selected
culture medium. The retentate may be frozen at −70°C for later
use or for retesting. The cultures are incubated at 27–30°C and
monitored by dark-field microscopy and/or by observing acidification of the medium. It is important to note that several nonsterol-requiring Acholeplasma species have also been isolated from
plant and insect material (Tully et al., 1994b).
Insect material, primarily from gut contents or hemolymph
obtained by dissection or by fine-pointed glass pipettes, should
be added to small volumes of SP-4 or M1D medium and filtered
through a 450 nm membrane filter. Serial tenfold dilutions of
the filtrate should be incubated at 27–30°C and observed for
a decrease in pH of the medium. After two to three serial passages, the organisms should be purified by conventional filtercloning techniques (Tully, 1983) and stocks of various clones
and early passage isolates frozen for further identification procedures (Whitcomb and Hackett, 1996).
Stock cultures of entomoplasmas can be maintained well in
SP-4 and/or M1D broth medium containing about 17% fetal
bovine serum. Most strains in the group can be adapted to
grow in a broth medium containing bovine serum. Stock cultures in broth medium can be stored at −70°C for indefinite
periods. For optimum preservation, the organisms should be
lyophilized as broth cultures in the early exponential phase of
growth and the dried cultures should be sealed under vacuum
and stored at 4°C.
Genus I. Entomoplasma
Differentiation of the genus Entomoplasma
from other genera
sequences. Aerobic or facultative anaerobic growth in artificial
media and the necessity for sterols for growth exclude assignment to the genera Anaeroplasma, Asteroleplasma, Acholeplasma,
Mesoplasma, or “Candidatus Phytoplasma”. Non-helical cellular
morphology and regular association with arthropod or plant
hosts support exclusion from the genera Spiroplasma or Mycoplasma. The inability to hydrolyze urea excludes assignment
to the genus Ureaplasma. However, the difficulty in assigning
novel species to this genus is well demonstrated by the earlier difficulties in establishing accurately the taxonomic status
of these organisms (Tully et al., 1993). The availability of 16S
rRNA gene sequence analyses was critical to the differentiation
of these organisms from other mollicutes. Although isolates
from vertebrates are very unlikely to be entomoplasmas, two
bona fide Mycoplasma species, Mycoplasma iowae and Mycoplasma
equigenitalium, have been isolated from plants [Grau et al., 1991;
J.C. Vignault, J.M. Bové and J.G. Tully, unpublished (see ATCC
49192)].
Taxonomic comments
The landmark studies of Weisburg et al. (1989), using 16S rRNA
gene sequences of about 50 species of mollicutes, were critical
in the resolution of certain taxonomic conflicts regarding the
species that became Entomoplasma. The first entomoplasmas to
be recognized were serologically related isolates from the flowers of Melaleuca and Grevillea trees (McCoy et al., 1979). Others,
found in a wide range of insect species (Tully et al., 1987),
included strain ELCN-1T from the hemolymph of the firefly
beetle Ellychnia corrusca (Tully et al., 1989) and three serologically distinct strains isolated from gut contents of Pyractomena
and Photinus beetles (Williamson et al., 1990). Although these
nonhelical, sterol-requiring mollicutes were initially placed in
the genus Mycoplasma, 16S rRNA gene sequence analysis clearly
indicated that strain M1T, previously designated Mycoplasma
melaleucae, and strain ELCN-1T, previously designated Mycoplasma ellychniae, were most closely affiliated with the Spiroplasma
lineage of helical organisms isolated primarily from arthropods. These findings prompted a proposal to reclassify the nonhelical mollicutes from arthropods and plants in a new order,
Entomoplasmatales, and new family, Entomoplasmataceae, with
the genus Entomoplasma reserved for sterol-requiring species
(Tully et al., 1993). Strains M1T and ELCN-1T were renamed
as Entomoplasma melaleucae and Entomoplasma ellychniae, respectively. Subsequent phylogenetic analysis of Mycoplasma freundtii,
later renamed Entomoplasma freundtii, confirmed the placement
(Tully et al., 1998).
647
The paraphyletic relationship between the genera Entomoplasma and Mesoplasma is currently an unresolved problem in
the systematics of this genus. It is possible that these genera, separated by the single criterion of sterol requirement, should be
combined into the single genus Entomoplasma. However, Knight
(2004) showed that Mesoplasma pleciae (Tully et al., 1994b) should
belong to the genus Acholeplasma based on 16S rRNA gene
sequence similarity and the preferred use of UGG rather than
UGA as the codon for tryptophan. Therefore, transfer of the currently remaining members of genus Mesoplasma to other genera
cannot be endorsed until similar analyses have been completed
for all of those species (D.V. Volokhov, unpublished).
Acknowledgements
We thank Karl-Erik Johansson for helpful comments and suggestions and Gail E. Gasparich for her landmark contributions
regarding the phylogenetics of the Entomoplasmatales. The major
contributions to the foundation of this material by Joseph G.
Tully are gratefully acknowledged.
Further reading
Tully, J.G. 1989. Class Mollicutes: new perspectives from plant
and arthropod studies. In The Mycoplasmas, vol. 5 (edited by
Whitcomb and Tully). Academic Press, San Diego, pp. 1–31.
Tully, J.G. 1996. Mollicute–host interrelationships: current concepts and diagnostic implications. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 2 (edited by Tully and
Razin). Academic Press, San Diego, pp. 1–21.
Entomoplasma
The primary technique for differentiation of Entomoplasma
species is 16S rRNA gene sequence comparisons, confirmed
by serology (Brown et al., 2007). Nonhelical mollicutes that
belong to a known species isolated from arthropods or plants
can be readily identified serologically provided that a battery
of potent antisera for classified species is available. Growth
inhibition tests, performed by placing paper discs saturated
with type-specific antisera on agar plates inoculated with
the organism, are perhaps the most convenient and rapid
serological technique to differentiate species (Clyde, 1983).
The agar plate immunofluorescence test is also a convenient
and rapid means of mollicute species identification. In the
absence of specific conjugated antiserum, an indirect immunofluorescence test can be performed with type-specific antiserum and a fluorescein-conjugated secondary antibody. The
metabolism inhibition test (Taylor-Robinson, 1983) has also
been applied to differentiation of Entomoplasma species (Tully
et al., 1998).
List of species of the genus Entomoplasma
1. Entomoplasma ellychniae (Tully, Rose, Hackett, Whitcomb,
Carle, Bové, Colflesh and Williamson 1989) Tully, Bové,
Laigret and Whitcomb 1993, 380VP (Mycoplasma ellychniae
Tully, Rose, Hackett, Whitcomb, Carle, Bové, Colflesh and
Williamson 1989, 288)
el.lych.ni¢ae. N.L. n. Ellychnia a genus of firefly beetles; N.L.
gen. n. ellychniae of Ellychnia, from which the organism was
first isolated.
This is the type species of the genus Entomoplasma. Cells are
nonhelical, pleomorphic filaments, with some branching;
648
small coccoid forms, ranging in diameter from 200 to 300 nm,
also occur. Passage of broth cultures through 450 and 300 nm
porosity membrane filters does not reduce viable cell numbers,
whereas passage through 220 nm porosity reduces cell populations by about 10%. Grows well in SP-4 medium with fetal
bovine serum supplements. Does not grow well in horse serumsupplemented broth or agar media. Optimum temperature
for broth growth is 30°C; can grow at 18–32°C. Colonies incubated at 30°C under anaerobic conditions have a fried-egg
appearance. Does not hemadsorb guinea pig erythrocytes.
No evidence for pathogenicity for insects.
Source: isolated from the hemolymph of the firefly beetle
Ellychniae corrusca.
Type strain: ELCN-1, ATCC 43707, NCTC 11714.
2. Entomoplasma freundtii Tully, Whitcomb, Hackett, Williamson,
Laigret, Carle, Bové, Henegar, Ellis, Dodge and Adams 1998,
1203VP
freund¢ti.i. N.L. masc. gen. n. freundtii of Freundt, named
after Eyvind Freundt, a Danish pioneer in the taxonomy and
classification of mollicutes.
Cells are predominantly coccoid in shape, ranging from
300 to 1200 nm in diameter. Organisms are readily filterable
through membranes with mean pore diameters of 450, 300,
and 220 nm; more than 90% of viable cells in broth culture
are able to pass 220 nm porosity membranes. The temperature range for growth is 10–32°C, with an optimum at 30°C.
Colonies under anaerobic conditions are granular and frequently exhibit multiple satellite forms although the organism is considered nonmotile. The organism grows well in
SP-4 broth medium or other media containing horse serum
supplements.
No evidence for pathogenicity for insects.
Source: isolated from the gut contents of a green tiger beetle (Coleoptera: Cicindelidae).
Type strain: BARC 318, ATCC 51999.
3. Entomoplasma lucivorax (Williamson, Tully, Rose, Hackett,
Henegar, Carle, Bové, Colflesh and Whitcomb 1990) Tully,
Bové, Laigret and Whitcomb 1993, 380VP (Mycoplasma lucivorax Williamson, Tully, Rose, Hackett, Henegar, Carle, Bové,
Colflesh and Whitcomb 1990, 164)
lu.ci.vo¢rax. L. fem. n. lux lucis light; L. neut. adj. vorax gluttonous, devouring; N.L. neut. adj. lucivorax light devouring,
referring to the predacious habit of the host insect, which
preys on other luminescent firefly species.
Cells are either pleomorphic coccoidal or subcoccoidal,
with a diameter of 200–300 nm, or are short, branched or
unbranched filaments. Cells are readily filterable through
membrane filters with mean pore diameters of 450, 300, and
220 nm, but do not pass 100 nm porosity membranes. Optimum temperature for growth is 30°C; can grow at 10–32°C.
Nonmotile. Colonies under anaerobic conditions usually have
a fried-egg appearance. Grows well in SP-4 broth medium or
other media containing horse serum supplements. Colonies
do not hemadsorb guinea pig erythrocytes.
No evidence of pathogenicity for insects or plants.
Source: first isolated from the gut of a firefly beetle (Photinus pyralis); also isolated from a flower (Spirea ulmaria;
C. Chastel, unpublished).
Type strain: PIPN-2, ATCC 49196, NCTC 11716.
4. Entomoplasma luminosum (Williamson, Tully, Rose, Hackett,
Henegar, Carle, Bové, Colflesh and Whitcomb 1990)
Tully, Bové, Laigret and Whitcomb 1993, 380VP (Mycoplasma
luminosum Williamson, Tully, Rose, Hackett, Henegar, Carle,
Bové, Colflesh and Whitcomb 1990, 163)
lu.mi.no¢sum. L. neut. adj. luminosum luminous, emitting
light, referring to the luminescence of the adult host from
which the organism was isolated.
Cells are pleomorphic and coccoidal or subcoccoidal
with a diameter of 200–300 nm. Cells also occur as short,
branched or unbranched filaments. The organisms are readily filterable through membranes with mean pore diameters
of 450, 300, and 220 nm, but do not pass 100 nm porosity
membranes. The temperature range for growth is 10–32°C,
with an optimum at 32°C. Nonmotile. Colonies under anaerobic conditions have a fried-egg appearance. The organism
grows well in SP-4 broth medium or other media containing
horse serum supplements. Colonies hemadsorb guinea pig
erythrocytes.
No evidence of pathogenicity for insects.
Source: isolated from the gut of the firefly beetle (Photinus
marginata).
Type strain: PIMN-1, ATCC 49195, NCTC 11717.
5. Entomoplasma melaleucae (Tully, Rose, McCoy, Carle, Bové,
Whitcomb and Weisburg 1990) Tully, Bové, Laigret and Whitcomb 1993, 380VP (Mycoplasma melaleucae Tully, Rose, McCoy,
Carle, Bové, Whitcomb and Weisburg 1990, 146)
me la.leu¢cae. N.L. n. Melaleuca a genus of tropical trees having white flowers with sweet fragrance; N.L. gen. n. melaleucae
of Melaleuca, the plant from which the type strain was isolated.
Cells are pleomorphic and coccoidal or subcoccoidal, with
few filamentous forms. Coccoidal forms have mean diameters
of 250–300 nm. Cells are readily filterable through 450 and
300 nm porosity membrane filters, with few cells passing 220
nm porosity membranes. The temperature range for growth
is 10–30°C, with an optimum at about 23°C. Nonmotile. Colonies under anaerobic conditions at 23–30°C display a friedegg appearance. Grows well in SP-4 broth or in modified
Edward medium containing fetal bovine serum. The organism does not grow well in horse serum-based broth medium.
Agar colonies do not adsorb guinea pig erythrocytes.
No evidence of pathogenicity for insects or plants.
Source: isolated from flower surfaces of a subtropical plant,
Melaleuca quinquenervia, in south Florida. Related strains
have been isolated from flowers of other subtropical trees
in Florida, Melaleuca decora and Grevillea robusta (silk oak),
and from an anthophorine bee (Xylocopa micans) in the same
geographic area.
Genus II. Mesoplasma
Type strain: M1, ATCC 49191, NCTC 11715.
Sequence accession nos (16S rRNA gene): M24478, AY345990.
Further comment: the 16S rRNA gene sequence is more similar to that of members of genus Mesoplasma than to others in
the genus Entomoplasma.
6. Entomoplasma somnilux (Williamson, Tully, Rose, Hackett,
Henegar, Carle, Bové, Colflesh and Whitcomb 1990) Tully,
Bové, Laigret and Whitcomb 1993, 380VP (Mycoplasma
somnilux Williamson, Tully, Rose, Hackett, Henegar, Carle,
Bové, Colflesh and Whitcomb 1990, 163)
som.ni¢lux. L. masc. n. somnus sleep; L. fem. n. lux light; N.L.
n. somnilux intended to mean sleeping light, referring to the
quiescent pupal stage of the host from which the organism
was isolated, which precedes the luminescent adult stage.
649
Cells are pleomorphic and coccoidal or subcoccoidal, with
a diameter of 200–300 nm; also occur as short, branched or
unbranched filaments. Readily filterable through membranes
with mean pore diameters of 450, 300, and 220 nm. The temperature range for growth is 10–32°C, with optimum growth
at 30°C. Nonmotile. Colonies incubated under anaerobic
conditions at 30°C have a fried-egg appearance. The organism grows well in SP-4 broth medium or other media containing horse serum supplements. Colonies do not adsorb
guinea pig erythrocytes.
Source: isolated from a pupal gut of the firefly beetle (Pyractomena angulata).
Type strain: PYAN-1, ATCC 49194, NCTC 11719.
Genus II. Mesoplasma Tully, Bové, Laigret and Whitcomb 1993, 380VP
Me.so.plas¢ma. Gr. adj. mesos middle; Gr. neut. n. plasma something formed or molded, a form; N.L. neut.
n. Mesoplasma middle form, name intended to denote a middle position with respect to sterol or cholesterol
requirement.
Cells are nonhelical and nonmotile, generally coccoid or short
filamentous forms. Coccoid cells are usually 220–300 nm in dia
meter, but some cells in some species can be as large as 400–500 nm.
Most strains ferment glucose and most, but not all, lack the ability
to hydrolyze arginine. Species possess the phosphoenolpyruvatedependent sugar-phosphotransferase system. Neither serum nor
cholesterol is required for growth, but strains show sustained
growth in a serum-free or cholesterol-free medium when the
medium is supplemented with 0.04% PES. The optimum temperature for growth is usually near 28–32°C, with some strains able to
grow well at temperatures as low as 23°C or as high as 37°C.
Genome sizes range from 825 to 930 kbp (PFGE).
Type species: Mesoplasma florum (McCoy, Basham, Tully, Rose,
Carle and Bové 1984) Tully, Bové, Laigret and Whitcomb 1993,
380VP (Acholeplasma florum McCoy, Basham, Tully, Rose, Carle
and Bové 1984, 14).
Cells are predominantly coccoid in the exponential phase of
growth when examined by dark-field microscopy. Cells from
broth cultures examined by transmission electron microscopy
are also coccoid, with individual cells usually 220–500 nm in
diameter and clearly defined by a single cytoplasmic membrane. Colony growth is best obtained on SP-4 agar medium.
Plates incubated under anaerobic conditions at about 30°C
usually display characteristic fried-egg type colonies after 5–7
d incubation.
Several mesoplasmas lack certain key metabolic activities
found in other mollicutes, especially PPi-dependent phosphofructokinase, dUTPase, and uracil DNA glycosylase activity
(Pollack et al., 1996). Most mesoplasmas were isolated in M1D
medium containing 15% fetal bovine serum (Whitcomb, 1983),
but adapt well to growth in SP-4 broth containing 15–17% fetal
bovine serum, or in broth medium containing a 1% bovine
serum fraction supplement (Tully, 1984; Tully et al., 1994a). All
species show strong fermentation of glucose with acid production (Table 141), with a rapid decline in pH of the medium and
loss of viability. Arginine hydrolysis has been observed only with
the type strain (PUPA-2T) of Mesoplasma photuris.
Antisera directed against whole-cell antigens of filter-cloned
mesoplasmas have been used extensively to establish species
and to provide species identifications. There is no evidence of
pathogenicity of any currently established species in the genus
for either an insect or plant host. Mesoplasmas are resistant to
500 U/ml penicillin.
Enrichment, isolation, and maintenance procedures
The culture media and procedures for isolation and maintenance of entomoplasmas from plant and insect sources can also
be effectively applied for mesoplasmas.
Differentiation of the genus Mesoplasma
from other genera
Properties that fulfill criteria for assignment to this genus are
the same as those for the genus Entomoplasma, with the exception that the genus Mesoplasma is currently reserved for species
that are able to grow in serum-free medium supplemented with
PES (Tully et al., 1993).
Taxonomic comments
The existence of a flora of nonhelical, wall-less prokaryotes
associated with arthropod or plant hosts was first documented
by T.B. Clark, S. Eden-Green, and R.E. McCoy and colleagues.
Some of the plant isolates were clearly related to previously
described Acholeplasma species, such as Acholeplasma oculi (EdenGreen and Tully, 1979), whereas others were established as novel
Acholeplasma species, able to grow well in broth media without
any cholesterol, serum, or fatty acid supplements. However, a
significant group of other similarly derived strains were able to
650
M. coleopterae
M. corruscae
M. entomophilum
M. grammopterae
M. lactucae
M. photuris
M. seiffertii
M. syrphidae
M. tabanidae
Arginine hydrolysis
Hemadsorption of guinea pig red blood cells
DNA G+C content (mol%)
M. chauliocola
Characteristic
M. florum
Table 141. Differential characteristics of species of the genus Mesoplasma a
+
−
−
27.3
+
−
+
28.3
+
−
−
27.7
+
−
+
26.4
+
−
+
30
+
−
−
29.1
+
−
+
30
+
+
−
28.8
+
−
+
30
+
−
+
27.6
+
−
−
28.3
Symbols: +, >85% positive; −, 0–15% positive.
a
grow in serum-free or cholesterol-free media only when small
amounts of PES were added to the medium. Because these
strains grew in the absence of cholesterol or serum, several of
them were initially described as Acholeplasma species, including
Acholeplasma florum (McCoy et al., 1984), Acholeplasma entomophilum (Tully et al., 1988), and Acholeplasma seiffertii (Bonnet et al.,
1991). Although the growth response to PES in serum-free or
cholesterol-free media suggested that there were fundamental
differences between such mollicutes and classic acholeplasmas,
conclusive taxonomic evidence was lacking. The subsequent
analysis of 16S rRNA gene sequences by Weisburg et al. (1989)
showed that the PES-requiring organisms were closely related
to the spiroplasma group of mollicutes and were phylogenetically distant from acholeplasmas. On the basis of these findings
and additional phylogenetic data, a proposal was made that the
plant- and insect-derived mollicutes with growth responses to
PES in serum-free or cholesterol-free media would be assigned
to a new family, Entomoplasmataceae, and a new genus, Mesoplasma
(Tully et al., 1993). Three of the plant-derived strains previously
described as Acholeplasma species (Acholeplasma florum, Acholeplasma entomophilum, and Acholeplasma seiffertii) were transferred
to the genus Mesoplasma, with retention of their species epithets.
A single plant-derived strain that had previously been described
as Mycoplasma lactucae, and later found to grow in serum-free or
cholesterol-free media supplemented with PES, was renamed
Mesoplasma lactucae. Later, eight novel Mesoplasma species were
described (Tully et al., 1994a).
The paraphyletic relationship between the genera Entomoplasma and Mesoplasma is a currently unresolved problem in
the systematics of this genus. It is possible that these genera,
s eparated by the single criterion of sterol requirement, should
be combined into the single genus Entomoplasma. However,
Knight (2004) showed that Mesoplasma pleciae (Tully et al.,
1994a) should belong to the genus Acholeplasma based on 16S
rRNA gene sequence similarity and the preferred use of UGG
rather than UGA as the codon for tryptophan. Therefore, transfer of the currently remaining members of the genus Mesoplasma to other genera cannot be endorsed until similar analyses
have been completed for all of those species (D.V. Volokhov,
unpublished).
Acknowledgements
We thank Karl-Erik Johansson for helpful comments and suggestions and Gail E. Gasparich for her landmark contributions
regarding the phylogenetics of the Entomoplasmatales. The major
contributions to the foundation of this material by Joseph G.
Tully are gratefully acknowledged.
Further reading
and arthropod studies. In The Mycoplasmas, vol. 5 (edited by
Whitcomb and Tully). Academic Press, San Diego, pp. 1–31.
Tully, J.G. 1996. Mollicute-host interrelationships: current concepts and diagnostic implications. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 2 (edited by Tully and
Razin). Academic Press, San Diego, pp. 1–21.
Differentiation of the species of the genus Mesoplasma
The techniques for differentiation of Mesoplasma species are the
same as those for genus Entomoplasma.
List of species of the genus Mesoplasma
1. Mesoplasma florum (McCoy, Basham, Tully, Rose, Carle and
Bové 1984) Tully, Bové, Laigret and Whitcomb 1993, 380VP
(Acholeplasma florum McCoy, Basham, Tully, Rose, Carle and
Bové 1984, 14)
flo¢rum. L. gen. pl. n. florum of flowers, indicating the recovery site of the organism.
This is the type species of the genus. Cells are oval or coccoid. The organism is readily filterable through membranes
with mean pore diameters of 450, 300, and 220 nm, but does
not pass a membrane with 100 nm porosity. Temperature
range for growth is 18–37°C, with an optimum at 28–30°C.
Colonies on agar medium containing horse serum supplements have a typical fried-egg appearance after anaerobic
incubation at 37°C. Colonies on agar do not hemadsorb
guinea pig erythrocytes.
The 16S rRNA gene sequence is identical to that of Mesoplasma entomophilum (GenBank accession no. AF305693),
but antiserum against Mesoplasma florum did not inhibit
growth of Mesoplasma entomophilum or label the surfaces of
Mesoplasma entomophilum colonies on agar (Tully et al., 1988).
There are additional phenotypic distinctions between the
two species.
No evidence of pathogenicity for plants or insects.
Source: first isolated from surface of flowers on a
lemon tree (Citrus limon) in Florida, with subsequent
isolations from floral surfaces of grapefruit (Citrus
651
p aradisi) and powderpuff trees (Albizia julibrissin) in
Florida (McCoy et al., 1979). Also isolated from a variety
of plants and from the gut tissues of numerous species of
insects (Clark et al., 1986; Tully et al., 1990; Whitcomb
et al., 1982).
DNA G+C content (mol%): 27.3 (Bd, whole genome
sequence).
Type strain: L1, ATCC 33453, NCTC 11704.
Sequence accession nos: AF300327 (16S rRNA gene),
NC_006055 (strain L1T genome sequence).
emperature range for growth is 10–32°C, with an optimum
T
of 30°C. Nonmotile. Colonies incubated anaerobically at
30°C usually have a fried-egg appearance. Colonies hemadsorb guinea pig erythrocytes.
Source: original isolation was from the gut of an adult
firefly (Ellychnia corrusca).
DNA G+C content (mol%): 26.4 (Bd, Tm, HPLC).
Type strain: ELCA-2, ATCC 49579.
2. Mesoplasma chauliocola Tully, Whitcomb, Hackett, Rose,
Henegar, Bové, Carle, Williamson and Clark 1994a, 691VP
5. Mesoplasma entomophilum (Tully, Rose, Carle, Bové,
Hackett and Whitcomb 1988) Tully, Bové, Laigret and
Whitcomb 1993, 380VP (Acholeplasma entomophilum Tully,
Rose, Carle, Bové, Hackett and Whitcomb 1988, 166)
chau.li.o¢co.la. N.L. n. chaulio first part of the genus name
of goldenrod beetle (Chauliognathus); L. suff. -cola (from L.
masc. or fem. n. incola) inhabitant; N.L. masc. n. chauliocola
inhabitant of the goldenrod beetle.
Cells are primarily coccoid, ranging in size from 300 to
500 nm in diameter. Cells are readily filterable through
membranes with mean pore diameters of 450, 300, and
220 nm, with a small number of cells able to pass through
100 nm porosity filters. Temperature range for growth is
10–37°C, with an optimum of 32–37°C. Nonmotile. Colonies incubated anaerobically at 32–37°C show fried-egg
morphology. Colonies hemadsorb guinea pig erythrocytes.
Source: originally isolated from gut fluid of an adult goldenrod soldier beetle (Chauliognathus pennsylvanicus).
Type strain: CHPA-2, ATCC 49578.
3. Mesoplasma coleopterae Tully, Whitcomb, Hackett, Rose,
co.le.op.te¢rae. N.L. fem. gen. n. coleopterae of Coleoptera,
referring to the order of insects (Coleoptera) from which
the organism was first isolated.
Cells are primarily coccoid, ranging in diameter from
300 to 500 nm. Organisms are readily filterable through
220 nm. Temperature range for growth is 10–37°C, with an
optimum of 30–37°C. Nonmotile. Colonies incubated
anaerobically at 30°C usually have a fried-egg appearance.
Agar colonies do not hemadsorb guinea pig erythrocytes.
soldier beetle (Chauliognathus sp.).
Sequence accession no. (16S rRNA gene): DQ514605 (partial
sequence).
4. Mesoplasma corruscae Tully, Whitcomb, Hackett, Rose,
cor.rus¢cae. N.L. fem. gen. n. corruscae of corrusca, referring to the species of firefly beetle (Ellychnia corrusca) from
300 to 500 nm. Cells are readily filterable through membranes with mean pore diameters of 450, 300, and 220 nm.
en.to.mo.phi¢lum. Gr. n. entomon insect; N.L. neut. adj.
philum (from Gr. neut. adj. philon) friend, loving; N.L. neut.
adj. entomophilum insect-loving.
Cells are pleomorphic, but primarily coccoid, ranging
from 300 to 500 nm in diameter. Cells are readily filterable
through 220 nm porosity membrane filters. The temperature range for growth is 23–32°C, with an optimum at 30°C.
Nonmotile. Colonies incubated under anaerobic conditions at 30°C usually have a fried-egg appearance. Colonies
hemadsorb guinea pig erythrocytes.
The 16S rRNA gene sequence is identical to that of
Mesoplasma florum (GenBank accession no. AF300327), but
antiserum against Mesoplasma florum did not inhibit growth
of Mesoplasma entomophilum or label the surfaces of Mesoplasma entomophilum colonies on agar (Tully et al., 1988).
There are additional phenotypic distinctions between the
two species.
Source: original isolation was from the gut contents of a
tabanid fly (Tabanus catenatus). Also isolated from a variety
of other species of insects.
Type strain: TAC, ATCC 43706, NCTC 11713.
6. Mesoplasma grammopterae Tully, Whitcomb, Hackett, Rose,
gram.mop.te¢rae. N.L. fem. gen. n. grammopterae of Grammoptera, referring to the genus of beetle (Grammoptera) from
300 to 500 nm. Cells are readily filterable through membrane filters with mean pore diameters of 450, 300, and
220 nm. Temperature range for growth is 10–37°C, with an
optimum at 30°C. Nonmotile. Colonies incubated under
anaerobic conditions at 30°C have a fried-egg appearance.
Colonies do not hemadsorb guinea pig erythrocytes.
Source: original isolation was from the gut contents of an
adult long-horned beetle (Grammoptera sp.). Other isolations were made from adult soldier beetle (Cantharidae sp.)
and from an adult mining bee (Andrena sp.).
Type strain: GRUA-1, ATCC 49580.
652
7. Mesoplasma lactucae (Rose, Kocka, Somerson, Tully,
Whitcomb, Carle, Bové, Colflesh and Williamson 1990)
Tully, Bové, Laigret and Whitcomb 1993, 380VP (Mycoplasma
lactucae Rose, Kocka, Somerson, Tully, Whitcomb, Carle,
Bové, Colflesh and Williamson 1990, 141)
lac.tu¢cae. L. fem. n. lactuca lettuce; L. gen. n. lactucae of
lettuce, referring to the plant from which the organism was
first isolated.
500 nm in diameter, with only occasional short, nonhelical,
pleomorphic filaments. Cells are readily filterable through
membrane filters with mean pore diameters of 450, 300,
and 220 nm, and a few cells are able to pass 100 nm porosity membranes. Temperature range for growth is 18–37°C,
with optimal growth at 30°C. Nonmotile. Colonies incubated under anaerobic conditions at 30°C have a fried-egg
appearance. Colonies hemadsorb guinea pig erythrocytes.
Source: original isolation was from lettuce (Lactuca
sativa).
Type strain: 831-C4, ATCC 49193, NCTC 11718.
Sequence accession no. (16S rRNA gene): AF303132. Has been
reported to possess three rRNA operons (Grau, 1991).
8. Mesoplasma photuris Tully, Whitcomb, Hackett, Rose,
pho.tu¢ris. N.L. gen. n. photuris of Photuris, referring to the
genus of firefly beetle (Photuris sp.) from which the organism was first isolated.
300 to 500 nm. Readily filterable through membrane filters
with mean pore diameters of 450, 300, and 220 nm. Temperature range for growth is 10–32°C, with optimum at
30°C. Nonmotile. Colonies incubated under anaerobic conditions at 30°C have a fried-egg appearance. Colonies do
not hemadsorb guinea pig erythrocytes.
Source: original isolation was from gut fluids of larval and
adult fireflies (Photuris lucicrescens and other Photuris spp.).
One isolate (BARC 1976) was obtained by F.E. French from
the gut of a horse fly (Tabanus americanus).
Type strain: PUPA-2, ATCC 49581.
9. Mesoplasma seiffertii (Bonnet, Saillard, Vignault, Garnier,
Carle, Bové, Rose, Tully and Whitcomb 1991) Tully, Bové,
Laigret and Whitcomb 1993, 380VP (Acholeplasma seiffertii
Bonnet, Saillard, Vignault, Garnier, Carle, Bové, Rose, Tully
and Whitcomb 1991, 48)
seif.fer¢ti.i. N.L. masc. gen. n. seiffertii of Seiffert, in honor
of Gustav Seiffert, a German microbiologist who performed
pioneering studies on mollicutes that occur in soil and compost and do not require sterols for growth.
Cells are primarily coccoid, ranging in diameter
from 300 to 500 nm. Cells are readily filterable through
220 nm. Temperature range for growth is 20–35°C,
with optimum at about 28–30°C. Nonmotile. Colonies
incubated under anaerobic conditions at 30°C have a
fried-egg appearance. Colonies hemadsorb guinea pig
erythrocytes.
Three insect isolates of Mesoplasma seiffertii, two from
mosquitoes and one from a horse fly, were compared to
strain F7T of plant origin. High relatedness values of 78–98%
DNA–DNA reassociation under high stringency conditions
were obtained (Gros et al., 1996).
Source: first isolated from floral surfaces of a sweet orange
tree (Citrus sinensis) and from wild angelica (Angelica sylvestris). Also isolated from insects.
Type strain: F7, ATCC 49495.
10. Mesoplasma syrphidae Tully, Whitcomb, Hackett, Rose,
syr.phi¢dae. N.L. fem. gen. n. syrphidae of a syrphid, referring to the syrphid fly family (Syrphidae), from which the
organism was first isolated.
500 nm in diameter. Cells readily pass membrane filters
with mean pore diameters of 450, 300, and 220 nm. Temperature range for growth is 10–32°C, with optimum at
23–25°C. Nonmotile. Colonies incubated under anaerobic
conditions at 23–25°C have a fried-egg appearance. Colonies hemadsorb guinea pig erythrocytes.
syrphid fly (Diptera: Syrphidae). Similar strains have been
isolated from a bumblebee (Bombus sp.) and a skipper
(Lepidoptera: Hesperiidae).
Type strain: YJS, ATCC 51578.
11.Mesoplasma tabanidae Tully, Whitcomb, Hackett, Rose,
ta.ba.ni.dae. N.L. fem. gen. n. tabanidae of a tabanid, referring to the horse fly family (Tabanidae), the host from
Cells are primarily coccoid, ranging in size from 300
to 500 nm in diameter. Cells readily pass membrane filters with mean pore diameters of 450, 300, and 220 nm.
Temperature range for growth is 10–37°C, with optimum
at 37°C. Nonmotile. Colonies incubated under anaerobic
conditions at 37°C display a fried-egg appearance. Colonies
do not hemadsorb guinea pig erythrocytes.
horse fly (Tabanus abactor).
References
Bonnet, F., C. Saillard, J.C. Vignault, M. Garnier, P. Carle, J.M. Bové, D.L.
Rose, J.G. Tully and R.F. Whitcomb. 1991. Acholeplasma seiffertii sp. nov.,
a mollicute from plant surfaces. Int. J. Syst. Bacteriol. 41: 45–49.
Brown, D.R., R.F. Whitcomb and J.M. Bradbury. 2007. Revised minimal
standards for description of new species of the class Mollicutes (division Tenericutes). Int. J. Syst. Evol. Microbiol. 57: 2703–2719.
Clark, T.B., J.G. Tully, D.L. Rose, R. Henegar and R.F. Whitcomb.
1986. Acholeplasmas and similar nonsterol-requiring mollicutes
from insects: missing link in microbial ecology. Curr. Microbiol. 13:
11–16.
Clyde, W.A., Jr. 1983. Growth inhibition tests. In Methods in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, New
York, pp. 405–410.
Eden-Green, S.J. and J.G. Tully. 1979. Isolation of Acholeplasma spp.
from coconut palms affected by lethal yellowing disease in Jamaica.
Curr. Microbiol. 2: 311–316.
Grau, O. 1991. Analyse des gènes ribosomiques des mollicutes, application à l’identification d’un mollicute non classé et conséquences
taxonomiques [thesis]. Bordeaux, France.
Grau, O., F. Laigret, P. Carle, J.G. Tully, D.L. Rose and J.M. Bové. 1991.
Identification of a plant-derived mollicute as a strain of an avian
pathogen, Mycoplasma iowae, and its implications for mollicute taxonomy. Int. J. Syst. Bacteriol. 41: 473–478.
Gros, O., C. Saillard, C. Helias, F. LeGoff, M. Marjolet, J.M. Bové and
C. Chastel. 1996. Serological and molecular characterization of Mesoplasma seiffertii strains isolated from hematophagous dipterans in
France. Int. J. Syst. Bacteriol. 46 : 112–115.
Hackett, K.J. and R.F. Whitcomb. 1995. Cultivation of spiroplasmas
in undefined and defined media. In Molecular and Diagnostic
Procedures in Mycoplasmology, vol. 1 (edited by Razin and Tully).
May, M., R.F. Whitcomb and D.R. Brown. 2008. Mycoplasma and related
organisms. In Practical Handbook of Microbiology (edited by Goldman and Green). CRC Press, Boca Raton, pp. 467–491.
McCoy, R.E., D.S. Williams and D.L. Thomas. 1979. Isolation of mycoplasmas from flowers. Proceedings of the Republic of China-United
States Cooperative Science Seminar, Symposium series 1, National
Science Council, Taipei, Taiwan, pp. 75–81.
McCoy, R.E., H.G. Basham, J.G. Tully, D.L. Rose, P. Carle and J.M. Bové.
1984. Acholeplasma florum, a new species isolated from plants. Int. J.
Pollack, J.D., M.V. Williams, J. Banzon, M.A. Jones, L. Harvey and J.G.
Tully. 1996. Comparative metabolism of Mesoplasma, Entomoplasma,
Mycoplasma, and Acholeplasma. Int. J. Syst. Bacteriol. 46: 885–890.
Rose, D.L., J.P. Kocka, N.L. Somerson, J.G. Tully, R.F. Whitcomb, P.
Carle, J.M. Bové, D.E. Colflesh and D.L. Williamson. 1990. Mycoplasma lactucae sp. nov., a sterol-requiring mollicute from a plant surface. Int. J. Syst. Bacteriol. 40: 138–142.
Taylor-Robinson, D. 1983. Metabolism inhibition tests. In Methods in
Tully, J.G. 1983. Cloning and filtration techniques for mycoplasmas.
In Methods in Mycoplasmology, vol. 1 (edited by Razin and Tully).
Tully, J.G. 1984. Genus Acholeplasma. In Bergey’s Manual of Systematic
Bacteriology, vol. 1 (edited by Krieg and Holt). Williams & Wilkins,
Baltimore, pp. 775–781.
653
Tully, J.G. 1995. Determination of cholesterol and polyoxyethylene
sorbitan growth requirements of mollicutes. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 1 (edited by Razin and
Tully). Academic Press, San Diego, pp. 381–389.
Tully, J.G., D.L. Rose, R.F. Whitcomb, K.J. Hackett, T.B. Clark,
R.B. Henegar, E. Clark, P. Carle and J.M. Bové. 1987. Characterization of some new insect-derived acholeplasmas. Isr. J. Med. Sci. 23:
699–703.
Tully, J.G., D.L. Rose, P. Carle, J.M. Bové, K.J. Hackett and R.F. Whitcomb. 1988. Acholeplasma entomophilum sp. nov. from gut contents of
a wide-range of host insects. Int. J. Syst. Bacteriol. 38: 164–167.
Tully, J.G., D.L. Rose, K.J. Hackett, R.F. Whitcomb, P. Carle, J.M. Bové,
D.E. Colflesh and D.L. Williamson. 1989. Mycoplasma ellychniae sp.
nov., a sterol-requiring mollicute from the firefly beetle Ellychnia corrusca. Int. J. Syst. Bacteriol. 39: 284–289.
Tully, J.G., D.L. Rose, R.E. McCoy, P. Carle, J.M. Bové, R.F. Whitcomb
and W.G. Weisburg. 1990. Mycoplasma melaleucae sp. nov., a sterolrequiring mollicute from flowers of several tropical plants. Int. J. Syst.
Bacteriol. 40: 143–147.
cluster of arthropod-associated mollicutes to ordinal rank (Entomoplasmatales ord. nov.), with provision for familial rank to separate
of the order Mycoplasmatales, family Mycoplasmataceae. Int. J. Syst. Bacteriol. 43: 378–385.
Tully, J.G., R.F. Whitcomb, K.J. Hackett, D.L. Rose, R.B. Henegar, J.M.
Bové, P. Carle, D.L. Williamson and T.B. Clark. 1994a. Taxonomic
Tully, J.G., R.F. Whitcomb, D.L. Rose, J.M. Bové, P. Carle, N.L. Somerson,
D.L. Williamson and S. Edengreen. 1994b. Acholeplasma brassicae sp.
nov. and Acholeplasma palmae sp. nov., two non-sterol-requiring mollicutes from plant surfaces. Int. J. Syst. Bacteriol. 44: 680–684.
Tully, J.G., D.L. Rose, C.E. Yunker, P. Carle, J.M. Bové, D.L. Williamson
and R.F. Whitcomb. 1995. Spiroplasma ixodetis sp. nov., a new species
from Ixodes pacificus ticks collected in Oregon. Int. J. Syst. Bacteriol.
45: 23–28.
Tully, J.G., R.F. Whitcomb, K.J. Hackett, D.L. Williamson, F. Laigret, P.
Carle, J.M. Bové, R.B. Henegar, N.M. Ellis, D.E. Dodge and J. Adams.
1998. Entomoplasma freundtii sp. nov., a new species from a green
tiger beetle (Coleoptera: Cicindelidae). Int. J. Syst. Bacteriol. 48:
1197–1204.
Weisburg, W.G., J.G. Tully, D.L. Rose, J.P. Petzel, H. Oyaizu, D. Yang, L.
Mandelco, J. Sechrest, T.G. Lawrence, J. Van Etten, J. Maniloff and
C.R. Woese. 1989. A phylogenetic analysis of the mycoplasmas: basis
for their classification. J. Bacteriol. 171: 6455–6467.
Whitcomb, R.F. 1983. Culture media for spiroplasmas. In Methods in
Whitcomb, R.F. and K.J. Hackett. 1996. Identification of mollicutes from
insects. In Molecular and Diagnostic Procedures in Mycoplasmology,
vol. 2 (edited by Tully and Razin). Academic Press, San Diego, pp.
313–322.
Whitcomb, R.F., J.G. Tully, D.L. Rose, E.B. Stephens, A. Smith, R.E.
McCoy and M.F. Barile. 1982. Wall-less prokaryotes from fall flowers in central United States and Maryland. Curr. Microbiol. 7:
285–290.
Williamson, D.L., J.G. Tully, D.L. Rose, K.J. Hackett, R. Henegar, P.
Carle, J.M. Bové, D.E. Colflesh and R.F. Whitcomb. 1990. Mycoplasma
somnilux sp. nov., Mycoplasma luminosum sp. nov., and Mycoplasma
lucivorax sp. nov., new sterol-requiring mollicutes from firefly beetles
(Coleoptera, Lampyridae). Int. J. Syst. Bacteriol. 40 : 160–164.
654
Family II. Spiroplasmataceae
Family II. Spiroplasmataceae Skripal 1983, 408VP
David L. Williamson, Gail E. Gasparich, Laura B. Regassa, Collette Saillard, Joël Renaudin,
Joseph M. Bové and Robert F. Whitcomb*
Spi.ro.plas.ma.ta.ce′ae. N.L. neut. n. Spiroplasma, -atos type genus of the family; -aceae ending to
denote a family; N.L. fem. pl. n. Spiroplasmataceae the Spiroplasma family.
Cells are helical during exponential growth, with rotatory,
flexional, and translational motility. Genome size is variable:
780–2220 kbp. Variable sterol requirement for growth. Procedures for determining sterol requirement are as described
for Family I (Entomoplasmataceae). Possess a phosphoenolpyruvate phosphotransferase system for glucose uptake. Reduced
nicotinamide adenine dinucleotide (NADH) oxidase activity is
located only in the cytoplasm. Unable to synthesize fatty acids
from acetate. Other characteristics are as described for the
class and order.
Type genus: Spiroplasma Saglio, L’Hospital, Laflèche, Dupont,
Bové, Tully and Freundt 1973, 201AL.
Genus I. Spiroplasma Saglio, L’Hospital, Laflèche, Dupont, Bové, Tully and Freundt 1973, 201AL
David L. Williamson, Gail E. Gasparich, Laura B. Regassa, Collette Saillard, Joël Renaudin, Joseph M. Bové
and Robert F. Whitcomb*
Spi.ro.plas¢ma. Gr. n. speira (L. transliteration spira) a coil, spiral; Gr. neut. n. plasma something formed or
molded, a form; N.L. neut. n. Spiroplasma spiral form.
Cells are pleomorphic, varying in size and shape from helical
and branched nonhelical filaments to spherical or ovoid. The
helical forms, usually 100–200 nm in diameter and 3–5 mm in
length, generally occur during the exponential phase of growth
and in some species persist during stationary phase. The cells
of some species are short (1–2 mm). In certain cases, helical
cells may be very tightly coiled, or the coils may show continuous variation in amplitude. Spherical cells ~300 nm in diameter
and nonhelical filaments are frequently seen in the stationary
phase, where they may not be viable, and in all growth phases
in suboptimal growth media, where they may or may not be
viable. In some species during certain phases, spherical forms
may be the replicating form. Helical filaments are motile, with
flexional and twitching movements, and often show an apparent rotatory motility. Fibrils are associated with the membrane,
but flagellae, periplasmic fibrils, or other organelles of locomotion are absent. Fimbriae and pili observed on the cell surface
of insect- and plant-pathogenic spiroplasmas are believed to
be involved in host-cell attachment and conjugation (Ammar
et al., 2004; Özbek et al., 2003), but not in locomotion. Cells
divide by binary fission, with doubling times of 0.7–37 h. Facultatively anaerobic. The temperature growth range varies
among species, from 5 to 41°C. Colonies on solid media are
frequently diffuse, with irregular shapes and borders, a condition that reflects the motility of the cells during active growth
(Figure 111). Colony type is strongly dependent on the agar
concentration. Colony sizes vary from 0.1 to 4.0 mm in diameter. Colonies formed by nonmotile variants or mutants, or by
cultures growing on inadequate media are typically umbonate
with diameters of 200 mm or less. Some species, such as Spiroplasma platyhelix, have barely visible helicity along most of their
length and display little rotatory or flexing motility. Colonies
of motile, fast-growing spiroplasmas are diffuse, often with satellite colonies developing from foci adjacent to the initial site
of colony development. Light turbidity may be produced in
liquid cultures. Chemo-organotrophic. Acid is produced from
glucose. Hydrolysis of arginine is variable. Urea, arbutin, and
FIGURE 111. Colonial morphology of Spiroplasma lampyridicola strain
PUP-lT grown on SP-4 agar under anaerobic conditions for 4 d at 30°C.
The diffuse appearance and indistinct margins reflect the motility of
spiroplasmas during active growth. Bar = 50 mm. (Reprinted with permission from Stevens et al., 1997. Int. J. Syst. Bacteriol. 47: 709–712.)
esculin are not hydrolyzed. Sterol requirements are variable.
An optimum osmolality, usually in the range of 300–800 mOsm,
has been demonstrated for some spiroplasmas. Media containing mycoplasma broth base, serum, and other supplements are
required for primary growth, but after adaptation, growth often
occurs in less complex media. Defined or semi-defined media
are available for some species. Resistant to 10,000 U/ml penicillin. Insensitive to rifampicin, sensitive to erythromycin and
tetracycline. Isolated from the surfaces of flowers and other
plant parts, from the guts and hemolymph of various insects
and crustaceans, and from tick triturates. Also isolated from
vascular plant fluids (phloem sap) and insects that feed on the
fluids. Specific host associations are common. The type species, Spiroplasma citri, is pathogenic for citrus (e.g., orange and
Genus I. Spiroplasma
grapefruit), producing “stubborn” disease. Experimental or
natural infections also occur in horseradish, periwinkle, radish,
broad bean, carrot, and other plant species. Spiroplasma kunkelii
is a maize pathogen. Some species are pathogenic for insects.
Certain species are pathogenic, under experimental conditions, for a variety of suckling rodents (rats, mice, hamsters and
rabbits) and/or chicken embryos. Genome sizes vary from 780
to 2220 kbp (PFGE).
DNA G+C content (mol%): 24–31 (Tm, Bd).
Type species: Spiroplasma citri Saglio, L’Hospital, Laflèche,
Dupont, Bové, Tully and Freundt 1973, 202AL.
Morphology. The morphology of spiroplasmas is most easily observed in suspensions with the light microscope under
dark-field illumination (Williamson and Poulson, 1979). In the
exponential phase in liquid media, most spiroplasma cells are
helical filaments 90–250 nm in diameter and of variable length
(Figure 112). Fixed and negatively stained cells usually show
a blunt and a tapered end (Williamson, 1969; Williamson and
Whitcomb, 1974). The tapered ends of the cells are a consequence of the constriction process preceding division (Garnier
et al., 1981, 1984). However, they are adapted as attachment
sites in some species (Ammar et al., 2004).
Motility. Helical spiroplasma cells exhibit flexing, twitching,
and apparent rotation about the longitudinal axis (Cole et al.,
1973; Davis and Worley, 1973). Spiroplasmas exhibit temperature-dependent chemotactic movement toward higher concentrations of nutrients, such as carbohydrates and amino acids
FIGURE 112. Electron micrograph of Drosophila willistoni strain B3SR
sex-ratio spiroplasmas. Hemolymph suspension in phosphate buffered
saline, glutaraldehyde vapor-fixed, and negatively stained with 1% phosphotungstic acid, pH 7.2. (Reprinted with permission from Whitcomb
et al., 2007. Biodiversity and Conservation 16: 3877–3894.)
655
(Daniels and Longland, 1984, 1980); but motility is random
in the absence of attractants (Daniels and Longland, 1984).
Both natural (Townsend et al., 1980b, 1977) and engineered
(Cohen et al., 1989; Duret et al., 1999; Jacob et al., 1997) motility mutants have been described. These mutants form perfectly
umbonate colonies on solid medium. Mutational analysis has
highlighted the involvement of the smc1 gene in motility. Jacob
et al. (1997) demonstrated that a Tn4001 insertion mutant with
reduced flexional motility and no rotational motility could be
complemented with the wild-type scm1 gene. The scm1 gene
encodes a 409 amino acid polypeptide having ten transmembrane domains but no significant homology with known proteins. In another study, the scm1 gene was inactivated through
homologous recombination, abolishing motility (Duret et al.,
1999). The disrupted scm1− mutant was injected into the leafhopper vector (Circulifer haematoceps); it multiplied actively in
the insect vector and was then transmitted to periwinkle plants.
The mutant induced symptoms that were indistinguishable
from those caused by the motile wild-type strain showing that
spiroplasma motility is not essential for phytopathogenicity and
transmission to the plant host (Duret et al., 1999).
Fibrils and motility. Microfibrils 3.6 nm in width have been
envisioned in the membranes of some spiroplasmas. These
structures have repeat intervals of 9 nm along their lengths
(Williamson, 1974) and form a ribbon that extends the entire
length of the helix (Charbonneau and Ghiorse, 1984; Williamson et al., 1984). The sequence of the fibril protein gene has
been determined (Williamson et al., 1991) and the calculated
mass of the fibril protein is 59 kDa. The flat, monolayered,
membrane-bound ribbon composed of several well-ordered
fibrils represents the internal spiroplasmal cytoskeleton. The
spiroplasmal cytoskeletal ribbon follows the shortest helical
line on the cellular coil. Recent studies have focused on the
detailed cellular and molecular organization of the cytoskeleton in Spiroplasma melliferum and Spiroplasma citri (Gilad et al.,
2003; Trachtenberg, 2004; Trachtenberg et al., 2003a, b; Trachtenberg and Gilad, 2001). Each cytoskeletal ribbon contains
seven fibril pairs (or 14 fibrils) and the functional unit is a pair
of aligned fibrils (Trachtenberg et al., 2003a). Paired fibrils can
be viewed as chains of tetramers composed of 59 kDa monomers. Cryo-electron tomography has been used to elucidate
the native state, cytoskeletal structure of Spiroplasma melliferum
and suggested the presence of three parallel ribbons under
the membrane: two appear to be composed of the fibril protein and the third is composed of the actin-like MreB protein
(Kürner et al., 2005). Subsequent studies suggest the presence
of a single ribbon structure (Trachtenberg et al., 2008). The
subunits in the fibrils undergo conformational changes from
circular to elliptical, which results in shortening of the fibrils
and helix contraction, or from elliptical to circular, leading to
a length increase of the fibrils and cell helix. The cytoskeleton,
which is bound to the spiroplasmal membrane over its entire
length, acts as a scaffold and controls the helical shape of the
cell. The cell shape is therefore dynamic. Movement appears to
be driven by the propagation of a pair of kinks that travel down
the length of the cell along the fibril ribbons (Shaevitz et al.,
2005; Wada and Netz, 2007; Wolgemuth and Charon, 2005).
The contractile cytoskeleton can thus be seen as a “linear
motor” in contrast to the common “rotary motor” that is part
of the flagellar apparatus in bacteria (Trachtenberg, 2006).
656
There are several adherent proteins that copurify with the
cytoskeleton, ranging in size from 26 to 170 kDa (Townsend
et al., 1980a; Trachtenberg, 2006; Trachtenberg and Gilad,
2001). These proteins are apparently membrane-associated
and may function as anchor proteins (Trachtenberg and Gilad,
2001). The structural organization of the cytoskeleton-associated proteins of Spiroplasma melliferum is beginning to be elucidated (Trachtenberg et al., 2008). The 59 kDa polypeptide
is the cytoskeletal fibril protein. The 26 kDa polypeptide is
probably spiralin, the major spiroplasmal membrane protein.
However, the involvement of spiralin in helicity and motility is
unlikely (see “Spiralin” section below), especially since spiralin is anchored on the outside surface of the cell (Bévén et al.,
1996; Bové, 1993; Brenner et al., 1995; Foissac et al., 1996) and
spiralin-deficient mutants maintain helicity and motility (Duret
et al., 2003). The 45 kDa protein may correspond to the product of the scm1 gene, shown to be essential for motility (Jacob
et al., 1997), and the 34 kDa protein may be the product of the
mreB1 gene (W. Maccheroni and J. Renaudin, unpublished).
MreB is the bacterial homolog to eukaryotic actin (Jones et al.,
2001; Van den Ent et al., 2001). Early work provided evidence
for the presence of actin-like proteins in spiroplasmas. Antisera
prepared against SDS-denatured invertebrate actin coupled to
horseradish peroxidase specifically stained cells of Spiroplasma citri
(Williamson et al., 1979a). Also, a protein with a molecular mass
similar to that of actin (protein P25) was isolated from Spiroplasma
citri and reacted with IgG directed against rabbit actin (Mouches
et al., 1982b, c, 1983b). Monospecific antibodies raised against
the P25 protein recognized not only P25 of Spiroplasma citri, but
also a homologous protein from Mycoplasma mycoides PG50 and
Ureaplasma urealyticum serotype V (Mouches et al., 1983b). More
recent work has focused on the molecular organization of the
genes. mreB genes are present in rod-shaped, filamentous, and
helical bacteria, but not in coccoid, spherical bacteria, regardless of whether or not they are Gram-stain-positive or Gramstain-negative. mreB genes are also absent from the pleomorphic
mycoplasmas. However, Spiroplasma citri contains five homologs
of Bacillus subtilis mreB genes (Maccheroni et al., 2002). Four of
these (mreB2, 3, 4, and 5) form a cluster on the genome and are
transcribed in two separate operons. Gene mreB1 is transcribed as
a monocistronic operon and at a much higher level.
Growth characteristics. Spiroplasma cells increase in length
and divide by constriction. Pulse labeling of the membrane with
tritiated amino acids revealed a polar growth of the helix. Polarity was also observed by tellurium-labeling of oxido-reduction
sites (Garnier et al., 1984). In the stationary or death phase, the
cells are usually distorted, often forming either subovoid bodies or nonhelical filaments. Within cultured insect cells, all the
spiroplasma cells were subovoid, but presumably viable (Wayadande and Fletcher, 1998). Thus, the ability of cells to grow and
divide is not linked inextricably to helicity.
Growth rate. Enumerated microscopically (Rodwell and
Whitcomb, 1983), spiroplasmas reach titers of 108–1011 cells/
ml in medium containing horse or fetal bovine serum. Growth
rates of related strains tend to be similar. Konai et al. (1996a)
calculated doubling times from the time required for medium
acidification. In general, spiroplasmas adapted to complex
cycles or single hosts had slower growth rates than spiroplasmas
known or suspected to be transmitted on plant surfaces.
Temperature. Konai et al. (1996a) determined temperature
ranges and optima for a large number of spiroplasma strains.
The ranges of some strains (e.g., Spiroplasma apis) were very
wide (5–41°C), but some group I strains from leafhoppers and
plants grew only at 25° and 30°C. Although some spiroplasmas
grew well at 41°C, none grew at 43°C.
Biochemical reactions. All tested spiroplasmas ferment
glucose with concomitant acid production, although the utilization rates may vary. Some strains of group I (e.g., members
of subgroups I-4 and I-6) and all strains of Spiroplasma mirum
ferment glucose slowly. With Spiroplasma citri, all strains tested
grew actively on fructose and strain GII3 grew on fructose, glucose, or trehalose. The ability of spiroplasmas to utilize arginine varies (Hackett et al., 1996a). Arginine hydrolysis by some
spiroplasmas can be observed only if glucose is also present
in the medium. In other cases, aggressive glucose metabolism
interferes with detection of arginine hydrolysis (Hackett et al.,
1996a).
Regulation of the fructose and trehalose operons of Spiroplasma
citri. The fructose operon of Spiroplasma citri (Gaurivaud
et al., 2000a) became of special interest when fructose utilization was implicated in Spiroplasma citri phytopathogenicity (see
“Mechanism of Spiroplasma citri phytopathogenicity” below).
In particular, the role of the first gene of the operon, fruR,
was investigated. In vivo transcription of the operon is greatly
enhanced by the presence of fructose in the growth medium,
whereas glucose has no effect. When fruR is not expressed
(fruR− mutants), transcription of the operon is not stimulated
by fructose and the rate of fructose fermentation is decreased,
indicating that FruR is an activator of the fructose operon (Gaurivaud et al., 2001). Trehalose is the major sugar in leafhoppers
and other insects. The trehalose operon of Spiroplasma citri has
a gene organization very similar to that of the fructose operon
and the first gene of the trehalose operon, treR, also encodes a
transcriptional activator of the operon (André et al., 2003).
Sterol utilization. It was originally thought that all spiroplasmas require sterol for growth. Subsequent screening by Rose
et al. (1993) showed that a minority of the spiroplasmas tested
were able to sustain growth in mycoplasma broth base medium
without sterols. The discovery that the sterol requirement in
Mollicutes is polyphyletic greatly diminished the significance of
sterol requirements in mollicute taxonomy (Tully et al., 1993).
Metabolic pathways and enzymes. The intermediary meta
bolism of Mollicutes has been reviewed (Miles, 1992; Pollack,
2002a, b; Pollack et al., 1997). Like all mollicutes, Spiroplasma
species apparently lack both cytochromes and, except for malate
dehydrogenase, the enzymes of the tricarboxylic acid cycle.
They do not have an electron-transport system and their respiration is characterized as being flavin-terminated. McElwain
et al. (1988) studied Spiroplasma citri and Pollack et al. (1989)
screened ten spiroplasma species for 67 enzyme activities. All
spiroplasmas were fermentative; their 6-phosphofructokinases
(6-PFKs) required ATP for substrate phosphorylation during
glycolysis. This enzymic requirement is common to all mollicutes except Acholeplasma and Anaeroplasma spp. The 6-PFKs of
the species in these genera require pyrophosphate and cannot
use ATP. Additionally, except for Spiroplasma floricola, all Spiroplasma species have dUTPase activity. Pollack et al. (1989) also
reported that all spiroplasmas except Spiroplasma floricola have
deoxyguanosine kinase activity. They found that deoxyguanosine, but no other nucleoside, could be phosphorylated to GMP
with ATP.
Spiroplasmal proteins with multiple functions have been
described. The CpG-specific methylase from Spiroplasma monobiae appears to also have topoisomerase activity (Matsuo et al.,
1994). Protein P46 of Spiroplasma citri is a bifunctional protein
in which the N-terminal domain represents ribosomal protein L29, whereas the C-terminal domain is capable of binding a specific inverted repeat sequence. It could be involved
in regulation (Le Dantec et al., 1998). Such protein multifunctionality may reflect genomic economy in the small mollicute
genome (Pollack, 2002b). However, functional redundancy has
also been reported; Spiroplasma citri apparently has two distinct
membrane ATPases (Simoneau and Labarère, 1991).
Genome size, genomic maps, and chromosomal rearrangements. PFGE revealed that the genome size range for spiroplasmas varied continuously (Pyle and Finch, 1988) from 780
kbp for Spiroplasma platyhelix to 2220 kbp for Spiroplasma ixodetis
(Carle et al., 1995, 1990). There is a general trend for genomic
simplification in Spiroplasma lineages. This trend culminated in
loss of helicity and motility in the Entomoplasmataceae and eventually to the host transfer events forming the mycoides group of
mycoplasmas (Gasparich et al., 2004).
The genome size of Spiroplasma citri varies among strains
from 1650 to 1910 kbp (Ye et al., 1995). It was found that the
relative positions of mapped loci were conserved in most of the
strains, but that differences in the sizes of certain fragments
permitted genome size variation. Genome size can fluctuate
rapidly in spiroplasma cultures after a relatively short number
of in vitro passages (Melcher and Fletcher, 1999; Ye et al., 1996).
The genome of Spiroplasma melliferum is 360 kbp shorter than
that of Spiroplasma citri strain R8-A2T, but DNA hybridization has
shown that the two spiroplasmas share extensive DNA hybridization (65%). Comparison of their genomic maps revealed that
the genome region, which is shorter in Spiroplasma melliferum,
corresponds to a variable region in the genomes of Spiroplasma
citri strains and that a large region of the Spiroplasma melliferum
genome is inverted in comparison with Spiroplasma citri. Therefore, chromosomal rearrangements and deletions were probably major events during evolution of the genomes of Spiroplasma
citri and Spiroplasma melliferum. In addition, a large amount of
noncoding DNA is present as repeat sequences (McIntosh et al.,
1992; Nur et al., 1986, 1987) and integrated viral DNA (Bébéar
et al., 1996) may also account for differences in genome sizes of
closely related species.
Base composition. The DNA G+C content for most spiroplasma groups and subgroups has been determined (Carle
et al., 1995, 1990; Williamson et al., 1998). Most group I spiroplasmas and Spiroplasma poulsonii have a G+C content of 25–27
mol%. However, the G+C content of subgroup I-6 Spiroplasma
insolitum is significantly higher, indicating that the base composition of spiroplasmal DNA may shift over relatively short evolutionary periods. The range of G+C content of 25–27 mol%
is modal for Spiroplasma and is also common in the Apis clade.
However, Spiroplasma mirum (group V), strains of Spiroplasma
apis (group IV), and group VIII strains have a G+C content of
about 29–31 mol%. Restriction sites containing only G and C
657
nucleotides are not uniformly distributed over the genome (Ye
et al., 1992).
Methylated bases. Methylated bases have been detected in
spiroplasmal DNA (Nur et al., 1985). The gene encoding the
CpG methylase in Spiroplasma monobiae has been cloned (Renbaum et al., 1990) and its mode of action studied (Renbaum
and Razin, 1992).
DNA restriction patterns. Restriction patterns of spiroplasmal DNA, as determined by polyacrylamide gel electrophoresis,
may be highly similar among strains of a given species (Bové
et al., 1989). Variations in restriction fragment length patterns
among strains of Spiroplasma corruscae correlated imperfectly
with serological variation, so their significance was uncertain
(Gasparich et al., 1998).
RNA genes. Some spiroplasmas, such as Spiroplasma citri,
have only one rRNA operon, whereas others, such as Spiroplasma apis, have two (Amikam et al., 1984, 1982; Bové, 1993;
Grau et al., 1988; Razin, 1985). The three rRNA genes are
linked in the classical order found in bacteria: 5¢-16S–23S-5S-3¢.
The sequence of the 16S rRNA gene (rDNA) of most spiroplasma species has been determined for phylogenetic studies
(Gasparich et al., 2004; Weisburg et al., 1989). A gene cluster of
ten tRNAs (Cys, Arg, Pro, Ala, Met, Ile, Ser, fMet, Asp, Phe) was
identified in Spiroplasma melliferum (Rogers et al., 1987). Similar tRNA gene clusters have been cloned and sequenced from
Spiroplasma citri (Citti et al., 1992).
Codon usage. In spiroplasmas, UGA is not a stop codon but
encodes tryptophan. The universal tryptophan codon, UGG, is
also used (Citti et al., 1992; Renaudin et al., 1986). Codon usage
also reflects the A+T richness of spiroplasmal DNA (usually
about 74 mol% A+T). For example, in Spiroplasma citri, UGA is
used to code for tryptophan eight times more frequently than
the universal tryptophan codon UGG (Bové, 1993; Citti et al.,
1992; Navas-Castillo et al., 1992). Also, synonymous codons with
U or A at the 5¢ or 3¢ ends are preferentially used over those
with a C or G in that position.
RNA polymerase and spiroplasmal insensitivity to rifampicin. Spiroplasmas are insensitive to rifampicin. DNAdependent RNA polymerases from Spiroplasma melliferum and
Spiroplasma apis were at least 1000 times less sensitive to rifampicin than the corresponding Escherichia coli enzyme (Gadeau
et al., 1986). Rifampicin insensitivity of Spiroplasma citri and
all other mollicutes tested was found to be associated with the
presence of an asparagine residue at position 526 in RpoB.
The importance of the asparagine residue was confirmed by
site-directed mutagenesis of the histidine codon (CAC) to
an asparagine codon (AAC) at position 526 of Escherichia coli
RpoB, resulting in a rifampicin-resistant mutant (Gaurivaud
et al., 1996). The genetic organization surrounding the rpoB
gene in spiroplasmas is also atypical. In many bacteria, rpoB is
part of the b operon in which the four genes rplK, rplA, rplJ,
and rplL, encoding ribosomal proteins L11, L1, L10, and L12,
respectively, are located immediately upstream of rpoB; rpoC is
immediately downstream of rpoB. In Spiroplasma citri, the gene
organization is different in that the hsdS gene, encoding a component of a type I restriction-modification system, is upstream
of rpoB. Sequences showing similarities with insertion elements
are found between hsdS and rpoB (Laigret et al., 1996).
658
DNA polymerases and other proteins involved in DNA replication and repair. From genomic studies, it appears that
Mycoplasma species carry the essential, multimeric enzyme for
genomic DNA replication, DNA polymerase III. The subunit
responsible for actual DNA biosynthesis is subunit a, encoded
by polC (dnaE). The polC gene has been identified in all
sequenced mollicute genomes, including Spiroplasma citri. The
genes encoding the other subunits, dnaN (subunit b) and dnaX
(subunits t and g), are also shared by the Spiroplasma and Mycoplasma species studied to date. So, it seems that spiroplasmas,
like other mollicutes, possess DNA polymerase III and that it is
probably the major DNA replication enzyme. However, there
is also evidence for two additional DNA polymerases. A second
gene for a DNA polymerase (enzyme B) was found in the Spiroplasma citri genome and there is evidence that the Spiroplasma
kunkelii polA gene may encode a full-length DNA polymerase
I protein (Bai and Hogenhout, 2002). DNA polymerase I is a
single polypeptide that has, in addition to DNA synthesis activity, two exonuclease activities: exo-3¢ to 5¢ as well as exo-5¢ to
3¢. At this stage, it is not possible to determine the equivalence
between the three spiroplasmal DNA polymerases identified by
sequencing (Pol III, enzyme B, and Pol I) and those originally
detected biochemically (ScA, ScB, ScC) (Charron et al., 1979,
1982). As the Spiroplasma citri genome sequencing project has
progressed, the following Spiroplasma citri genes involved in
DNA replication have been detected: dnaA, dnaB, polA, dnaE,
polC, dnaN, dnaX, holB, dinB (truncated), dnaJ, dnaK, gyrA, gyrB,
parC, parE, topA, rnhB, rnhC, rnpA, rnR, rnc, yrrc, xseA, xseB, and ssb
(Carle et al., 2010; accession numbers AM285301–AM285339).
Genes encoding DNA replication proteins have also been identified in Spiroplasma kunkelii (Bai and Hogenhout, 2002). Spiroplasma citri is highly sensitive to UV irradiation (Labarère and
Barroso, 1989) and the organism has no functional recA gene,
since a significant portion of the C-terminal part of the gene is
lacking (Marais et al., 1996).
region and an amino acid sequence repetition, including a
VTKXE consensus sequence, are present in all spiralins analyzed
(Foissac et al., 1997a). Spiralin confers a significant amount of
the antigenic activity in group I spiroplasmas (Whitcomb et al.,
1983) and has a high degree of species specificity, although
minor cross-reactions have been detected (Zaaria et al., 1990).
The spiralin genes of Spiroplasma citri and Spiroplasma melliferum
species, which have about 65% overall DNA–DNA hybridization,
shared 89% nucleotide sequence identity and 75% deduced
amino acid sequence similarity (Bové et al., 1993).
Spiralin mutants were constructed through homologous
recombination in Spiroplasma citri to examine the role of spiralin in vivo (Duret et al., 2003). Phenotypic characterization
of mutant 9a2 showed that, in spite of a total lack of spiralin, it
maintained helicity and motility similar to the wild-type strain
GII3 (Duret et al., 2003). When injected into the leafhopper
vector, Circulifer haematoceps, the mutant multiplied to a high
titer, but transmission efficiency to periwinkle plants was very
low compared to the wild-type strain. In the infected plants,
however, the spiralin-deficient mutant multiplied well and produced the typical symptoms of the disease. In addition, preliminary results indicated that the mutant could not be acquired
by insects feeding on 9a2-infected plants, suggesting that spiralin may mediate spiroplasma invasion of insect tissues (Duret
et al., 2003). In order to test this possibility, Circulifer haematoceps leafhopper proteins were screened as putative Spiroplasma
citri-binding molecules using Far-Western analysis (Killiny et al.,
2005).These experiments showed that spiralin is a lectin capable of binding to insect 50 and 60 kDa mannose glycoproteins.
Hence, spiralin could play a key role in insect transmission of
Spiroplasma citri by mediating spiroplasma adherence to epithelial cells of the insect vector gut or salivary gland (Killiny et al.,
2005). This would also explain why the spiralin-negative mutant
9a2 is poorly transmitted by the vector and is not acquired by
insects feeding on 9a2-infected plants.
Origin of DNA replication. Even before the Spiroplasma
citri genome project was initiated, some fragments with multiple open reading frames had been completely sequenced.
For example, Ye et al. (1994b) sequenced a 5.6 kbp fragment
containing genes for the replication initiation protein (dnaA),
the beta subunit of DNA polymerase III (dnaN), and the DNA
gyrase subunits A and B (gyrA and gyrB). Several dnaA-box consensus sequences were found upstream and downstream of the
dnaA gene. From these data, it was established that the dnaA
region was the origin of replication in Spiroplasma citri (Ye et al.,
1994b). Zhao et al. (2004a) cloned a cell division gene cluster
from Spiroplasma kunkelii and functionally characterized the key
division gene, ftsZsk, and showed that it encodes a cell division
protein similar to FtsZ proteins from other bacteria.
Viruses. Four different virus types have been found in Spiroplasma, SpV1-SpV4. Use of SpV1 viruses for recombinant DNA
studies in Spiroplasma citri is described later in the section on
“Tools for molecular genetics of Spiroplasma citri ”.
Cells of many spiroplasma species contain filamentous/rodshaped viruses (SVC1 = SpV1) that are associated with nonlytic
infections (Bové et al., 1989; Ranhand et al., 1980; Renaudin
and Bové, 1994). They belong to the Plectrovirus group within
the Inoviridae. SpV1 viruses have circular, single-stranded DNA
genomes (7.5 to 8.5 kbp), some of which have been sequenced
(Renaudin and Bové, 1994). SpV1 sequences also occur as
prophages in the genome of the majority of Spiroplasma citri
strains studied (Renaudin and Bové, 1994). These insertions
take place at numerous sites in the chromosomes of Spiroplasma
citri (Ye et al., 1992) and Spiroplasma melliferum (Ye et al., 1994a).
The SpV1-ORF3 and the repeat sequences could be part of an
IS-like element of chromosomal origin. Resistance of spiroplasmas to virus infection may be associated with integration of
viral DNA sequences in the chromosome or extrachromosomal
elements (Sha et al., 1995). The evolutionary history of these
viruses is unclear, but there is some evidence for virus and plasmid co-evolution in the group I Spiroplasma species (Gasparich
et al., 1993a) and indications of potentially widespread horizontal transmission (Vaughn and de Vos, 1995). Virus infection of
spiroplasma cells can pose problems in cultures. For example,
Spiralin. Spiralin, encoded by the spi gene, is the major
membrane protein of Spiroplasma citri (Wróblewski et al., 1977,
1989). The deduced amino acid sequence of the protein (Bové
et al., 1993; Chevalier et al., 1990; Saillard et al., 1990) corresponds well with the experimentally determined amino acid
composition (Wróblewski et al., 1984). In particular, spiralin
lacks tryptophan and, thus, has no UGG and/or UGA codons,
which facilitates gene expression in Escherichia coli. Detailed
analyses showed that all Spiroplasma citri spiralins were 241–242
amino acids long (Foissac et al., 1996). A conserved central
lyophilized early passages of Spiroplasma citri R8-A2T proved
difficult to grow and electron microscopy revealed that these
cells carried large numbers of virions of virus SpV1-R8A2 (Cole
et al., 1974). Likewise, SpV1 viruses have been found in Spiroplasma poulsonii (Cohen et al., 1987) and Spiroplasma melliferum
(Liss and Cole., 1981); the Spiroplasma melliferum SpV1-KC3
virus forms plaques on various strains of Spiroplasma melliferum,
including the type strain BC-3T.
A second virus, reminiscent of a type B tailed bacterial virus,
occurs in a small number of Spiroplasma citri strains (Cole et al.,
1973). This SCV2 (= SpV2) virus is a polyhedron with a long,
noncontractile tail. It may be associated with lytic infection.
Infections in which large numbers of virions of SpV2 viruses are
produced tend to be irregular and difficult to maintain under
experimental conditions, so this is the least studied of the spiroplasma viruses.
A third virus (SpV3) forms polyhedral virions with short
tails and has been found in many strains of Spiroplasma citri
(Cole, 1979, 1977, 1974). The SpV3 genome is a linear double-stranded DNA molecule of 16 kbp, which can circularize
to form a covalently closed molecule with single-stranded gaps,
indicating that the linear molecule has cohesive ends. There
is significant diversity among SpV3 viruses, extending even to
major differences in genome sizes. Virus SpV3-AV9/3 was isolated from Spiroplasma citri strain ASP-9 (Stephens, 1980). Dickinson and Townsend (1984) isolated the SpV3 virus from plants
infected with Spiroplasma citri. This virus, when plated on cells of
Spiroplasma citri, had a plaque morphology typical of temperate
phages. In spiroplasma cells that have been lysogenized, complete virus genomes may be integrated into the spiroplasma
chromosome. These cells are then immune to superinfection
by the lysogenizing virus, but susceptible to other SpV3 viruses.
It is possible that lysogenization of Spiroplasma citri by SpV3-ai
affects spiroplasma pathogenicity, particularly with respect to
attenuation. Drosophila spiroplasmas, male-lethal or nonlethal,
usually carry SpV3 viruses. Each strain of Drosophila spiroplasma
carries an associated virus that is lytic to certain other strains
(Oishi et al., 1984).
A fourth virus (SpV4), with a naked, icosahedral nucleocapsid 25 nm in diameter, was discovered (Ricard et al., 1982)
in the B63 strain of Spiroplasma melliferum. SpV4 has a circular, single-stranded DNA genome (Renaudin and Bové, 1994;
Renaudin et al., 1984a, b) and is a lytic Spiromicrovirus within
the Microviridae (Chipman et al., 1998). Infection with this virus
results in very clear plaques, indicating a lytic process. Host
range studies (Renaudin et al., 1984a, b) have shown that only
Spiroplasma melliferum is susceptible to SpV4. Two strains of Spiroplasma melliferum, including the type strain BC-3T and B63, are
not susceptible, as no plaques were formed on lawns of these
spiroplasmas. These strains could be infected by transfection
suggesting that resistance to the whole virus occurred at the
level of adsorption or penetration of the virus (Renaudin and
Bové, 1994; Renaudin et al., 1984b).
Genome sequencing. Genomic DNA sequencing efforts
for two Spiroplasma species are in progress. For Spiroplasma citri
GII3 (Carle et al., 2010; Saillard et al., 2008), assembly of 20,000
sequencing reads obtained from shotgun and chromosome
specific libraries yielded: (1) 39 chromosomal contigs totalling
1525 kbp of the 1820 kbp Spiroplasma citri GII3 chromosome as
well as (2) 8 circular contigs, which proved to represent seven
659
plasmids: pSciA (7.8 kbp), pSci1 to pSci6 (12.9 to 35.3 kbp),
and one viral RF DNA (SVTS2). The chromosomal contigs contained 1905 putative genes or coding sequences (CDS). Of the
CDS-encoded proteins, 29% are involved in cellular processes,
cell metabolism, or cell structure. CDS for viral proteins and
mobile elements represented 24% of the total, whereas 47% of
the CDS were for hypothetical proteins with no known function;
21% of the total CDS appeared truncated as compared to their
bacterial orthologs. Families of paralogs were mainly clustered
in a large region of the chromosome opposite the origin of replication. Eighty-four CDS were assigned to transport functions,
including phosphoenolpyruvate phosphotransferase systems
(PTS), ATP binding cassette (ABC) transporters, and ferritin. In
addition to the general enzymes EI and HPr, glucose- fructoseand trehalose-specific PTS permeases, and glycolytic and ATP
synthesis pathways, Spiroplasma citri possesses a Sec-dependent
protein export system and a nearly complete pathway for terpenoid biosynthesis. The sequencing of the Spiroplasma kunkelii
CR2-3x genome (1.55 Mb) is also nearing completion (http://
www.genome.ou.edu/spiro.html); the physical and genetic
maps have been published (Dally et al., 2006). Several studies
have begun to focus on gene content and genomic organization (Zhao et al., 2003, 2004a, b). Results show that, in addition
to virus SpV1 DNA insertions, the Spiroplasma kunkelii genome
harbors more purine and amino acid biosynthesis, transcriptional regulation, cell envelope, and DNA transport/binding
genes than Mycoplasmataceae (e.g., Mycoplasma genitalium and
Mycoplasma pneumoniae) genomes (Bai and Hogenhout, 2002).
Plasmids. Several plasmids have been discovered in spiroplasmas (Archer et al., 1981; Gasparich and Hackett, 1994;
Gasparich et al., 1993a; Mouches et al., 1984a; Ranhand et al.,
1980). They are especially common in spiroplasmas of group
I. Eight extrachromosomal elements, including seven plasmids, were discovered during the Spiroplasma citri GII3 genome
sequencing project. The six largest plasmids, pSci1 to pSci6,
range from 12.9 to 35.3 kb (Saillard et al., 2008). In silico
analyses of plasmid sequences revealed that they share extensive regions of homology and display a mosaic gene organization. Genes encoding proteins of the TraD-TraG, TrsE-TraE,
and Soj-ParA protein families, were predicted in most of the
pSci sequences. The presence of such genes, usually involved
in chromosome integration, cell to cell DNA transfer, or DNA
element partitioning; suggests that these molecules could be
inherited vertically as well as horizontally. The largest plasmid,
pSci6, encodes P32 (Killiny et al., 2006), a membrane-associated protein interestingly absent in all insect non-transmissible
strains tested so far. The five remaining plasmids (pSci1 to
pSci5) encode eight different Spiroplasma citri adhesion-related
proteins. The complete sequences of plasmids pSKU146 from
Spiroplasma kunkelii CR2-3x and pBJS-O from Spiroplasma citri
BR3 have been reported (Davis et al., 2005; Joshi et al., 2005).
These large plasmids, like the above Spiroplasma citri plasmids,
encode an adhesin and components of a type IV translocationrelated conjugation system. Characterizing the replication and
stability regions of Spiroplasma citri plasmids resulted in the identification of a novel replication protein, suggesting that Spiroplasma citri plasmids belong to a new plasmid family and that
the soj gene is involved in segregational stability of these plasmids (Breton et al., 2008a). Similar replicons were detected in
various spiroplasmas of group I, such as Spiroplasma melliferum,
660
Spiroplasma kunkelii, Spiroplasma sp. 277F, and Spiroplasma phoeniceum, showing that they are not restricted to plant pathogenic
spiroplasmas.
Tools for molecular genetics of Spiroplasma citri. Recent
recombinant DNA tools are described in this section.
Several reports have been published concerning the use
of SpV1 viruses as tools to introduce recombinant DNA into
spiroplasmas, including optimization of transfection conditions
(Gasparich et al., 1993b). The replicative form of SpV1 was
used to clone and express the Escherichia coli-derived chloramphenicol acetyltransferase (cat) gene in Spiroplasma citri. Both
the replicative form (RF) and the virion DNA produced by the
transfected cells contained the cat gene sequences (Stamburski et al., 1991). The G fragment of the Mycoplasma pneumoniae
cytadhesin P1 gene could also be expressed in Spiroplasma citri
(Marais et al., 1993) using a similar method. However, the
recombinant RF proved unstable, resulting in the loss of the
DNA insert (Marais et al., 1996).
Recombinant plasmids have also been developed to introduce genes into Spiroplasma citri cells. The introduced genes
include antibiotic resistance markers and wild-type genes to
complement auxotrophic mutants. Most recombinant plasmids contain the origin of DNA replication (oriC) of the Spiroplasma citri chromosome (Ye et al., 1994b). One such plasmid
is pBOT1 (Renaudin, 2002; Renaudin et al., 1995). This plasmid contains a 2 kbp oriC region, a tetracycline resistance gene
(tetM) from Tn916, and the linearized Escherichia coli plasmid
pBS with a colE1 origin of replication. Because of its two origins of replication, oriC and colE1, pBOT1 is able to shuttle
between Spiroplasma citri and Escherichia coli. When introduced
into Spiroplasma citri, pBOT1 replicates first as a free extrachromosomal element, but later integrates into the chromosome via
homologous recombination involving a single crossover event
in the oriC region. Once integrated into the host chromosome,
the whole plasmid is stably maintained. Recent studies suggest
that the broad host range Spiroplasma citri GII3 plasmids and
their shuttle derivatives may have significant advantages over
oriC plasmids for gene transfer and expression in spiroplasmas
(Breton et al., 2008a). They transform Spiroplasma citri (as well
as Spiroplasma kunkelii and Spiroplasma phoeniceum) strains at
relatively high efficiencies, the growth of the transformants is
not significantly affected, they do not integrate into the chromosome, and their stability/loss can be modulated depending
upon the presence/absence of the soj gene.
Spiroplasma citri mutants have been produced by random and
targeted approaches. The transposon Tn4001 has been used
successfully for random mutagenesis of Spiroplasma citri (Foissac
et al., 1997c). For targeted gene inactivation, plasmids derived
from pBOT1 have been used to disrupt genes (e.g., fructose
operon, motility gene scm1) through homologous recombination involving a single crossover event (Duret et al., 1999;
Gaurivaud et al., 2000c). More recently, Lartigue et al. (2002)
developed vector pC2, in which the oriC fragment was reduced
to the minimal sequence needed to promote plasmid replication; this vector increases recombination frequency at the target gene. To avoid the extensive passaging that was required for
recombination prior to transformant screening, vector pC55
was designed using a selective tetracycline resistance marker
that is only expressed after the plasmid has integrated into the
chromosome at the target gene. This approach was used to
inactivate the spiralin gene (spi) and the gene encoding the
IICB component of the glucose phosphotransferase system
permease (ptsG) (André et al., 2005; Duret et al., 2003; Lartigue et al., 2002). Another series of recombinant plasmids, the
pGOT vectors, allow for selection of rare recombination events
by using two distinct selective markers. First, transformants are
screened for their resistance to gentamicin and next, site-specific recombinants are selected for based on their resistance to
tetracycline, which can only be expressed through recombination at the target gene. In this way, inactivation of the crr gene,
encoding the glucose phosphotransferase permease IIA component, was obtained (Duret et al., 2005). The use of the transposon gd TnpR/res recombination system to produce unmarked
mutations (i.e., without insertion of antibiotic markers) in Spiroplasma citri was demonstrated by the production of a disrupted
arcA mutant (Duret et al., 2005); arcA encodes arginine deiminase. In this system, the target gene is disrupted by integration
of a plasmid containing target gene sequences along with the
tetM gene flanked by binding-specific recombination (res) sites.
After integration of the plasmid, a second plasmid is introduced
that encodes the resolvase TnpR. TnpR mediates the resolution
of the cointegrate at the res sites, thereby removing tetM but
leaving behind a mutated version of the target gene. The TnpRencoding plasmid is lost spontaneously when selective pressure
is removed.
Antigenic structure. Growth inhibition tests (Whitcomb
et al., 1982) were used in the early years to identify spiroplasma
species or groups, but metabolism inhibition (Williamson et al.,
1979b; Williamson and Whitcomb, 1983) and deformation tests
(Williamson et al., 1978) are now used almost exclusively (see
below).
Antigenic variability, which has been described for some
Mycoplasma species (Rosengarten and Wise, 1990; Yogev et al.,
1991), has not been demonstrated in spiroplasmas (R. Rosengarten, personal communication).
Group classification. The classification of spiroplasmas was
first proposed by Junca et al. (1980) and has been revised periodically (Tully et al., 1987; Williamson et al., 1998). These classifications are based on serological reactions of the organisms
in growth inhibition, deformation and metabolism inhibition
tests and/or characteristics of their genomes. Development
of a classification scheme has resulted in the delineation of
spiroplasma groups and subgroups (Table 142). In the scheme,
“groups” have been defined as clusters of similar organisms,
all of which possess negligible DNA–DNA hybridization with
representatives of other groups, but moderate to high levels of
hybridization (20–100%) with each other. Groups are, therefore, putative species. This level of genomic differentiation correlates well with substantial differences in serology. Thirty-four
groups were presented in a revised classification of spiroplasmas in 1998 (Williamson et al., 1998). Four additional groups
(XXXV–XXXVIII) were proposed recently as the result of a
global spiroplasma environmental survey (Whitcomb et al.,
2007) and more are anticipated (Jandhyam et al., 2008). Subgroups have been defined by the International Committee on
Systematics of Bacteria (ICSB) Subcommittee on the Taxonomy
of Mollicutes (ICSB, 1984) as clusters of spiroplasma strains
showing intermediate levels of intragroup DNA–DNA hybridization (10–70%) and possessing corollary serological relationships. Three spiroplasma groups [group I (Junca et al., 1980;
Saillard et al., 1987), group VIII (Gasparich et al., 1993c), and
661
Table 142. Biological properties of spiroplasmasa
Group
Spiroplasma
Strain
ATCC no.
Morphologyb
Genomec
G+Cd
Arge
Dtf
OptTg
Host
I-1
I-2
I-3
I-4
I-5
I-6
I-7
I-8
I-9
S. citri
S. melliferum
S. kunkelii
Spiroplasma sp.
Spiroplasma sp.
S. insolitum
Spiroplasma sp.
S. phoeniceum
S. penaei
R8-A2
BC-3T
E275T
277F
LB-12
M55T
N525
P40T
SHRIMPT
Long helix
Long helix
Long helix
Long helix
Long helix
Long helix
Long helix
Long helix
Helix
1820
1460
1610
1620
1020
1810
1780
1860
nd
26
26
26
26
26
28
26
26
29
+
+
+
+
−
−
+
+
+
4.1
1.5
27.3
2.3
26.3
7.2
4.7
16.8
nd
32
37
30
32
30
30
32
30
28
Phloem/leafhopper
Honey bee
Phloem/leafhopper
Rabbit tick
Plant bug
Flower surface
Green June beetle
Phloem/vector
Pacific white shrimp
DW-1T
27556
33219T
29320T
29761
33649
33502T
33287
43115T
BAA-1082T
(CAIM 1252T)
43153T
II
S. poulsonii
Long helix
1040
26
nd
15.8
30
S. floricola
S. apis
S. mirum
S. ixodetis
S. monobiae
S. syrphidicola
Spiroplasma sp.
S. chrysopicola
S. clarkii
S. culicicola
S. velocicrescens
S. diabroticae
S. sabaudiense
S. corruscae
Spiroplasma sp.
S. cantharicola
Spiroplasma sp.
Spiroplasma sp.
S. turonicum
S. litorale
S. lampyridicola
S. leptinotarsae
OMBG
B31T
SMCAT
Y32T
MQ-1T
EA-1T
TAAS-1
DF-1T
CN-5T
AES-1T
MQ-4T
DU-1T
Ar-1343T
EC-1T
I-25
CC-1T
CB-1
Ar-1357
Tab4cT
TN-1T
PUP-1T
LD-1T
29989T
33834T
29335T
33835T
33825T
33826T
51123
43209T
33827T
35112T
35262T
43210T
43303T
43212T
43262
43207T
43208
51126
700271T
43211T
43206T
43213T
Helix
Helix
Helix
Tight coil
Helix
Minute helix
Minute helix
Minute helix
Helix
Short helix
Short helix
Helix
Helix
Helix
Wave-coil
Helix
Helix
Helix
Helix
Helix
Unstable helix
Motile funnel
1270
1300
1300
2220
940
1230
1170
1270
1720
1350
1480
1350
1175
nd
1380
nd
1320
nd
1305
1370
1375
1085
26
30
30
25
28
30
31
29
29
26
26
25
29
26
26
26
26
26
25
25
25
25
−
+
+
−
−
+
+
+
+
−
−
+
+
−
−
−
−
−
−
−
−
+
0.9
1.1
7.8
9.2
1.9
1.0
1.4
6.4
4.3
1.0
0.6
0.9
4.1
1.5
3.4
2.6
2.6
3.4
nd
1.7
9.8
7.2
37
34.5
37
30
32
32
37
30
30
37
37
32
30
32
30
32
32
30
30
32
30
30
XXI
XXII
XXIII
XXIV
XXV
XXVI
XXVII
XXVIII
XXIX
XXX
XXXI
XXXII
XXXIII
XXXIV
XXXV
XXXVI
XXXVII
XXXVIII
Nd
Spiroplasma sp.
S. taiwanense
S. gladiatoris
S. chinense
S. diminutum
S. alleghenense
S. lineolae
S. platyhelix
Spiroplasma sp.
Spiroplasma sp.
S. montanense
S. helicoides
S. tabanidicola
Spiroplasma sp.
Spiroplasma sp.
Spiroplasma sp.
Spiroplasma sp.
Spiroplasma sp.
S. atrichopogonis
W115
CT-1T
TG-1T
CCHT
CUAS-1T
PLHS-1T
TALS-2T
PALS-1T
TIUS-1
BIUS-1
HYOS-1T
TABS-2T
TAUS-1T
B1901
BARC 4886
BARC 4900
BARC 4908
GSU5450
GNAT3597T
980
1195
nd
1530
1080
1465
1390
780
840
nd
1225
nd
1375
1295
nd
nd
nd
nd
nd
24
26
26
29
26
31
25
29
28
28
28
27
26
25
nd
nd
nd
nd
28
−
−
−
−
−
+
−
+
−
−
+
−
−
−
−
−
−
−
+
4.0
4.8
4.1
0.8
1.0
6.4
5.6
6.4
3.6
0.9
0.7
3.0
3.7
nd
0.6
1.0
1.2
1.5
nd
30
30
31
37
32
30
30
30
30
37
32
32
30
nd
32
30
32
32
30
Drosophila
hemolymph
Plant surface
Honey bee
Rabbit tick
Ixodid tick
Monobia wasp
Syrphid fly
Horse fly
Deer fly
Green June beetle
Mosquito
Monobia wasp
Beetle
Mosquito
Horse fly/beetle
Leafhopper
Cantharid beetle
Cantharid beetle
Mosquito
Horse fly
Horse fly
Firefly
Colorado potato
beetle
Flower surface
Mosquito
Horse fly
Flower surface
Mosquito
Scorpion fly
Horse fly
Dragonfly
Tiphiid wasp
Flower surface
Horse fly
Horse fly
Horse fly
Horse fly
Horse fly
Horse fly
Horse fly
Horse fly
Biting midge
III
IV
V
VI
VII
VIII-1
VIII-3
VIII-2
IX
X
XI
XII
XIII
XIV
XV
XVI-1
XVI-2
XVI-3
XVII
XVIII
XIX
XX
Nd
S. leucomae
nd
24
+
nd
30
Satin moth
T
SMAT
nd, Not determined.
a
For descriptions of morphotypes, see text.
b
Genome size (kbp).
c
DNA G+C content (mol%).
d
, Catabolizes arginine.
e+
Doubling time (h) (Konai et al., 1996a).
f
Optimum growth temperature (°C).
g
T
43260
Helix
43302T
Helix
43525T
Helix
43960T
Helix
49235T
Short helix
51752T
Helix
51749T
Helix
51748T
Wave-coil
51751
Rare helices
51750
Late helices
51745T
Helix
51746T
Helix
51747T
Helix
700283
Helix
BAA-1183
Helix
BAA-1184
Helix
BAA-1187
Helix
BAA-1188
Helix
BAA-520T (NBRC
Helix
100390T)
BAA-521T (NBRC
Helix
100392T)
662
group XVI (Abalain-Colloc et al., 1993)] have been divided into
a total of 15 subgroups. “Serovars” have been defined as genotypic clusters varying substantially in metabolism inhibition and
deformation serology, but that are insufficiently differentiated
from members of existing groups or subgroups to warrant separation. However, with the discovery of a large number of strains
for some groups (e.g., group VIII), the serovar/subgroup picture has become very confused (Regassa et al., 2004; see Phylogeny, below).
Procedures for species descriptions and minimal standards. pecies descriptions of spiroplasmas have been in accord with
S
recommendations of minimum standards proposed by the ICSP
(International Committee on Systematics of Prokaryotes) Subcommittee on the Taxonomy of Mollicutes (Brown et al., 2007).
Cloning. Production of spiroplasma lineages produced
from a single cell or clonings are performed largely by serial
dilution of filtered cultures using 96-well microtiter plates
(Whitcomb et al., 1986; Whitcomb and Hackett, 1987). At a
certain dilution, which varies from plating to plating, the mean
number of cells per well decreases so that fewer than about 8 of
the 96-wells support growth of a spiroplasma clone. Very probably, such clones arise from a single spiroplasmal cell.
Cellular morphology. Using dark-field microscopy, cultures
should appear helical and motile during at least one growth
phase (see “Morphology” and “Motility” above). However, morphological exceptions do occur (see “Differentiating characters” below and reviewed by Gasparich et al., 2004).
16S rRNA gene sequence analysis. Preliminary identification is performed by PCR amplification using universal 16S
rRNA (Gasparich et al., 2004) or other described primers (e.g.,
Fukatsu and Nikoh, 1998; Jandhyam, 2008). DNA sequence
analysis using a blast search provides preliminary placement
within the genus Spiroplasma. Those strains showing close phylogenetic relationships based on 16S rRNA gene sequence analyses should then be screened using serological tests.
Serological tests. The deformation test (Williamson et al.,
1978) is used routinely for serological analyses. Reciprocal titers
of ³320 are generally required for definitive group placement.
Deformation is defined as entire or partial loss of helicity. At the
end point, cells are often seen in which an unaffected part of the
helical filament exhibits flexing motility despite the presence of
a bleb on another part of the cell. The deformation titer is the
reciprocal of the final antiserum dilution that exhibits deformation of ³50% of the cells. Antiserum should be produced
for any strain thought to represent a novel serogroup and any
positive test against characterized groups requires a recriprocal
test using the newly prepared antiserum.
The high levels of specificity and sensitivity of the metabolism inhibition test make it especially useful for defining groups
and subgroups (Williamson et al., 1979b; Williamson and Whitcomb, 1983). Other serological tests have also been employed
for characterization of spiroplasmas. Growth inhibition tests
were used for delineation of spiroplasma groups I through XI
(Whitcomb et al., 1982), but were not used thereafter. Growth
inhibition tests are problematic for spiroplasmas because they
require development of procedures for obtaining colonies. The
spiroplasma motility inhibition test (Hackett et al., 1997) has
proved useful for determination of intraspecific variation in
Spiroplasma leptinotarsae. ELISA has been used for detection of
Spiroplasma kunkelii (Gordon et al., 1985) and Spiroplasma citri
(Saillard and Bové, 1983).
Optimum growth temperature. Optimal growth temperatures between 10 and 41°C have been determined (Konai et al.,
1996a).
Substrate metabolism. The ability to ferment glucose and
produce acid must be examined (Aluotto et al., 1970). The
ability to hydrolyze arginine and produce ammonia should be
assessed (Barile, 1983). See the section on “Biochemical reactions” above for more details.
Ecology. The species description must include ecological
information such as isolation site within the host and cultivation conditions, common and binomial host name, geographical location of host (with GPS), any known interaction between
the spiroplasma and its host, and, in the case of a pathogen,
disease symptoms observed.
Antibiotic sensitivities. In early studies (Bowyer and Calavan, 1974; Liao and Chen, 1981b), spiroplasmas proved to be
especially sensitive in vitro to tetracycline, erythromycin, tylosin,
tobramycin, and lincomycin. Strains have been isolated that are
permanently resistant to kanamycin, neomycin, gentamicin,
erythromycin, and several tetracycline antibiotics (Liao and
Chen, 1981b). Insensitivity to rifampicin has been studied in
relation to its inhibition of transcription (see “RNA polymerase
and spiroplasmal insensitivity to rifampicin” above) and penicillin insensitivity is seen for all spiroplasmas due to the lack of a
cell wall. Natural amphipathic peptides such as Gramicidin S
alter the membrane potential of spiroplasma cells and induce
the loss of cell motility and helicity (Bévén and Wróblewski,
1997). The toxicity of the lipopeptide antibiotic globomycin was
found to be correlated with an inhibition of spiralin processing
(Bévén et al., 1996). As with Gramicidin S, the antibiotic was
effective against spiroplasmas, but not Mycoplasma mycoides. Natural 18-residue peptaibols (trichorzins PA) are bacteriocidal to
spiroplasmas (Bévén et al., 1998). The mode of action appears
to be permeabilization of the host cell membrane.
Hosts, ecology, and pathogenicity
Hosts. Almost all spiroplasmas have been found to be associated with arthropods or an arthropod connection is strongly
suspected. Hackett et al. (1990) searched for mollicutes in a
wide variety of insect orders. Isolates were obtained from six
orders and 14 insect families. Only one of these orders, Odonata (dragonflies), was primitive (heterometabolous) and it was
speculated that the spiroplasma from a dragonfly host might
have been acquired via predation. Hackett et al. (1990) suggested that the Spiroplasma/Entomoplasma clade may have arisen
in a paraneopteran-holometabolan ancestor, coevolved with
these orders, and never adapted to more primitive insect orders.
Some insect families have an especially rich spiroplasma, entomoplasma, and mesoplasma flora.
Insect gut. The majority of spiroplasmas appear to be maintained in an insect gut/plant surface cycle. Clark (1984) hypothesized several types of gut infection in which persistence in the
gut and the ability to invade hemolymph varied among spiroplasma species. It has been hypothesized (Hackett and Clark,
1989) that the gut cycle was primitive and that other cycles were
derived from it. Spiroplasmas have been isolated from guts of
tabanids (Diptera: Tabanidae) worldwide (French et al., 1997,
1990, 1996; Jandhyam et al., 2008; Le Goff et al., 1991, 1993;
Regassa and Gasparich, 2006; Vazeille-Falcoz et al., 1997; Whitcomb et al., 1997a). Examination of diversity trends among the
tabanid isolates suggests that spiroplasma diversity increases
with temperature, resulting in more diversity in southern
climes in the Northern Hemisphere (Whitcomb et al., 2007).
Although evidence points strongly to multiple cycles of horizontal transmission, the sites where such transmission occurs
remain unknown. However, some tabanids utilize honeydew
(excreta of sucking insects) deposited on leaf surfaces, suggesting a possible transmission mechanism. Mosquitoes (Chastel
and Humphery-Smith, 1991) are also common spiroplasma
hosts (Lindh et al., 2005). Additionally, spiroplasmas inhabit the
gut of ground beetles (Harpalus pensylvanicus and Anisodactylus
sanctaecrucis) as evidenced by 16S rRNA gene sequence analysis
of the digestive tract bacterial flora (Lundgren et al., 2007).
Plant surfaces. Flowers and other plant surfaces represent a
major site where spiroplasmas and other microbes are transmitted from insect to insect (Clark, 1978; Davis, 1978; McCoy et al.,
1979). Members of several spiroplasma groups have been isolated only from flowers and strains of several other spiroplasmas
have been isolated from both insects and flowers. Biological
evidence suggests that mosquito spiroplasmas are transmitted
from insect to insect on flowers (Chastel et al., 1990; Le Goff
et al., 1990). It is not known whether any of the so-called “flower
spiroplasmas” can exist as true epiphytes. Isolations of spiroplasmas from a variety of insects (Clark, 1982; Hackett et al., 1990)
suggest that it is likely that many or most of these flower isolates
are deposited passively by visiting arthropods.
Plant phloem and sucking insects. Several spiroplasmas possess a life cycle that involves infection of plant phloem and
homopterous insects (Bové, 1997; Fletcher et al., 1998; Garnier
et al., 2001; Saglio and Whitcomb, 1979). In the course of passage through the insect, spiroplasmas pass through, accumulate, or multiply in gut epithelial cells and salivary cells. They
also accumulate in the insect neurolemma. Large accumulations of spiroplasma cells occur frequently in the hemolymph,
where they undoubtedly multiply (Whitcomb and Williamson,
1979). Spiroplasmas may multiply in a number of sucking insect
species that have been exposed to diseased plants, but often
only a single vector or several vector species transmit spiroplasmal pathogens from plant to plant (summarized in Calavan and
Bové, 1989; Whitcomb, 1989; Kersting and Sengonca, 1992).
Sex ratio organisms. Once thought to be a genetic factor, the
sex ratio trait in Drosophila was shown by Poulson and Sakaguchi
(1961) to be induced by a micro-organism, Spiroplasma poulsonii
(Williamson et al., 1999). A number of other spiroplasmas in a
variety of insect hosts have been identified that also cause sex
ratio distortions, including isolates from the chrysomelid beetle
Adalia bipunctata (Hurst and Jiggins, 2000; Hurst et al., 1999)
and the butterfly Danaus chrysippus (Jiggins et al., 2000). In
addition, 16S rRNA gene sequence analysis identified spiroplasmas as the causative agent for male-killing: in a population of
Harmonia axyridis (ladybird beetle) in Japan (Nakamura et al.,
2005); in populations of Drosophila neocardini, Drosophila ornatifrons and Drosophila paraguayensis from Brazil (Montenegro
et al., 2006, 2005); in populations of Anisosticta novemdecimpunctata (ladybird beetle) in Britain (Tinsley and Majerus, 2006); in
663
a population of Adalia bipunctata (Sokolova et al., 2002); in several strains from the Tucson Drosophila stock culture collection
(Mateos et al., 2006); and in Drosophila melanogaster populations
from Uganda and Brazil (Pool et al., 2006). Other organisms
closely associated with their insect hosts were discovered inferentially by PCR studies (Fukatsu and Nikoh, 2000, 2001) and
also appear to be related to Spiroplasma mirum. They also cause
preferential male killing in an infected Drosophila population
(Anbutsu and Fukatsu, 2003). Natural infection rates of malekilling spiroplasmas in Drosophila melanogaster are about 2.3%,
as determined for a Brazilian population (Montenegro et al.,
2005), and vary between 0.1 and 3% for Japanese populations
of Drosophila hydei (Kageyama et al., 2006). The male-killing
spiroplasma strain isolated from Adalia bipunctata was used to
artificially infect eight different coccinellid beetle species. The
data suggest that host range could serve to limit horizontal
transfer to closely related host species (Tinsley and Majerus,
2007). Supporting this hypothesis was the study that showed
the interspecific lateral transmission of spiroplasmas from
Drosophila nebulosa to Drosophila willistoni via ectoparasitic mites
(Jaenike et al., 2007). A recent multilocus analysis by Haselkorn
et al. (2009) showed that Drosophila species are infected with at
least four distinct spiroplasma haplotypes.
Studies on Drosophila infections by the sex-ratio organism
showed that it did not induce the innate immunity of the insect
(Hurst et al., 2003). The sex-ratio spiroplasmas have been shown
to be vertically transmitted through female hosts, with spiroplasmas present during oogenesis (Anbutsu and Fukatsu, 2003).
Although the exact mechanism of male-killing has not been
determined, studies have shown that male killing occurs shortly
after formation of the host dosage compensation complex (Bentley et al., 2007) and that male Drosophila melanogaster mutants
lacking any of the five genes involved in the dosage compensation complex are not killed (Veneti et al., 2005). In the Kenyan
butterfly Danaus chrysippus, a correlation between male killing
and a recessive allele for a gene controlling infection susceptibility has been reported. Moreover, infections seemed to have a
negative effect on body size (Herren et al., 2007).
Ticks. Three Spiroplasma species have been isolated from
ticks. Two of these, Spiroplasma mirum and Spiroplasma sp. 277F,
are from the rabbit tick Haemaphysalis leporispalustris (Tully et al.,
1982; Williamson et al., 1989). The third species was isolated
from Ixodes pacificus ticks and named Spiroplasma ixodetis (Tully
et al., 1995). 16S rRNA gene sequence analysis of spiroplasmas
originally isolated from Ixodes ticks and growing in a Buffalo
Green Monkey mammalian cell culture line showed a high
degree of identity with the Spiroplasma ixodetis 16S rRNA gene
(Henning et al., 2006). Analysis of the 16S rRNA gene sequence
from DNA extracted from unfed Ixodes ovatus from Japan indicated the presence of spiroplasmas that were also closely related
to Spiroplasma ixodetis (Taroura et al., 2005). The ability of tick
spiroplasmas, including Spiroplasma ixodetis, to multiply at 37°C
reflects the role of vertebrates as tick hosts. The ability of Spiroplasma ixodetis to grow at 32°C as well as 37°C (Tully et al., 1982)
may reflect the ecology of some of the cold-blooded vertebrate
hosts of these ticks. There is no evidence that any of these spiroplasmas are transmitted to vertebrate hosts of the ticks.
Crustaceans. Spiroplasma sp. have recently been isolated in
both freshwater and salt-water crustaceans.
664
Spiroplasma penaei (strain SHRIMPT) was isolated from the
hemolymph of Pacific white shrimp (Penaeus vannamei) after
high mortalities were observed in an aquaculture pond in
Columbia, South America (Nunan et al., 2004). The pathogenic agent was the spiroplasma (Nunan et al., 2005). Although
not cultivated, 16S rRNA gene sequence analysis also revealed
the presence of spiroplasmas in the gut of the hydrothermal
shrimp Rimicaris exoculata (Zbinden and Cambon-Bonavita,
2003). In another outbreak, Chinese mitten crab (Eriocheir sinensis) reared in aquaculture ponds in China became infected
with tremor disease. The causative agent was determined to be
a spiroplasma with 99% 16S rRNA gene sequence identity to
Spiroplasma mirum (Wang et al., 2004a, b). However, recent studies suggest that the infective agent may be a species similar to,
but distinct from, Spiroplasma mirum (Bi et al., 2008). The same
organism also infects red swamp crayfish (Procambarus clarkii)
that are co-reared with the Chinese mitten crab (Bi et al., 2008;
Wang et al., 2005) as well as the shrimp Penaeus vannamei (Bi
et al., 2008).
Other hosts. Spiroplasmas have been identified in a variety of
other hosts, although not necessarily linked to the gut habitat.
The first spiroplasma isolated from a lepidopteran came from
the hemolymph of white satin moth larvae (Leucoma salicis L.)
from Poland (designated strain SMAT) and was serologically distinct from any previously described spiroplasma group (Oduori
et al., 2005). Another novel spiroplasma (designated strain
GNAT3597T) was isolated from biting midges from the genus
Atrichopogon (Koerber et al., 2005). Spiroplasmas that are closely
related to the male-killing spiroplasmas in ladybird beetles
(Majerus et al., 1999; Tinsley and Majerus, 2006) have also been
identified in the predatory mite Neoseiulus californicus using 16S
rRNA gene sequence analysis (Enigl and Schausberger, 2007).
A broad survey of 16 spider families for the presence of endosymbionts using 16S rRNA gene sequence analysis revealed
that six families contained spiroplasmas, including Agelenidae,
Araneidae, Gnaphosidae, Linyphiidae, Lycosidae, and Tetragnathidae
(Goodacre et al., 2006).
Biogeography. Spiroplasmas have been identified from hosts
in Africa, Asia, Australia, Europe, South America, and North
America. While they are worldwide in distribution, studies suggest that biodiversity may be greatest in warm climates (Whitcomb et al., 2007). Because spiroplasmas are host-associated, it
seems reasonable that Spiroplasma species distribution would be
limited by host biogeography. Early studies indicated that some
spiroplasmas have discrete geographic distributions (Whitcomb
et al., 1990). As the diversity of sampling sites increases, the
view of spiroplasma biogeography is likely to shift (Regassa and
Gasparich, 2006). Distinct distributions may exist, but probably
on a larger geographic scale. While it is not clear what factors
account for spiroplasma ranges, the level of host specificity and
host overwintering ranges may contribute to the biogeography
of Spiroplasma species (Whitcomb et al., 2007).
Pathogenicity. Symptoms of infection and confirmation of
Koch’s postulates have been reported for the etiologic roles of:
Spiroplasma citri in “stubborn” disease of citrus (Calavan and
Bové, 1989; Markham et al., 1974); corn stunt spiroplasma
(Chen and Liao, 1975; Nault and Bradfute, 1979; Williamson and Whitcomb, 1975); Spiroplasma phoeniceum in aster, an
experimental host (Saillard et al., 1987); Spiroplasma poulsonii
in Drosophila pseudoobscura (Williamson et al., 1989); Spiroplasma
penaei in Penaeus vannamei (Nunan et al., 2005); and Spiroplasma
eriocheiris (Wang et al., 2010) in the Chinese mitten crab, Eriocheir
sinensis (Wang et al., 2004b). Recent studies have focused on
spiroplasma infection and replication in the midgut and Malpighian tubules of leafhoppers (Özbek et al., 2003). The use
of immunofluorescence confocal laser scanning microscopy
has revealed the presence of Spiroplasma kunkelii in the midgut,
filter chamber, Malpighian tubules, hindgut, fat tissues, hemocytes, muscle, trachea, and salivary glands of leafhopper hosts,
but not in the nerve cells of the brain or nerve ganglia (Ammar
and Hogenhout., 2005). Plant spiroplasmas may also be pathogenic for unusual vectors (Whitcomb and Williamson, 1979),
but are much less so for their usual host (Madden and Nault,
1983; Nault et al., 1984). In fact, some spiroplasmas are beneficial to their leafhopper hosts (Ebbert and Nault, 1994) and it
has been hypothesized that infection plays an important role in
the host’s overwintering strategies (Moya-Raygoza et al., 2007a,
b; Summers et al., 2004).
Spiroplasma mirum is experimentally pathogenic for a variety
of suckling animals, causing cataract and other ocular symptoms, neural pathology (Clark and Rorke, 1979), and malignant transformation in cultured cells (Kotani et al., 1990).
Spiroplasma melliferum also persists and causes pathology in suckling mice (Chastel et al., 1990, 1991). Spiroplasma eriocheiris is
neurotropic to brain tissue in experimentally injected chicken
embryos (Wang et al., 2003). There are two recent reports of
spiroplasmas in aquatic invertebrates. Nunan et al. (2005) characterized a spiroplasma in commercially raised shrimp that led
to a lethal disease. Spiroplasma melliferum and Spiroplasma apis
cause disease in honey bees (Clark, 1977; Mouches et al., 1982a,
1983a). Intrathoracic inoculation of Spiroplasma taiwanense
reduced the survival and impaired the flight capacity of inoculated mosquitoes (Humphery-Smith et al., 1991a), and inoculation of Spiroplasma taiwanense per os decreased the survival of
mosquito larvae in laboratory trials (Humphery-Smith et al.,
1991b). Spiroplasma poulsonii causes sex ratio abnormalities
(male-killing) in Drosophila (Williamson and Poulson, 1979).
Male-killing spiroplasma strains related to Spiroplasma poulsonii
cause necrosis in neuroblastic and fibroblastic cells (Kuroda
et al., 1992). The significance of some biological properties of
spiroplasmas is incompletely understood. For example, membranes of Spiroplasma monobiae are potent inducers of tumor
necrosis factor alpha secretion and of blast transformation
(Sher et al., 1990a, b) in insect cell culture.
Spiroplasmas are implicated by circumstantial evidence, in
the view of some workers, to be associated with human disease.
Bastian first claimed in 1979 that spiroplasmas were associated with Creutzfeldt–Jakob Disease (CJD), an extremely rare
scrapie-like disease of humans (Bastian, 1979). Bastian and
Foster (2001) reported finding spiroplasma 16S rRNA genes in
CJD- and scrapie-infected brains that were not observed in controls. More recent studies (Bastian et al., 2004) presented evidence to show that spiroplasma 16S rRNA genes were found in
brain tissue samples from scrapie-infected sheep, chronic wasting disease-infected cervids, and CJD-infected humans. All the
brain tissues from non-infected controls were negative for spiroplasmal DNA. These authors further showed that the sequence
of the PCR products from the infected brains was 96% identical to the Spiroplasma mirum 16S rRNA gene. However, these
results could not be replicated in an independent blind study of
uninfected and Scrapie-infected hamster brains using the same
primers (Alexeeva et al., 2006). A recent study to fulfill Koch’s
postulate reported the transfer of spiroplasma from transmissible spongiform encephalopathy (TSE) brains and Spiroplasma
mirum to induce spongiform encephalopathy in ruminants
(Bastian et al., 2007). The current status of the involvement of
spiroplasmas in TSE is the subject of recent reviews (Bastian,
2005; Bastian and Fermin, 2005). Other proposed connections
between mollicutes and human disease have been evaluated by
Baseman and Tully (1997).
Mechanism of Spiroplasma citri phytopathogenicity. Transposon (Tn4001) mutants have been examined extensively to elucidate the molecular mechanisms associated with Spiroplasma
citri phytopathogenicity. One of these mutants, GMT553, highlighted the involvement of selective carbohydrate utilization in
Spiroplasma citri pathogenicity (see review by Bové et al., 2003).
When introduced into periwinkle plants via injected leafhoppers (Circulifer haematoceps), GMT553 multiplied in the plants
as actively as wild-type Spiroplasma citri strain GII3, but did not
induce symptoms (Foissac et al., 1997b, c; Gaurivaud et al.,
2000b). In this mutant, the transposon was found to be inserted
in fruR, a transcriptional activator of the fructose operon (fruRAK; Gaurivaud et al., 2000a). The second gene of the operon,
fruA, encodes fructose permease, which enables uptake of
fructose; and the third gene, fruK, encodes 1-phosphofructokinase. In mutant GMT553, transcription of the fructose operon
is abolished and, hence, the mutant cannot utilize fructose as
a carbon or energy source (Gaurivaud et al., 2000a). Mutant
GMT553 was functionally complemented for fructose utilization and phytopathogenicity in trans by a recombinant fruR–
fruA–fruK operon, fruA–fruK partial operon, or fruA alone, but
not fruR or fruR–fruA (Gaurivaud et al., 2000a, b). It should be
pointed out that both fructose+ and fructose− spiroplasmas are
able to utilize glucose.
Further insight into Spiroplasma citri phytopathogenicity in relation to sugar metabolism comes from the production of a spiroplasma mutant unable to use glucose (André et al., 2005). The
import of glucose into Spiroplasma citri cells involves a phosphotransferase (PTS) system composed of two distinct polypeptides
encoded by (1) crr (glucose PTS permease IIAGlc component)
and (2) ptsG (glucose PTS permease IICBGlc component). A ptsG
mutant (GII3-glc1) proved unable to import glucose. When
introduced into periwinkle (Catharanthus roseus) plants through
leafhopper transmission, the mutant induced severe symptoms
similar to those obtained with wild-type GII3, in strong contrast
to the fructose operon mutant, GMT553, which was virtually
non-pathogenic. These results indicated that fructose and glucose utilization were not equally involved in pathogenicity and
are consistent with biochemical data showing that, in the presence of both sugars, Spiroplasma citri preferentially used fructose.
NMR analyses of carbohydrates in plant extracts revealed the
accumulation of soluble sugars, particularly glucose, in plants
infected by wild-type Spiroplasma citri GII3 or GII3-glc1, but not
in those infected by GMT553. In the infected plant, Spiroplasma
citri cells are restricted to the sieve tubes. In the companion cell,
sucrose is cleaved by invertase to fructose and glucose. In the
sieve tube, wild-type Spiroplasma citri cells will use fructose preferentially over glucose leading to a decreased fructose concentration and, consequently, to an increase of invertase activity, which
665
in turn results in glucose accumulation. Glucose accumulation
is known to induce stunting and repression of photosynthesis
genes in Arabidopsis thaliana. Such symptoms are precisely those
observed in periwinkle plants infected by wild-type Spiroplasma
citri (André et al., 2005).
Genes that are up- or down-regulated in plants following
infection with Spiroplasma citri have been studied by differential
display analysis of mRNAs in healthy and symptomatic periwinkle plants (Jagoueix-Eveillard et al., 2001). Expression of the
transketolase gene was inhibited in plants infected by the wildtype spiroplasma, but not by the non-phytopathogenic mutant
GMT553, further indicating that sugar metabolism and transport are important factors in pathogenicity. Sugar PTS system
permeases have been shown to be important in rapid adaptation to sugar differences between plant host and insect vector
(André et al., 2003).
Leafhopper transmission of Spiroplasma citri. Spiroplasmas
are acquired by leafhopper vectors that imbibe sap from the
sieve tubes of infected plants. However, in order to be transmitted to a plant, the mollicutes need first to multiply in the
insect vector after crossing the gut barrier (Wayadande and
Fletcher, 1995). They multiply to high titers (106–107/ml) in
the insect hemolymph, but only when they have reached the
salivary glands can they be inoculated into a plant. One gene
required for efficient transmission, sc76, was inactivated in a
transposon mutant (G76) with reduced transmissibility (Boutareaud et al., 2004); sc76 encodes a putative lipoprotein. Plants
infected with the G76 mutant showed symptoms 4–5 weeks
later than those infected with wild-type GII3, but when they
appeared, the symptoms induced were severe. Mutant G76 multiplied in plants and leafhoppers as efficiently as the wild-type
strain. However, leafhoppers injected with the wild-type spiroplasma transmitted the spiroplasma to 100% of exposed plants.
In contrast, those injected with mutant G76 infected only 50%
of the plants. This inefficiency was shown to be associated with
a numerical decrease in spiroplasma cells in the salivary glands
that correlated with reduced output from the stylets of transmitting leafhoppers; the number of mutant cells transmitted
through Parafilm membranes was less than 5% of numbers of
wild-type cells transmitted based on colony-forming units. Functional complementation of the G76 mutant with the sc76 gene
restored the wild-type phenotype. Because both wild-type and
mutant cells multiplied to equally high titers in the hemolymph,
the results suggest that the mutant is inefficiently passed from
the hemolymph into the salivary glands or that it may multiply
to a lower titer in the glands.
Transmission of Spiroplasma citri by leafhopper vectors must
involve adherence to and invasion of insect host cells. Electron microscopic studies of leafhopper midgut by Ammar
et al. (2004) have demonstrated the attachment of Spiroplasma
kunkelii cells by a tip structure to the cell membrane between
microvilli of epithelial cells. Spiroplasma citri surface protein P89
was shown to mediate adhesion of the spiroplasma to cells of
the vector Circulifer tenellus and was designated SARP1 (Berg
et al., 2001; Yu et al., 2000). The gene encoding SARP1, arp1,
was cloned and characterized from Spiroplasma citri BR3-T. The
putative gene product SARP1 contains a novel domain at the
N terminus, called “sarpin” (Berg et al., 2001). The arp1 gene
is located on plasmid pBJS-O in Spiroplasma citri (Joshi et al.,
2005). The Spiroplasma kunkelii plasmid pSKU146 encodes an
666
adhesin that is a homolog of SARP1 (Davis et al., 2005). Other
spiroplasma plasmids encode additional adhesin-related proteins. As indicated above (see Plasmids), Spiroplasma citri GII3
contains six large plasmids, pSci1 to pSci6 (Saillard et al.,
2008). Although plasmids pSci1 to pSci5 encode eight different Spiroplasma citri adhesin-related proteins (ScARPs), they
are not required for insect transmission (Berho et al., 2006b).
One of the ScARPs, protein P80, shared 63% similarity and
45% identity with SARP1. Protein P80 is carried by plasmid
pSci4 and has been named ScARP4a. The ScARP-encoding
genes could not be detected in DNA from non-transmissible
strains (Berho et al., 2006b). Sequence alignments of ScARP
proteins revealed that they share common features including
a conserved signal peptide followed by six to eight repeats of
39–42 amino acids each, a central conserved region of 330
amino acids, and a transmembrane domain at the C terminus
(Saillard et al., 2008).
Plasmid pSci6 carries the gene for protein P32, which is present in all Spiroplasma citri strains capable of being transmitted by
the leafhopper vector Circulifer haematoceps, but absent from all
non-transmissible strains (Killiny et al., 2006). Complementation studies with P32 alone did not restore transmissibility (Killiny et al., 2006). However, if the pSci6 plasmid was transferred
to an insect-non-transmissible Spiroplasma citri strain, then the
phenotype could be converted to insect-transmissible, indicating the likely presence of additional transmissibility factors on
pSci6 (Berho et al., 2006a). Indeed, recent data indicates that
factors essential for transmissibility are encoded by a 10 kbp
fragment of pSci6 (Breton et al., 2010). The finding that the
insect-transmissible strain Spiroplasma citri Alc254 contains only
a single plasmid, pSci6 (S. Richard and J. Renaudin, unpublished) also reinforces the hypothesis that pSci6-encoded determinants play a key role in insect transmission of Spiroplasma citri
by its leafhopper vector.
Isolation. Success in the isolation of fastidious spiroplasmas is
influenced strongly by the titer of the inoculum. Spiroplasmas
have been isolated from salivary glands, gut, and nerve tissues
of their insect hosts. Many spiroplasmas envisioned by darkfield microscopy have proved to be noncultivable (Hackett and
Clark, 1989). Initial insect extracts in growth media are passed
through a 0.45 mm filter. The filtrate is then observed daily for
pH indicator change. An alternative to filtration involves the use
of antibiotics or other inhibitors (Grulet et al., 1993; Markham
et al., 1983; Whitcomb et al., 1973). Spiroplasma isolations from
infected plants are best obtained from sap expressed from vascular bundles of hosts showing early disease symptoms. Plant
sap often contains spiroplasmal substances (Liao et al., 1979)
whose presence in primary cultures may necessitate blind passage or serial dilution.
Isolation media. M1D medium (Whitcomb, 1983) has been
used for primary isolations of the large proportion of spiroplasma species. SP-4 medium, a rich formulation derived from
experiments with M1D, is necessary for isolation of Spiroplasma
mirum from fluids of the embryonated egg (Tully et al., 1982).
SP-4 medium is also required for isolation of Spiroplasma ixodetis (Tully et al., 1981). Some very fastidious spiroplasmas such
as Spiroplasma poulsonii (Hackett et al., 1986) and Spiroplasma
leptinotarsae (Hackett and Lynn, 1985) were isolated by cocultivation with insect cells. However, the requirement for cocultivation of Spiroplasma leptinotarsae can be circumvented by
placing the primary cultures in BBL anaerobic GasPak jar systems with low redox potential and enhanced CO2 atmosphere
(Konai et al., 1996b). By lowering the pH of the growth medium
from 7.4 to 6.2 and using bromocresol purple as a pH indicator
(pH 5.2 yellow to pH 6.8 purple), it was possible to perform
metabolism inhibition tests involving Spiroplasma leptinotarsae as
the antigen. The same low-pH medium containing 2.0% Noble
agar permitted the growth of colonies (Williamson, unpublished data). Cohen and Williamson (1988) reported that a
fortuitous contamination of H-2 medium by a slow-growing,
pink-colored yeast (Rhodotorula rubra) permitted primary isolation of the non-male-lethal variant of the Dorsophila willistoni
spiroplasma. After 10–12 passages with yeast, the spiroplasmas
were able to grow in yeast-free H-2 medium.
Maintenance procedures. Adaptation. Most spiroplasmas can be adapted to a wide variety of media formulations.
Spiroplasmas commonly grow more slowly upon transfer to new
media. Initial reduction in growth rate is probably related to
a combination of differences in nutrients, pH, osmolality, etc.
Isolates may grow at only slightly reduced rates during the first
1–5 passages in a new medium. However, if the new medium
is markedly deficient, the growth rate may decrease precipitously after 5–10 passages. Continuous careful passaging may
result in growth rate recovery to levels similar to that in the
initial medium. For such adaptations, best results are achieved
by starting with a 1:1 ratio of old and new media and gradually
withdrawing the old formulation. Spiroplasma clarkii, after continuous passage for hundreds of generations, finally adapted to
extremely simple media (Hackett et al., 1994). Adaptation may
involve mutation and/or activation of adaptive enzymes, or,
possibly, other mechanisms. Growth rates in such simple media
were much slower than those in rich media.
Maintenance media. Spiroplasma citri can be cultivated in
a relatively simple medium that utilizes sorbitol to maintain
osmolality (Saglio et al., 1971). A modification of this medium
(BSR) has been used extensively for Spiroplasma citri (Bové and
Saillard, 1979), in which the horse serum content was lowered
to 10% and the fresh yeast extract was omitted. Other simple
media, such as C-3G (Liao and Chen, 1977), are suitable for
maintenance or large-batch cultivation of fast-growing spiroplasmas. This medium is also adequate for primary isolation of
Spiroplasma kunkelii (Alivizatos, 1988). However, cultivation of
more fastidious spiroplasmas is best achieved in M1D medium
(Hackett and Whitcomb, 1995; Whitcomb, 1983; Williamson
and Whitcomb, 1975) if they derive from plant or insect habitats. SP-4 medium (Tully et al., 1977) is very suitable if spiroplasmas derive from tick habitats. SM-1 medium (Clark, 1982) has
also been successfully employed for many insect spiroplasmas.
Defined media. Spiroplasma floricola and some strains of
Spiroplasma apis have been cultivated in chemically defined
media (Chang, 1989, 1982).
Preservation. Spiroplasmas are routinely preserved by
lyophilization (FAO/WHO, 1974). Most spiroplasmas can be
maintained at −70°C indefinitely. Preservation success at −20°C
is irregular and uncertain.
Differentiation of the genus Spiroplasma
from other closely related taxa
Spiroplasmas can be clearly differentiated from all other microorganisms by their unique properties of helicity and motility,
combined with the complete absence of periplasmic fibrils,
cell walls, or cell wall precursors. However, spiroplasmas may
be nonhelical under some environmental conditions or when
cultures are in the stationary phase of growth. Morphological
study of the organisms in the exponential phase of growth usually reveals characteristic helical forms. However, the existence
of spiroplasmas that appear entirely or largely as nonhelical
forms (e.g., Spiroplasma ixodetis and group XXIII strain TIUS1) raises the theoretical possibility that an organism situated
at an apomorphic (advanced) position on the spiroplasma
phylogenetic tree could totally lack helicity or motility. In fact,
the clade containing Mycoplasma mycoides and the Entomoplasmataceae has apparently done exactly that. Spiroplasma floricola
produces nonhelical, but viable, cells early in stationary phase,
which can begin within 24 h of medium inoculation. For reasons such as this, it is necessary to examine cultures throughout
the growth cycle to ensure that an adequate search for helical
cells has been made.
Taxonomic comments
Early history. The term “spiroplasma” was first coined as a
trivial term to describe helical organisms shown to be associated with corn stunt disease (Davis et al., 1972a, b) that could
not, at that time, be cultivated (Davis and Worley, 1973). Shortly
thereafter, when similar organisms associated with citrus stubborn disease were characterized (Saglio et al., 1973), the trivial term was adopted as the generic name and the stubborn
organism was named Spiroplasma citri. This species was the
first cultured spiroplasma and the first cultured mollicute of
plant origin. Shortly after the stubborn agent was named, the
genus Spiroplasma was elevated to the status of a family (Skripal, 1974) and added to the Approved Lists of Bacterial Names
(Skripal, 1983). The organism that was eventually named Spiroplasma mirum (Tully et al., 1982) was isolated by Clark (1964)
in embryonated chicken eggs soon after the discovery of the
organism later named Spiroplasma poulsonii. Because Spiroplasma
mirum readily passed through filters, it was first mistaken for
a virus. The subgroup I-4 277F spiroplasma was cultivated in
1968, but was mistaken for a spirochete (Pickens et al., 1968).
The first organism to be initially recognized as a spiroplasma
was Spiroplasma kunkelii, which was envisioned by dark-field and
electron microscopy in 1971–1972 and cultivated in 1975 (Liao
and Chen, 1977; Williamson and Whitcomb, 1975). More than
a decade passed before Clark (1982) showed that spiroplasmas,
many of them fast-growing, occurred principally in insects.
Species concept. The species concept in spiroplasmas, as in
all bacteria, was based on DNA–DNA reassociation (ICSB Subcommittee on the Taxonomy of Mollicutes, 1995; Johnson, 1994;
Rosselló-Mora and Amann, 2001; Stackebrandt et al., 2002;
Wayne et al., 1987). In practice, DNA–DNA reassociation results
with spiroplasmas have proven difficult to standardize. Estimates
of reassociation between Spiroplasma citri (subgroup I-1) and
Spiroplasma kunkelii (subgroup I-3) varied between 30 and 70%,
depending on the method employed and the degree of stringency (Bové and Saillard, 1979; Christiansen et al., 1979; Lee
667
and Davis, 1980; Liao and Chen, 1981a; Rahimian and Gumpf,
1980). Given these challenges, an alternative method was identified in serology. Surface serology of spiroplasmas has proven to
be a robust surrogate for DNA–DNA hybridization assays.
Phylogeny. Phylogenetic studies of Spiroplasma became
possible when Carl Woese and colleagues, searching for a
molecular chronometer by which microbial evolution could be
reconstructed, found that rRNA met most or all of the desired
criteria (reviewed by Woese, 1987). Today, sequencing of rRNA
genes has become a universal tool for phylogenetic reconstruction. Early phylogenetic analyses involved distance estimates
(DeSoete, 1983). Later, neighbor-joining (Saitou and Nei, 1987)
was introduced into mollicute phylogeny (Maniloff, 1992)
and several mollicute workers have used maximum-likelihood
(Felsenstein, 1993). The extensive and classical studies of K.-E.
Johansson’s group (Johansson et al., 1998; Pettersson et al.,
2000) were completed using neighbor-joining, but selectively
confirmed by maximum-likelihood and maximum-parsimony
(Swofford, 1998). Gasparich et al. (2004) studied the phylogeny
of Spiroplasma and its nonhelical descendants using parsimony,
maximum-likelihood, distance, and neighbor-joining analyses,
which generated 24 phylogenetic inferences that were common to all, or almost all, of the trees. More recently, Bayesian
analysis [MrBayes (http://mrbayes.csit.fsu.edu/index.php)]
was used to examine an expanded Spiroplasma Apis clade based
on 16S rRNA and 16S–23S ITS sequences; the analyses showed
congruency between Bayesian and maximum-parsimony trees
(Jandhyam et al., 2008).
Woese et al. (1980) presented a 16S rRNA gene-based phylogenetic tree for Mollicutes, including Spiroplasma, indicating that
these wall-less bacteria were related to members of the phylum
Firmicutes such as Lactobacillus spp. and Clostridium innocuum.
The tree suggested that Mollicutes might be monophyletic.
However, a later study by Weisburg et al. (1989) with 40 additional species of Mollicutes including ten spiroplasmas, failed
to confirm the monophyly of Mollicutes at the deepest branching orders. The Woese et al. (1980) model also suggested that
the genus Mycoplasma might not be monophyletic, in that the
type species, Mycoplasma mycoides, and two related species, Mycoplasma capricolum and Mycoplasma putrefaciens, appeared to be
more closely related to the Apis clade of Spiroplasma than to
the other Mycoplasma species. This conclusion was supported
by analyses of the 5S rRNA genes (Rogers et al., 1985). All trees
so far obtained indicate that the acholeplasma-anaeroplasma
(Acholeplasmatales–Anaeroplasmatales) and spiroplasma-mycoplasma (Mycoplasmatales–Entomoplasmatales) lineages are monophyletic, but are separated by an ancient divergence.
In-depth analysis of characterized spiroplasmas and their nonhelical descendants indicates the existence of four major clades
within the monophyletic spiroplasma-mycoplasma lineage (Gasparich et al., 2004; Figure 113). One of the four clades consists
of the nonhelical species of the mycoides group (as defined
by Johansson, 2002) as well as the six species of Entomoplasma
and twelve species of Mesoplasma (the Entomoplasmataceae); this
assemblage was designated the Mycoides-Entomoplasmataceae
clade. The analyses indicated that the remaining three clades
represented Spiroplasma species. One of these clades, the Apis
clade, was found to be a sister to the Mycoides-Entomoplasmataceae clade. The Apis clade contains a large number of species from diverse insect hosts, many of which possess life cycles
Spiroplasma chrysopicola
* Spiroplasma syrphidicola
*
*
*
Scale:
Spiroplasma sp. TAAS-1
Spiroplasma mirum
Spiroplasma sp. LB-12
Spiroplasma sp. 277F
Spiroplasma sp. N525
Spiroplasma poulsonii
* Spiroplasma penaei
Spiroplasma insolitum
Spiroplasma phoeniceum P40
Spiroplasma kunkelii CR2-3x
Spiroplasma citri
**
Spiroplasma melliferum
Entomoplasma freundtii
*
Mycoplasma mycoides
Mesoplasma seiffertii
Spiroplasma monobiae
** Spiroplasma diabroticae
Spiroplasma floricola
Spiroplasma sp. BIUS-1
Spiroplasma sp. W115
*
Spiroplasma cantharicola CC-1
* Spiroplasma sp. CB-1
Spiroplasma sp. Ar-1357
Spiroplasma diminutum
Spiroplasma taiwanense
Spiroplasma gladiatoris
Spiroplasma lineolae TALS-2
*
Spiroplasma sp. BARC 1901
Spiroplasma helicoides
Spiroplasma clarkii
Spiroplasma apis
*
** Spiroplasma montanense
Spiroplasma litorale
Spiroplasma turonicum
*
Spiroplasma corruscae
Spiroplasma culicicola
Spiroplasma velocicrescens
Spiroplasma chinense
Spiroplasma leptinotarsae
*
Spiroplasma lampyridicola
Spiroplasma sabaudiense
Spiroplasma alleghenense
Spiroplasma ixodetis
’Candidatus Phytoplasma’ sp. vigna Il
Anaeroplasma bactoclasticum
Clostridium innocuum
Bacillus subtilis TB11
Asteroleplasma anaerobium
Escherichia coli
0.1 substitutions/site
FIGURE 113. Phylogenetic relationships of members of the class Mollicutes and selected members of the phylum Firmicutes. The phylogram was
based on a Jukes-Cantor corrected distance matrix and weighted neighbor-joining analysis of the 16S rRNA gene sequences of the type strains,
except where noted. Escherichia coli was the outgroup. Bootstrap values (100 replicates) <50% are indicated (*). The GenBank accession numbers
for 16S rRNA gene sequences used are: Mycoplasma mycoides (U26039); Mycoplasma pneumoniae (M29061); Entomoplasma freundtii (AF036954);
Mesoplasma seiffertii (AY351331); Spiroplasma apis (M23937); Spiroplasma clarkii (M 24474); Spiroplasma gladiatoris (M24475); Spiroplasma taiwanense (M24476); Spiroplasma monobiae (M24481); Spiroplasma diabroticae (M24482); Spiroplasma melliferum (AY325304); Spiroplasma citri (M23942);
Spiroplasma mirum (M24662); Spiroplasma ixodetis (M24477); Spiroplasma sp. strain N525 (DQ186642); Spiroplasma poulsonii (M24483); Spiroplasma
penaei (AY771927); Spiroplasma phoeniceum (AY772395); Spiroplasma kunkelii (DQ319068); Spiroplasma cantharicola (DQ861914); Spiroplasma lineolae
(DQ860100); Spiroplasma sp. strain 277F (AY189312); Spiroplasma sp. strain LB-12 (AY189313); Spiroplasma insolitum (AY189133); Spiroplasma floricola (AY189131); Spiroplasma syrphidicola (AY189309); Spiroplasma chrysopicola (AY189127); Spiroplasma sp. strain TAAS-1 (AY189314); Spiroplasma
culicicola (AY189129); Spiroplasma velocicrescens (AY189311); Spiroplasma sabaudiense (AY189308); Spiroplasma corruscae (AY189128); Spiroplasma sp.
strain CB-1 (AY189315); Spiroplasma sp. strain Ar-1357 (AY189316); Spiroplasma turonicum (AY189310); Spiroplasma litorale (AY189306); Spiroplasma
lampyridicola (AY189134); Spiroplasma leptinotarsae (AY189305); Spiroplasma sp. strain W115 (AY189317); Spiroplasma chinense (AY189126); Spiroplasma diminutum (AY189130); Spiroplasma alleghenense (AY189125); Spiroplasma sp. strain BIUS-1 (AY189319); Spiroplasma montanense (AY189307);
Spiroplasma helicoides (AY189132); Spiroplasma sp. strain BARC 1901 (AY189320); Ureaplasma urealyticum (M23935); “Candidatus Phytoplasma” sp.
Vigna II (AJ289195); Acholeplasma laidlawii (M23932); Anaeroplasma bactoclasticum (M25049); Clostridium innocuum (M23732); Asteroleplasma anaerobium (M22351); Bacillus subtilis (AF058766); Escherichia coli (J01859).
involving transmission between the guts of insects and plant
surfaces. One of these species, Spiroplasma sp. TIUS-1 (group
XXVIII) has very poor helicity and a genome size of 840 kbp,
smaller than that of most other spiroplasmas. This species
diverged from the spiroplasma lineage close to the node of
entomoplasmal divergence and can be envisioned as a “missing
link” in the evolutionary development of the Mycoides-Entomoplasmataceae clade. The other two Spiroplasma clades are the
monospecific Ixodetis clade (group VI) and the Citri-Chrysopicola-Mirum clade (with representatives from groups I, II, V,
and VIII). The Citri-Chrysopicola-Mirum clade contains Spiroplasma mirum, Spiroplasma poulsonii, the three subgroups of the
Chrysopicola (group VIII) clade, and the nine subgroups of the
Citri (group I) clade. Members of group I and group VIII show
close intragroup relationships, as indicated by the similarities of
their 16S rRNA gene sequences (Gasparich et al., 2004). DNA–
DNA reassociation studies for group I (Bové et al., 1983, 1982;
Junca et al., 1980) spiroplasmas supported the subgroup cluster. The Chrysopicola clade (group VIII) subgroups have met a
different fate. Although their DNA–DNA similarities in reassociation procedures were slightly less than 70%, their 16S rRNA
gene sequence similarities were >99% (Gasparich et al., 1993c).
The strains of this group, including not only the subgroups, but
a plethora of isolates from the same ecological context, appear
to form a matrix of interrelated strains. Boundaries that seemed
clear when the subgroups were initially described, eventually
eroded beyond recognition. The 16S rRNA gene sequence similarities are too high to permit cladistic analysis and even 16S–
23S rRNA spacer region sequence analysis failed to resolve the
669
existing subgroups (Regassa et al., 2004). Over time, the concept of the microbial species has undergone a subtle change.
It is now recognized (Rosselló-Mora and Amann, 2001; Stackebrandt et al., 2002) that microbial species must at times consist
of strain clusters that may contain species with <70% similarity
as determined by DNA–DNA reassociation. Group VIII spiroplasmas may comprise such a cluster and efforts to subdivide
this cluster may have been inadvisable.
Character mapping of non-genetic features has been completed in conjunction with phylogenetic analyses (Gasparich
et al., 2004). Serological classifications of spiroplasmas are generally supported by the trees, but the resolution of genetic analyses appears to be much greater than that of serology. Genome
size and G+C content were moderately conserved among closely
related strains. Apparent conservation of slower growth rates
in some clades was most likely attributable to host affiliation;
spiroplasmas of all groups that were well adapted to a specific
host had slower growth rates. Sterol requirements were polyphyletic, as was the ability to grow in the presence of PES, but
not serum.
Acknowledgements
We gratefully acknowledge J. Dennis Pollack for assistance on
sections concerning intermediary metabolism.
Further reading
Whitcomb, R.F. and J.G. Tully (editors). 1989. The Mycoplasmas, vol. 5, Spiroplasmas, acholeplasmas, and mycoplasmas
of plants and arthropods. Academic Press, New York.
List of species of the genus Spiroplasma
1. Spiroplasma citri Saglio, L’Hospital, Laflèche, Dupont,
Bové, Tully and Freundt 1973, 202AL
cit¢ri. L. masc. n. citrus the citrus; N.L. masc. n. Citrus generic
name; N.L. gen. n. citri of Citrus, to denote the plant host.
Cells are helices that divide in mid-exponential phase
when they have four turns. Helical filaments are usually
100–200 nm in diameter and 2–4 mm in length. Cells
are longer in late exponential phase and early stationary
phase. Nonviable cells in late exponential phase are nonhelical.
Colonies on solid media containing 20% horse serum
and 0.8% Noble agar (Difco) are umbonate, 60–150 mm in
diameter. Moderate turbidity is produced in liquid cultures.
Biological properties are listed in Table 142.
Serologically distinct from other Spiroplasma species,
groups, and subgroups, but shares some cross-reactivity
with members of other group I subgroups. Has close phylogenetic affinities with other group I members, and with
Spiroplasma poulsonii in trees constructed using 16S rRNA
gene sequences.
Pathogenic for citrus plants and a variety of plant hosts
(aster, periwinkle, broad bean) following transmission by
infected insects (leafhoppers).
DNA–DNA renaturation experiments confirm serological data that indicate that the differences between the type
strain (subgroup I-1) and other subgroups of group I are
great enough to warrant its designation as a distinct species.
The genome size is 1820 kbp (PFGE).
Source: isolated from leaves, seed coats, and fruits of
c itrus plants (orange and grapefruit) infected with stubborn disease, and from other naturally infected plants (e.g.,
periwinkle, horseradish or brassicaceous weeds) or insects.
Known from Mediterranean and other warm climates of
Europe, North Africa, Near and Middle East, and the Western United States (California and Arizona).
Type strain: ATCC 27556, Morocco strain, R8-A2.
2. Spiroplasma alleghenense Adams, Whitcomb, Tully, Clark,
Rose, Carle, Konai, Bové, Henegar and Williamson 1997,
762VP
al.le.ghen.en¢se. N.L. neut. adj. alleghenense of the Allegheny
Mountains, referring to the geographic origin of the type
strain, the range of the Appalachian Mountains from which
it was derived.
Cells are motile helical filaments, 100–300 nm in
iameter. Under many growth conditions, cells in medium
d
are deformed. Colonies on solid medium containing
3.0% Noble agar are small and granular and never have
a fried-egg appearance. Biological properties are listed in
Table 142.
groups, and subgroups. When tested as an antigen, crossreacts broadly with many nonspecific sera (one-way reaction). Has close phylogenetic relationship to Spiroplasma
670
sabaudiense (group XIII) and strain TIUS-1 (group XXVIII)
in trees constructed using 16S rRNA gene sequences. The
genome size is 1,465 kbp (PFGE).
Source: isolated from the hemolymph of a common scorpion fly, Panorpa helena in West Virginia, USA.
DNA G+C content (mol%): 31 ± 1 (Tm, Bd).
Type strain: ATCC 51752, PLHS-1.
3. Spiroplasma apis Mouches, Bové, Tully, Rose, McCoy, CarleJunca, Garnier and Saillard 1984b, 91VP (Effective publication: Mouches, Bové, Tully, Rose, McCoy, Carle-Junca, Garnier and Saillard 1983a, 383.)
a¢pis. L. fem. n. apis, -is a bee, and also the genus name of
the honey bee, Apis mellifera; L. gen. n. apis of a bee, of Apis
mellifera, the insect host for this species.
The morphology is as described for the genus. Helical
filaments are usually 100–150 nm in diameter and 3–10 mm
in length. Colonies on solid medium containing 20% fetal
bovine serum and 0.8% Noble agar (Difco) are usually diffuse, rarely exhibiting central zones of growth into the agar.
Colonies on solid medium with 2.25% Noble agar and 1–5%
bovine serum fraction are smaller, but exhibit central zones
of growth into the agar and some peripheral growth on the
agar surface around the central zones. Marked turbidity is
produced during growth in most spiroplasma media (BSR,
M1A, SP-4). Biological properties are listed in Table 142.
groups, and subgroups. Many strains show partial crossreactions when tested against sera to strain B31T (Tully
et al., 1980). These strains show more than 80% DNA–DNA
reassociation with strain B31T, but their exact taxonomic
status is unclear. Some strains show a very low level reciprocal cross-reaction with Spiroplasma montanense in deformation serology. In accordance with serology, Spiroplasma apis
and Spiroplasma montanense are sister species in phylogenetic
trees constructed using 16S rRNA gene sequences. The
genome size is 1300 kbp (PFGE).
Etiologic agent of May disease of honey bees in southwestern France. Various strains of the organism exhibit
experimental pathogenicity for young honey bees in feeding experiments.
Source: isolated from honey bees (Apis mellifera) and from
flower surfaces in widely separated geographic regions
(France, Corsica, Morocco, USA).
Type strain: ATCC 33834, B31.
4. Spiroplasma atrichopogonis Koerber, Gasparich, Frana and
Grogan 2005, 291VP
a.tri.cho.po.go¢nis. N.L. gen. n. atrichopogonis of Atrichopogon,
systematic genus name of a biting midge (Diptera: Ceratopogonidae).
The morphology is as described for the genus. Cells are
helical and motile. Biological properties are listed in Table
142.
Serologically distinct from previously established Spiroplasma species, groups, and subgroups. The genome size
has not been determined.
Source: isolated from a pooled sample of two nearly identical species of biting midges (Atrichopogon geminus and
Atrichopogon levis).
DNA G+C content (mol%): 28.8 ± 1 (Tm).
Type strain: ATTC BAA-520, NBRC 100390, GNAT3597.
Sequence accession no. (16S rRNA gene): not available.
5. Spiroplasma cantharicola Whitcomb, Chastel, AbalainColloc, Stevens, Tully, Rose, Carle, Bové, Henegar, Hackett,
Clark, Konai and Williamson 1993a, 423VP
can.thar.i¢co.la. Gr. kantharos scarab beetle; L. suff. -cola
(from L. n. incola) inhabitant, dweller; N.L. n. cantharicola
an inhabitant of a family of beetles.
helical and motile. Colonies on solid medium containing
0.8% Noble agar are diffuse, without fried-egg morphology.
groups, and subgroups, but shares some reciprocal crossreactivity with members of other group XVI subgroups.
Not yet classified phylogenetically, but no doubt closely
related to subgroups XVI-2 and XVI-3, which are sisters forming a clade related to Spiroplasma diminutum in phylogenetic
trees constructed using 16S rRNA gene sequences. Moreover, DNA–DNA renaturation experiments confirm that the
differences between the type strain and other subgroups of
group XVI are great enough to warrant its designation as a
distinct species. The genome size is 1320 kbp (PFGE).
Source: isolated from the gut of an adult cantharid beetle
(Cantharis carolinus) in Maryland, USA. Based on its residence in the gut of a flower-visiting insect, this species is
thought to be transmitted on flowers.
DNA G+C content (mol%): 26 ± 1 (Tm, Bd, HPLC).
Type strain: ATCC 43207, CC-1.
6. Spiroplasma chinense Guo, Chen, Whitcomb, Rose, Tully,
Williamson, Ye and Chen 1990, 424VP
chi.nen¢se. N.L. neut. adj. chinense of China, the location
where the organism was first isolated.
motile helical filaments ~160 nm in diameter. Colonies on
solid medium containing 0.8–1.0% Noble agar are diffuse
with many small satellite colonies; growth on 2.25% agar
produces smaller rough or granular colonies and fewer satellite forms. Biological properties are listed in Table 142.
groups, and subgroups. Phylogenetically, this species is
closely related to Spiroplasma velocicrescens in phylogenetic
trees constructed using 16S rRNA gene sequences. The
Source: isolated from flower surfaces of bindweed (Calystegia hederacea) in Jiangsu, People’s Republic of China.
DNA G+C content (mol%): 29 ± 1 (Tm).
Type strain: ATCC 43960, CCH.
7. Spiroplasma chrysopicola Whitcomb, French, Tully, Gasparich, Rose, Carle, Bové, Henegar, Konai, Hackett, Adams,
Clark and Williamson 1997b, 718VP
chry.so.pi¢co.la. N.L. n. Chrysops a genus of deer flies in
the Tabanidae; L. suff. -cola (from L. n. incola) inhabitant,
dweller; N.L. n. chrysopicola inhabiting Chrysops spp.
Helical motile filaments are short and thin, passing a 220
nm filter quantitatively. Grows to titers as high as 1011/ml.
Colonies on solid medium containing 2.25% Noble agar
have dense centers and smooth edges (a fried-egg appearance) and do not have satellites. Biological properties are
listed in Table 142.
groups, and subgroups, but exhibits some reciprocal or
one-way cross-reactivity with members of other group VIII
subgroups. Some strains of group VIII spiroplasmas may
be difficult to identify to subgroup. Shares less than 70%
DNA–DNA reassociation with Spiroplasma syrphidicola and
strain TAAS-1 (subgroup VIII-3). Phylogenetically, this species is closely related to other group VIII strains in trees
constructed using 16S rRNA gene sequences. The 16S
rRNA gene similarity coefficients of group VIII spiroplasmas are >0.99, so this gene is insufficient for distinguishing
species in group VIII. The genome size is 1270 kbp (PFGE).
Pathogenicity for insects has not been determined.
Source: isolated from the gut of a deer fly (Chrysops sp.)
in Maryland, USA. Other strains from deer flies have been
collected from as far west as Wyoming, from New England,
and very rarely, as far south as Georgia, USA.
DNA G+C content (mol%): 30 ± 1 (Bd).
Type strain: ATCC 43209, DF-1.
8. Spiroplasma clarkii Whitcomb, Vignault, Tully, Rose, Carle, Bové, Hackett, Henegar, Konai and Williamson 1993c,
264VP
clar¢ki.i. N.L. masc. gen. n. clarkii of Clark, in honor of Truman B. Clark, a pioneer spiroplasma ecologist.
The morphology is as described for the genus. The helical motile filaments remain stable throughout exponential
growth. Colonies on solid medium containing 0.8% Noble
agar are diffuse, without fried-egg morphology. Biological
properties are listed in Table 142.
Serologically distinct from other Spiroplasma species, groups,
and subgroups. Phylogenetically, this species is placed in the
classical Apis cluster of spiroplasmas, but it does not have an
especially close neighbor in trees constructed using 16S rRNA
gene sequences. The genome size is 1720 kbp (PFGE). Pathogenicity for insects has not been determined.
Source: isolated from the gut of a larval scarabaeid beetle
(Cotinus nitida) in Maryland, USA.
Type strain: ATCC 33827, CN-5.
9. Spiroplasma corruscae Hackett, Whitcomb, French, Tully,
Gasparich, Rose, Carle, Bové, Henegar, Clark, Konai, Clark
and Williamson 1996c, 949VP
cor.rus¢cae. N.L. gen. n. corruscae of corrusca, referring to the
species of firefly beetle (Ellychnia corrusca) from which the
organism was first isolated.
helical and motile. Colonies on solid medium containing
671
2.25% Noble agar are slightly diffuse to discrete and generally without the characteristic fried-egg morphology. Biological properties are listed in Table 142.
Serologically distinct from previously established Spiroplasma species, groups, and subgroups. Phylogenetically,
closely related to Spiroplasma turonicum and Spiroplasma litorale in trees constructed using 16S rRNA gene sequences.
The genome size has not been determined.
Source: isolated from the gut of an adult lampyrid beetle
(Ellychnia corrusca) in Maryland in early spring, but found
much more frequently in horse flies in summer months.
Other strains have been collected from Canada and Georgia, Connecticut, South Dakota, and Texas, USA.
Type strain: ATCC 43212, EC-1.
10. Spiroplasma culicicola Hung, Chen, Whitcomb, Tully and
Chen 1987, 368VP
cu.li.ci′co.la. L. n. culex, -icis a gnat, midge, and also a genus
of mosquitoes (Culex, family Culicidae); L. suffix -cola (from
L. n. incola) inhabitant, dweller; N.L. n. culicicola intended
to mean an inhabitant of the Culicidae.
Cells are pleomorphic, but are commonly very short
motile helices, 1–2 mm in length. Colonies on solid medium
containing 1% Noble agar have a fried-egg appearance with
satellites. Biological properties are listed in Table 142.
placed in the classical Apis cluster of spiroplasmas, but does
not have an especially close neighbor in trees constructed
using 16S rRNA gene sequences. The genome size is 1350
kbp (PFGE).
Source: isolated from a triturate of a salt marsh mosquito
(Aedes sollicitans) collected in New Jersey, USA.
Type strain: ATCC 35112, AES-1.
11. Spiroplasma diabroticae Carle, Whitcomb, Hackett, Tully,
Rose, Bové, Henegar, Konai and Williamson 1997, 80VP
di.a.bro.ti′cae. N.L. gen. n. diabroticae of Diabrotica, referring
to Diabrotica undecimpunctata, the chrysomelid beetle from
which the organism was isolated.
helical, motile filaments, 200–300 nm in diameter. Colonies
on solid medium containing 0.8% Noble agar are diffuse,
without fried-egg morphology. Biological properties are
Serologically distinct from other established Spiroplasma
species, groups, and subgroups. Phylogenetically, closely
related to Spiroplasma floricola in trees constructed using
16S rRNA gene sequences. The genome size is 1350 kbp
(PFGE).
Source: isolated from the hemolymph of an adult chrysomelid beetle, Diabroticae undecimpunctata howardi.
Type strain: ATCC 43210, DU-1.
672
12. Spiroplasma diminutum Williamson, Tully, Rosen, Rose,
Whitcomb, Abalain-Colloc, Carle, Bové and Smyth 1996,
232VP
di.min.u¢tum. L. v. deminuere to break into small pieces,
make smaller; L. neut. part. adj. diminutum made smaller,
reflecting a smaller size.
short (1–2 mm), helical filaments, 100–200 nm in diameter
that appear to be rapidly moving, irregularly spherical bodies when exponential phase broth cultures are examined
under dark-field illumination. Colonies on solid medium
containing 1.6% Noble agar have dense centers, granular
perimeters, and nondistinct edges with satellite colonies.
groups, and subgroups. Phylogenetically, closely related
to group XVI spiroplasmas in trees constructed using
(PFGE).
Source: isolated from a frozen triturate of adult female
Culex annulus mosquitoes collected in Taishan, Taiwan.
Type strain: ATCC 49235, CUAS-1.
13. Spiroplasma floricola Davis, Lee and Worley 1981, 462VP
flor.i¢co.la. L. n. flos, -oris a flower; L. suff. -cola (from L. n.
incola) inhabitant, dweller; N.L. n. floricola flower-dweller.
cells are 150–200 nm in diameter and 2–5 mm in length.
Colonies on solid media have granular central regions
surrounded by satellite colonies that probably form after
migration of cells from the central focus. Biological properties are listed in Table 142.
groups, and subgroups. Phylogenetically, closely related
to Spiroplasma diabroticae and various flower spiroplasmas
genome size of strain OBMG is 1270 kbp. Experimentally
pathogenic for insects and embryonated chicken eggs.
Source: isolated from flowers of tulip tree and magnolia
trees in Maryland, USA. Other strains have been collected
from coleopterous insects.
Type strain: ATCC 29989, 23-6.
14. Spiroplasma gladiatoris Whitcomb, French, Tully, Gasparich, Rose, Carle, Bové, Henegar, Konai, Hackett, Adams,
gla.di.a¢to.ris. L. gen. n. gladiatoris of a gladiator, reflecting the initial isolation of the organism from the horse fly
Tabanus gladiator.
Morphology is as described for the genus. Cells are
motile helical filaments. Colonies on solid medium containing 3% Noble agar are granular with dense centers and diffuse edges, do not have satellites, and never have a fried-egg
appearance. Biological properties are listed in Table 142.
groups, and subgroups. This species has a specific antigen,
common to several spiroplasmal inhabitants of horse flies,
that confers a high level of one way cross-reactivity when it
is used as an antigen. Phylogenetically, closely related to two
other tabanid spiroplasmas, Spiroplasma helicoides and group
XXXIV strain B1901, in phylogenetic trees constructed
using 16S rRNA gene sequences. The genome size has not
been determined.
Source: isolated from the gut of a horse fly (Tabanus gladiator) in Maryland, USA. Other strains have been collected
at various locations in the southeastern United States.
Type strain: ATCC 43525, TG-1.
15. Spiroplasma helicoides Whitcomb, French, Tully, Gasparich, Rose, Carle, Bové, Henegar, Konai, Hackett, Adams,
he.li.co.i¢des. Gr. n. helix spiral; Gr. suff. -oides like, resembling, similar; N.L. neut. adj. helicoides spiral-like.
motile helical filaments that lack a cell wall. Colonies on
solid medium containing 2.25% Noble agar have dense
centers and smooth edges, do not have satellites, and have
a perfect fried-egg appearance. Biological properties are
groups, and subgroups. This species has a specific antigen,
common to several spiroplasmal inhabitants of horse flies,
that confers a high level one-way cross-reaction when it is
used as antigen. Phylogenetically, closely related to two
other tabanid spiroplasmas, Spiroplasma gladiatoris and Spiroplasma sp. BARC 1901, in trees constructed using 16S rRNA
gene sequences. Genome size has not been determined.
Source: isolated from the gut of a horse fly Tabanus abactor
collected in Oklahoma, USA. Other strains have been collected in Georgia, USA.
Type strain: ATCC 51746, TABS-2.
16. Spiroplasma insolitum Hackett, Whitcomb, Tully, Rose,
Carle, Bové, Henegar, Clark, Clark, Konai, Adams and Williamson 1993, 276VP
in.so′li.tum. L. neut. adj. insolitum unusual or uncommon,
to denote unusual base composition.
Cells in exponential phase are long, motile, helical cells
that lack true cell walls and periplasmic fibrils. Colonies on
solid SP-4 medium containing 0.8 or 2.25% Noble agar are
diffuse, with small central zones of growth surrounded by
small satellite colonies. Colonies on solid SP-4 medium containing horse serum and 0.8% Noble agar show fried-egg
morphology. Biological properties are listed in Table 142.
Serologically distinct from other Spiroplasma species and
groups, but cross-reacts reciprocally in complex patterns
of relatedness with group I subgroups and Spiroplasma
poulsonii. DNA–DNA renaturation experiments confirm
that the differences between the type strain and other subgroups of group I are great enough to warrant its designation as a distinct species. Has close phylogenetic affinities
with other group I members and with Spiroplasma poulsonii
genome size is 1850 kbp (PFGE). Pathogenicity for insects
has not been determined.
Source: the type strain was isolated from a fall flower (Asteraceae: Bidens sp.) collected in Maryland, USA. Similar isolates have been found in the hemocoel of click beetles. Also
isolated from other composite and onagracead flowers and
from the guts of many insects visiting these flowers, including cantharid and meloid beetles; syrphid flies; andrenid
and megachilid bees; and four families of butterflies.
Type strain: ATCC 33502, M55.
17. Spiroplasma ixodetis Tully, Rose, Yunker, Carle, Bové, Williamson and Whitcomb 1995, 27VP
ix.o.de¢tis. N.L. gen. n. ixodetis of Ixodes, the genus name
of Ixodes pacificus ticks, from which the organism was first
isolated.
Cells are coccoid forms, 300–500 nm in diameter, straight
and branched filaments, or tightly coiled helical organisms.
Motility is flexional, but not translational. Colonies on solid
medium containing 2.25% Noble agar usually have the
appearance of fried eggs. Biological properties are listed in
Table 142.
groups, and subgroups. Phylogenetically unique; occurs
at base of spiroplasma lineage in trees constructed using
(PFGE).
Source: isolated from macerated tissue suspensions prepared from pooled adult Ixodes pacificus ticks (Ixodidae)
collected in Oregon, USA.
Type strain: ATCC 33835, Y32.
18. Spiroplasma kunkelii Whitcomb, Chen, Williamson, Liao,
Tully, Bové, Mouches, Rose, Coan and Clark 1986, 175VP
kun.kel¢i.i. N.L. masc. gen. n. kunkelii of Kunkel, named
after Louis Otto Kunkel (1884–1960), to honor his major
and fundamental contributions to the study of plant mollicutes.
Cells in exponential phase are helical, motile filaments,
100–150 nm in diameter and 3–10 mm long to nonhelical
filaments or spherical cells, 300–800 nm in diameter. Colonies on solid medium containing 0.8% Noble agar are usually diffuse, rarely exhibiting central zones of growth into
agar. Colonies on solid C-3G medium containing 5% horse
serum or on media containing 2.25% Noble agar frequently
have a fried-egg morphology. Biological properties are
groups, and subgroups, but shares complex patterns of
reciprocal cross-reactivity with members of other group I
subgroups and Spiroplasma poulsonii. DNA–DNA renaturation experiments confirm that the serological differences
between the type strain and other subgroups of group I are
great enough to warrant its designation as a distinct species. Has close phylogenetic affinities with other group I
673
members and with Spiroplasma poulsonii in trees constructed
kbp (PFGE). Pathogenicity for plants and insects has been
experimentally verified.
Source: isolated from maize displaying symptoms of corn
stunt disease and from leafhoppers associated with diseased
maize, largely in the neotropics.
Type strain: ATCC 29320, E275.
Sequence accession no. (16S rRNA gene): DQ319068 (strain
CR2-3x).
19. Spiroplasma lampyridicola Stevens, Tang, Jenkins, Goins,
Tully, Rose, Konai, Williamson, Carle, Bové, Hackett,
French, Wedincamp, Henegar and Whitcomb 1997, 711VP
lam.py.ri.di¢co.la. N.L. n. Lampyridae the firefly beetle family;
L. suff. -cola (from L. n. incola) inhabitant, dweller; N.L. n.
lampyridicola an inhabitant of members of the Lampyridae.
motile helical filaments. Colonies on solid medium containing 3.0% Noble agar are small and granular with dense centers, but do not have a true fried-egg appearance. Biological
groups, and subgroups. When tested as antigen, cross-reacts
(one-way) with many specific spiroplasma antisera. Phylogenetically, a sister to Spiroplasma leptinotarsae in trees constructed using 16S rRNA gene sequences. The genome size
is 1375 kbp (PFGE).
Source: isolated from the gut fluids of a firefly beetle
(Photuris pennsylvanicus) collected in Maryland, USA. Also
known from Georgia and New Jersey, USA.
Type strain: ATCC 43206, PUP-1.
20. Spiroplasma leptinotarsae Hackett, Whitcomb, Clark, Henegar, Lynn, Wagner, Tully, Gasparich, Rose, Carle, Bové,
Konai, Clark, Adams and Williamson 1996b, 910VP
lep.ti.no.tar¢sae. N.L. gen. n. leptinotarsae of Leptinotarsa,
referring to Leptinotarsa decemlineata, the Colorado potato
beetle.
Cells in vivo are usually seen in the resting stage, in which
they consist of coin-like compressed coils. When placed in
fresh medium, these bodies turn immediately into “spring”or “funnel”-shaped spirals, which are capable of very rapid
translational motility. After a relatively small number of
passes in vitro, this spectacular morphology is lost and the
cells return to the modal morphology as described for the
genus. Colonies on solid medium containing 2.0% Noble
agar are slightly diffuse to discrete and produce numerous
groups, and subgroups. When tested as antigen, cross-reacts
with many spiroplasma antisera (one-way). Phylogenetically, a sister to Spiroplasma lampyridicola in trees constructed
using 16S rRNA gene sequences. The genome size is 1,085
kbp (PFGE).
Source: isolated from the gut of Colorado potato beetle
(Leptinotarsa decemlineata) larvae in Maryland, USA. Also
674
isolated from beetles collected in Maryland, Michigan, New
Mexico, North Carolina, Texas, Canada, and Poland.
Type strain: ATCC 43213, LD-1.
21. Spiroplasma leucomae Oduori, Lipa and Gasparich 2005,
2449VP
leu.co¢mae. N.L. gen. n. leucomae of Leucoma, systematic
genus name of the white satin moth (Lepidoptera: Lymantriidae), the source of the type strain.
Morphology is as described for the genus. Cells are filamentous, helical, motile, and approximately 150 nm in
diameter. They freely pass through filters with pores of 450
and 220 nm, but do not pass through filters with 100 nm
pores. Biological properties are listed in Table 142.
Serologically distinct from previously established Spiroplasma species, groups, and subgroups. The genome size
has not been determined. Pathogenicity for the moth larvae is not known.
Source: isolated from fifth instar satin moth larvae (Leucoma salicis).
Type strain: ATCC BAA-521, NBRC 100392, SMA.
22. Spiroplasma lineolae French, Whitcomb, Tully, Carle, Bové,
Henegar, Adams, Gasparich and Williamson 1997, 1080VP
lin.e.o¢lae. N.L. n. lineola a species of tabanid fly; N.L. gen.
n. lineolae of Tabanus lineola, from which the organism was
isolated.
motile, helical filaments, 200–300 nm in diameter. Colonies on solid medium containing 3% Noble agar are small,
granular, and never have a fried-egg appearance. Biological
groups, and subgroups. Phylogenetic position has not been
determined, but its other taxonomic properties suggest that
it may be related to other tabanid spiroplasmas of the Apis
cluster. The genome size is 1390 kbp (PFGE).
Source: type strain isolated from the viscera of the tabanid
fly Tabanus lineola collected in coastal Georgia. A strain from
Tabanus lineola has been collected in Costa Rica (Whitcomb
et al., 2007).
Type strain: ATCC 51749, TALS-2.
23. Spiroplasma litorale Konai, Whitcomb, French, Tully, Rose,
Carle, Bové, Hackett, Henegar, Clark and Williamson 1997,
361VP
li.to.ra¢le. L. neut. adj. litorale of the shore or coastal area.
motile, helical filaments. Colonies on solid medium containing 2.25% Noble agar are granular with dense centers, uneven
margins, and multiple satellites, and never have fried-egg
groups, and subgroups. Phylogenetically, closely related to
two other tabanid spiroplasmas, Spiroplasma turonicum and
Spiroplasma litorale, in trees constructed using 16S rRNA
gene sequences. The genome size is 1370 kbp (PFGE).
Source: isolated from the gut of a female green-eyed horse
fly (Tabanus nigrovittatus) from the Outer Banks of North
Carolina. Also collected from coastal Georgia and both
Atlantic and Pacific coasts of Costa Rica.
Type strain: ATCC 34211, TN-1.
24. Spiroplasma melliferum Clark, Whitcomb, Tully, Mouches,
Saillard, Bové, Wróblewski, Carle, Rose, Henegar and Williamson 1985, 305VP
mel.li′fe.rum. L. adj. mellifer, -fera, -ferum honey-bearing,
honey-producing; L. neut. adj. melliferum intended to mean
isolated from the honey bee (Apis mellifera).
Morphology is as described for the genus. Cells are pleomorphic, varying from helical filaments that are 100–150
nm in diameter and 3–10 mm in length to nonhelical filaments or spherical cells that are 300–800 nm in diameter.
The motile cells lack true cell wells and periplasmic fibrils.
Colonies on solid medium supplemented with 0.8% Noble
agar are usually diffuse, rarely exhibiting central zones of
growth into agar. Colonies on solid medium containing
2.25% Noble agar are smaller, but frequently have a friedegg morphology. Physiological and genomic properties are
groups, and subgroups, but shares complex patterns of
reciprocal cross-reactivity with members of other group I
subgroups and Spiroplasma poulsonii. Has close phylogenetic affinities with other group I members and with Spiroplasma poulsonii in trees constructed using 16S rRNA gene
sequences. DNA–DNA renaturation experiments confirm
that the serological differences between the type strain and
other subgroups of group I are great enough to warrant its
designation as a distinct species. The genome size is 1460
kbp (PFGE). Pathogenic for honey bees in natural and
experimental oral infections.
Source: isolated from hemolymph and gut of honey bees
(Apis mellifera) in widely separated geographic regions. Also
recovered from hemolymph of bumble bees, leafcutter
bees, and a robber fly, and the intestinal contents of sweat
bees, digger bees, bumble bees, and a butterfly. Also recovered from a variety of plant surfaces (flowers) in widely
separated geographic regions.
Type strain: ATCC 33219, BC-3.
25. Spiroplasma mirum Tully, Whitcomb, Rose and Bové 1982,
99VP
mi′rum. L. neut. adj. mirum extraordinary.
filaments measure 100–200 nm in diameter and 3–8 mm
in length. Colonies on solid media containing fetal bovine
serum and 0.8–2.25% Noble agar (Difco) are diffuse and
without central zones of growth into the agar. Solid media
prepared with 1.25% agar and in which fetal bovine serum
has been replaced with bovine serum fraction yield colonies
with central zones of growth into the agar and no peripheral
growth on the surface of the medium. Moderate turbidity is
produced during growth in liquid media. Biological properties are listed in Table 142. This species has been cultivated
in a defined medium.
groups, and subgroups. Phylogenetically, in trees constructed using 16S rRNA gene sequences, this species is
basal to group I and group VIII spiroplasmas on the one
hand, and to the Apis cluster and Entomoplasmataceae on the
other. It is the most primitive (plesiomorphic) spiroplasma
with modal helicity. The genome size is 1300 kbp (PFGE).
Produces experimental ocular and nervous system disease
and death in intracerebrally inoculated suckling animals
(rats, mice, hamsters, and rabbits). Pathogenic for chicken
embryos via yolk sac inoculation. Experimentally pathogenic for the wax moth (Galleria mellonella).
Source: the type strain was isolated from rabbit ticks (Haemaphysalis leporispalustris) collected in Georgia, USA. Other
strains have been collected in Georgia, Maryland, and New
York, USA.
DNA G+C content (mol%): 30–31 (Tm).
Type strain: ATCC 29335, SMCA.
26. Spiroplasma monobiae Whitcomb, Tully, Rose, Carle, Bové,
Henegar, Hackett, Clark, Konai, Adams and Williamson
1993b, 259VP
mo.no.bi′ae. N.L. n. Monobia a genus of vespid wasps; N.L.
gen. n. monobiae of the genus Monobia, from which the
organism was isolated.
The morphology is as described for the genus, with
motile helical filaments. Colonies on solid medium containing 2.25% Noble agar are diffuse and never have a fried-egg
groups, and subgroups. Phylogenetically, a member of the
Apis clade, but with no especially close neighbors in trees
constructed using 16S rRNA gene sequences. The genome
size is 940 kbp (PFGE).
Source: isolated from the hemolymph of an adult vespid
wasp (Monobia quadridens) collected in Maryland, USA.
Based on its residence in the gut of a flower-visiting insect,
this species is thought to be transmitted on flowers.
Type strain: ATCC 33825, MQ-1.
27. Spiroplasma montanense Whitcomb, French, Tully, Rose,
Carle, Bové, Clark, Henegar, Konai, Hackett, Adams and
Williamson 1997c, 722VP
mon.ta.nen¢se. N.L. neut. adj. montanense pertaining to
Montana, where the species was first isolated.
The morphology is as described for the genus. Cells
are motile, helical filaments that lack a cell wall. Colonies on solid medium containing 2.25% Noble agar are
granular and have dense centers, irregular margins, and
numerous small satellites. Biological properties are listed
in Table 142.
675
groups, and subgroups. Reacts reciprocally in deformation
serology at very low levels in deformation tests with Spiroplasma apis. “Bridge strains” have been isolated in Georgia
with substantial cross-reactivity with both Spiroplasma montanense and Spiroplasma apis. Sister to Spiroplasma apis in trees
constructed using 16S rRNA gene sequences. The genome
size is 1225 kbp (PFGE).
Source: isolated from the gut of the tabanid fly Hybomitra
opaca, in southwestern Montana. Other isolates have been
obtained from New England, Connecticut, and southeastern Canada.
Type strain: ATCC 51745, HYOS-1.
28. Spiroplasma penaei Nunan, Lightner, Oduori and Gasparich 2005, 2320VP
pe.na′e.i. N.L. n. Penaeus a species of shrimp; N.L. gen.
penaei of Penaeus, referring to Penaeus vannamei, from which
the organism was isolated.
The morphology is as described for the genus. Cells are helical and motile. Biological properties are listed in Table 142.
Serologically distinct from previously characterized Spiroplasma species, groups, and subgroups, but shares some
cross-reactivity with members of other group I subgroups.
Has close phylogenetic affinities with other group I members and with Spiroplasma poulsonii in trees constructed using
16S rRNA gene sequences. The genome size has not been
determined. Pathogenicity has been indicated by injection
into Penaeus vannamei.
Source: isolated from the hemolymph of the Pacific white
shrimp, Penaeus vannamei.
Type strain: CAIM 1252, SHRIMP, ATCC BAA-1082.
29. Spiroplasma phoeniceum Saillard, Vignault, Bové, Raie,
Tully, Williamson, Fos, Garnier, Gadeau, Carle and Whitcomb 1987, 113VP
phoe.ni¢ce.um. N.L. neut. adj. phoeniceum (from L. neut.
adj. phonicium) of Phoenice, an ancient country that was
located on today’s Syrian coast, referring to the geographical origin of the isolates.
Morphology is as described for the genus. Colonies on
solid medium containing 0.8% Noble agar show fried-egg
morphology. Physiological and genomic properties are
groups, and subgroups, but shares some cross-reactivity with
members of other group I subgroups and Spiroplasma poulsonii. Has close phylogenetic affinities with other group I
members and with Spiroplasma poulsonii in trees constructed
using 16S rRNA gene sequences. Has been shown to be
transmissible to leafhoppers by injection and experimentally pathogenic to aster inoculated by the injected leafhoppers. DNA–DNA renaturation experiments confirm that the
differences between the type strain and other subgroups of
group I are great enough to warrant its designation as a distinct species. The genome size is 1860 kbp (PFGE).
676
Source: isolated from periwinkles that were naturally
infected in various locations along the Syrian coastal area.
Type strain: ATCC 43115, P40.
32. Spiroplasma sabaudiense Abalain-Colloc, Chastel, Tully,
Bové, Whitcomb, Gilot and Williamson 1987, 264VP
30. Spiroplasma platyhelix Williamson, Adams, Whitcomb, Tully, Carle, Konai, Bové and Henegar 1997, 766VP
helical filaments, 100–160 nm in diameter and 3.1–3.8 mm
long. Motile. Colonies on solid medium containing 1.6%
Noble agar are diffuse, rarely exhibiting fried-egg morphology, with numerous satellite colonies. Physiological and
genomic properties are listed in Table 142.
groups, and subgroups. Phylogenetically, related to Spiroplasma alleghenense and Spiroplasma sp. TIUS-1 in trees constructed using 16S rRNA gene sequences. The genome size
is 1175 kbp (PFGE).
Source: isolated from a triturate of female Aedes spp. mosquitoes in Savoy, France.
Type strain: ATCC 43303, Ar-1343.
pla.ty.he¢lix. Gr. adj. platys flat; Gr. fem. n. helix a coil or spiral; N.L. fem. n. platyhelix flat coil, referring to the flattened
nature of the helical filament.
Cells are flattened, helical filaments, 200–300 nm in
diameter. They show no rotatory or translational motility,
but exhibit contractile movements in which tightness of coiling moves along the axis of the filament. Colonies on solid
medium containing 2.25% Noble agar form perfect friedegg colonies with dense centers, smooth edges, and without
Serologically distinct from other Spiroplasma species, groups, and subgroups. The genome size is 780 kbp
(PFGE).
Source: isolated from the gut of a dragonfly, Pachydiplax
longipennis, collected in Maryland, USA.
Type strain: ATCC 51748, PALS-1.
31. Spiroplasma poulsonii Williamson, Sakaguchi, Hackett,
Whitcomb, Tully, Carle, Bové, Adams, Konai and Henegar
1999, 616VP
poul.so′ni.i. N.L. masc. gen. n. poulsonii of Poulson, named
in memory of Donald F. Poulson, in whose laboratory at
Yale University this spiroplasma was discovered and studied
intensively.
Morphology is as described for the genus. Long, motile,
helical filaments, 200–250 nm in diameter occur in vivo
in Drosophila hemolymph and in vitro. Colonies on solid
medium containing 1.8% Noble agar are small (60–70 mm
in diameter), have dense centers and uneven edges, and
are without satellites. Biological properties are listed in
Table 142.
groups, and subgroups, but shares some reciprocal crossreactivity with members of group I subgroups. Phylogenetically related to group I spiroplasmas in trees constructed
kbp (PFGE). Spiroplasmas causing sex-ratio abnormalities
occur naturally in Drosophila spp. collected in Brazil, Colombia, Dominican Republic, Haiti, Jamaica, and the West
Indies. Non-male-lethal spiroplasmas also occur in natural populations of Drosophila hydei in Japan. Pathogenicity
(lethality to male progeny) has been confirmed by injection
into Drosophila pseudoobscura female flies. Vertical transmissibility is lost after cultivation and cloning.
Source: isolated from the hemolymph of Drosophila pseudoobscura females infected by hemolymph transfer of the
Barbados-3 strain of Drosophila willistoni SR organism.
Type strain: ATCC 43153, DW-1.
sa.bau.di.en¢se. L. neut. adj. sabaudiense of Sabaudia, an
ancient country of Gaul, corresponding to present day
Savoy, referring to the geographic origin of the isolate.
33. Spiroplasma syrphidicola Whitcomb, Gasparich, French,
Tully, Rose, Carle, Bové, Henegar, Konai, Hackett, Adams,
Clark and Williamson 1996, 799VP
syr.phi.di¢co.la. N.L. pl. n. Syrphidae a family of flies; L. suff.
-cola (from L. masc. or fem. n. incola) inhabitant, dweller;
N.L. masc. n. syrphidicola inhabitant of syrphid flies, the
insects from which the organism was isolated.
Helical motile filaments are short and thin, passing a
220 nm filter quantitatively. Grows to titers as high as 1011/
ml. These short, thin, abundant cells are provisionally diagnostic for group VIII. Colonies on solid medium containing 2.25% Noble agar are irregular with satellites, diffuse,
and never have a fried-egg appearance. Growth on solid
medium containing 1.6% Noble agar is diffuse. Biological
groups, and subgroups, but shares some reciprocal crossreactivity with members of other group VIII subgroups.
Placement of group VIII strains into subgroups has become
increasingly difficult as more strains have accumulated.
Phylogenetically, this species is closely related to other
group VIII strains in trees constructed using 16S rRNA
gene sequences. The 16S rRNA gene sequence similarity
coefficients of group VIII spiroplasmas are >0.99, so this
gene is insufficient for species separations in group VIII.
DNA–DNA renaturation experiments confirm that the differences between the type strain and other subgroups of
group VIII are great enough to warrant its designation as a
distinct species. Genome size is 1230 kbp (PFGE).
Source: isolated from the hemolymph of the syrphid fly
Eristalis arbustorum in Maryland, USA. Strains that are provisionally identified as Spiroplasma syrphidicola have been
obtained from horse flies collected from several locations
in the southeastern United States.
Type strain: ATCC 33826, EA-1.
677
34. Spiroplasma tabanidicola Whitcomb, French, Tully, Gasparich, Rose, Carle, Bové, Henegar, Konai, Hackett, Adams,
36. Spiroplasma turonicum Hélias, Vazeille-Falcoz, Le Goff, Abalain-Colloc, Rodhain, Carle, Whitcomb, Williamson, Tully,
Bové and Chastel 1998, 460VP
ta.ba.ni.di¢co.la. N.L. n. Tabanidae family name for horse
flies; L. suff. -cola (from L. n. incola) inhabitant, dweller;
N.L. n. tabanidicola an inhabitant of horse flies.
tu.ro¢ni.cum. L. neut. adj. turonicum of Touraine, the
province in France from which the organism was first isolated.
motile, helical filaments that lack a cell wall. Colonies on
solid medium containing 3% Noble agar are uneven and
granular with dense centers and irregular edges, do not
have satellites, and never have a fried-egg appearance. Physiological and genomic properties are listed in Table 142.
groups, and subgroups. However, some strains may show
a very low level reciprocal serological cross-reaction in
deformation serology with Spiroplasma gladiatoris. This species has a specific antigen, common to several spiroplasmal
inhabitants of horse flies, that confers a high level one-way
cross-reaction when it is used as antigen. The genome size
is 1375 kbp (PFGE).
Source: isolated from the gut of a horse fly belonging to
the Tabanus abdominalis-limbatinevris complex.
Type strain: ATCC 51747, TAUS-1.
motile, helical filaments. Colonies on solid medium containing 3% Noble agar exhibit a “cauliflower-like” appearance and do not have a fried-egg morphology. Biological
Serologically distinct from previously established Spiroplasma species. Phylogenetically, related to two other tabanid
spiroplasmas, Spiroplasma corruscae and Spiroplasma litorale,
Source: isolated from a triturate of a single horse fly (Haematopota pluvialis) collected in France.
Type strain: ATCC 700271, Tab4c.
35. Spiroplasma taiwanense Abalain-Colloc, Rosen, Tully, Bové,
Chastel and Williamson 1988, 105VP
tai.wan.en¢se. N.L. neut. adj. taiwanense of or belonging to
Taiwan, referring to the geographic origin of the isolate.
motile, helical filaments, 100–160 nm in diameter and 3.1–
3.8 mm long. Colonies on solid medium containing 1.6%
Noble agar have fried-egg morphology. Biological properties are listed in Table 142.
groups, and subgroups. Phylogenetically, this species is in the
classical Apis cluster of spiroplasmas, but does not have an
especially close neighbor in trees constructed using 16S rRNA
gene sequences. The genome size is 1195 kbp (PFGE).
Source: isolated from a triturate of female mosquitoes (Culex
tritaeniorhynchus) at Taishan, Taiwan, Republic of China.
Type strain: ATCC 43302, CT-1.
References
Abalain-Colloc, M.L., C. Chastel, J.G. Tully, J.M. Bové, R.F. Whitcomb,
B. Gilot and D.L. Williamson. 1987. Spiroplasma sabaudiense sp. nov.
from mosquitos collected in France. Int. J. Syst. Bacteriol. 37: 260–
265.
Abalain-Colloc, M.L., L. Rosen, J.G. Tully, J.M. Bové, C. Chastel and
D.L. Williamson. 1988. Spiroplasma taiwanense sp. nov. from Culex tritaeniorhynchus mosquitos collected in Taiwan. Int. J. Syst. Bacteriol.
38: 103–107.
Abalain-Colloc, M.L., D.L. Williamson, P. Carle, J.H. Abalain, F. Bonnet, J.G. Tully, M. Konai, R.F. Whitcomb, J.M. Bové and C. Chastel.
1993. Division of group XVI spiroplasmas into subgroups. Int. J. Syst.
Bacteriol. 43: 342–346.
37. Spiroplasma velocicrescens Konai, Whitcomb, Tully, Rose,
Carle, Bové, Henegar, Hackett, Clark and Williamson 1995,
205VP
ve.lo.ci.cres¢cens. L. adj. velox, -ocis fast, quick; L. part.
adj. crescens growing; N.L. n. part. adj. velocicrescens fastgrowing.
helical, motile filaments, 200–300 nm in diameter. Colonies
on solid medium containing 0.8% Noble agar are diffuse
and never have a fried-egg appearance. Biological properties are listed in Table 142.
sister to Spiroplasma chinense in trees constructed using
(PFGE).
Source: isolated from the gut of a vespid wasp, Monobia
quadridens, collected in Maryland, USA. Based on its residence in the gut of a flower-visiting insect, this species is
thought to be transmitted on flowers.
Type strain: ATCC 35262, MQ-4.
Adams, J.R., R.F. Whitcomb, J.G. Tully, E.A. Clark, D.L. Rose, P. Carle,
M. Konai, J.M. Bové, R.B. Henegar and D.L. Williamson. 1997.
Spiroplasma alleghenense sp. nov., a new species from the scorpion fly
Panorpa helena (Mecoptera: Panorpidae). Int. J. Syst. Bacteriol. 47:
759–762.
Alexeeva, I., E.J. Elliott, S. Rollins, G.E. Gasparich, J. Lazar and R.G.
Rohwer. 2006. Absence of Spiroplasma or other bacterial 16S rRNA
genes in brain tissue of hamsters with scrapie. J. Clin. Microbiol. 44:
91–97.
Alivizatos, A.S. 1988. Isolation and culture of corn stunt Spiroplasma in
serum-free medium. J. Phytopathol. 122: 68–75.
Aluotto, B.B., R. G. Wittler, C.O.Williams and J. E. Faber. 1970. Standardized bacteriologic techniques for characterization of Mycoplasma
species. Int. J. Syst. Bacteriol. 20: 35–58.
678
Amikam, D., S. Razin and G. Glaser. 1982. Ribosomal RNA genes in
Mycoplasma. Nucleic Acids Res. 10: 4215–4222.
Amikam, D., G. Glaser and S. Razin. 1984. Mycoplasmas (Mollicutes)
have a low number of rRNA genes. J. Bacteriol. 158: 376–378.
Ammar, E. and S.A. Hogenhout. 2005. Use of immunofluorescence confocal laser scanning microscopy to study distribution of the bacterium
corn stunt spiroplasma in vector leafhoppers (Hemiptera: Cicadellidae) and in host plants. Ann. Entomol. Soc. Am. 98: 820–826.
Ammar, E.D., D. Fulton, X. Bai, T. Meulia and S.A. Hogenhout. 2004. An
attachment tip and pili-like structures in insect- and plant-pathogenic
spiroplasmas of the class Mollicutes. Arch. Microbiol. 181: 97–105.
Anbutsu, H. and T. Fukatsu. 2003. Population dynamics of male-killing
and non-male-killing spiroplasmas in Drosophila melanogaster. Appl.
Environ. Microbiol. 69: 1428–1434.
André, A., W. Maccheroni, F. Doignon, M. Garnier and J. Renaudin. 2003.
Glucose and trehalose PTS permeases of Spiroplasma citri probably share
a single IIA domain, enabling the spiroplasma to adapt quickly to carbohydrate changes in its environment. Microbiology 149: 2687–2696.
André, A., M. Maucourt, A. Moing, D. Rolin and J. Renaudin. 2005.
Sugar import and phytopathogenicity of Spiroplasma citri: glucose and
fructose play distinct roles. Mol. Plant. Microbe. Interact. 18: 33–42.
Archer, D.B., J. Best and C. Barber. 1981. Isolation and restriction mapping of a spiroplasma plasmid. J. Gen. Microbiol. 126: 511–514.
Bai, X. and S.A. Hogenhout. 2002. A genome sequence survey of the
mollicute corn stunt spiroplasma Spiroplasma kunkelii. FEMS Microbiol. Lett. 210: 7–17.
Barile, M.F. 1983. Arginine hydrolysis. In Methods in Mycoplasmology, vol.
1 (edited by Razin and Tully). Academic Press, New York, pp. 345–349.
Baseman, J.B. and J.G. Tully. 1997. Mycoplasmas: sophisticated, re-emerging, and burdened by their notoriety. Emerg. Infect. Dis. 3: 21–32.
Bastian, F.O. 1979. Spiroplasma-like inclusions in Creutzfeldt-Jakob disease. Arch. Pathol. Lab. Med. 103: 665–669.
Bastian, F.O. 2005. Spiroplasma as a candidate agent for the transmissible spongiform encephalopathies. J. Neuropathol. Exp. Neurol. 64:
833–838.
Bastian, F.O. and J.W. Foster. 2001. Spiroplasma sp. 16S rDNA in
Creutzfeldt-Jakob disease and scrapie as shown by PCR and DNA
sequence analysis. J. Neuropathol. Exp. Neurol. 60: 613–620.
Bastian, F.O., S. Dash and R.F. Garry. 2004. Linking chronic wasting disease to scrapie by comparison of Spiroplasma mirum ribosomal DNA
sequences. Exp. Mol. Pathol. 77: 49–56.
Bastian, F.O. and C.D. Fermin. 2005. Slow virus disease: deciphering
conflicting data on the transmissible spongiform encephalopathies
(TSE) also called prion diseases. Microsc. Res. Tech. 68: 239–246.
Bastian, F.O., D.E. Sanders, W.A. Forbes, S.D. Hagius, J.V. Walker, W.G.
Henk, F.M. Enright and P.H. Elzer. 2007. Spiroplasma spp. from transmissible spongiform encephalopathy brains or ticks induce spongiform
encephalopathy in ruminants. J. Med. Microbiol. 56: 1235–1242.
Bébéar, C.M., P. Aullo, J.M. Bove and J. Renaudin. 1996. Spiroplasma citri
virus SpV1: characterization of viral sequences present in the spiroplasma host chromosome. Curr. Microbiol. 32: 134–140.
Bentley, J.K., Z. Veneti, J. Heraty and G.D. Hurst. 2007. The pathology
of embryo death caused by the male-killing Spiroplasma bacterium in
Drosophila nebulosa. BMC Biol. 5: 9.
Berg, M., U. Melcher and J. Fletcher. 2001. Characterization of Spiroplasma citri adhesion related protein SARP1, which contains a domain
of a novel family designated sarpin. Gene 275: 57–64.
Berho, N., S. Duret, J.L. Danet and J. Renaudin. 2006a. Plasmid pSci6
from Spiroplasma citri GII-3 confers insect transmissibility to the nontransmissible strain S. citri 44. Microbiology 152: 2703–2716.
Berho, N., S. Duret and J. Renaudin. 2006b. Absence of plasmids encoding adhesion-related proteins in non-insect-transmissible strains of
Spiroplasma citri. Microbiology 152: 873–886.
Bévén, L., M. LeHenaff, C. Fontenelle and H. Wróblewski. 1996. Inhibition of spiralin processing by the lipopeptide antibiotic globomycin.
Bévén, L. and H. Wróblewski. 1997. Effect of natural amphipathic peptides on viability, membrane potential, cell shape and motility of mollicutes. Res. Microbiol. 148: 163–175.
Bévén, L., D. Duval, S. Rebuffat, F.G. Riddell, B. Bodo and H. Wróblewski. 1998. Membrane permeabilisation and antimycoplasmic
activity of the 18-residue peptaibols, trichorzins PA. Biochim. Biophys. Acta 1372: 78–90.
Bi, K., H. Huang, W. Gu, J. Wang and W. Wang. 2008. Phylogenetic
analysis of Spiroplasmas from three freshwater crustaceans (Eriocheir
sinensis, Procambarus clarkia and Penaeus vannamei) in China. J. Invertebr. Pathol. 99: 57–65.
Boutareaud, A., J.L. Danet, M. Garnier and C. Saillard. 2004. Disruption
of a gene predicted to encode a solute binding protein of an ABC
transporter reduces transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps. Appl. Environ. Microbiol. 70: 3960–3967.
Bové, J.M. and C. Saillard. 1979. Cell biology of spiroplasmas. In The
Mycoplasmas, vol. 3 (edited by Whitcomb and Tully). Academic
Bové, J.M., C. Saillard, P. Junca, J.R. DeGorce-Dumas, B. Ricard, A.
Nhami, R.F. Whitcomb, D. Williamson and J.G. Tully. 1982. Guanineplus-cytosine content, hybridization percentages, and EcoRI restriction enzyme profiles of spiroplasmal DNA. Rev. Infect. Dis. 4 Suppl:
S129–136.
Bové, J.M., C. Mouches, P. Carle-Junca, J.R. Degorce-Dumas, J.G. Tully
and R.F. Whitcomb. 1983. Spiroplasmas of Group I: the Spiroplasma
citri cluster. Yale J. Biol. Med. 56: 573–582.
Bové, J.M., P. Carle, M. Garnier, F. Laigret, J. Renaudin and C. Saillard.
1989. Molecular and cellular biology of spiroplasmas. In The Mycoplasmas, vol. 5 (edited by Whitcomb and Tully). Academic Press,
Bové, J.M. 1993. Molecular features of mollicutes. Clin. Infect. Dis. 17
Suppl 1: S10–31.
Bové, J.M., X. Foissac and C. Saillard. 1993. Spiralins. In Subcellular
Biochemistry. Mycoplasma Cell Membranes (edited by Rottem and
Kahane). Plenum Press, New York, pp. 203–223.
Bové, J.M. 1997. Spiroplasmas: infectious agents of plants, arthropods
and vertebrates. Wien. Klin. Wochenschr. 109: 604–612.
Bové, J.M., J. Renaudin, C. Saillard, X. Foissac and M. Garnier. 2003.
Spiroplasma citri, a plant pathogenic mollicute: relationships with its
two hosts, the plant and the leafhopper vector. Annu. Rev. Phytopathol. 41: 483–500.
Bowyer, J.W. and E.C. Calavan. 1974. Antibiotic sensitivity in vitro of the
mycoplasmalike organism associated with citrus stubborn disease.
Phytopathology 64: 346–349.
Brenner, C., H. Duclohier, V. Krchnak and H. Wroblewski. 1995. Conformation, pore-forming activity, and antigenicity of synthetic peptide
analogues of a spiralin putative amphipathic alpha helix. Biochim.
Biophys. Acta 1235: 161–168.
Breton, M., S. Duret, N. Arricau-Bouvery, L. Beven and J. Renaudin.
2008a. Characterizing the replication and stability regions of Spiroplasma citri plasmids identifies a novel replication protein and
expands the genetic toolbox for plant-pathogenic spiroplasmas.
Microbiology 154: 3232–3244.
Breton, M., S. Duret, J.L. Danet, M.P. Dubrana and J. Renaudin. 2010.
Sequences essential for transmission of Spiroplasma citri leafhopper vector, Circulifer haematoceps, revealed by plasmid curing and replacement
based on incompatibility. Appl. Environ. Microbiol. 76: 3198–3205.
Brown, D.R., R.F. Whitcomb and J.M. Bradbury. 2007. Revised minimal
standards for description of new species of the class Mollicutes (division Tenericutes). Int. J. Syst. Evol. Microbiol. 57: 2703–2719.
Calavan, E.C. and J.M. Bové. 1989. Molecular and cellular biology of
spiroplasmas. In The Mycoplasmas, vol. 5 (edited by Whitcomb and
Carle, P., J.G. Tully, R.F. Whitcomb and J.M. Bové. 1990. Size of the
spiroplasmal genome and guanosine plus cytosine content of spiroplasmal DNA. Zentralbl. Bakteriol. Suppl. 20: 926–931.
Carle, P., F. Laigret, J.G. Tully and J.M. Bové. 1995. Heterogeneity of
genome sizes within the genus Spiroplasma. Int. J. Syst. Bacteriol. 45:
178–181.
Carle, P., C. Saillard, N. Carrere, S. Carrere, S. Duret, S. Eveillard, P.
Gaurivaud, G. Gourgues, J. Gouzy, P. Salar, E. Verdin, M. Breton, A.
Blanchard, F. Laigret, J.M. Bové, J. Renaudin and X. Foissac. 2010.
Partial chromosome sequence of Spiroplasma citri reveals extensive
viral invasion and important gene decay. Appl. Environ. Microbiol.
76: 3420–3446.
Carle, P., R.F. Whitcomb, K.J. Hackett, J.G. Tully, D.L. Rose, J.M. Bové,
R.B. Henegar, M. Konai and D.L. Williamson. 1997. Spiroplasma diabroticae sp. nov., from the southern corn rootworm beetle, Diabrotica
undecimpunctata (Coleoptera: Chrysomelidae). Int. J. Syst. Bacteriol.
47: 78–80.
Chang, C.J. and T.A. Chen. 1982. Spiroplasmas: cultivation in chemically
defined medium. Science 215: 1121–1122.
Chang, C.J. 1989. Nutrition and cultivation of spiroplasmas. In The
Charbonneau, D.L. and W.C. Ghiorse. 1984. Ultrastructure and location of cytoplasmic fibrils in Spiroplasma-Floricola OBMG. Curr. Microbiol. 10: 65–71.
Charron, A., C. Bébéar, G. Brun, P. Yot, J. Latrille and J.M. Bové. 1979.
Separation and partial characterization of two deoxyribonucleic acid
polymerases from Spiroplasma citri. J. Bacteriol. 140: 763–768.
Charron, A., M. Castroviejo, C. Bébéar, J. Latrille and J.M. Bové. 1982. A
third DNA polymerase from Spiroplasma citri and two other spiroplasmas. J. Bacteriol. 149: 1138–1141.
Chastel, C., B. Gilot, F. Le Goff, B. Divau, G. Kerdraon, I. HumpherySmith, R. Gruffax and A.M. Simitzis-Le Flohic. 1990. New developments in the ecology of mosquito spiroplasmas. Zentralbl. Bakteriol.
Suppl. 20: 445–460.
Chastel, C. and I. Humphery-Smith. 1991. Mosquito spiroplasmas. Adv.
Dis. Vector Res. 7: 149–205.
Chastel, C., F. Le Goff and I. Humphery-Smith. 1991. Multiplication
and persistence of Spiroplasma melliferum strain A56 in experimentally
infected suckling mice. Res. Microbiol. 142: 411–417.
Chen, T.A. and C.H. Liao. 1975. Corn stunt spiroplasma: Isolation, cultivation and proof of pathogenesis. Science 188: 1015–1017.
Chevalier, C., C. Saillard and J.M. Bové. 1990. Organization and nucleotide sequences of the Spiroplasma citri genes for ribosomal protein
S2, elongation factor Ts, spiralin, phosphofructokinase, pyruvate
kinase, and an unidentified protein. J. Bacteriol. 172: 2693–2703.
Chipman, P.R., M. Agbandje-McKenna, J. Renaudin, T.S. Baker and R.
McKenna. 1998. Structural analysis of the Spiroplasma virus, SpV4:
implications for evolutionary variation to obtain host diversity among
the Microviridae. Structure 6: 135–145.
Christiansen, C., G. Askaa, E.A. Freundt and R.F. Whitcomb. 1979.
Nucleic-acid hybridization experiments with Spiroplasma citri and the
corn stunt and suckling mouse cataract spiroplasmas. Curr. Microbiol. 2: 323–326.
Citti, C., L. Marechal-Drouard, C. Saillard, J.H. Weil and J.M. Bové.
1992. Spiroplasma citri UGG and UGA tryptophan codons: sequence
of the two tryptophanyl-tRNAs and organization of the corresponding genes. J. Bacteriol. 174: 6471–6478.
Clark, HF. 1964. Suckling mouse cataract agent. J. Infect. Dis. 114:
476–487.
Clark, HF. and L.B. Rorke. 1979. Spiroplasmas of tick origin and their
pathogenicity. In The Mycoplasmas, vol. 3 (edited by Whitcomb and
Clark, T.B. 1977. Spiroplasma sp., a new pathogen in honey bees. J. Invertebr. Pathol. 29: 112–113.
Clark, T.B. 1978. Honey bee spiroplasmosis, a new problem for beekeepers. Am. Bee J. 118: 18–19.
Clark, T.B. 1982. Spiroplasmas: diversity of arthropod reservoirs and
host-parasite relationships. Science 217: 57–59.
679
Clark, T.B. 1984. Diversity of spiroplasma host-parasite relationships.
Isr. J. Med. Sci. 20: 995–997.
Clark, T.B., R.F. Whitcomb, J.G. Tully, C. Mouches, C. Saillard, J.M.
Bové, H. Wroblewski, P. Carle, D.L. Rose, R.B. Henegar and D.L. Williamson. 1985. Spiroplasma melliferum, a new species from the honeybee (Apis mellifera). Int. J. Syst. Bacteriol. 35: 296–308.
Cohen, A.J., D.L. Williamson and K. Oishi. 1987. SpV3 viruses of Drosophila spiroplasmas. Isr. J. Med. Sci. 23: 429–433.
Cohen, A.J. and D.L. Williamson. 1988. Yeast supported growth of Drosophila species spiroplasmas. Proceedings of the 7th International Congress of the International Organization for Mycoplasmology, Vienna,
Austria.
Cohen, A.J., D.L. Williamson and P.R. Brink. 1989. A motility mutant
of Spiroplasma melliferum induced with nitrous acid. Curr. Microbiol.
18: 219–222.
Cole, R.M., J.G. Tully, T.J. Popkin and J.M. Bové. 1973. Morphology,
ultrastructure, and bacteriophage infection of the helical mycoplasma-like organism (Spiroplasma citri gen. nov., sp. nov.) cultured
from “stubborn” disease of citrus. J. Bacteriol. 115: 367–384.
Cole, R.M., J.G. Tully and T.J. Popkin. 1974. Virus-like particles in Spiroplasma citri. Colloq. Inst. Natl. Santé Rech. Med. 33: 125–132.
Cole, R.M., W.O. Mitchell and C.F. Garon. 1977. Spiroplasma citri 3:
propagation, purification, proteins, and nucleic acid. Science 198:
1262–1263.
Cole, R.M. 1979. Mycoplasma and Spiroplasma viruses: ultrastructure.
In The Mycoplasmas, vol. 1 (edited by Barile and Razin). Academic
Dally, E.L., T.S. Barros, Y. Zhao, S. Lin, B.A. Roe and R.E. Davis. 2006.
Physical and genetic map of the Spiroplasma kunkelii CR2–3x chromosome. Can. J. Microbiol. 52: 857–867.
Daniels, M.J., J.M. Longland and J. Gilbart. 1980. Aspects of motility
and chemotaxis in spiroplasmas. J. Gen. Microbiol. 118: 429–436.
Daniels, M.J. and J.M. Longland. 1984. Chemotactic behavior of spiroplasmas. Curr. Microbiol. 10: 191–193.
Davis, R.E., J.F. Worley, R.F. Whitcomb, T. Ishijima and R.L. Steere.
1972a. Helical filaments produced by a mycoplasma-like organism
associated with corn stunt disease. Science 176: 521–523.
Davis, R.E., R.F. Whitcomb, T.A. Chen and R.R. Granados. 1972b.
Current status of the aetiology of corn stunt disease. In Pathogenic
Mycoplasmas (edited by Elliott and Birch). Elsevier-Excerpta Medica-North-Holland, Amsterdam, pp. 205–214.
Davis, R.E. and J.F. Worley. 1973. Spiroplasma: Motile, helical microorganism associated with corn stunt disease. Phytopathology 63: 403–408.
Davis, R.E. 1978. Spiroplasma associated with flowers of tulip tree (Liriodendron tulipifera L). Can. J. Microbiol. 24: 954–959.
Davis, R.E., I.M. Lee and J.F. Worley. 1981. Spiroplasma floricola, a new
species isolated from surfaces of flowers of the tulip tree, Liriodendron
tulipifera L. Int. J. Syst. Bacteriol. 31: 456–464.
Davis, R.E., E.L. Dally, R. Jomantiene, Y. Zhao, B. Roe, S. Lin and J. Shao.
2005. Cryptic plasmid pSKU146 from the wall-less plant pathogen
Spiroplasma kunkelii encodes an adhesin and components of a type IV
translocation-related conjugation system. Plasmid 53: 179–190.
DeSoete, G. 1983. A least square algorithm for fitting additive trees to
proximity data. Psychometrika 48: 621–626.
Dickinson, M.J. and R. Townsend. 1984. Characterization of the
genome of a rod-shaped virus Infecting Spiroplasma citri. J. Gen. Virol.
65: 1607–1610.
Duret, S., J.L. Danet, M. Garnier and J. Renaudin. 1999. Gene disruption through homologous recombination in Spiroplasma citri:
an scm1-disrupted motility mutant is pathogenic. J. Bacteriol. 181:
7449–7456.
Duret, S., N. Berho, J.L. Danet, M. Garnier and J. Renaudin. 2003.
Spiralin is not essential for helicity, motility, or pathogenicity but
is required for efficient transmission of Spiroplasma citri by its leafhopper vector Circulifer haematoceps. Appl. Environ. Microbiol. 69:
6225–6234.
680
Duret, S., A. Andre and J. Renaudin. 2005. Specific gene targeting
in Spiroplasma citri: improved vectors and production of unmarked
mutations using site-specific recombination. Microbiology 151:
2793–2803.
Ebbert, M. and L.R. Nault. 1994. Improved overwintering ability in
Dalbulus maidis (Homoptera: Cicadellidae) vectors infected with
Spiroplasma kunkelii (Mycoplasmatales: Spiroplasmataceae). Environ.
Entomol. 23: 634–644.
Enigl, M. and P. Schausberger. 2007. Incidence of the endosymbionts
Wolbachia, Cardinium and Spiroplasma in phytoseiid mites and associated prey. Exp. Appl. Acarol. 42: 75–85.
FAO/WHO. 1974. Preservation of mycoplasmas by lyophilization.
World Health Organization working document VPH/MIC/741.
FAO/WHO Programme on Comparative Mycoplasmology Working
Group. World Health Organization, Geneva.
Felsenstein, J. 1993. PHYLIP (Phylogeny Inference Package) 3.57 edn.
Department of Genetics, University of Washington, Seattle.
Fletcher, J., A. Wayadande, U. Melcher and F.C. Ye. 1998. The phytopathogenic mollicute–insect vector interface: a closer look. Phytopathology 88: 1351–1358.
Foissac, X., C. Saillard, J. Gandar, L. Zreik and J.M. Bové. 1996. Spiralin
polymorphism in strains of Spiroplasma citri is not due to differences
in posttranslational palmitoylation. J. Bacteriol. 178: 2934–2940.
Foissac, X., J.M. Bové and C. Saillard. 1997a. Sequence analysis of Spiroplasma phoeniceum and Spiroplasma kunkelii spiralin genes and comparison with other spiralin genes. Curr. Microbiol. 35: 240–243.
Foissac, X., J.L. Danet, C. Saillard, P. Gaurivaud, F. Laigret, C. Paré
and J.M. Bové. 1997b. Mutagenesis by insertion of Tn4001 into the
genome of Spiroplasma citri: Characterization of mutants affected in
plant pathogenicity and transmission to the plant by the leafhopper vector Circulifer haematoceps. Mol. Plant Microbe Interact. 10:
454–461.
Foissac, X., C. Saillard and J.M. Bové. 1997c. Random insertion of transposon Tn4001 in the genome of Spiroplasma citri strain GII3. Plasmid
37: 80–86.
French, F.E., R.F. Whitcomb, J.G. Tully, K.J. Hackett, E.A. Clark, R.B.
Henegar, A.G. Wagner and D.L. Rose. 1990. Tabanid spiroplasmas of
the southeast USA: new groups and correlation with host life history
strategy. Zentralbl. Bakteriol. Suppl. 20: 919–922.
French, F.E., R.F. Whitcomb, J.G. Tully, D.L. Williamson and R.B. Henegar. 1996. Spiroplasmas of Tabanus lineola. IOM Lett. 4: 211–212.
French, F.E., R.F. Whitcomb, J.G. Tully, P. Carle, J.M. Bové, R.B. Henegar, J.R. Adams, G.E. Gasparich and D.L. Williamson. 1997. Spiroplasma lineolae sp. nov., from the horsefly Tabanus lineola (Diptera:
Tabanidae). Int. J. Syst. Bacteriol. 47: 1078–1081.
Fukatsu, T. and N. Nikoh. 1998. Two intracellular symbiotic bacteria
from the mulberry psyllid Anomoneura mori (Insecta, Homoptera).
Appl. Environ. Microbiol. 64: 3599–3606.
Fukatsu, T. and N. Nikoh. 2000. Endosymbiotic microbiota of the bamboo pseudococcid Antonina crawii (Insecta, Homoptera). Appl. Environ. Microbiol. 66: 643–650.
Fukatsu, T., T. Tsuchida, N. Nikoh and R. Koga. 2001. Spiroplasma symbiont of the pea aphid, Acyrthosiphon pisum (Insecta: Homoptera).
Appl. Environ. Microbiol. 67: 1284–1291.
Gadeau, A.P., C. Mouches and J.M. Bové. 1986. Probable insensitivity
of mollicutes to rifampin and characterization of spiroplasmal DNAdependent RNA polymerase. J. Bacteriol. 166: 824–828.
Garnier, M., M. Clerc and J.M. Bové. 1981. Growth and division of
spiroplasmas: morphology of Spiroplasma citri during growth in liquid
medium. J. Bacteriol. 147: 642–652.
Garnier, M., M. Clerc and J.M. Bové. 1984. Growth and division of
Spiroplasma citri: elongation of elementary helices. J. Bacteriol. 158:
23–28.
Garnier, M., X. Foissac, P. Gaurivaud, F. Laigret, J. Renaudin, C. Saillard
and J.M. Bové. 2001. Mycoplasmas, plants, insect vectors: a matrimonial triangle. C. R. Acad. Sci. III 324: 923–928.
Gasparich, G.E., K.J. Hackett, E.A. Clark, J. Renaudin and R.F. Whitcomb. 1993a. Occurrence of extrachromosomal deoxyribonucleic
acids in spiroplasmas associated with plants, insects, and ticks. Plasmid 29: 81–93.
Gasparich, G.E., K.J. Hackett, C. Stamburski, J. Renaudin and J.M.
Bové. 1993b. Optimization of methods for transfecting Spiroplasma
citri strain R8A2 HP with the spiroplasma virus SpV1 replicative form.
Plasmid 29: 193–205.
Gasparich, G.E., C. Saillard, E.A. Clark, M. Konai, F.E. French, J.G.
Tully, K.J. Hackett and R.F. Whitcomb. 1993c. Serologic and genomic
relatedness of group-VIII and group-XVII spiroplasmas and subdivision of spiroplasma group-VIII into subgroups. Int. J. Syst. Bacteriol.
43: 338–341.
Gasparich, G.E. and K.J. Hackett. 1994. Characterization of a cryptic
extrachromosomal element isolated from the mollicute Spiroplasma
taiwanense. Plasmid 32: 342–343.
Gasparich, G.E., K.J. Hackett, F.E. French and R.F. Whitcomb. 1998.
Serologic and genomic relatedness of group XIV spiroplasma isolates from a lampyrid beetle and tabanid flies: an ecologic paradox.
Gasparich, G.E., R.F. Whitcomb, D. Dodge, F.E. French, J. Glass and
D.L. Williamson. 2004. The genus Spiroplasma and its non-helical
descendants: phylogenetic classification, correlation with phenotype
and roots of the Mycoplasma mycoides clade. Int. J. Syst. Evol. Microbiol. 54: 893–918.
Gaurivaud, P., F. Laigret and J.M. Bové. 1996. Insusceptibility of members of the class Mollicutes to rifampin: studies of the Spiroplasma citri
RNA polymerase b-subunit gene. Antimicrob. Agents Chemother. 40:
858–862.
Gaurivaud, P., J.L. Danet, F. Laigret, M. Garnier and J.M. Bové. 2000a.
Fructose utilization and phytopathogenicity of Spiroplasma citri. Mol.
Plant Microbe Interact. 13: 1145–1155.
Gaurivaud, P., F. Laigret, M. Garnier and J.M. Bové. 2000b. Fructose
utilization and pathogenicity of Spiroplasma citri: characterization of
the fructose operon. Gene 252: 61–69.
Gaurivaud, P., F. Laigret, E. Verdin, M. Garnier and J.M. Bové. 2000c.
Fructose operon mutants of Spiroplasma citri. Microbiology 146:
2229–2236.
Gaurivaud, P., F. Laigret, M. Garnier and J.M. Bove. 2001. Characterization of FruR as a putative activator of the fructose operon of Spiroplasma citri. FEMS Microbiol. Lett. 198: 73–78.
Gilad, R., A. Porat and S. Trachtenberg. 2003. Motility modes of Spiroplasma melliferum BC3: a helical, wall-less bacterium driven by a linear
motor. Mol. Microbiol. 47: 657–669.
Goodacre, S.L., O.Y. Martin, C.F. Thomas and G.M. Hewitt. 2006. Wolbachia and other endosymbiont infections in spiders. Mol. Ecol. 15:
517–527.
Gordon, D.T., L.R. Nault, N.H. Gordon and S.E. Heady. 1985. Serological detection of corn stunt spiroplasma and maize rayado fino
virus in field-collected Dalbulus spp. from Mexico. Plant Dis. 69:
108–111.
Grau, O., F. Laigret and J.M. Bové. 1988. Analysis of ribosomal RNA
genes in two spiroplasmas, one acholeplasma and one unclassified
mollicute. Zentralbl. Bakteriol. Suppl. 20: 895–897.
Grulet, O., I. Humphery-Smith, C. Sunyach, F. Le Goff and C. Chastel.
1993. Spiromed: a rapid and inexpensive spiroplasma isolation technique. J. Microbiol. Methods 17: 123–128.
Guo, Y.H., T.A. Chen, R.F. Whitcomb, D.L. Rose, J.G. Tully, D.L. Williamson, X.D. Ye and Y.X. Chen. 1990. Spiroplasma chinense sp. nov.
from flowers of Calystegia hederacea in China. Int. J. Syst. Bacteriol.
40: 421–425.
Hackett, K.J. and D.E. Lynn. 1985. Cell-assisted growth of a fastidious
spiroplasma. Science 230: 825–827.
Hackett, K.J., D.E. Lynn, D.L. Williamson, A.S. Ginsberg and R.F. Whitcomb. 1986. Cultivation of the Drosophila sex-ratio Spiroplasma. Science 232: 1253–1255.
Hackett, K.J. and T.B. Clark. 1989. Ecology of Spiroplasmas. In The
Hackett, K.J., R.F. Whitcomb, R.B. Henegar, A.G. Wagner, E.A. Clark,
J.G. Tully, F. Green, W.H. McKay, P. Santini, D.L. Rose, J.J. Anderson and D.E. Lynn. 1990. Mollicute diversity in arthropod hosts.
Zentralbl. Bakteriol. Suppl. 20: 441–454.
Hackett, K.J., R.F. Whitcomb, J.G. Tully, D.L. Rose, P. Carle, J.M. Bové,
R.B. Henegar, T.B. Clark, E.A. Clark, M. Konai, J.R. Adams and D.L.
Williamson. 1993. Spiroplasma insolitum sp. nov., a new species of
group-I spiroplasma with an unusual DNA base composition. Int. J.
Hackett, K.J., R.H. Hackett, E.A. Clark, G.E. Gasparich, J.D. Pollack and
R.F. Whitcomb. 1994. Development of the first completely defined
medium for a spiroplasma, Spiroplasma clarkii strain CN-5. IOM Lett.
3: 446–447.
Hackett, K.J. and R.F. Whitcomb. 1995. Cultivation of spiroplasmas in
undefined and defined media. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, San Diego, pp. 41–53.
Hackett, K.J., E.A. Clark, R.F. Whitcomb, M. Camp and J.G. Tully. 1996a.
Amended data on arginine utilization by Spiroplasma species. Int. J.
Hackett, K.J., R.F. Whitcomb, T.B. Clark, R.B. Henegar, D.E. Lynn, A.G.
Wagner, J.G. Tully, G.E. Gasparich, D.L. Rose, P. Carle, J.M. Bové,
M. Konai, E.A. Clark, J.R. Adams and D.L. Williamson. 1996b. Spiroplasma leptinotarsae sp. nov., a mollicute uniquely adapted to its host,
the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera:
Chrysomelidae). Int. J. Syst. Bacteriol. 46: 906–911.
Hackett, K.J., R.F. Whitcomb, F.E. French, J.G. Tully, G.E. Gasparich,
D.L. Rose, P. Carle, J.M. Bové, R.B. Henegar, T.B. Clark, M. Konai,
E.A. Clark and D.L. Williamson. 1996c. Spiroplasma corruscae sp. nov.,
from a firefly beetle (Coleoptera: Lampyridae) and tabanid flies
(Diptera: Tabanidae). Int. J. Syst. Bacteriol. 46: 947–950.
Hackett, K.J., J.J. Lipa, G.E. Gasparich, D.E. Lynn, M. Konai, M. Camp
and R.F. Whitcomb. 1997. The spiroplasma motility inhibition test, a
new method for determining intraspecific variation among Colorado
potato beetle spiroplasmas. Int. J. Syst. Bacteriol. 47: 33–37.
Haselkorn, T.S., T.A. Markow and N.A. Moran. 2009. Multiple introductions of the Spiroplasma bacterial endosymbiont into Drosophila. Mol
Ecol 18: 1294–1305.
Hélias, C., M. Vazeille-Falcoz, F. Le Goff, M.L. Abalain-Colloc, F. Rodhain, P. Carle, R.F. Whitcomb, D.L. Williamson, J.G. Tully, J.M. Bové
and C. Chastel. 1998. Spiroplasma turonicum sp. nov. from Haematopota
horse flies (Diptera: Tabanidae) in France. Int. J. Syst. Bacteriol. 48:
457–461.
Henning, K., S. Greiner-Fischer, H. Hotzel, M. Ebsen and D. Theegarten. 2006. Isolation of Spiroplasma sp. from an Ixodes tick. Int. J. Med.
Microbiol. 296 Suppl 40: 157–161.
Herren, J.K., I. Gordon, P. W. H. Holland and D. Smith. 2007. The butterfly Danaus chrysippus (Lepidoptera: Nymphalidae) in Kenya is variably infected with respect to genotype and body size by a maternally
transmitted male-killing endosymbiont (Spiroplasma). Int. J. Trop.
Ins. Sc.: 62–69.
Humphery-Smith, I., O. Grulet and C. Chastel. 1991a. Pathogenicity of
Spiroplasma taiwanense for larval Aedes aegypti mosquitoes. Med. Vet.
Entomol. 5: 229–232.
Humphery-Smith, I., O. Grulet, F. Le Goff and C. Chastel. 1991b. Spiroplasma (Mollicutes: Spiroplasmataceae) pathogenic for Aedes aegypti and
Anopheles stephensi (Diptera: Culicidae). J. Med. Entomol. 28: 219–222.
Hung, S.H.Y., T.A. Chen, R.F. Whitcomb, J.G. Tully and Y.X. Chen. 1987.
Spiroplasma culicicola sp. nov. from the salt-marsh mosquito Aedes sollicitans. Int. J. Syst. Bacteriol. 37: 365–370.
Hurst, G.D.D., H. Anbutsu, M. Kutsukake and T. Fukatsu. 2003. Hidden
from the host: Spiroplasma bacteria infecting Drosophila do not cause
an immune response, but are suppressed by ectopic immune activation. Insect Mol. Biol. 12: 93–97.
681
Hurst, G.D.D., J.H.G. von der Schulenburg, T.M.O. Majerus, D. Bertrand,
I.A. Zakharov, J. Baungaard, W. Volkl, R. Stouthamer and M.E.N.
Majerus. 1999. Invasion of one insect species, Adalia bipunctata, by two
different male-killing bacteria. Insect Mol. Biol. 8: 133–139.
Hurst, G.D.D. and F.M. Jiggins. 2000. Male-killing bacteria in insects: mechanisms, incidence, and implications. Emerg. Infect. Dis. 6: 329–336.
International Committee on Systematics of Bacteria. 1984. Minutes of
the interim meeting. 30 August and 6 September 1982, Tokyo, Japan
International Committee on Systematics of Bacteria Subcommittee on
the Taxonomy of Mollicutes. 1995. Revised minimum standards for
description of new species of the class Mollicutes (division Tenericutes).
Int J. Syst. Bacteriol 45: 605–612.
Jacob, C., F. Nouzieres, S. Duret, J.M. Bové and J. Renaudin. 1997. Isolation, characterization, and complementation of a motility mutant of
Spiroplasma citri. J. Bacteriol. 179: 4802–4810.
Jaenike, J., M. Polak, A. Fiskin, M. Helou and M. Minhas. 2007. Interspecific transmission of endosymbiotic Spiroplasma by mites. Biol.
Lett. 3: 23–25.
Jagoueix-Eveillard, S., F. Tarendeau, K. Guolter, J.L. Danet, J.M. Bové and
M. Garnier. 2001. Catharanthus roseus genes regulated differentially by
mollicute infections. Mol. Plant Microbe Interact. 14: 225–233.
Jandhyam, H., C. R. Bates, T. E. Young, L. Beatti, G. E. Gasparich, F. E.
French and L. B. Regassa. 2008. Global spiroplasma biodiversity in a
single host. Presented at the 17th Congress of International Organization for Mycoplasmology, Beijing, China. Abstract no. 206, p. 130.
Jiggins, F.M., G.D. Hurst, C.D. Jiggins, J.H. von der Schulenburg and
M.E. Majerus. 2000. The butterfly Danaus chrysippus is infected by a
male-killing Spiroplasma bacterium. Parasitology 120: 439–446.
Johansson, K.-E., M.U.K. Heldtander and B. Pettersson. 1998. Characterization of mycoplasmas by PCR and sequence analysis with universal 16S rDNA primers. In Methods in Molecular Biology: Mycoplasma
protocols, vol. 104 (edited by Miles and Nicholas). Humana Press,
Totawa, NJ, pp. 145–165.
Johansson, K.-E. and B. Pettersson. 2002. Taxonomy of Mollicutes. In
Molecular Biology and Pathogenicity of Mycoplasmas (edited by
Razin and Herrmann). Kluwer Academic/Plenum Publishers,
London, pp. 1–31.
Johnson, J.L. 1994. Similarity analysis of DNAs. In Methods for General
and Molecular Bacteriology (edited by Gerhardt, Murray, Wood and
Krieg). ASM Press, Washington, D.C., pp. 656–682.
Jones, L.J., R. Carballido-Lopez and J. Errington. 2001. Control of cell
shape in bacteria: helical, actin-like filaments in Bacillus subtilis. Cell
104: 913–922.
Joshi, B.D., M. Berg, J. Rogers, J. Fletcher and U. Melcher. 2005.
Sequence comparisons of plasmids pBJS-O of Spiroplasma citri and
pSKU146 of S. kunkelii: implications for plasmid evolution. BMC
Genomics 6: 175.
Junca, P., C. Saillard, J. Tully, O. Garcia-Jurado, J.R. Degorce-Dumas, C.
Mouches, J.C. Vignault, R. Vogel, R. McCoy, R. Whitcomb, D. Williamson, J. Latrille and J.M. Bové. 1980. Characterization of spiroplasmas isolated from insects and flowers in continental France,
Corsica and Morocco. Proposals for a taxonomical classification of
spiroplasmas. [transl. from. Fr.] C. R. Hebd. Des Seances Acad. Sci.
Ser. D Sci. Nat. 290: 1209–1211.
Kageyama, D., H. Anbutsu, M. Watada, T. Hosokawa, M. Shimada and T.
Fukatsu. 2006. Prevalence of a non-male-killing spiroplasma in natural
populations of Drosophila hydei. Appl Environ Microbiol 72: 6667–6673.
Kersting, U. and C. Sengonca. 1992. Detection of insect vectors of the
citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the
citrus growing area of south Turkey. J. Appl. Entomol. 113: 356–364.
Killiny, N., M. Castroviejo and C. Saillard. 2005. Spiroplasma citri spiralin
acts in vitro as a lectin binding to glycoproteins from its insect vector
Circulifer haematoceps. Phytopathology 95: 541–548.
Killiny, N., B. Batailler, X. Foissac and C. Saillard. 2006. Identification of
a Spiroplasma citri hydrophilic protein associated with insect transmissibility. Microbiology 152: 1221–1230.
682
Koerber, R.T., G.E. Gasparich, M.F. Frana and W.L. Grogan, Jr. 2005.
Spiroplasma atrichopogonis sp. nov., from a ceratopogonid biting
midge. Int. J. Syst. Evol. Microbiol. 55: 289–292.
Konai, M., R.F. Whitcomb, J.G. Tully, D.L. Rose, P. Carle, J.M. Bové, R.B.
Henegar, K.J. Hackett, T.B. Clark and D.L. Williamson. 1995. Spiroplasma velocicrescens sp. nov., from the vespid wasp Monobia quadridens.
Konai, M., E.A. Clark, M. Camp, A.L. Koeh and R.F. Whitcomb. 1996a.
Temperature ranges, growth optima, and growth rates of Spiroplasma
(Spiroplasmataceae, class Mollicutes) species. Curr. Microbiol. 32: 314–
319.
Konai, M., K.J. Hackett, D.L. Williamson, J.J. Lipa, J.D. Pollack, G.E. Gasparich, E.A. Clark, D.C. Vacek and R.F. Whitcomb. 1996b. Improved
cultivation systems for isolation of the Colorado potato beetle spiroplasma. Appl. Environ. Microbiol. 62: 3453–3458.
Konai, M., R.F. Whitcomb, F.E. French, J.G. Tully, D.L. Rose, P. Carle,
J.M. Bové, K.J. Hackett, R.B. Henegar, T.B. Clark and D.L. Williamson. 1997. Spiroplasma litorale sp. nov., from tabanid flies (Tabanidae:
Diptera) in the southeastern United States. Int. J. Syst. Bacteriol. 47:
359–362.
Kotani, H., G.H. Butler and G.J. McGarrity. 1990. Malignant transformation by Spiroplasma mirum. Zentralbl. Bakteriol. Suppl. 20: 145–152.
Kürner, J., A.S. Frangakis and W. Baumeister. 2005. Cyro-electron
tomography reveals the cytoskeletal structure of Spiroplasma melliferum. Science 307: 436–438.
Kuroda, Y., Y. Shimada, B. Sakaguchi and K. Oishi. 1992. Effects of sexratio (SR)-spiroplasma infection on Drosophila primary embryonic
cultured cells and on embryogenesis. Zool. Sci. 9: 283–291.
Labarère, J. and G. Barroso. 1989. Lethal and mutation frequency
responses of Spiroplasma citri cells to UV irradiation. Mutat. Res. 210:
135–141.
Laigret, F., P. Gaurivaud and J.M. Bové. 1996. The unique organization
of the rpoB region of Spiroplasma citri: a restriction and modification
system gene is adjacent to rpoB. Gene 171: 95–98.
Lartigue, C., S. Duret, M. Garnier and J. Renaudin. 2002. New plasmid vectors for specific gene targeting in Spiroplasma citri. Plasmid
48: 149–159.
Le Dantec, L., M. Castroviejo, J.M. Bové and C. Saillard. 1998. Purification, cloning, and preliminary characterization of a Spiroplasma citri
ribosomal protein with DNA binding capacity. J. Biol. Chem. 273:
24379–24386.
Le Goff, F., M. Marjolet, J. Guilloteau, I. Humphery-Smith and C. Chastel. 1990. Characterization and ecology of mosquito spiroplasmas
from Atlantic biotopes in France. Ann. Parasitol. Hum. Comp. 65:
107–110.
Le Goff, F., I. Humphery-Smith, M. Leclercq and C. Chastel. 1991. Spiroplasmas from European Tabanidae. Med. Vet. Entomol. 5: 143–144.
Le Goff, F., M. Marjolet, I. Humphery-Smith, M. Leclercq, C. Hélias,
F. Suplisson and C. Chastel. 1993. Tabanid spiroplasmas from France:
characterization, ecology and experimental study. Ann. Parasitol.
Hum. Comp. 68: 150–153.
Lee, I.M. and R.E. Davis. 1980. DNA homology among diverse spiroplasma strains representing several serological groups. Can. J. Microbiol. 26: 1356–1363.
Liao, C.H. and T.A. Chen. 1977. Culture of corn stunt spiroplasma in a
simple medium. Phytopathology 67: 802–807.
Liao, C.H., C.J. Chang and T.A. Chen. 1979. Spiroplasmastatic action of
plant tissue extracts. Proceedings of the R. O. C. U. S. Coop. Science
Seminar Mycoplasma Diseases of Plants, Taipei, pp. 99–103.
Liao, C.H. and T.A. Chen. 1981a. Deoxyribonucleic acid hybridization between Spiroplasma citri and the corn stunt spiroplasma. Curr.
Liao, C.H. and T.A. Chen. 1981b. In vitro susceptibility and resistance of
two spiroplasmas to antibiotics. Phytopathology 71: 442–445.
Lindh, J.M., O. Terenius and I. Faye. 2005. 16S rRNA gene-based identification of midgut bacteria from field-caught Anopheles gambiae sensu
lato and A. funestus mosquitoes reveals new species related to known
insect symbionts. Appl. Environ. Microbiol. 71: 7217–7223.
Liss, A. and R.M. Cole. 1981. Spiroplasmavirus group I: isolation,
growth, and properties. Curr. Microbiol. 5: 357–362.
Lundgren, J.G., R. M. Lehman and J. Chee-Sanford. 2007. Bacterial
communities within digestive tracts of ground beetles (Coleoptera:
Carabidae). Ann. Entomol. Soc. Am. 100: 275–282.
Maccheroni, W., J. L. Danet, S. Duret-Nurbel, J. M. Bové, M. Garnier
and J. Renaudin. 2002. Cell shape determination in Spiroplasma citri:
organization of mreB genes and effect of mreB1 disruption on insect
transmission and pathogenicity. Proceedings of the 15th Conference
of the International Organization of Citrus Virologists (edited by
Duran-Vila, Milne and da Graça), Riverside, California, p. 443.
Madden, L.V. and L.R. Nault. 1983. Differential pathogenicity of corn
stunting mollicutes to leafhopper vectors in Dalbulus and Baldulus
species. Phytopathology 73: 1608–1614.
Majerus, T.M., J.H. Graf von der Schulenburg, M.E. Majerus and G.D.
Hurst. 1999. Molecular identification of a male-killing agent in the
ladybird Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae).
Insect. Mol. Biol. 8: 551–555.
Maniloff, J. 1992. Phylogeny of mycoplasmas. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch
D.C., pp. 549–559.
Marais, A., J.M. Bové, S.F. Dallo, J.B. Baseman and J. Renaudin. 1993.
Expression in Spiroplasma citri of an epitope carried on the G fragment of the cytadhesin P1 gene from Mycoplasma pneumoniae. J. Bacteriol. 175: 2783–2787.
Marais, A., J.M. Bové and J. Renaudin. 1996. Spiroplasma citri virus
SpV1-derived cloning vector: deletion formation by illegitimate and
homologous recombination in a spiroplasmal host strain which probably lacks a functional recA gene. J. Bacteriol. 178: 862–870.
Markham, P.G., R. Townsend, M. Bar Joseph, M.J. Daniels, A. Plaskitt
and B.M. Meddins. 1974. Spiroplasmas are causal agents of citrus
little-leaf disease. Ann. Appl. Biol. 78: 49–57.
Markham, P.G., T.B. Clark and R.F. Whitcomb. 1983. Culture techniques
for spiroplasmas from arthropods. In Methods in Mycoplasmology, vol.
2 (edited by Tully and Razin). Academic Press, New York, pp. 217–223.
Mateos, M., S.J. Castrezana, B.J. Nankivell, A.M. Estes, T.A. Markow and
N.A. Moran. 2006. Heritable endosymbionts of Drosophila. Genetics
174: 363–376.
Matsuo, K., J. Silke, K. Gramatikoff and W. Schaffner. 1994. The CpGspecific methylase SssI has topoisomerase activity in the presence of
Mg2+. Nucleic Acids Res. 22: 5354–5359.
McCoy, R.E., D.S. Williams and D.L. Thomas. 1979. Isolation of mycoplasmas from flowers. Proceedings of the Republic of China-United
States Cooperative Science Seminar, Symposium series 1, National
Science Council, Taipei, Taiwan, pp. 75–81.
McElwain, M.C., D.K.F. Chandler, M.F. Barile, T.F. Young, V.V. Tryon,
J.W. Davis, J.P. Petzel, C.J. Chang, M.V. Williams and J.D. Pollack.
1988. Purine and pyrimidine metabolism in Mollicutes species. Int. J.
McIntosh, M.A., G. Deng, J. Zheng and R.V. Ferrell. 1992. Repetitive
DNA sequences. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch and Baseman). American
Melcher, U. and J. Fletcher. 1999. Genetic variation in Spiroplasma citri.
Eur. J. Plant Pathol. 105: 519–533.
Miles, R.J. 1992. Catabolism in Mollicutes. J. Gen. Microbiol. 138: 1773–
1783.
Montenegro, H., V.N. Solferini, L.B. Klaczko and G.D. Hurst. 2005.
Male-killing Spiroplasma naturally infecting Drosophila melanogaster.
Insect. Mol. Biol. 14: 281–287.
Montenegro, H., L. M. Hatadani, H. F. Medeiros and L.B. Klaczko. 2006.
Male killing in three species of the tripunctata radiation of Drosophila
(Diptera: Drosophilidae). J. Zoo. Syst. Evol. Res. 44: 130–135.
Mouches, C., J.M. Bové, J. Albisetti, T.B. Clark and J.G. Tully. 1982a.
A spiroplasma of serogroup IV causes a May-disease-like disorder of
honeybees in southwestern France. Microb. Ecol. 8: 387–399.
Mouches, C., A. Menara, B. Geny, D. Charlemagne and J.M. Bové.
1982b. Synthesis of Spiroplasma citri protein specifically recognized by
rabbit immunoglobulin to rabbit actin. Rev. Infect. Dis. 4: S277.
Mouches, C., A. Menara, J.G. Tully and J.M. Bové. 1982c. Polyacrylamide gel analysis of spiroplasmas proteins and its contribution to the
taxonomy of spiroplasmas. Rev. Infect. Dis. 4 Suppl: S141–147.
Mouches, C., J.M. Bové, J.G. Tully, D.L. Rose, R.E. McCoy, P. CarleJunca, M. Garnier and C. Saillard. 1983a. Spiroplasma apis, a new species from the honey bee Apis mellifera. Ann. Microbiol. (Paris) 134A:
383–397.
Mouches, C., T. Candresse, G.J. McGarrity and J.M. Bové. 1983b. Analysis of spiroplasma proteins: contribution to the taxonomy of group
IV spiroplasmas and the characterization of spiroplasma protein antigens. Yale J. Biol. Med. 56: 431–437.
Mouches, C., G. Barroso, A. Gadeau and J.M. Bové. 1984a. Characterization of two cryptic plasmids from Spiroplasma citri and occurrence
of their DNA sequences among various spiroplasmas. Ann. Microbiol. (Paris) 135A: 17–24.
Mouches, C., J. M. Bové, J. G. Tully, D. L. Rose, R. E. McCoy, P. CarleJunca, M. Garnier and C. Saillard. 1984b. In Validation of the publication of new names and new combinations previously effectively
published outside the IJSB. List no. 13. Int. J. Syst. Bacteriol. 34:
91–92.
Moya-Raygoza, G., S.A. Hogenhout and L.R. Nault. 2007a. Habitat of
the corn leafhopper (Hemiptera: Cicadellidae) during the dry (winter) season in Mexico. Environ Entomol 36: 1066–1072.
Moya-Raygoza, G., V. Palomera-Avalos and C. Galaviz-Mejia. 2007b.
Field overwintering biology of Spiroplasma kunkelii (Mycoplasmatales:
Spiroplasmataceae) and its vector Dalbulus maidis (Hemiptera: Cicadellidae). Ann. Appl. Biol. 151: 373–379.
Nakamura, K., H. Ueno and K. Miura. 2005. Prevalence of inherited
male-killing microorganisms in Japanese populations of ladybird
beetle Harmonia axyridis (Coleoptera: Coccinellidae). Ann. Ent. Soc.
Am. 98: 96–99.
Nault, L.R. and O.E. Bradfute. 1979. Corn stunt: involvement of a
complex of leafhopper-borne pathogens. In Leafhopper Vectors
and Plant Disease Agents (edited by Maramorosch and Harris). Academic Press, New York, pp. 561–586.
Nault, L.R., L.V. Madden, W.E. Styer, B.W. Triplehorn, G.F. Shambaugh
and S.E. Heady. 1984. Pathogenicity of corn stunt spiroplasma and
maize bushy stunt mycoplasma to their vector, Dalbulus longulus. Phytopathology 74: 977–979.
Navas-Castillo, J., F. Laigret, J.G. Tully and J.M. Bové. 1992. The mollicute Acholeplasma florum possesses a gene of phosphoenolpyruvatesugar phosphotransferase system and it uses UGA as tryptophan
codon. C. R. Acad. Sci. Ser. III Life Sci. 315: 43–48.
Nunan, L.M., C.R. Pantoja, M. Salazar, F. Aranguren and D.V. Lightner. 2004. Characterization and molecular methods for detection
of a novel spiroplasma pathogenic to Penaeus vannamei. Dis. Aquat.
Organ. 62: 255–264.
Nunan, L.M., D.V. Lightner, M.A. Oduori and G.E. Gasparich. 2005. Spiroplasma penaei sp. nov., associated with mortalities in Penaeus vannamei,
Pacific white shrimp. Int. J. Syst. Evol. Microbiol. 55: 2317–2322.
Nur, I., M. Szyf, A. Razin, G. Glaser, S. Rottem and S. Razin. 1985. Procaryotic and eucaryotic traits of DNA methylation in spiroplasmas
(mycoplasmas). J. Bacteriol. 164: 19–24.
Nur, I., G. Glaser and S. Razin. 1986. Free and integrated plasmid DNA
in spiroplasmas. Curr. Microbiol. 14: 169–176.
Nur, I., D.J. LeBlanc and J.G. Tully. 1987. Short, interspersed, and repetitive DNA sequences in Spiroplasma species. Plasmid 17: 110–116.
Oduori, M.A., J.J. Lipa and G.E. Gasparich. 2005. Spiroplasma leucomae
sp. nov., isolated in Poland from white satin moth (Leucoma salicis L.)
larvae. Int. J. Syst. Evol. Microbiol. 55: 2447–2450.
683
Oishi, K., D.F. Poulson and D.L. Williamson. 1984. Virus-mediated
change in clumping properties of Drosophila SR spiroplasmas. Curr.
Microbiol. 10: 153–158.
Özbek, E., S.A. Miller, T. Meulia and S.A. Hogenhout. 2003. Infection
and replication sites of Spiroplasma kunkelii (Class: Mollicutes) in
midgut and Malpighian tubules of the leafhopper Dalbulus maidis. J.
Invertebr. Pathol. 82: 167–175.
Pickens, E.G., R.K. Gerloff and W. Burgdorfer. 1968. Spirochete from
the rabbit tick, Haemaphysalis leporispalustris (Packard). I. Isolation
and preliminary characterization. J. Bacteriol. 95: 291–299.
Pollack, J.D., M.C. McElwain, D. Desantis, J.T. Manolukas, J.G. Tully,
C.J. Chang, R.F. Whitcomb, K.J. Hackett and M.V. Williams. 1989.
Metabolism of members of the Spiroplasmataceae. Int. J. Syst. Bacteriol. 39: 406–412.
Pollack, J.D., M.V. Williams and R.N. McElhaney. 1997. The comparative metabolism of the mollicutes (mycoplasmas): the utility for taxonomic classification and the relationship of putative gene annotation
and phylogeny to enzymatic function in the smallest free-living cells.
Crit. Rev. Microbiol. 23: 269–354.
Pollack, J.D. 2002a. The necessity of combining genomic and enzymatic
data to infer metabolic function and pathways in the smallest bacteria: amino acid, purine and pyrimidine metabolism in mollicutes.
Front. Biosci. 7: d1762–1781.
Pollack, J.D. 2002b. Central carbohydrate pathways: Metabolic flexibility and the extra role of some “housekeeping enzymes”. In Molecular Biology and Pathogenicity of Mycoplasmas (edited by Razin and
Herrmann). Kluwer Academic/Plenum Publishers, New York, pp.
163–199.
Pool, J.E., A. Wong and C.F. Aquadro. 2006. Finding of male-killing
Spiroplasma infecting Drosophila melanogaster in Africa implies transatlantic migration of this endosymbiont. Heredity 97: 27–32.
Poulson, D.F. and B. Sakaguchi. 1961. Nature of “sex-ratio” agent in
Drosophila. Science 133: 1489–1490.
Pyle, L.E. and L.R. Finch. 1988. A physical map of the genome of
Mycoplasma mycoides subspecies mycoides Y with some functional loci.
Nucleic Acids Res. 16: 6027–6039.
Rahimian, H. and D.J. Gumpf. 1980. Deoxyribonucleic acid relationship between Spiroplasma citri and the corn stunt spiroplasma. Int. J.
Ranhand, J.M., W.O. Mitchell, T.J. Popkin and R.M. Cole. 1980. Covalently closed circular deoxyribonucleic acids in spiroplasmas. J. Bacteriol. 143: 1194–1199.
Razin, S. 1985. Molecular biology and genetics of mycoplasmas (Mollicutes). Microbiol. Rev. 49: 419–455.
Regassa, L.B., K.M. Stewart, A.C. Murphy, F.E. French, T. Lin and R.F.
Whitcomb. 2004. Differentiation of group VIII Spiroplasma strains
with sequences of the 16S–23S rDNA intergenic spacer region. Can.
J. Microbiol. 50: 1061–1067.
Regassa, L.B. and G.E. Gasparich. 2006. Spiroplasmas: evolutionary
relationships and biodiversity. Front. Biosci. 11: 2983–3002.
Renaudin, J., M.C. Pascarel, M. Garnier, P. Carle-Junca and J.M. Bové.
1984a. SpV4, a new Spiroplasma virus with circular, single-stranded
DNA. Ann. Virol. 135E: 163–168.
Renaudin, J., M.C. Pascarel, M. Garnier, P. Carle and J.M. Bové. 1984b.
Characterization of spiroplasma virus group 4 (SV4). Isr. J. Med. Sci.
20: 797–799.
Renaudin, J., M.C. Pascarel, C. Saillard, C. Chevalier and J.M. Bové.
1986. Chez les spiroplasmes le codon UGA n’est pas non-sens et semble coder pour le tryptophane. C. R. Acad. Sci. Ser. III 303: 539–540.
Renaudin, J. and J.M. Bové. 1994. SpV1 and SpV4, spiroplasma viruses
with circular, single-stranded DNA genomes, and their contribution to
the molecular biology of spiroplasmas. Adv. Virus Res. 44: 429–463.
684
Renaudin, J., A. Marais, E. Verdin, S. Duret, X. Foissac, F. Laigret and
J.M. Bové. 1995. Integrative and free Spiroplasma citri oriC plasmids:
Expression of the Spiroplasma phoeniceum spiralin in Spiroplasma citri.
J. Bacteriol. 177: 2870–2877.
Renaudin, J. 2002. Extrachromosomal elements and gene transfer.
In Molecular Biology and Pathogenicity of Mycoplasmas (edited
by Razin and Herrmann). Academic/Plenum Press, New York, pp.
347–370.
Renbaum, P., D. Abrahamove, A. Fainsod, G.G. Wilson, S. Rottem and
A. Razin. 1990. Cloning, characterization, and expression in Escherichia coli of the gene coding for the CpG DNA methylase from Spiroplasma sp. strain MQ1(M-SssI). Nucleic Acids Res. 18: 1145–1152.
Renbaum, P. and A. Razin. 1992. Mode of action of the Spiroplasma CpG
methylase M-SssI. FEBS Lett. 313: 243–247.
Ricard, B., M. Garnier and J.M. Bové. 1982. Characterization of spiroplasmal virus 3 from spiroplasmas and discovery of a new spiroplasmal virus (SpV4). Rev. Infect. Dis. 4: S275.
Rodwell, A.W. and R.F. Whitcomb. 1983. Methods of direct and indirect
measurement of mycoplasma growth. In Methods in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, New York,
pp. 185–196.
Rogers, M.J., J. Simmons, R.T. Walker, W.G. Weisburg, C.R. Woese, R.S.
Tanner, I.M. Robinson, D.A. Stahl, G. Olsen, R.H. Leach and J. Maniloff.
1985. Construction of the mycoplasma evolutionary tree from 5S rRNA
sequence data. Proc. Natl. Acad. Sci. U. S. A. 82: 1160–1164.
Rogers, M.J., A.A. Steinmetz and R.T. Walker. 1987. Organization and
structure of tRNA genes in Spiroplasma melliferum. Isr. J. Med. Sci. 23:
357–360.
Rose, D.L., J.G. Tully, J.M. Bové and R.F. Whitcomb. 1993. A test for
Rosengarten, R. and K.S. Wise. 1990. Phenotypic switching in mycoplasmas: phase variation of diverse surface lipoproteins. Science 247:
315–318.
Rosselló-Mora, R. and R. Amann. 2001. The species concept for prokaryotes. FEMS Microbiol. Rev 25: 39–67.
Saglio, P., D. Laflèche, C. Bonissol and J.M. Bové. 1971. Isolation, culture
and electronmicroscopy of mycoplasma-like structures associated
with stubborn disease of citrus and their comparison with structures
observed in citrus plants affected by greening disease. [transl. from
Fr.] Physiol. Vég. 9: 569–582.
Saglio, P., M. L’Hospital, D. Laflèche, G. Dupont, J.M. Bové, J.G. Tully
and E.A. Freundt. 1973. Spiroplasma citri gen. and sp. nov.: a mycoplasma-like organism associated with stubborn disease of citrus. Int.
Saglio, P.H.M. and R.F. Whitcomb. 1979. Diversity of wall-less prokaryotes in plant vascular tissue, fungi and invertebrate animals. In The
Saillard, C. and J.M. Bové. 1983. Application of ELISA to spiroplasma
detection and classification. In Methods in Mycoplasmology, vol. 1
(edited by Razin and Tully). Academic Press, New York, pp. 471–476.
Saillard, C., J.C. Vignault, J.M. Bové, A. Raie, J.G. Tully, D.L. Williamson,
A. Fos, M. Garnier, A. Gadeau, P. Carle and R.F. Whitcomb. 1987.
Spiroplasma phoeniceum sp. nov., a new plant-pathogenic species from
Syria. Int. J. Syst. Bacteriol. 37: 106–115.
Saillard, C., C. Chevalier and J.M. Bové. 1990. Structure and organization of the spiralin gene. Zentralbl. Bakteriol. Suppl. 20: 897–901.
Saillard, C., P. Carle, S. Duret-Nurbel, R. Henri, N. Killiny, S. Carrere,
J. Gouzy, J.M. Bové, J. Renaudin and X. Foissac. 2008. The abundant
extrachromosomal DNA content of the Spiroplasma citri GII3–3X
genome. BMC Genomics 9: 195.
Saitou, N. and M. Nei. 1987. The neighbor-joining method: a new
method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4:
406–425.
Sha, Y., U. Melcher, R.E. Davis and J. Fletcher. 1995. Resistance of Spiroplasma citri lines to the virus SVTS2 is associated with integration of
viral DNA sequences into host chromosomal and extrachromosomal
DNA. Appl. Environ. Microbiol. 61: 3950–3959.
Shaevitz, J.W., J.Y. Lee and D.A. Fletcher. 2005. Spiroplasma swim by a
processive change in body helicity. Cell 122: 941–945.
Sher, T., A. Yamin, M. Matzliach, S. Rottem and R. Gallily. 1990a. Partial
biochemical characterization of spiroplasma membrane component
inducing tumor necrosis factor alpha. Anticancer Drugs 1: 83–87.
Sher, T., A. Yamin, S. Rottem and R. Gallily. 1990b. In vitro induction
of tumor necrosis factor alpha, tumor cytolysis, and blast transformation by Spiroplasma membranes. J. Natl. Cancer Inst. 82: 1142–
1145.
Simoneau, P. and J. Labarère. 1991. Evidence for the presence of two
distinct membrane ATPases in Spiroplasma citri. J. Gen. Microbiol.
137: 179–185.
Skripal, I.G. 1974. On improvement of taxonomy of the class Mollicutes
and establishment in the order Mycoplasmatales of the new family
Spiroplasmataceae fam. nov. Mikrobiol. Zh. (Kiev). 36: 462–467.
Skripal, I.G. 1983. Revival of the name Spiroplasmataceae fam. nov., nom.
rev., omitted from the 1980 Approved Lists of Bacterial Names. Int.
J. Syst. Bacteriol. 33: 408.
Sokolova, M.I., N.S. Zinkevich and I.A. Zakharov. 2002. Bacteria in ovarioles of females from maleless families of ladybird beetles Adalia
bipunctata L. (Coleoptera: Coccinellidae) naturally infected with
Rickettsia, Wolbachia, and Spiroplasma. J. Invertebr. Pathol. 79: 72–79.
Stackebrandt, E., W. Frederiksen, G.M. Garrity, P.A. Grimont, P. Kämpfer, M.C. Maiden, X. Nesme, R. Rosselló-Mora, J. Swings, H.G. Trüper,
L. Vauterin, A.C. Ward and W.B. Whitman. 2002. Report of the ad
hoc committee for the re-evaluation of the species definition in bacteriology. Int. J. Syst. Evol. Microbiol. 52: 1043–1047.
Stamburski, C., J. Renaudin and J.M. Bove. 1991. First step toward a
virus-derived vector for gene cloning and expression in spiroplasmas, organisms which read UGA as a tryptophan codon: synthesis
of chloramphenicol acetyltransferase in Spiroplasma citri. J. Bacteriol.
173: 2225–2230.
Stephens, M.A. 1980. Studies on Spiroplasma viruses. PhD thesis, University of East Anglia, Norwich, UK.
Stevens, C., A.Y. Tang, E. Jenkins, R.L. Goins, J.G. Tully, D.L. Rose, M.
Konai, D.L. Williamson, P. Carle, J. Bove, K.J. Hackett, F.E. French,
J. Wedincamp, R.B. Henegar and R.F. Whitcomb. 1997. Spiroplasma
lampyridicola sp. nov., from the firefly beetle Photuris pennsylvanicus.
Summers, C.G., A. S. Newton and D.C. Opgenorth. 2004. Overwintering of corn leafhopper, Dalbulus maidis (Homoptera: Cicadellidae),
and Spiroplasma kunkelii (Mycoplasmatales: Spiroplasmataceae) in California’s San Joaquin Valley. Environ. Entomol. 33: 1644–1651.
Swofford, D.L. 1998. PAUP: Phylogenetic analysis using parsimony and
other methods, 4 edn. Sinauer Associates, Sunderland, MA.
Taroura, S., Y. Shimada, Y. Sakata, T. Miyama, H. Hiraoka, M. Watanabe,
K. Itamoto, M. Okuda and H. Inokuma. 2005. Detection of DNA
of ‘Candidatus Mycoplasma haemominutum’ and Spiroplasma sp. in
unfed ticks collected from vegetation in Japan. J. Vet. Med. Sci. 67:
1277–1279.
Tinsley, M.C. and M.E. Majerus. 2006. A new male-killing parasitism:
Spiroplasma bacteria infect the ladybird beetle Anisosticta novemdecimpunctata (Coleoptera: Coccinellidae). Parasitology 132: 757–765.
Tinsley, M.C. and M.E. Majerus. 2007. Small steps or giant leaps for
male-killers? Phylogenetic constraints to male-killer host shifts. BMC
Evol. Biol. 7: 238.
Townsend, R., P.G. Markham, K.A. Plaskitt and M.J. Daniels. 1977. Isolation and characterization of a nonhelical strain of Spiroplasma citri. J.
Gen. Microbiol. 100: 15–21.
Townsend, R., D.B. Archer and K.A. Plaskitt. 1980a. Purification and
preliminary characterization of Spiroplasma fibrils. J. Bacteriol. 142:
694–700.
Townsend, R., J. Burgess and K.A. Plaskitt. 1980b. Morphology and
ultrastructure of helical an nonhelical strains of Spiroplasma citri. J.
Bacteriol. 142: 973–981.
Trachtenberg, S. and R. Gilad. 2001. A bacterial linear motor: cellular and
molecular organization of the contractile cytoskeleton of the helical
bacterium Spiroplasma melliferum BC3. Mol. Microbiol. 41: 827–848.
Trachtenberg, S., S.B. Andrews and R.D. Leapman. 2003a. Mass distribution and spatial organization of the linear bacterial motor of Spiroplasma citri R8A2. J. Bacteriol. 185: 1987–1994.
Trachtenberg, S., R. Gilad and N. Geffen. 2003b. The bacterial linear
motor of Spiroplasma melliferum BC3: from single molecules to swimming cells. Mol. Microbiol. 47: 671–697.
Trachtenberg, S. 2004. Shaping and moving a Spiroplasma. J. Mol. Microbiol. Biotechnol. 7: 78–87.
Trachtenberg, S. 2006. The cytoskeleton of Spiroplasma: a complex linear motor. J. Mol. Microbiol. Biotechnol. 11: 265–283.
Trachtenberg, S., L.M. Dorward, V.V. Speransky, H. Jaffe, S.B. Andrews
and R.D. Leapman. 2008. Structure of the cytoskeleton of Spiroplasma
melliferum BC3 and its interactions with the cell membrane. J. Mol.
Biol. 378: 778–789.
Tully, J.G., R.F. Whitcomb, H.F. Clark and D.L. Williamson. 1977. Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new
Spiroplasma. Science 195: 892–894.
Tully, J.G., D.L. Rose, O. Garciajurado, J.C. Vignault, C. Saillard, J.M.
Bové, R.E. McCoy and D.L. Williamson. 1980. Serological analysis of
a new group of spiroplasmas. Curr. Microbiol. 3: 369–372.
Tully, J.G., D.L. Rose, C.E. Yunker, J. Cory, R.F. Whitcomb and D.L.
Williamson. 1981. Helical mycoplasmas (spiroplasmas) from Ixodes
ticks. Science 212: 1043–1045.
Tully, J.G., R.F. Whitcomb, D.L. Rose and J.M. Bové. 1982. Spiroplasma
mirum, a new species from the rabbit tick (Haemaphysalis leporispalustris). Int. J. Syst. Bacteriol. 32: 92–100.
Tully, J.G., D.L. Rose, E. Clark, P. Carle, J.M. Bové, R.B. Henegar, R.F.
Whitcomb, D.E. Colflesh and D.L. Williamson. 1987. Revised group
classification of the genus Spiroplasma (class Mollicutes), with proposed new groups XII to XXIII. Int. J. Syst. Bacteriol. 37: 357–364.
cluster of arthropod-associated mollicutes to ordinal rank (Entomoplasmatales ord. nov.), with provision for familial rank to separate species
with nonhelical morphology (Entomoplasmataceae fam. nov.) from
helical species (Spiroplasmataceae), and emended descriptions of the
order Mycoplasmatales, family Mycoplasmataceae. Int. J. Syst. Bacteriol.
43: 378–385.
Tully, J.G., D.L. Rose, C.E. Yunker, P. Carle, J.M. Bové, D.L. Williamson
and R.F. Whitcomb. 1995. Spiroplasma ixodetis sp. nov., a new species
from Ixodes pacificus ticks collected in Oregon. Int. J. Syst. Bacteriol.
45: 23–28.
Van den Ent, F., L. A. Amos and J. Löwe. 2001. Prokaryotic origin of the
actin cytoskeleton. Nature: 39–44.
Vaughn, E.E. and W.M. de Vos. 1995. Identification and characterization of the insertion element IS1070 from Leuconostoc lactis NZ6009.
Gene 155: 95–100.
Vazeille-Falcoz, M., C. Hélias, F. Le Goff, F. Rodhain and C. Chastel.
1997. Three spiroplasmas isolated from Haematopota sp. (Diptera:
Tabanidae) in France. J. Med. Entomol. 34: 238–241.
Veneti, Z., J.K. Bentley, T. Koana, H.R. Braig and G.D.D. Hurst. 2005. A
functional dosage compensation complex required for male killing
in Drosophila. Science 307: 1461–1463.
Wada, H. and R.R. Netz. 2007. Model for self-propulsive helical filaments: kink-pair propagation. Phys. Rev. Lett. 99: 108102.
Wang, W., L. Rong, W. Gu, K. Du and J. Chen. 2003. Study on experimental infections of Spiroplasma from the Chinese mitten crab in crayfish, mice and embryonated chickens. Res. Microbiol. 154: 677–680.
Wang, W., J. Chen, K. Du and Z. Xu. 2004a. Morphology of spiroplasmas
in the Chinese mitten crab Eriocheir sinensis associated with tremor
disease. Res Microbiol 155: 630–635.
Wang, W., B. Wen, G.E. Gasparich, N. Zhu, L. Rong, J. Chen and Z. Xu.
2004b. A Spiroplasma associated with tremor disease in the Chinese
mitten crab (Eriocheir sinensis). Microbiology 150: 3035–3040.
685
Wang, W., W. Gu, Z. Ding, Y. Ren, J. Chen and Y. Hou. 2005. A novel
Spiroplasma pathogen causing systemic infection in the crayfish Procambarus clarkii (Crustacea: Decapod), in China. FEMS Microbiol.
Lett. 249: 131–137.
Wang, W., W. Gu, G.E. Gasparich, K. Bi, J. Ou, Q. Meng, T. Liang,
Q. Feng, J. Zhang and Y. Zhang. 2010. Spiroplasma eriocheiris sp. nov.,
a novel species associated with mortalities in Eriocheiris sinensis,
Chinese mitten crab. Int. J. Syst. Evol. Microbiol. ijs.0.020529-Ov1ijs.0.020529-0.
Wayadande, A.C. and J. Fletcher. 1995. Transmission of Spiroplasma citri
lines and their ability to cross gut and salivary gland barriers within
the leafhopper vector Circulifer tenellus. Phytopathology 85: 1256–
1259.
Wayadande, A.C. and J. Fletcher. 1998. Development and use of an
established cell line of the leafhopper Circulifer tenellus to characterize
Spiroplasma citri-vector interactions. J. Invertebr. Pathol. 72: 126–131.
Wayne, L.G., D.J. Brenner, R.R. Colwell, P.A.D. Grimont, O. Kandler,
M.I. Krichevsky, L.H. Moore, W.E.C. Moore, R.G.E. Murray, E. Stackebrandt, M.P. Starr and H.G. Trüper. 1987. Report of the ad hoc committee on the reconciliation of approaches to bacterial systematics.
Weisburg, W.G., J.G. Tully, D.L. Rose, J.P. Petzel, H. Oyaizu, D. Yang, L.
Mandelco, J. Sechrest, T.G. Lawrence, J. Van Etten, J. Maniloff and
C.R. Woese. 1989. A phylogenetic analysis of the mycoplasmas: basis
for their classification. J. Bacteriol. 171: 6455–6467.
Whitcomb, R.F., J.G. Tully, J.M. Bové and P. Saglio. 1973. Spiroplasmas
and acholeplasmas: multiplication in insects. Science 182: 1251–
1253.
Whitcomb, R.F. and D.L. Williamson. 1979. Pathogenicity of mycoplasmas for arthropods. Zentralbl. Bakteriol. Orig. A 245: 200–221.
Whitcomb, R.F., J. G. Tully, P. McCawley and D.L. Rose. 1982. Application of the growth-inhibition test to Spiroplasma taxonomy. Int. J. Syst.
Bacteriol. 32: 387–394.
Whitcomb, R.F. 1983. Culture media for spiroplasmas. In Methods in
Whitcomb, R.F., T.A. Chen, D.L. Williamson, C. Liao, J.G. Tully, J.M.
Bové, C. Mouches, D.L. Rose, M.E. Coan and T.B. Clark. 1986. Spiroplasma kunkelii sp. nov.: characterization of the etiologic agent of
corn stunt disease. Int. J. Syst. Bacteriol. 36: 170–178.
Whitcomb, R.F. and K.J. Hackett. 1987. Cloning by limiting dilution in
liquid media: an improved alternative for cloning mollicute species.
Isr. J. Med. Sci. 23: 517.
Whitcomb, R.F. 1989. The biology of Spiroplasma kunkelii. In The Mycoplasmas, vol. 5 (edited by Whitcomb and Tully). Academic Press,
Whitcomb, R.F., K. J. Hackett, J. G. Tully, E. A. Clark, F. E. French, R. B.
Henegar, D. L. Rose and A.G. Wagner. 1990. Tabanid spiroplasmas
as a model for mollicute biogeography. Zentrabl. Bakteriol. Suppl.:
931–934.
Whitcomb, R.F., C. Chastel, M. Abalain-Colloc, C. Stevens, J.G. Tully,
D.L. Rose, P. Carle, J.M. Bové, R.B. Henegar, K.J. Hackett, T.B. Clark,
M. Konai and D.L. Williamson. 1993a. Spiroplasma cantharicola sp.
nov., from cantharid beetles (Coleoptera: Cantharidae). Int. J. Syst.
Bacteriol. 43: 421–424.
Whitcomb, R.F., J.G. Tully, D.L. Rose, P. Carle, J.M. Bové, R.B. Henegar,
K.J. Hackett, T.B. Clark, M. Konai, J. Adams and D.L. Williamson.
1993b. Spiroplasma monobiae sp. nov. from the vespid wasp Monobia
quadridens (Hymenoptera, Vespidae). Int. J. Syst. Bacteriol. 43: 256–
260.
Whitcomb, R.F., J.C. Vignault, J.G. Tully, D.L. Rose, P. Carle, J.M. Bové,
K.J. Hackett, R.B. Henegar, M. Konai and D.L. Williamson. 1993c.
Spiroplasma clarkii sp. nov. from the green June beetle (Coleoptera,
Scarabaeidae). Int. J. Syst. Bacteriol. 43: 261–265.
Whitcomb, R.F., G.E. Gasparich, F.E. French, J.G. Tully, D.L. Rose, P.
Carle, J.M. Bové, R.B. Henegar, M. Konai, K.J. Hackett, J.R. Adams,
T.B. Clark and D.L. Williamson. 1996. Spiroplasma syrphidicola sp.
686
nov., from a syrphid fly (Diptera: Syrphidae). Int. J. Syst. Bacteriol.
46: 797–801.
Whitcomb, R.F., F. E. French, J. G. Tully, P. Carle, R. Henegar, K. J.
Hackett, G. E. Gasparich and D.L. Williamson. 1997a. Spiroplasma
species, groups, and subgroups from North American Tabanidae.
Whitcomb, R.F., F.E. French, J.G. Tully, G.E. Gasparich, D.L. Rose, P.
Carle, J. Bové, R.B. Henegar, M. Konai, K.J. Hackett, J.R. Adams, T.B.
Clark and D.L. Williamson. 1997b. Spiroplasma chrysopicola sp. nov.,
Spiroplasma gladiatoris sp. nov., Spiroplasma helicoides sp. nov., and Spiroplasma tabanidicola sp. nov., from tabanid (Diptera: Tabanidae) flies.
Whitcomb, R.F., F.E. French, J.G. Tully, D.L. Rose, P.M. Carle, J.M. Bove,
E.A. Clark, R.B. Henegar, M. Konai, K.J. Hackett, J.R. Adams and
D.L. Williamson. 1997c. Spiroplasma montanense sp. nov., from Hybomitra horseflies at northern latitudes in north America. Int. J. Syst. Bacteriol. 47: 720–723.
Whitcomb, R.F., J. G. Tully, G. E. Gasparich, L. B. Regassa, D. L. Williamson and F.E. French. 2007. Spiroplasma species in the Costa Rican
highlands: implications for biogeography and biodiversity. Biodivers.
Conserv. 16: 3877–3894.
Williamson, D.L. 1969. The sex ratio spirochete in Drosophila robusta.
Jpn. J. Genet. 44: 36–41.
Williamson, D.L. 1974. Unusual fibrils from the spirochete-like sex ratio
organism. J. Bacteriol. 117: 904–906.
Williamson, D.L. and R.F. Whitcomb. 1974. Helical wall-free prokaryotes in Drosophila, leafhoppers and plants. Colloq. Inst. Natl. Santé
Rech. Med. 33: 283–290.
Williamson, D.L. and R.F. Whitcomb. 1975. Plant mycoplasmas: a cultivable spiroplasma causes corn stunt disease. Science 188: 1018–1020.
Williamson, D.L., R. F. Whitcomb and J.G. Tully. 1978. The Spiroplasma
deformation test, a new serological method. Curr. Microbiol. 1: 203–
207.
Williamson, D.L., D.I. Blaustein, R.J.C. Levine and M.J. Elfvin. 1979a.
Anti-actin-peroxidase staining of the helical wall-free prokaryote
Spiroplasma citri. Curr. Microbiol. 2: 143–145.
Williamson, D.L., J. G. Tully and R.F. Whitcomb. 1979b. Serological
relationships of spiroplasmas as shown by combined deformation
and metabolism inhibition tests. Int. J. Syst. Bacteriol. 29: 345–351.
Williamson, D.L. and D.F. Poulson. 1979. Sex ratio organisms (spiroplasmas) of Drosophila. In The Mycoplasmas, vol. 3 (edited by Whitcomb and Tully). Academic Press, New York, pp. 175–208.
Williamson, D.L. and R.F. Whitcomb. 1983. Special serological tests for
spiroplasma identification. In Methods in Mycoplasmology, vol. 2
(edited by Razin and Tully). Academic Press, New York, pp. 249–259.
Williamson, D.L., P.R. Brink and G.W. Zieve. 1984. Spiroplasma fibrils. Isr.
J. Med. Sci. 20: 830–835.
Williamson, D.L., J. G. Tully and R.F. Whitcomb. 1989. The genus Spiroplasma. In The Mycoplasmas, vol. 5 (edited by Whitcomb and Tully).
Williamson, D.L., J. Renaudin and J.M. Bové. 1991. Nucleotide
sequence of the Spiroplasma citri fibril protein gene. J. Bacteriol. 173:
4353–4362.
Williamson, D.L., J.G. Tully, L. Rosen, D.L. Rose, R.F. Whitcomb, M.L.
Abalain-Colloc, P. Carle, J.M. Bové and J. Smyth. 1996. Spiroplasma
diminutum sp. nov., from Culex annulus mosquitoes collected in Taiwan. Int. J. Syst. Bacteriol. 46: 229–233.
Williamson, D.L., J.R. Adams, R.F. Whitcomb, J.G. Tully, P. Carle, M.
Konai, J.M. Bove and R.B. Henegar. 1997. Spiroplasma platyhelix sp.
nov., a new mollicute with unusual morphology and genome size
from the dragonfly Pachydiplax longipennis. Int. J. Syst. Bacteriol. 47:
763–766.
Williamson, D.L., R.F. Whitcomb, J.G. Tully, G.E. Gasparich, D.L. Rose,
P. Carle, J.M. Bové, K.J. Hackett, J.R. Adams, R.B. Henegar, M. Konai,
C. Chastel and F.E. French. 1998. Revised group classification of the
genus Spiroplasma. Int. J. Syst. Bacteriol. 48: 1–12.
Williamson, D.L., B. Sakaguchi, K.J. Hackett, R.F. Whitcomb, J.G. Tully,
P. Carle, J.M. Bové, J.R. Adams, M. Konai and R.B. Henegar. 1999.
Spiroplasma poulsonii sp. nov., a new species associated with malelethality in Drosophila willistoni, a neotropical species of fruit fly. Int. J.
Woese, C.R., J. Maniloff and L.B. Zablen. 1980. Phylogenetic analysis of
the mycoplasmas. Proc. Natl. Acad. Sci. U. S. A. 77: 494–498.
Woese, C.R. 1987. Bacterial evolution. Microbiol. Rev. 51: 221–271.
Wolgemuth, C.W. and N.W. Charon. 2005. The kinky propulsion of
Spiroplasma. Cell 122: 827–828.
Wróblewski, H., K.E. Johansson and S. Hjérten. 1977. Purification and
characterization of spiralin, the main protein of the Spiroplasma citri
membrane. Biochim. Biophys. Acta 465: 275–289.
Wróblewski, H., S. Nyström, A. Blanchard and A. Wieslander. 1989.
Topology and acylation of spiralin. J. Bacteriol. 171: 5039–5047.
Wróblewski, H., D. Robic, D. Thomas and A. Blanchard. 1984. Comparison of the amino acid compositions and antigenic properties of
spiralins purified from the plasma membranes of different spiroplasmas. Ann. Microbiol. (Paris) 135A: 73–82.
Ye, F., F. Laigret, J.C. Whitley, C. Citti, L.R. Finch, P. Carle, J. Renaudin
and J.M. Bové. 1992. A physical and genetic map of the Spiroplasma
citri genome. Nucleic Acids Res. 20: 1559–1565.
Ye, F., F. Laigret and J.M. Bové. 1994a. A physical and genomic map
of the prokaryote Spiroplasma melliferum and its comparison with the
Spiroplasma citri map. C. R. Acad. Sci. Ser. III 317: 392–398.
Ye, F., J. Renaudin, J.M. Bové and F. Laigret. 1994b. Cloning and
sequencing of the replication origin (oriC) of the Spiroplasma citri
chromosome and construction of autonomously replicating artificial
plasmids. Curr. Microbiol. 29: 23–29.
Ye, F., F. Laigret, P. Carle and J.M. Bové. 1995. Chromosomal heterogeneity among various strains of Spiroplasma citri. Int. J. Syst. Bacteriol.
45: 729–734.
Ye, F., U. Melcher, J.E. Rascoe and J. Fletcher. 1996. Extensive chromosome aberrations in Spiroplasma citri strain BR3. Biochem. Genet. 34:
269–286.
Yogev, D., R. Rosengarten, R. Watson-McKown and K.S. Wise. 1991.
Molecular basis of Mycoplasma surface antigenic variation: a novel set
of divergent genes undergo spontaneous mutation of periodic coding regions and 5¢ regulatory sequences. EMBO J. 10: 4069–4079.
Yu, J., A.C. Wayadande and J. Fletcher. 2000. Spiroplasma citri surface
protein P89 implicated in adhesion to cells of the vector Circulifer
tenellus. Phytopathology 90: 716–722.
Zaaria, A., C. Fontenelle, M. Le Henaff and H. Wróblewski. 1990. Antigenic relatedness between the spiralins of Spiroplasma citri and Spiroplasma melliferum. J. Bacteriol. 172: 5494–5496.
Zbinden, M. and M.A. Cambon-Bonavita. 2003. Occurrence of Deferribacterales and Entomoplasmatales in the deep-sea Alvinocarid shrimp
Rimicaris exoculata gut. FEMS Microbiol. Ecol. 46: 23–30.
Zhao, Y., R.W. Hammond, R. Jomantiene, E.L. Dally, I.M. Lee, H. Jia,
H. Wu, S. Lin, P. Zhang, S. Kenton, F.Z. Najar, A. Hua, B.A. Roe,
J. Fletcher and R.E. Davis. 2003. Gene content and organization
of an 85-kb DNA segment from the genome of the phytopathogenic mollicute Spiroplasma kunkelii. Mol. Genet. Genomics 269:
592–602.
Zhao, Y., R.W. Hammond, I.M. Lee, B.A. Roe, S. Lin and R.E. Davis.
2004a. Cell division gene cluster in Spiroplasma kunkelii: functional
characterization of ftsZ and the first report of ftsA in mollicutes. DNA
Cell Biol. 23: 127–134.
Zhao, Y., H. Wang, R.W. Hammond, R. Jomantiene, Q. Liu, S. Lin, B.A.
Roe and R.E. Davis. 2004b. Predicted ATP-binding cassette systems
in the phytopathogenic mollicute Spiroplasma kunkelii. Mol. Genet.
Genomics 271: 325–338.
Family I. Acholeplasmataceae
687
Order III. Acholeplasmatales Freundt, Whitcomb, Barile, Razin and Tully 1984, 348VP
Daniel R. Brown, Janet M. Bradbury and Karl-Erik Johansson
A.cho.le.plas.ma.ta¢les. N.L. neut. n. Acholeplasma type genus of the order; -ales ending to
denote an order; N.L. fem. pl. n. Acholeplasmatales the Acholeplasma order.
This order in the class Mollicutes is assigned to a group of wallless prokaryotes that do not require sterol for growth and occur
in a wide variety of habitats, including many vertebrate hosts,
insects, and plants. A single family, Acholeplasmataceae, and a single genus, Acholeplasma, recognize the prominent and distinct
characteristics of the assigned organisms.
Type genus: Acholeplasma Edward and Freundt 1970, 1AL.
The trivial name acholeplasma(s) is commonly used when
reference is made to species of this order. The initial proposal for elevation of the acholeplasmas to ordinal rank
(Freundt et al., 1984) was based primarily on the universal
lack of a sterol requirement for growth of Acholeplasma
species, in addition to other major genetic, nutritional, biochemical, and physiological characteristics that distinguish
them from other members of the class Mollicutes. A subsequent proposal for an additional order, Entomoplasmatales
(Tully et al., 1993), within the class to distinguish a group of
mollicutes that are phylogenetically more closely related to
the Mycoplasmatales than to acholeplasmas necessitated
further revisions within the class.
References
Edward, D.G. 1971. Determination of sterol requirement for Mycoplasmatales. J. Gen. Microbiol. 69 : 205–210.
Freundt, E.A., R.F. Whitcomb, M.F. Barile, S. Razin and J.G. Tully. 1984.
Proposal for elevation of the family Acholeplasmataceae to ordinal
rank: Acholeplasmatales. Int. J. Syst. Bacteriol. 34: 346–349.
IRPCM Phytoplasma/Spiroplasma Working Team – Phytoplasma
Taxonomy Group. 2004. Description of the genus ‘Candidatus Phytoplasma’, a taxon for the wall-less non-helical prokaryotes that
colonize plant phloem and insects. Int. J. Syst. Evol. Microbiol. 54:
1243–1255.
Lim, P.O. and B.B. Sears. 1992. Evolutionary relationships of a plantpathogenic mycoplasmalike organism and Acholeplasma laidlawii
deduced from two ribosomal protein gene sequences. J. Bacteriol.
174: 2606–2611.
Razin, S. and J.G. Tully. 1970. Cholesterol requirement of mycoplasmas.
J. Bacteriol. 102: 306–310.
Although most mollicutes require exogenous cholesterol or
serum for growth, all species within the genus Acholeplasma and
some assigned to the genera Asteroleplasma, Spiroplasma, and Mesoplasma do not have that requirement. The species that do not
have a sterol requirement can easily be excluded from the sterolrequiring taxa by tests that measure growth responses to cholesterol or to a number of serum-free broth preparations (Edward,
1971; Razin and Tully, 1970; Rose et al., 1993; Tully, 1995). For
instance, the Acholeplasmatales grow through end-point dilutions
in serum-containing medium and in serum-free preparations,
indicating the absence of a growth requirement for cholesterol.
Analyses of rRNA and other genes have shown that a large
group of uncultured, plant-pathogenic organisms referred to by
the trivial name phytoplasmas (Sears and Kirkpatrick, 1994) are
closely related to acholeplasmas (Lim and Sears, 1992; Toth
et al., 1994). The 16S rRNA gene sequences for members of the
genus Acholeplasma that have been determined so far show that
the acholeplasmas form two clades, one of which is a sister lineage
to the phytoplasmas, although the formal taxonomic assignment
of “Candidatus Phytoplasma” proposed gen. nov. (IRPCM Phytoplasma/Spiroplasma Working Team – Phytoplasma Taxonomy
Group, 2004) currently remains incertae sedis.
Rose, D.L., J.G. Tully, J.M. Bové and R.F. Whitcomb. 1993. A test for
Sears, B.B. and B.C. Kirkpatrick. 1994. Unveiling the evolutionary relationships of plant pathogenic mycoplasmalike organisms. ASM News
60: 307–312.
Toth, K.F., N. Harrison and B.B. Sears. 1994. Phylogenetic relationships among members of the class Mollicutes deduced from rps3 gene
sequences. Int. J. Syst. Bacteriol. 44: 119–124.
Tully, J.G., J.M. Bové, F. Laigret and R.F. Whitcomb. 1993. Revised taxonomy
of the class Mollicutes - proposed elevation of a monophyletic cluster of
arthropod-associated mollicutes to ordinal rank (Entomoplasmatales ord.
nov.), with provision for familial rank to separate species with nonhelical
morphology (Entomoplasmataceae fam. nov.) from helical species (Spiroplasmataceae), and emended descriptions of the order Mycoplasmatales,
family Mycoplasmataceae. Int. J. Syst. Bacteriol. 43: 378–385.
Family I. Acholeplasmataceae Edward and Freundt 1970, 1AL
Daniel
R. Brown, Janet M. Bradbury and Karl-Erik Johansson
A.cho.le.plas.ma.ta.ce¢ae. N.L. neut. n. Acholeplasma, -atos type genus of the family; -aceae ending
to denote a family; N.L. fem. pl. n. Acholeplasmataceae the Acholeplasma family.
Type genus: Acholeplasma Edward and Freundt 1970, 1AL.
This family is monotypic, so its properties are essentially those
of the genus Acholeplasma.
688
Genus I. Acholeplasma Edward and Freundt 1970, 1AL
A.cho.le.plas¢ma. Gr. pref. a not; Gr. n. chole bile; Gr. neut. n. plasma something formed or molded, a
form; N.L. neut. n. Acholeplasma name intended to indicate that cholesterol, a constituent of bile, is not
required.
Cells are spherical, with a diameter of about 300 nm, or filamentous, 2–5 µm long. Nonmotile. Colonies have a “fried-egg”
appearance and may reach 2–3 mm in diameter. Facultatively
anaerobic; most strains grow readily in simple media. All members lack a sterol requirement for growth. Chemo-organotrophic,
most species utilizing glucose and other sugars as the major
energy sources. Many strains are capable of fatty acid biosynthesis from acetate. Arginine and urea are not hydrolyzed.
Pigmented carotenoids occur in some species. All species are
resistant, or only slightly susceptible, to 1.5% digitonin. Saprophytes found in soil, compost, wastewaters, or commensals of
vertebrates, insects, or plants. None are known to be a primary
pathogen, but they may cause cytopathic effects in tissue cultures. The genome sizes range from about 1500 to 2100 kbp.
All species examined utilize the universal genetic code in which
UGA is a stop codon.
Type species: Acholeplasma laidlawii (Sabin 1941) Edward and
Freundt 1970, 1AL (Sapromyces laidlawi Sabin 1941, 334).
Cells of acholeplasmas typically appear as pleomorphic coccoid, coccobacillary, or short filamentous forms when grown in
mycoplasma broth containing 20% horse serum or 1% bovine
serum fraction. Viable spherical cells usually have a minimum
diameter of about 300 nm. Filaments may be as much as 500 nm
in length, but some longer filaments and branching filaments
occur in some strains. Filaments often show beading with eventual development of coccoid forms. Cellular morphology may
also depend upon the ratio of unsaturated to saturated fatty
acids in the medium. Adjustment of preparative materials to
the osmolarity of the culture medium is necessary for proper
morphological examination.
Most acholeplasmas exhibit heavy turbidity when grown aerobically in broth containing 5–20% serum, usually of horse or
fetal bovine origin, or when grown in 1% bovine serum fraction broth at 37°C. Less turbidity is evident when most acholeplasmas are cultured in serum-free broth and some species may
be inhibited in media containing 20% horse serum. Strains of
some acholeplasmas (Acholeplasma morum, Acholeplasma modicum, and Acholeplasma axanthum) may not grow well in serumfree medium unless glucose and some fatty acids (Tween 80 and
palmitic acid) are included. Colonies on solid medium containing serum or bovine serum fraction are usually large (100–
200 nm in diameter) with the classical “fried-egg” appearance
after 24–72 h at 37°C (Figure 114). Colonies of Acholeplasma
axanthum and several other acholeplasmas may show only central zones of growth into the agar or other unusual colony forms,
such as mulberry-like colonies. Most acholeplasmas display
optimum growth at 37°C. Growth is much slower at 25–27°C
and strains may require 7–10 d to reach the turbidity observed
after 24 h at 37°C. Species of plant origin (Acholeplasma brassicae
and Acholeplasma palmae) have an optimum growth temperature
of 30°C.
FIGURE 114. Colonies of Acholeplasma laidlawii PG8T (=NCTC 10116T;
diameter 0.15–0.25 mm) after 3 d growth on Mycoplasma Experience
Solid Medium at 36°C in 95% nitrogen/5% carbon dioxide. Original
magnification = 25×. Image provided by Helena Windsor and David
Windsor.
Most species in the genus are strong fermenters and produce
acid from glucose metabolism, although a few species such as
Acholeplasma parvum may not ferment glucose or other carbohydrates (Table 143). Fermentation of mannose is usually negative, although several species do catabolize this carbohydrate.
All Acholeplasma species examined possess a fructose 1,6-diphosphate-activated lactate dehydrogenase, which is a property
shared with certain streptococci.
Gourlay (1970) found that a fresh isolate of Acholeplasma
laidlawii from a bovine source was infected with a filamentous, single-stranded DNA virus designated L1 (Bruce et al.,
1972; Maniloff, 1992). Later, L2 and L3 viruses were also isolated from Acholeplasma laidlawii (Gourlay, 1971, 1972, 1973;
Gourlay et al., 1973). L2 virus is a quasi-spherical, doublestranded DNA virus (Maniloff et al., 1977), and L3 is a shorttailed phage with double-stranded DNA (Garwes et al., 1975;
Gourlay, 1974; Haberer et al., 1979; Maniloff et al., 1977).
Another virus isolated from Acholeplasma laidlawii is L172, a
single-stranded DNA, quasi-spherical virus that is different
from L1 (Liska, 1972). Two viruses have been isolated from
other acholeplasmas, including one from Acholeplasma modicum, designated M1 (Congdon et al., 1979), and from Acholeplasma oculi strain PG49 (designated O1) (Ichimaru and
Nakamura, 1983). The nucleic acid structure of the last two
viruses has not been defined.
Antisera to filter-cloned whole-cell antigens are utilized in several serological techniques to assess the antigenic structure of
acholeplasmas and to provide identification of the organism to
the species level (Tully, 1979). The three most useful techniques
are growth inhibition (Clyde, 1983), plate immunofluorescence
689
Genus I. Acholeplasma
A. brassicae
A. cavigenitalium
A. equifetale
A. granularum
A. hippikon
A. modicum
A. morum
A. multilocale
A. oculi
A. palmae
A. parvum
A. pleciae
A. vituli
Mannose fermentation
Arbutin hydrolysis
Esculin hydrolysis
Film and spots
Benzyl viologen
reduction
DNA G+C content
(mol%)
A. axanthum
Characteristic
A. laidlawii
TABLE 143. Differential characteristics of the species of the genus Acholeplasma a
+
−
+
+
+
+
+
−
+
+
−
+
+
−
−
nd
nd
+
+
−
−
nd
−
+
+
+
nd
nd
+
+
+
−
−
−
−
+
+
+
nd
nd
+
+
+
−
−
−
−
+
+
−
+
+
−
+
+
+
nd
nd
+
−
+
−
+
+
−
+
+
−
−
nd
nd
+
−
nd
nd
−
nd
+
+
nd
nd
nd
nd
nd
+
+
−
−
−
nd
31–36
31
35.5
36
30.5
30–32
33
29
34
31
27
30
29
31.6
37.6–38.3
Symbols: +, >85% positive; −, 0–15% positive; nd, not determined.
a
(Gardella et al., 1983; Tully, 1973), and metabolism inhibition
(Taylor-Robinson, 1983).
Acholeplasmas may be the most common mollicutes in vertebrate animals and they are found frequently in the upper
respiratory tract and urogenital tract of such hosts (Tully, 1979,
1996). Eukaryotic cells in continuous culture are frequently contaminated with acholeplasmas, primarily from the occurrence of
acholeplasmas in animal serum used in tissue culture media. At
least five Acholeplasma species have been identified on plant surfaces (Acholeplasma axanthum, Acholeplasma brassicae, Acholeplasma
laidlawii, Acholeplasma oculi, and Acholeplasma palmae), possibly
representing contamination from insects. However, with the
exception of Acholeplasma pleciae (Knight, 2004), the only acholeplasmas identified from insects have been from mosquitoes.
Acholeplasma laidlawii was identified in a pool of Anopheles sinensis,
and a strain of Acholeplasma morum was present in a pool of Armigeres subalbatus (D.L. Williamson and J.G. Tully, unpublished).
Little evidence exists for a pathogenic role of acholeplasmas in
natural diseases. The widespread distribution of acholeplasmas
in both healthy and diseased animal tissues and of antibodies
against acholeplasmas in most animal sera complicates experimental pathogenicity studies. However, Acholeplasma axanthum
was pathogenic for goslings and young goose embryos (Kisary
et al., 1975, 1976). Inoculation into leafhoppers, including
those known to be vectors of plant mycoplasma diseases, shows
multiplication and prolonged persistence of acholeplasmas in
host tissues (Eden-Green and Markham, 1987; Whitcomb et al.,
1973; Whitcomb and Williamson, 1975), but there is no evidence that the few Acholeplasma species found on plant surfaces
play any role in plant or insect disease.
A few recent reports are available on the antibiotic sensitivity
of acholeplasmas and whether the actions of these drugs are
inhibitory to growth or kill cells. Acholeplasmas are sensitive
to the following antibiotics (minimum inhibitory concentration
range in µg/ml): tetracycline, 0.5–25.0; erythromycin, 0.03–1.0;
lincomycin, 0.25–1.0; tylosin tartarate, 0.1–12.5; and kanamycin, 20–200 (Kato et al., 1972; Lewis and Poland, 1978; Ogata
et al., 1971).
Typical steps in isolation of all mollicutes were outlined in the
recently revised minimal standards for descriptions of novel
species (Brown et al., 2007). Techniques for isolation of acholeplasmas from animal tissues and from cell cultures have been
described (Tully, 1983). Although colonies are occasionally
first detected on blood agar, complex undefined media such
as American Type Culture Collection medium 988 (SP-4) are
usually required for primary isolation and maintenance. Cell
wall-targeting antibiotics are included to discourage growth of
other bacteria. Phenol red facilitates detection of species that
excrete acidic or alkaline metabolites. Commonly used alternatives such as Frey’s, Hayflick’s and Friis’ media differ from SP-4
mainly in the proportions of inorganic salts, amino acids, serum
sources, and types of antibiotics included. Broths are incubated
aerobically at 37°C for 14 d and examined periodically for turbidity or pH changes, either to acid or alkaline levels. Tubes
showing turbidity are plated to agar prepared from the same
medium formulation, and the plates are incubated at 37°C in
an atmosphere of 95% N2, 5% CO2, as in the GasPak system.
Tubes without obvious turbidity should be plated at the end of
the 14-d incubation period. Initial isolates may contain a mixture of species, so cloning by repeated filtration through membrane filters with a pore size of 450 or 220 nm is essential. The
initial filtrate and dilutions of it are cultured on solid medium
and an isolated colony is subsequently picked from a plate on
which only a few colonies have developed. This colony is used
to found a new cultural line, which is then expanded, filtered,
plated, and picked two additional times. Identification is confirmed by additional biochemical and serological tests.
Stock acholeplasma cultures can be maintained in either
mycoplasma broth medium containing 5–20% serum or in
the serum-fraction broth formulation at room temperature
(25–30°C) with only weekly transfer (Tully, 1995). Maintenance
is best in broth medium devoid of glucose, since excess acid
production reduces viability. Stock cultures can also be maintained indefinitely when frozen at −70°C. Agar colonies can
also be maintained for 1–2 weeks at 25°C if plates are sealed
to prevent drying. For optimum preservation, acholeplasmas
should be lyophilized directly in the culture medium when the
broth cultures reach a mid-exponential phase, usually 1–2 d at
37°C. Lyophilized cultures should be sealed under vacuum and
stored at 4°C (Leach, 1983).
690
Differentiation of the genus Acholeplasma
from other genera
wall, filterability, and the presence of conserved 16S rRNA
gene sequences. They usually possess two 16S rRNA operons.
Aerobic or facultative anaerobic growth in artificial media
and the absence of a requirement for sterols or cholesterol
for growth exclude assignment to the genera Anaeroplasma,
Asteroleplasma, “Candidatus Phytoplasma”, Mycoplasma, or Ureaplasma. Absence of a spiral cellular morphology, regular association with a vertebrate host or fluids of vertebrate origin, and
regular use of the codon UGG to encode tryptophan (Knight,
2004) and UGA as a stop codon (Tanaka et al., 1989, 1991)
support exclusion from the genera Spiroplasma, Entomoplasma,
or Mesoplasma. Reduction of the redox indicator benzyl viologen has been reported to be fairly specific for differentiation
of the genus Acholeplasma from other mollicutes (Pollack et al.,
1996a). Only Acholeplasma multilocale failed to give a positive
reaction, although several Mesoplasma and Entomoplasma species yielded variable responses to the test (Pollack et al., 1996a).
Most acholeplasmas have membrane-localized NADH oxidase
activity, in comparison to the NADH oxidase activity located
in the cytoplasm of other genera within the class. Another
special characteristic is the occurrence in most acholeplasmas
of unique pyrimidine enzymic activities, especially a dUTPase
enzyme, with the possible exception again of Acholeplasma multilocale (Pollack et al., 1996b). Acholeplasmas may possess a
number of other biological characteristics that may distinguish
them from other genera within the class Mollicutes, including
polyterpenol synthesis (Smith and Langworthy, 1979), positional distribution of fatty acids (Rottem and Markowitz, 1979),
the presence of superoxide dismutase (Kirby et al., 1980; Lee
and Kenny, 1984; Lynch and Cole, 1980; O’Brien et al., 1981),
and the presence of spacer tRNA (Nakagawa et al., 1992).
However, most of these features have not been established for
even a majority of Acholeplasma species.
Taxonomic comments
Acholeplasma genome sizes range from 1215 to 2095 kbp by
pulsed-field gel electrophoresis or complete DNA sequencing,
but most are in a more narrow range of 1215–1610 kbp (Carle
et al., 1993; Neimark et al., 1992) that overlaps with genome
sizes of many Spiroplasma species. Tests of eight Acholeplasma
species showed less than 8% DNA–DNA hybridization between
type strains and surprisingly extensive genomic heterogeneity within species (Aulakh et al., 1983; Stephens et al., 1983a,
b). The highest level of relatedness, 21% DNA–DNA hybridization, was between the type strains of Acholeplasma laidlawii
and Acholeplasma granularum. Some strain pairs, such as within
Acholeplasma laidlawii, shared as little as 40% DNA–DNA hybridization, differences that in other genera would have justified
subdivision of an apparently diverse strain complex into component species. However, no polyphasic taxonomic basis was
found to support such designations. Restriction endonuclease
digest patterns also reflect heterogeneity within some species
(Razin et al., 1983). The DNA–DNA hybridization and restriction digest patterns of eight Acholeplasma axanthum strains isolated from a variety of hosts and habitats differed markedly
from each other and some heterogeneity occurred among six
different Acholeplasma oculi strains.
Mesoplasma pleciae was first isolated from the hemolymph of
a larva of a Plecia corn root maggot and assigned to the genus
Mesoplasma because sustained growth occurred in serum-free
mycoplasma broth only when the medium contained 0.04%
Tween 80 fatty acid mixture (Tully et al., 1994a). However, 16S
rRNA gene sequence similarities and its preferred use of UGG
rather than UGA to encode tryptophan support proper reclassification as Acholeplasma pleciae comb. nov. (Knight, 2004); the
type strain is PS-1T (Tully et al., 1994a).
Mycoplasma feliminutum was first described during a time
when the only named genus of mollicutes was Mycoplasma.
Its publication coincided with the first proposal of the genus
Acholeplasma (Edward and Freundt, 1969, 1970), with which
Mycoplasma feliminutum is properly affiliated through established phenotypic (Heyward et al., 1969) and 16S rRNA gene
sequence (Brown et al., 1995; Johansson and Pettersson, 2002)
similarities. This explains the apparent inconsistencies with its
assignment to the genus Mycoplasma. The name Mycoplasma
feliminutum should therefore be revised to Acholeplasma feliminutum comb. nov.; the type strain is BenT (=ATCC 25749T; Heyward et al., 1969).
The lack of signature enzymic activities cast serious doubt
on the status of Acholeplasma multilocale PN525T as an authentic
member of the genus Acholeplasma (Pollack et al., 1996b). It
may be affiliated with an unrecognized metabolic subgroup,
but it seems more likely to be a strain of Mycoplasma or Entomoplasma.
Acknowledgements
The major contributions to the foundation of this material by
Joseph G. Tully are gratefully acknowledged.
Further reading
Taylor-Robinson, D. and J.G. Tully. 1998. Mycoplasmas, ureaplasmas, spiroplasmas, and related organisms. In Topley
and Wilson, Principles and Practice of Microbiology, 9th edn,
vol. 2 (edited by Balows and Duerden). Arnold Publishers,
London, pp. 799–827.
and arthropod studies. In The Mycoplasmas (edited by Whitcomb and Tully). Academic Press, San Diego, pp. 1–31.
Acholeplasma
Esculin hydrolysis by a b-d-glucosidase and arbutin hydrolysis
are sometimes useful diagnostic tests for differentiation of
some acholeplasmas (Bradbury, 1977; Rose and Tully, 1983).
The production of carotenoid pigments, principally neurosporene, has been used to differentiate some acholeplasmas,
especially Acholeplasma axanthum and Acholeplasma modicum
(Mayberry et al., 1974; Smith and Langworthy, 1979; Tully
and Razin, 1970). Carotenoids are also synthesized in some
strains of Acholeplasma laidlawii under certain growth conditions (Johansson, 1974). The “film and spots” reaction, which
occurs in a number of Mycoplasma and several Acholeplasma species, relates to the production of crystallized calcium soaps of
fatty acids on the surface of agar plates (Edward, 1954; Fabricant
and Freundt, 1967). Fatty acids are liberated from the serum or
supplemental egg yolk (Fabricant and Freundt, 1967; Thorns
and Boughton, 1978) in the agar medium by the lipolytic activity of the organisms. Failure to cross-react with antisera against
previously recognized species provides evidence for species
novelty. For this reason, deposition of antiserum against a
novel type strain into a recognized collection is still mandatory for new species descriptions (Brown et al., 2007). Prelimi-
691
nary differentiation can be by PCR and DNA sequencing using
primers specific for bacterial 16S rRNA genes or the 16S–23S
intergenic region. A similarity matrix relating the candidate
strain to its closest neighbors, usually species with >0.94 16S
rRNA gene sequence similarity, will suggest an assemblage of
related species that should be examined for serological crossreactivities.
List of species of the genus Acholeplasma
1. Acholeplasma laidlawii (Sabin 1941) Edward and Freundt
1970, 1AL (Sapromyces laidlawi Sabin 1941, 334)
laid.law¢i.i. N.L. gen. masc. n. laidlawii of Laidlaw, named
after Patrick P. Laidlaw, one of the microbiologists who first
isolated this species.
This is the type species of the genus Acholeplasma. Filaments, usually relatively short, although much longer
branched filaments may develop in media with certain
ratios of saturated to unsaturated fatty acids. Coccoid forms
may predominate in certain cultures including co-culture
with eukaryotic cells. Agar colonies are large for a mollicute
and exhibit well-developed central zones and peripheral
growth on horse serum agar. On serum-free agar, colonies
are smaller and may show only the central zone of growth
into the agar. Relatively strong turbidity is produced during
growth in broth containing serum. Temperature range for
growth is 20–41°C with optimum at 37°C, even for strains
recovered from plant or non-animal sources. Usually produces large amounts of carotenoids when cultivated in the
presence of PPLO serum fraction (Difco).
Serologically distinct from most established species in
the genus, but partial cross-reactions may occur with Acholeplasma granularum strains. DNA–DNA hybridization between
strains of this species range from 40 to >80%. Acholeplasma
granularum strain BTS-39T showed 20% hybridization with
Acholeplasma laidlawii strain PG8T. Pathogenicity has not
been established.
Source: isolated from sewage, manure, humus, soil, and
many animal hosts and their tissues, including some isolates
from the human oral cavity, vagina, and wounds. Has been
recovered from the surfaces of some plants, although few
isolations have been reported from insect hosts. Frequent
contaminant of eukaryotic cell cultures.
DNA G+C content (mol%): 31.7–35.7 (Bd, Tm).
Type strain: ATCC 23206, PG8, NCTC 10116, CIP 75.27,
NBRC 14400.
Further comment: on the Approved Lists of Bacterial Names
and on the Approved Lists of Bacterial Names (Amended
Edition), this taxon is incorrectly cited as Acholeplasma laidlawii [Freundt 1955 (sic)] Edward and Freundt (1970).
2. Acholeplasma axanthum Tully and Razin 1970, 754AL
a.xan¢thum. Gr. pref. a not, without; Gr. adj. xanthos -ê -on
yellow; N.L. neut. adj. axanthum without yellow (pigment).
Predominantly coccobacillary and coccoid with a few
short myceloid elements. Large colonies with clearly
marked centers form on horse serum agar; colonies on
serum-free agar are smaller and usually lack the peripheral
growth around their center. Agar colonies produce zones
of b-hemolysis by the overlay technique. Growth in media
devoid of serum or serum fraction is much poorer than for
other acholeplasmas. Minimal nutritional requirements are
poorly defined, but marked stimulation of growth with polyoxyethylene sorbitan (Tween 80) suggests a requirement
for fatty acids. Temperature range for growth is 22–37°C
with optimum growth at 37°C. Synthesis of carotenoid pigments can be demonstrated only when large volume cultures are tested. Produces sphingolipids. No evidence for
pathogenicity.
Source: originally isolated from murine leukemia tissue culture cells, but numerous subsequent isolations of the organism from bovine serum and a variety of bovine tissue sites
(nasal cavity, lymph nodes, kidney) suggest cell-culture contamination was of bovine serum origin. Also isolated from
variety of other animals and surfaces of some plants.
Type strain: S-743, ATCC 25176, NCTC 10138.
3. Acholeplasma brassicae Tully, Whitcomb, Rose, Bové, Carle,
Somerson, Williamson and Eden-Green 1994b, 683VP
bras.si¢cae. L. fem. gen. n. brassicae of cabbage, referring to
the plant origin of the organism.
Cells are primarily coccoid. Temperature range for
growth is 18–37°C. Optimal growth occurs at 30°C. No evidence for pathogenicity.
Source: isolated as a surface contaminant from broccoli
(Brassica oleracea var. italica).
4. Acholeplasma cavigenitalium Hill 1992, 591VP
ca.vi.ge.ni.ta¢li.um. N.L. n. cavia guinea pig (Cavia cobaya);
L. pl. n. genitalia -ium the genitals; N.L. pl. gen. n. cavigenitalium of guinea pig genitals.
Pleomorphic cells, mostly coccoid. Grows on broth or
agar medium under aerobic conditions, with optimum temperature between 35 and 37°C. Colonies on agar medium
have typical fried-egg appearance. Originally described as a
non-fermenter, but the type strain ferments glucose. Does
not grow well on SP-4 broth or in horse serum broth, but
grows well on simple base medium with additions of 10–15%
fetal bovine serum. No evidence for pathogenicity.
Source: isolated from the vagina of guinea pigs.
Type strain: GP3, NCTC 11727, ATCC 49901.
692
5. Acholeplasma entomophilum Tully, Rose, Carle, Bové,
Hackett and Whitcomb 1988, 166VP
en.to.mo.phi¢lum. Gr. n. entomon insect; N.L. neut. adj. philum (from Gr. neut. adj. philon) friend, loving; N.L. neut.
adj. entomophilum insect-loving.
Cells are pleomorphic, but primarily coccoid. Colonies
on solid medium usually have a fried-egg appearance. Acid
is produced from glucose, but not mannose. Carotenoids
are not produced. “Film and spot” reaction is negative.
Agar colonies hemadsorb guinea pig erythrocytes. Strains
require 0.4% Tween 80 or fatty acid supplements for growth
in serum-free media. Temperature range for growth is
23–32°C, with optimum growth at about 30°C. Pathogenicity has not been established.
Source: isolated from gut contents of tabanid flies, beetles,
butterflies, honey bees, and moths, and from flowers.
Type strain: TAC, ATCC 43706.
Further comment: with the proposal of the order Entomoplasmatales (Tully et al., 1993), Acholeplasma entomophilum was transferred to the family Entomoplasmataceae. The
name Acholeplasma entomophilum was therefore revised to
Mesoplasma entomophilum comb. nov. The type strain is TACT
(=ATCC 43706T; Tully et al., 1988).
6. Acholeplasma equifetale Kirchhoff 1978, 81AL
eq.ui.fe.ta¢le. L. n. equus horse; N.L. adj. fetalis -is -e pertaining to the fetus; N.L. neut. adj. equifetale pertaining to the
horse fetus.
Cells are pleomorphic, but predominantly coccoid. Colonies on solid medium containing serum usually have a
fried-egg appearance; on serum-free medium, colonies are
similar, but usually smaller. Growth temperature range is
22–37°C. Pathogenicity has not been established.
Source: isolated from the lung and liver of aborted horse
fetuses. Also recovered from the respiratory tract of apparently normal horses and the respiratory tract and cloacae of
broiler chickens (Bradbury, 1978).
Type strain: C112, ATCC 29724, NCTC 10171.
Further comment: Kirchhoff is incorrectly cited as “Kirchoff”
on the Approved Lists of Bacterial Names.
7. Acholeplasma florum McCoy, Basham, Tully, Rose, Carle
and Bové 1984, 14VP
flo¢rum. L. gen. p1. n. florum of flowers, indicating the
recovery site of the organism.
Cells are ovoid. Colonies on agar are umbonate. Films
and spots are produced on serum-containing media. Glucose is utilized, but mannose is not. Carotenes are not produced, nor is b-d-glucosidase. Pathogenicity has not been
established.
Source: the known strains were isolated from flower surfaces.
Type strain: L1, ATCC 33453.
Sequence accession nos: AF300327 (16S rRNA gene),
NC_006055 (strain L1T complete genome).
Further comment: with the proposal of the order Entomoplasmatales (Tully et al., 1993), Acholeplasma florum was
transferred to the family Entomoplasmataceae. The name
Acholeplasma florum was therefore revised to Mesoplasma
florum comb. nov. The type strain is L1T (=ATCC 33453T;
McCoy et al., 1984).
8. Acholeplasma granularum (Switzer 1964) Edward and Freundt 1970, 2AL (Mycoplasma granularum Switzer 1964, 504)
gra.nu.la¢rum. N.L. fem. n. granula (from L. neut. n. granulum) a small grain, a granule; N.L. gen. pl. n. granularum of
small grains, made up of granules, granular.
Cells are pleomorphic, with short filaments and coccoid cells. Colonies on solid medium are large with clearly
marked central zones and a fried-egg appearance. Colonies
on serum-free medium are smaller and may lack the peripheral zone of growth around central core. Temperature
range for growth is 22–37°C, with optimum around 37°C.
Agar colonies produce a zone of b-hemolysis by the overlay
technique using sheep erythrocytes. DNA–DNA hybridization studies showed 20–22% hybridization with Acholeplasma
laidlawii, but none with other acholeplasmas. Pathogenicity has not been established. Aerosol challenge of specific
pathogen-free pigs did not induce clinical or histological
evidence of disease.
Source: isolated frequently from the nasal cavity of swine,
with occasional isolates from swine lung and feces. Also
isolated from the conjunctivae and nasopharynx of horses,
and the genital tract of guinea pigs. Occasional contaminant of eukaryotic cell cultures.
Type strain: BTS-39, ATCC 19168, NCTC 10128.
9. Acholeplasma hippikon Kirchhoff 1978, 81AL
hip.pi¢kon. Gr. neut. adj. hippikon pertaining to the horse.
Cells are pleomorphic with predominantly coccoid
forms. Colonies on solid medium containing horse serum
typically have a fried-egg appearance, with smaller colonies
on serum-free agar medium. Growth occurs over a temperature range of 22–37°C, with optimal growth at 35–37°C. Agar
colonies produce b-hemolysis with the overlay technique,
using a variety of animal red blood cells. Pathogenicity has
Source: isolated from the lung of aborted horse fetuses.
Type strain: C1, ATCC 29725, NCTC 10172.
Further comment: Kirchhoff is incorrectly cited as “Kirchoff”
on the Approved Lists of Bacterial Names.
10. Acholeplasma modicum Leach 1973, 147AL
mo¢di.cum. L. neut. adj. modicum moderate, referring to
moderate growth.
Cells are pleomorphic, with spherical, ring-shaped, and
coccobacillary forms. Colonies on solid medium are distinctly smaller than those of most other acholeplasmas.
Very small colonies without peripheral zones of growth are
noted on serum-free solid medium. Very light turbidity is
observed in serum-free broth, but more turbidity is found
in broth containing serum. Growth temperature range is
22–37°C, with optimum growth around 35–37°C. Can be
shown to produce carotenoids when large volumes of cells
are examined. Agar colonies produce a- or b-hemolysis by
the overlay technique using sheep, ox, or guinea pig red
blood cells. Pathogenicity has not been established.
Source: isolated from various tissues of cattle, including
blood, bronchial lymph nodes, thoracic fluids, lungs, and
semen. Also isolated from nasal secretions of pigs, and occasionally from chickens, turkeys, and ducks.
11. Acholeplasma morum Rose, Tully and Del Giudice 1980,
653VP
mor¢um. L. n. morum a mulberry, denoting the mulberrylike appearance of agar colonies of the organism.
Cells are pleomorphic, predominantly coccoid or coccobacillary forms, but with some beaded filaments. Colonies
on solid medium without serum supplements are very small
in size and have only central zones without any peripheral
growth. Optimal growth on solid medium occurs with a
10% serum concentration and colony growth appears to be
suppressed in a medium with 20% serum. Optimal growth
in broth is apparent when 5–10% serum is added or when
1% bovine serum fraction supplements are added, but
poor growth occurs in broth containing 20% horse serum.
Growth in serum-free broth usually requires some fatty acid
supplements, such as palmitic acid or polyoxyethylene sorbitan (Tween 80). Temperature range for growth is 23–37°C,
with optimum growth at about 35–37°C.
Pathogenicity has not been established. Calf kidney cell cultures containing the organism show cytopathogenic effects.
Source: originally recovered from commercial fetal bovine
serum and from calf kidney cultures containing fetal bovine
serum. One isolation, in broth containing horse serum, was
from a pool of Armigeres subalbatus mosquitoes collected by
Leon Rosen in Taiwan in 1978 (strain SP7; D.L. Williamson
and J.G. Tully, unpublished).
Type strain: 72-043, ATCC 33211, NCTC 10188.
12. Acholeplasma multilocale Hill, Polak-Vogelzang and Angulo
1992, 516VP
mul.ti.lo.ca¢le. L. adj. multus many, numerous; L. adj. localis
-is -e of or belonging to a place, local; N.L. neut. adj. multilocale referring to more than one location.
Pleomorphic cells. Colonies on agar medium have a
typical fried-egg appearance. Organisms grow well in broth
medium at 35–37°C. No evidence for pathogenicity.
Source: isolated from the nasopharynx of a horse and the
feces of a rabbit.
Type strain: PN525, NCTC 11723, ATCC 49900.
13. Acholeplasma oculi corrig. al-Aubaidi, Dardiri, Muscoplatt
and McCauley 1973, 126AL
o¢cu.li. L. n. oculus the eye; L. gen. n. oculi of the eye.
693
Cells are pleomorphic, including spherical, ring-shaped,
and coccobacillary forms. Medium-sized colonies with typical fried-egg appearance are formed on horse serum agar.
Colonies on serum-free agar are smaller and may lack the
peripheral growth around the central core. Growth occurs
at temperatures of 25–37°C. Agar colonies produce zones of
hemolysis by the overlay technique using sheep red blood
cells.
Pathogenicity is not well established. Intravenous inoculation of goats produced signs of pneumonia and death
within 6 d. Conjunctival inoculation of goats produced mild
conjunctivitis.
Source: isolated from the conjunctiva of goats with keratoconjunctivitis; porcine nasal secretions; equine nasopharynx, lung, spinal fluid, joint, and semen; the urogenital
tract of cattle; and the external genitalia of guinea pigs.
Present in amniotic fluid of pregnant women (Waites et al.,
1987). Occasionally isolated from ducks and turkeys, with
unreported isolations from an ostrich. Also several isolations from palm trees and other plants (Eden-Green and
Tully, 1979; Somerson et al., 1982). Isolations from eukaryotic cell cultures may represent contamination of bovine
origin.
Type strain: 19-L, ATCC 27350, NCTC 10150.
Further comment: originally named Acholeplasma oculusi
by al-Aubaidi et al. (1973); the orthographic error was corrected by al-Aubaidi (1975).
14. Acholeplasma palmae Tully, Whitcomb, Rose, Bové, Carle,
Somerson, Williamson and Eden-Green 1994b, 683VP
pal¢mae. L. fem. gen. n. palmae of a palm tree, referring to
the plant from which the organism was isolated.
Cells are primarily coccoid. Colonies on solid medium
usually have a fried-egg appearance. The temperature
range for growth is 18–37°C, with optimal growth occurring
at 30°C. No evidence for pathogenicity. It is one of the closest phylogenetic relatives of the phytoplasmas.
Source: isolated from the crown tissues of a palm tree
(Cocos nucifera) with lethal yellowing disease.
DNA G+C content (mol%): 30 (Bd, Tm, HPLC).
Type strain: J233, ATCC 49389.
15. Acholeplasma parvum Atobe, Watabe and Ogata 1983, 348VP
par¢vum. L. neut. adj. parvum small, intended to refer to
the poor biochemical activities and tiny agar colonies of the
organism.
Pleomorphic coccobacillary cells. Colonies on agar
medium present a typical fried-egg appearance under both
aerobic and anaerobic conditions. Initial reports of growth
in the absence of cholesterol or serum have been made, but
growth on serum-free medium is not well confirmed. The
organism does not grow in most standard media for acholeplasmas or in most other medium formulations for sterolrequiring mycoplasmas. Needs special growth factor of 1%
phytone or soytone peptone supplements; growth is sometimes better with the addition of 15% fetal bovine serum.
Organisms grow on agar better than in broth; growth is
694
etter under aerobic conditions than under anaerobic
b
conditions and better at 22–30°C than at 37°C. No evidence
of fermentation of any carbohydrate, including glucose,
salicin, and esculin. No evidence for pathogenicity.
Source: isolated from the oral cavities and vagina of
healthy horses.
Type strain: H23M, ATCC 29892, NCTC 10198.
16. Acholeplasma pleciae (Tully, Whitcomb, Hackett, Rose,
Henegar, Bové, Carle, Williamson and Clark 1994a) Knight
2004, 1952VP (Mesoplasma pleciae Tully, Whitcomb, Hackett, Rose,
Henegar, Bové, Carle, Williamson and Clark 1994a, 690)
ple.ci¢ae. N.L. gen. n. pleciae of Plecia, referring to the genus
of corn maggot (Plecia sp.) from which the organism was
first isolated.
Cells are primarily coccoid. Colonies on solid media
incubated under anaerobic conditions at 30°C have a friedegg appearance. Supplements of 0.04% polyoxyethylene
sorbitan (Tween 80) are required for growth in serum-free
media. Temperature range for growth is 18–32°C, with
optimal growth at 30°C. Agar colonies do not hemadsorb
guinea pig erythrocytes. No evidence for pathogenicity.
Source: originally isolated from the hemolymph of a larva
of the corn root maggot (Plecia sp.).
Type strain: PS-1, ATCC 49582.
17. Acholeplasma seiffertii Bonnet, Saillard, Vignault, Garnier,
Carle, Bové, Rose, Tully and Whitcomb 1991, 48VP
seif.fer¢ti.i. N.L. gen. masc. n. seiffertii of Seiffert, in honor
of Gustav Seiffert, a German microbiologist who performed
References
al-Aubaidi, J.M. 1975. Orthographic errors in the name Acholeplasma
oculusi. Int. J. Syst. Bacteriol. 25: 221.
al-Aubaidi, J.M., A.H. Dardiri, C.C. Muscoplatt and E.H. McCauley.
1973. Identification and characterization of Acholeplasma oculusi spec.
nov. from the eyes of goats with keratoconjunctivitis. Cornell Vet. 63:
117–129.
Angulo, A.F., R. Reijgers, J. Brugman, I. Kroesen, F.E.N. Hekkens,
P. Carle, J.M. Bové, J.G. Tully, A.C. Hill, L.M. Schouls, C.S. Schot,
P.J.M. Roholl and A.A. Polak-Vogelzang. 2000. Acholeplasma vituli sp.
nov., from bovine serum and cell cultures. Int. J. Syst. Evol. Microbiol. 50: 1125–1131.
Atobe, H., J. Watabe and M. Ogata. 1983. Acholeplasma parvum, a new
species from horses. Int. J. Syst. Bacteriol. 33: 344–349.
Aulakh, G.S., E.B. Stephens, D.L. Rose, J.G. Tully and M.F. Barile. 1983.
Nucleic acid relationships among Acholeplasma species. J. Bacteriol.
153: 1338–1341.
Bonnet, F., C. Saillard, J.C. Vignault, M. Garnier, P. Carle, J.M. Bové,
D.L. Rose, J.G. Tully and R.F. Whitcomb. 1991. Acholeplasma seiffertii sp.
nov., a mollicute from plant surfaces. Int. J. Syst. Bacteriol. 41: 45–49.
Bradbury, J. 1977. Rapid biochemical tests for characterization of the
Mycoplasmatales. J. Clin. Microbiol. 5: 531–534.
Bradbury, J.M. 1978. Acholeplasma equifetale in broiler chickens. Vet. Rec.
102: 516.
pioneering studies of sterol-nonrequiring mollicutes that
occur in soil and compost.
Cells are primarily coccoid. Colonies on solid medium
sually have the appearance of fried-eggs. Acid produced
u
from glucose and mannose. Colonies on agar hemadsorb
guinea pig erythrocytes. Temperature range for growth is
20–35°C; optimum growth occurs at 28°C. No evidence for
pathogenicity.
Source: isolated from floral surfaces of a sweet orange
(Citrus sinensis) and wild angelica (Angelica sylvestris).
Type strain: F7, ATCC 49495.
Further comment: with the proposal of the order Entomoplasmatales (Tully et al., 1993), Acholeplasma seiffertii was
transferred to the family Entomoplasmataceae. The name
Acholeplasma seiffertii was therefore revised to Mesoplasma
seiffertii comb. nov. The type strain is F7T (=ATCC 49495T;
Bonnet et al., 1991).
18. Acholeplasma vituli Angulo, Reijgers, Brugman, Kroesen,
Hekkens, Carle, Bové, Tully, Hill, Schouls, Schot, Roholl
and Polak-Vogelzang 2000, 1130VP
vi.tu¢li. L. n. vitulus calf; L. gen. n. vituli of calf, referring to the
provenance or occurrence of the organism in fetal calf serum.
Cells are predominantly coccoid in shape. Colonies on
solid media demonstrate a fried-egg appearance under
both aerobic and anaerobic conditions. Temperature range
for growth is 25–37°C. No evidence for pathogenicity.
Source: isolated from fetal bovine serum or contaminated
eukaryotic cell cultures containing serum.
DNA G+C content (mol%): 38.3 (Bd), 37.6 (Tm).
Type strain: FC 097-2, ATCC 700667, CIP 107001.
Brown, D., G. McLaughlin and M. Brown. 1995. Taxonomy of the feline
mycoplasmas Mycoplasma felifaucium, Mycoplasma feliminutum, Mycoplasma felis, Mycoplasma gateae, Mycoplasma leocaptivus, Mycoplasma
leopharyngis, and Mycoplasma simbae by 16S rRNA gene sequence comparisons. Int. J. Syst. Bacteriol. 45: 560–564.
Bruce, J., R.N. Gourlay, R. Hull and D.J. Garwes. 1972. Ultrastructure of
Mycoplasmatales virus laidlawii I. J. Gen. Virol. 16: 215–221.
Carle, P., D.L. Rose, J.G. Tully and J.M. Bové. 1993. The genome size of
spiroplasmas and other mollicutes. Int. Org. Mycoplasmol. Lett. 2: 263.
Clyde, W.A., Jr. 1983. Growth inhibition tests. In Methods in Mycoplasmology, vol. 1 (edited by Razin and Tully). Academic Press, New
York, pp. 405–410.
Congdon, A.L., E.S. Boatman and G.E. Kenny. 1979. Mycoplasmatales
virus MV-M1: discovery in Acholeplasma modicum and preliminary
characterization. Curr. Microbiol. 3: 111–115.
Eden-Green, S.J. and P.G. Markham. 1987. Multiplication and persistence of Acholeplasma spp. in leafhoppers. J. Invertebr. Pathol. 49:
235–241.
Eden-Green, S.J. and J.G. Tully. 1979. Isolation of Acholeplasma spp.
from coconut palms affected by lethal yellowing disease in Jamaica.
Edward, D.G. 1954. The pleuropneumonia group of organisms: a
review, together with some new observations. J. Gen. Microbiol. 10:
27–64.
Edward, D.G. and E.A. Freundt. 1969. Proposal for classifying organisms
related to Mycoplasma laidlawii in a family Sapromycetaceae, genus Sapromyces, within the Mycoplasmatales. J. Gen. Microbiol. 57: 391–395.
Fabricant, J. and E.A. Freundt. 1967. Importance of extension and
standardization of laboratory tests for the identification and classification of mycoplasma. Ann. N. Y. Acad. Sci. 143: 50–58.
Gardella, R.S., R.A. Del Giudice and J.G. Tully. 1983. Immunofluorescence. In Methods in Mycoplasmology, vol. 1 (edited by Razin and
Garwes, D., B. Pike, S. Wyld, D. Pocock and R. Gourlay. 1975. Characterization of Mycoplasmatales virus-laidlawii 3. J. Gen. Virol. 29: 11–24.
Gourlay, R.N. 1970. Isolation of a virus infecting a strain of Mycoplasma
laidlawii. Nature 225: 1165.
Gourlay, R.N. 1971. Mycoplasmatales virus-laidlawii 2, a new virus isolated
from Acholeplasma laidlawii. J. Gen. Virol. 12: 65–67.
Gourlay, R.N. 1972. Isolation and characterization of mycoplasma
viruses. Proceedings of the CIBA Found. Symp., pp. 145–156.
Gourlay, R.N. 1973. Mycoplasma viruses: isolation, physicohemical, and
biological properties. Ann. N. Y. Acad. Sci. 225: 144–148.
Gourlay, R.N. 1974. Mycoplasma viruses: isolation, physicochemical, and
biological properties. CRC Crit. Rev. Microbiol. 3: 315–331.
Gourlay, R.N., J. Brownlie and C.J. Howard. 1973. Isolation of
T-mycoplasmas from goats, and the production of subclinical mastitis
in goats by the intramammary inoculation of human T-mycoplasmas.
J. Gen. Microbiol. 76: 251–254.
Haberer, K., G. Klotz, J. Maniloff and A. Kleinschmidt. 1979. Structural
and biological properties of mycoplasmavirus MVL3: an unusual
virus-procaryote interaction. J. Virol. 32: 268–275.
Heyward, J.T., M.Z. Sabry and W.R. Dowdle. 1969. Characterization of
Mycoplasma species of feline origin. Am. J. Vet. Res. 30: 615–622.
Hill, A. 1992. Acholeplasma cavigenitalium sp. nov., isolated from the
vagina of guinea pigs. Int. J. Syst. Bacteriol. 42: 589–592.
Hill, A., A. Polak-Vogelzang and A. Angulo. 1992. Acholeplasma multilocale sp. nov., isolated from a horse and a rabbit. Int. J. Syst. Bacteriol.
42: 513–517.
Ichimaru, H. and M. Nakamura. 1983. Biological properties of a plaqueinducing agent obtained from Acholeplasma oculi. Yale J. Biol. Med.
56: 761–763.
Johansson, K-E. 1974. Fractionation of membrane proteins from Acholeplasma laidlawii by preparative agarose suspension electrophoresis.
In Protides of the Biological Fluids – 21st Colloquium (edited by
Peeters). Pergamon Press, Oxford, pp. 151–156.
Johansson, K.E., Pettersson B. 2002. Taxonomy of Mollicutes. In Molecular Biology and Pathogenicity of Mycoplasmas (edited by Razin and
Herrmann). Kluwer Academic/Plenum Press, New York, pp. 1–30.
Kato, H., T. Murakami, S. Takase and K. Ono. 1972. Sensitivities in vitro
to antibiotics of Mycoplasma isolated from canine sources. Jpn. J. Vet.
Sci. 34: 197–206.
Kirby, T., J. Blum, I. Kahane and I. Fridovich. 1980. Distinguishing
between manganese-containing and iron-containing superoxide
dismutases in crude extracts of cells. Arch. Biochem. Biophys. 20:
551–555.
Kirchhoff, H. 1978. Acholeplasma equifetale and Acholeplasma hippikon,
two new species from aborted horse fetuses. Int. J. Syst. Bacteriol.
28: 76–81.
Kisary, J. and L. Stipkovits. 1975. Effect of Mycoplasma gallinarum on the
replication in vitro of goose parvovirus strain “B”. Acta Microbiol.
Acad. Sci. Hung. 22: 305–307.
Kisary, J., A. El-Ebeedy and L. Stipkovits. 1976. Mycoplasma infection of
geese. II. Studies on pathogenicity of mycoplasmas in goslings and
goose and chicken embryos. Avian Pathol. 5: 15–20.
695
Leach, R.H. 1973. Further studies on classification of bovine strains of
Mycoplasmatales, with proposals for new species, Acholeplasma modicum
and Mycoplasma alkalescens. J. Gen. Microbiol. 75: 135–153.
Lee, G.Y. and G.E. Kenny. 1984. Immunological heterogeneity of superoxide dismutases in the Acholeplasmataceae. Int. J. Syst. Bacteriol. 34:
74–76.
Lewis, J. and J. Poland. 1978. Sensitivity of mycoplasmas of the respiratory tract of pigs and horses to erythromycin and its use in selective
media. Res. Vet. Sci. 24: 121–123.
Liska, B. 1972. Isolation of a new Mycoplasmatales virus. Stud. Biophys.
34: 151–155.
Lynch, R.E. and B.C. Cole. 1980. Mycoplasma pneumoniae: a prokaryote which consumes oxygen and generates superoxide but which
lacks superoxide dismutase. Biochem. Biophys. Res. Commun. 96:
98–105.
Maniloff, J. 1992. Mycoplasma viruses. In Mycoplasmas: Molecular Biology and Pathogenesis (edited by Maniloff, McElhaney, Finch and
Baseman). American Society for Microbiology, Washington, DC,
pp. 41–59.
Maniloff, J., J. Das and J.R. Christensen. 1977. Viruses of mycoplasmas
and spiroplasmas. Adv. Virus Res. 21: 343–380.
Mayberry, W.R., P.F. Smith and T.A. Langworthy. 1974. Heptose-containing pentaglycosyl diglyceride among the lipids of Acholeplasma modicum. J. Bacteriol. 118: 898–904.
McCoy, R.E., H.G. Basham, J.G. Tully, D.L. Rose, P. Carle and J.M. Bové.
1984. Acholeplasma florum, a new species isolated from plants. Int. J.
Nakagawa, T., T. Uemori, K. Asada, I. Kato and R. Harasawa. 1992.
Acholeplasma laidlawii has tRNA genes in the 16S–23S spacer of the
rRNA operon. J. Bacteriol. 174: 8163–8165.
Neimark, H.C., J.G. Tully, D. Rose and C. Lange. 1992. Chromosome size
polymorphism among mollicutes. Int. Org. Mycoplasmol. Lett. 2: 261.
O’Brien, S.J., J.M. Simonson, M.W. Grabowski and M.F. Barile. 1981.
Analysis of multiple isoenzyme expression among twenty-two species
of Mycoplasma and Acholeplasma. J. Bacteriol. 146: 222–232.
Ogata, M., H. Atobe, H. Kushida and K. Yamamoto. 1971. In vitro sensitivity of mycoplasmas isolated from various animals and sewage to
antibiotics and nitrofurans. J. Antibiot. (Tokyo) 24: 443–451.
Pollack, J.D., J. Banzon, K. Donelson, J.G. Tully, Jr, J.W. Davis, K.J.
Hackett, C. Agbanyim and R.J. Miles. 1996a. Reduction of benzyl
viologen distinguishes genera of the class Mollicutes. Int. J. Syst. Bacteriol. 46: 881–884.
Pollack, J.D., M.V. Williams, J. Banzon, M.A. Jones, L. Harvey and
J.G. Tully. 1996b. Comparative metabolism of Mesoplasma, Entomoplasma, Mycoplasma, and Acholeplasma. Int. J. Syst. Bacteriol. 46:
885–890.
Razin, S., J. Tully, D. Rose and M. Barile. 1983. DNA cleavage patterns as
indicators of genotypic heterogeneity among strains of Acholeplasma
and Mycoplasma species. J. Gen. Microbiol. 129: 1935–1944.
Rose, D.L. and J.G. Tully. 1983. Detection of b-d-glucosidase: hydrolysis
of esculin and arbutin. In Methods in Mycoplasmology (edited by
Rose, D.L., J.G. Tully and R.A. Del Giudice. 1980. Acholeplasma morum,
a new non-sterol-requiring species. Int. J. Syst. Bacteriol. 30: 647–
654.
Rottem, S. and O. Markowitz. 1979. Unusual positional distribution of
fatty acids in phosphatidylglycerol of sterol-requiring mycoplasmas.
FEBS Lett. 107: 379–382.
Sabin, A.B. 1941. The filterable microorganisms of the pleuropneumonia group. Bacteriol. Rev. 5: 1–66.
696
Smith, P. and T. Langworthy. 1979. Existence of carotenoids in Acholeplasma axanthum. J. Bacteriol. 137: 185–188.
Somerson, N., J. Kocka, D. Rose and R. Del Giudice. 1982. Isolation
of acholeplasmas and a mycoplasma from vegetables. Appl. Environ.
Microbiol. 43: 412–417.
Stephens, E.B., G.S. Aulakh, D.L. Rose, J.G. Tully and M.F. Barile.
1983a. Intraspecies genetic relatedness among strains of Acholeplasma
laidlawii and of Acholeplasma axanthum by nucleic acid hybridization.
J. Gen. Microbiol. 129: 1929–1934.
Stephens, E.B., G.S. Aulakh, D.L. Rose, J.G. Tully and M.F. Barile. 1983b.
Interspecies and intraspecies DNA homology among established species of Acholeplasma: a review. Yale J. Biol. Med. 56: 729–735.
Switzer, W.P. 1964. Mycoplasmosis. In Diseases of Swine, 2nd edn (edited
by Dunne). Iowa State University Press, Ames, IA, pp. 498–507.
Tanaka, R., A. Muto and S. Osawa. 1989. Nucleotide sequence of
tryptophan tRNA gene in Acholeplasma laidlawii. Nucleic Acids Res. 17:
5842.
Tanaka, R., Y. Andachi and A. Muto. 1991. Evolution of tRNAs and tRNA
genes in Acholeplasma laidlawii. Nucleic Acids Res. 19: 6787–6792.
Taylor-Robinson, D. 1983. Metabolism inhibition tests. In Methods in
Thorns, C. and E. Boughton. 1978. Studies on film production and
its specific inhibition, with special reference to Mycoplasma bovis
(M. agalactiae var. bovis). Zentralbl. Veterinarmed. B 25: 657–667.
Tully, J.G. 1973. Biological and serological characteristics of the acholeplasmas. N. Y. Acad. Sci. 225: 74–93.
Tully, J.G. 1979. Special features of the acholeplasmas. In The Mycoplasmas, vol. 1 (edited by Barile and Razin). Academic Press, New York,
pp. 431–449.
Tully, J.G. 1983. Methods in mycoplasmology, vol. 2, Diagnostic Mycoplasmology. Academic Press, New York.
Tully, J.G. 1996. Mollicute-host interrelationships: current concepts and
diagnostic implications. In Molecular and Diagnostic Procedures in
San Diego, pp. 1–21.
Tully, J. and S. Razin. 1970. Acholeplasma axanthum, sp. n.: a new sterol-nonrequiring member of the Mycoplasmatales. J. Bacteriol. 103:
751–754.
Tully, J.G., D.L. Rose, P. Carle, J.M. Bové, K.J. Hackett and R.F. Whitcomb. 1988. Acholeplasma entomophilum sp. nov. from gut contents of
a wide-range of host insects. Int. J. Syst. Bacteriol. 38: 164–167.
Tully, J.G., J.M. Bove, F. Laigret and R.F. Whitcomb. 1993. Revised taxonomy of the class Mollicutes – proposed elevation of a monophyletic
cluster of arthropod-associated mollicutes to ordinal rank (Entomoplasmatales ord. nov.), with provision for familial rank to separate
of the order Mycoplasmatales, family Mycoplasmataceae. Int. J. Syst.
Bacteriol. 43: 378–385.
Tully, J.G., R.F. Whitcomb, K.J. Hackett, D.L. Rose, R.B. Henegar,
J.M. Bové, P. Carle, D.L. Williamson and T.B. Clark. 1994a. Taxonomic
Tully, J.G., R.F. Whitcomb, D.L. Rose, J.M. Bové, P. Carle, N.L. Somerson, D.L. Williamson and S. Eden-Green. 1994b. Acholeplasma
brassicae sp. nov. and Acholeplasma palmae sp. nov., two non-sterolrequiring mollicutes from plant surfaces. Int. J. Syst. Bacteriol. 44:
680–684.
Waites, K.B., J.G. Tully, D.L. Rose, P.A. Marriott, R.O. Davis and
G.H. Cassell. 1987. Isolation of Acholeplasma oculi from human amniotic fluid in early pregnancy. Curr. Microbiol. 15: 325–327.
Whitcomb, R.F. and D.L. Williamson. 1975. Helical wall-free prokaryotes in insects: multiplication and pathogenicity. Ann. N. Y. Acad.
Sci. 266: 260–275.
Whitcomb, R.F., J.G. Tully, J.M. Bové and P. Saglio. 1973. Spiroplasmas and acholeplasmas: multiplication in insects. Science 182:
1251–1253.
This family includes the phytoplasma strains of the order
Acholeplasmatales. Although never cultured in cell-free media,
these plant pathogens and symbionts have been well studied by
culture-independent methods.
Genus I. “Candidatus Phytoplasma” gen. nov. IRPCM Phytoplasma/Spiroplasma Working Team 2004, 1244
Nigel A. Harrison, Dawn Gundersen-Rindal and Robert E. Davis
Phy.to.plas¢ma. Gr. masc. n. phytos a plant; Gr. neut. n. plasma something formed or molded, a form.
Phytoplasmas (Sears and Kirkpatrick, 1994) are wall-less, nutritionally fastidious, phytopathogenic prokaryotes 0.2–0.8 µm
in diameter that morphologically resemble members of the
Mollicutes. Sequencing of nearly full-length PCR-amplified
16S rRNA genes (Gundersen et al., 1994; Namba et al., 1993;
Seemüller et al., 1994), combined with earlier studies (Kuske
and Kirkpatrick, 1992b; Lim and Sears, 1989), provided the
first comprehensive phylogeny of the organisms and showed
that they constitute a unique, monophyletic clade within the
Mollicutes. These organisms are most closely related to members of the genus Acholeplasma within the Anaeroplasma clade as
defined by Weisburg et al. (1989). Sustained culture in cell-free
media has not yet been demonstrated for any phytoplasma.
Their genome sizes have been estimated to range from 530 to
1350 kb, and the G+C content of phytoplasma DNA is about
23–30 mol%. The presence of a characteristic oligonucleotide
sequence in the 16S rRNA gene, CAA GAY BAT KAT GTK TAG
CYG GDC T, and standard codon usage indicate that phytoplasmas represent a distinct taxon for which the name “Candidatus
Phytoplasma” has been adopted by specialists in the molecular
biology and pathogenicity of these and similar phytopathogenic organisms (IRPCM Phytoplasma/Spiroplasma Working
Team – Phytoplasma Taxonomy Group, 2004). At present, the
designation “Candidatus” must still be used for new types.
Genus I. “Candidatus Phytoplasma”
Phytoplasma cells typically have a diameter less than 1 µm
and are polymorphic. Viewed in ultra-thin section by electron
microscopy Figure 115, they appear ovoid, oblong, or filamentous in plant and insect hosts (Doi et al., 1967; Hearon et al.,
1976). Transmission electron microscopy of semi-thick (0.3 µm)
sections (Thomas, 1979) and serial sections (Chen and Hiruki,
1978; Florance and Cameron, 1978; Waters and P. Hunt., 1980),
and scanning electron microscopy studies (Bertaccini et al.,
1999; Haggis and Sinha, 1978; Marcone et al., 1996) have done
much to clarify the gross cellular morphology of phytoplasmas.
They range from spherical to filamentous, often with extensive branching reminiscent of that seen in Mycoplasma mycoides.
Small dense rounded forms ~0.1 µm in diameter, formerly considered to be “elementary bodies” when seen in thin section,
were shown to represent constrictions in filamentous forms.
Dumbbell-shaped forms once thought to be “dividing cells”
are actually branch points of filamentous forms, whereas forms
thought to have internal vesicles have been shown to have involuted membranes oriented such that the plane of the sections
cut through the cell membrane twice (McCoy, 1979). Phytoplasma cell membranes are resistant to digitonin and sensitive
to hypotonic salt solutions, and, as such, are similar to those of
non-sterol requiring mollicutes (Lim et al., 1992).
Phytoplasmas are consistently observed within phloem sieve
elements (Christensen et al., 2004; McCoy et al., 1989; Oshima
et al., 2001b; Webb et al., 1999) and occasionally have been
reported in both companion cells (Rudzinska-Langwald and
Kaminska., 1999; Sears and Klomparens, 1989) and parenchyma cells (Esau et al., 1976; Siller et al., 1987) of infected
plants. Sieve elements are specialized living cells that lack nuclei
when mature and transport photosynthate from leaves not only
to growing tissues, but also to other tissues unable to photosynthesize (Oparka and Turgeon, 1999; Sjölund, 1997). This
applies particularly to roots that require considerable energy
697
for the uptake of water and nutrients (Flores et al., 1999).
Phloem sap is unique in that it contains from 12 to 30% sucrose
and is under high hydrostatic (turgor) pressure (Evert, 1977).
Sieve elements have pores in their end plates and lateral walls,
allowing passage of photosynthate to adjacent sieve tube elements. The sieve pores, which have an average diameter of ~0.2
µm, are of sufficient size to allow passage of spherical and filamentous phytoplasma cells from one sieve element to another
(McCoy, 1979). The chemical composition of sieve sap is complex, containing sugars, minerals, free amino acids, proteins,
and ATP (Van Helden et al., 1994). This rich milieu, with its
high osmotic and hydrostatic pressures, serves to support extensive multiplication of phytoplasmas in planta. Phytoplasmas also
multiply in the internal tissues and organs of their insect vectors
(Kirkpatrick et al., 1987; Lefol et al., 1994; Marzachi et al., 2004;
Nasu et al., 1970), which are primarily leafhoppers, planthoppers, and psyllids (D’Arcy and Nault, 1982; Jones, 2002; Weintraub and Beanland, 2006). In many respects, the composition
of insect hemolymph is similar to that of plant phloem sap, as
both contain high levels of complex and simple organic compounds (Moriwaki et al., 2003; Saglio and Whitcomb, 1979).
Physical maps of several phytoplasma genomes have been
constructed (Firrao et al., 1996; Lauer and Seemüller, 2000;
Marcone and Seemüller, 2001; Padovan et al., 2000). The presence of extrachromosomal DNAs or plasmids in numerous
phytoplasmas has also been reported (Davis et al., 1988; Denes
and Sinha, 1991; Kuboyama et al., 1998; Kuske and Kirkpatrick, 1990; Liefting et al., 2004; Lin et al., 2009; Nakashima and
Hayashi, 1997; Nishigawa et al., 2003; Oshima et al., 2001a; TranNguyen and Gibb, 2006) and suggested as a potential means of
intermolecular recombination (Nishigawa et al., 2002b). Phytoplasma-associated extrachromosomal DNAs have been shown
to contain genes encoding a putative geminivirus-related replication (Rep) protein (Liefting et al., 2006; Nishigawa et al.,
2001; Rekab et al., 1999) and a single-stranded DNA-binding
FIGURE 115. Electron micrographs of ultrathin sections of leaf petiole from a sunnhemp (Crotalaria juncea L.) plant
displaying Crotalaria phyllody disease symptoms. (a) Polymorphic phytoplasma cells occluding the lumen of adjacent
leaf phloem sieve tube elements. Bar = 2 µm. (b) Ultratructural morphology indicates phytoplasma cells are bounded
by a unit membrane and contain DNA fibrils and ribosomes. Bar = 200 nm. Images provided by Phil Jones.
698
protein (Nishigawa et al., 2002a), as well as a putative gene similar to DNA primase of other bacterial chromosomes (Liefting
et al., 2004) and still other genes of as yet unknown identity.
Moreover, heterogeneity in extrachromosomal DNAs has been
associated with reduced pathogenicity and loss of insect vector
transmissibility (Denes and Sinha, 1992; Nishigawa et al., 2002a,
2003).
Onion yellows mild strain (OY-M) was the first phytoplasma
genome to be completely sequenced. The genome of this
aster yellows group strain consists of a circular chromosome
of 860,632 bp. It also contains two extrachromosomal DNAs,
EcOYM (5025 bp) and plasmid pOYM (3932 bp) (Nishigawa
et al., 2003; Oshima et al., 2002), representing two different
classes based on the type of replication protein encoded. While
EcOYM contains a rep gene homologous to that of the geminiviruses, pOYM has a rep gene that encodes a unique protein
with characteristics of both viral-rep and plasmid-rep (Namba,
2002). The chromosome is a circular DNA molecule with a G+C
content of 28 mol% and contains 754 open reading frames
(ORFs), comprising 73% of the chromosome. Of these, 66% of
ORFs exhibit significant homology to gene sequences currently
archived in the GenBank database. Putative proteins encoded
by ORFs could be assigned to one of six different functional
categories: (1) information storage and processing (260 ORFs);
(2) metabolism (107 ORFs); (3) cellular processes (77 ORFs);
(4) poorly characterized, i.e., with homology to uncharacterized proteins of other organisms (50 ORFs); or (5) others, i.e.,
without homology to any known proteins (260 ORFs). Like
mycoplasmal genomes, the OY-M phytoplasma genome lacks
many genes related to amino acid and fatty acid biosynthesis,
the tricarboxylic acid cycle, and oxidative phosphorylation.
However, OY-M phytoplasma differs from mycoplasma in that it
lacks genes for the phosphotransferase system and for metabolizing UDP-galactose to glucose 1-phosphate, suggesting that
it possesses a unique sugar intake and metabolic system. Furthermore, OY-M phytoplasma lacks most of the genes needed
to synthesize nucleotides and ATP suggesting that it probably
assimilates these and other necessary metabolites from host
cytoplasm. Many genes, such as those for glycolysis, are present as multiple redundant copies representing 18% of the total
genome. Twenty-seven genes encoding transporter systems
such as malate, metal-ion and amino acid transporters, some
of which have multiple copies, were identified, suggesting that
phytoplasmas aggressively import many metabolites from the
host cell. Other than genes encoding glucanase and hemolysinlike proteins, no other genes presently known to be related to
bacterial pathogenicity were evident in the OY-M phytoplasma
genome, suggesting novel mechanisms for virulence.
Annotation of the OY-M phytoplasma genome has been
followed by three other phytoplasma genome annotations.
Aster yellows witches’-broom phytoplasma (“Candidatus Phytoplasma asteris”-related strain AY-WB) possesses a circular 706,569 nucleotide chromosome and plasmids AYWB-pI
(3872 bp), -pII (4009 bp), -pIII (5104 bp), and -pIV (4316
bp) (Bai et al., 2006). Australian tomato big bud phytoplasma
(“Candidatus Phytoplasma australiense”-related strain TBB)
has a circular 879,324 bp chromosome and a 3700 bp plasmid (Tran-Nguyen et al., 2008), whereas apple proliferation
phytoplasma (“Candidatus Phytoplasma mali”-related strain
AT) has a linear 601,943 bp chromosome (Kube et al., 2008).
The chromosome of “Candidatus Phytoplasma mali” is characterized by large terminal inverted repeats and covalently closed
hairpin ends. Analysis of protein-coding genes revealed that glycolysis, the major energy-yielding pathway supposed for OY-M
phytoplasma, is incomplete in AT phytoplasma. It also differs
from OY-M and AY-WB phytoplasmas by a lower G+C content
(21.4 mol%), fewer paralogous genes, a strongly reduced number of ABC transporters for amino acids, and an extended set
of genes for homologous recombination, excision repair, and
SOS response.
Comparative genomics have also recently identified ORFs
shared by AY-WB phytoplasma and the distantly-related corn
stunt pathogen Spiroplasma kunkelii that are absent from obligate
animal and human pathogenic mollicutes. These proteins were
identified as polynucleotide phosphorylase (PNPase), cmpbinding factor (CBF), cytosine deaminase, and Y1xR protein
and could be important for insect transmission or plant pathogenicity. Also identified were four additional proteins, ppGpp
synthetase, HAD hydrolase, AtA (AAA type ATPase), and P-type
Mg2+ transport ATPase, that seemed to be more closely related
between AY-WB and Spiroplasma kunkelii than to their mycoplasmal counterparts (Bai et al., 2004).
Phytoplasmas possess a unique genome architecture that is
characterized by multiple, nonrandomly distributed sequencevariable mosaics (SVMs) of clustered genes, originally recognized in a study of closely related “Candidatus Phytoplasma
asteris”-related strains CPh and OY-M (Jomantiene and Davis,
2006). Targeted genome sequencing and comparative genomics
indicated that this genome architecture is a common characteristic among phytoplasmas, leading to the proposal that the
origin of SVMs was an ancient event in the evolution of the phytoplasma clade (Jomantiene et al., 2007), perhaps as a result of
recurrent targeted attacks by mobile elements such as phages
(Wei et al., 2008a). Jomantiene and Davis (2006) proposed that
sizes and numbers of SVMs could account in part for the known
variation in genome size among phytoplasma strains; this concept was independently suggested by Bai et al. (2006) on the
basis of results from a comparative study of two completely
sequenced phytoplasma genomes. Nucleotide sequences within
SVMs included full length or pseudogene forms of fliA, an ATPdependent Zn protease gene, tra5, smc, himA, tmk, and ssb (encoding single-stranded DNA-binding protein), genes potentially
encoding hypothetical proteins of unknown function, genes
exhibiting similarities to transposase, and a phage-related gene
(Jomantiene et al., 2007). A similar set of nucleotide sequences
occurs in AY-WB genomic regions termed potential mobile units
(PMUs) by Bai et al. (2006). The presence of sequences encoding putatively secreted and/or transmembrane, cell surfaceinteracting proteins indicates that these genomic features are
likely to be significant for phytoplasma/host interactions (Bai
et al., 2006; Jomantiene and Davis, 2006; Jomantiene et al.,
2007).
Short (17–35 bases) conserved, imperfect palindromic DNA
sequences (PhREPs) that are present in SVMs possibly play a
role in phytoplasma genome plasticity and targeting of mobile
genetic elements. SVMs can be viewed as composites formed
by the acquisition of genes through horizontal transfer, recombination, and rearrangement, and capture of mobile elements
recurrently targeted to SVMs, leading Jomantiene et al. (2007)
to suggest that SVMs provide loci for acquisition of new genes
and targeting of mobile genetic elements to specific regions in
phytoplasma chromosomes.
The chromosomes of avirulent, mildly, moderately, and
highly virulent strains of “Candidatus Phytoplasma mali”
(Seemüller and Schneider, 2007) differ from one another in
size and exhibit distinct restriction endonuclease patterns when
cleaved with rare cutting enzymes. PCR-based DNA amplifications, primed separately by eight primer pairs, revealed
target sequence heterogeneity among all “Candidatus Phytoplasma mali”-related strains tested, but no correlations linked
molecular markers with strain virulence or the maximum titer
obtained upon infection of apple trees. In a separate study, a
comparison of mild (OY-M) and severe (OY-W) strains of onion
yellows (OY) phytoplasma indicated that severe symptoms were
associated with higher populations of OY-W in infected host
plants (Oshima et al., 2001b). A cluster of eight genes, considered essential for glycolysis, were subsequently identified within
a similar 30 kb genomic region of both strains (Oshima et al.,
2007). Of these, five genes (smtA, greA, osmC, eno, and pfkA) were
randomly duplicated in OY-W, possibly influencing glycolytic
activitiy. A higher consumption of metabolites such as sugars in
the intracellular environment of the phloem may explain differences between OY-W and OY-M in growth rate, which in turn
may be linked, directly or indirectly, to symptom severity.
Cloned fragments of phytoplasma DNA have been widely
employed as probes in dot and Southern blot hybridization
assays to identify and characterize phytoplasmas (reviewed by
Lee and Davis, 1992; Lee et al. (2000). Southern blot restriction
fragment length polymorphism (RFLP) analysis has enabled
investigations of genetic relationships among phytoplasmas
associated with similar hosts or with symptomatologically similar
diseases (Kison et al., 1997, 1994; Kuske et al., 1991; Schneider
and Seemüller, 1994b). Several discrete phytoplasma groups,
each comprising strains that shared extensive sequence homology and little or no apparent homology with other phytoplasmas, were identified by this type of analysis. Lee and co-workers
(1992) coined the term “genomic strain cluster” to denote
each of seven discrete genotypic groups resolved by employing
a selection of phytoplasma genomic DNA probes (reviewed by
Lee and Davis, 1992). Of these, aster yellows (AY) was the largest
group, represented by 15 genetically variable strains that were
further delineated into three distinct genomic types (types I, II,
and III) or subclusters (Lee et al., 1992). Significantly, major
groupings later revealed by RFLP analysis of 16S rRNA genes
were consistent with those defined by monoclonal antibody
typing (Lee et al., 1993a) and other molecular methods (Lee
et al., 1998b), but differed from distinctions made in traditional
classification based solely on biological properties such as plant
host range, symptomatology, and insect vector specificity
(Chiykowski and Sinha, 1990).
Polyclonal antibodies (PAbs) have been produced against
phytoplasma-enriched extracts (intact organisms or membrane
fractions) partially purified from plants (reviewed by Chen
et al., 1989) and against vector leafhopper-derived immunogens
(Errampelli and Fletcher, 1993; Kirkpatrick et al., 1987). Most
PAbs exhibit relatively high background reactions with healthy
host antigens; thus, generation of useful polyclonal antisera has
been limited so far to a few phytoplasmas maintained at high
titer in host tissues. Phytoplasmas can be differentiated on the
basis of their antigenic properties through the use of PAbs in
699
enzyme-linked immunosorbent (ELISA), immunofluorescence,
or Western blot assays. Antigenic similarity revealed among phytoplasmas by these assays is often in agreement with relationships
demonstrated by vector transmission studies. Detection of antigenically distinct phytoplasmas in plants exhibiting very similar
disease symptoms attests to the unreliability of symptom expression alone as a means of differentiating phytoplasmas.
Improvements in phytoplasma extraction methods have
provided a source of immunogens for monoclonal antibody
(MAb) production (Chang et al., 1995; Hsu et al., 1990; Jiang
et al., 1989; Loi et al., 2002, 1998; Shen and Lin, 1993; Tanne
et al., 2001). Used in ELISA, dot or tissue blot immunoassay,
immunocapture PCR (Rajan et al., 1995), immunofluorescence
microscopy, or immunosorbent electron microscopy (ISEM)
(Clark, 1992; Shen and Lin, 1994), MAbs have demonstrated
considerable promise for detection and differentiation of phytoplasmas infecting a broad range of host plants, including
woody perennials (Guo et al., 1998). Due to their high degree
of specificity, monoclonals seem most suited for differentiating
very closely related strains (Lee et al., 1993a).
Isolation, cloning, and expression of immunodominant protein genes have identified putative proteins that account for a
major portion of the membrane proteins of several phytoplasmas (Arashida et al., 2008; Barbara et al., 2002; Berg et al., 1999;
Blomquist et al., 2001; Galetto et al., 2008; Kakizawa et al., 2004,
2009; Morton et al., 2003; Suzuki et al., 2006; Yu et al., 1998).
When these purified proteins were used as immunogens, the
resulting polyclonal antisera exhibited high specific titers and low
background reactions in ELISA and Western blot analyses that
were designed to detect phytoplasma proteins in infected hosts.
Similarly, the secA gene was cloned from an onion yellows (OY-M)
strain of aster yellows phytoplasma (Kakizawa et al., 2001) and
used to raise an anti-SecA rabbit antibody against a purified partial SecA protein expressed in Escherichia coli. Light microscopy
of thin sections of garland chrysanthemum (Chrysanthemum coronarium) treated by immunohistochemical straining revealed that
the SecA protein was present in phloem of OY-M-infected but not
healthy host plants. In addition, antisera against both OY-M phytoplasma SecA protein and GyrA protein of Acholeplasma laidlawii
reacted with proteins of several unrelated phytoplasmas extracted
from plant tissues (Koui et al., 2002; Wei et al., 2004a).
Phytoplasmas are the apparent etiological agents of diseases
of at least 1000 plant species worldwide (McCoy et al., 1989;
Seemüller et al., 1998). Although they can be transmitted from
infected to healthy plants by scion or root grafts, most plant
to plant spread occurs naturally via phloem-feeding insect vectors primarily of the family Cicadellidae (leafhoppers) and, less
commonly, by planthoppers (Fulgoroidea) of the family Ciixidae
and psyllids (Psylloidea) (D’Arcy and Nault, 1982; Tsai, 1979;
Weintraub and Beanland, 2006). Phytoplasmas are transmitted in a circulative-propagative manner that typically involves
a transmission latent period from 2 to 8 weeks (Carraro et al.,
2001; Webb et al., 1999). The insect vector becomes infected
upon ingesting phytoplasmas in phloem sap of infected plants.
After an incubation period of one to several weeks, the phytoplasma multiplies to high titer in the salivary glands and the
insect becomes capable of infecting the phloem of the healthy
plants on which it feeds (Kunkel, 1926; Lee et al., 1998a; Nasu
et al., 1970). Generally, phytoplasma infection does not appear
to significantly affect the activity, weight, longevity, or fecundity
700
of vector insects (Garnier et al., 2001). Some phytoplasmas can
be vectored by many species of leafhoppers (McCoy et al., 1989;
Nielson, 1979) and different insect species may serve as vectors
in different geographic regions. Several vectors also have the
ability to transmit more than one type of phytoplasma, whereas
other phytoplasmas are transmitted by one or a few vector species to a narrow range of plant species (Lee et al., 1998a). There
is mounting evidence also for transovarial transmission of some
phytoplasmas (Alma et al., 1997; Hanboonsong et al., 2002;
Kawakita et al., 2000; Tedeschi et al., 2006).
Plants may serve as both natural and experimental hosts to
several different phytoplasmas. Dual or mixed infections involving related or unrelated phytoplasmas are known to occur
naturally in plants and appear to be more common in perennial than annual plants (Bianco et al., 1993; Lee et al., 1995).
Also, closely related phytoplasma strains are capable of inducing dissimilar symptoms on the same plant species (White et al.,
1998), whereas similar symptoms on the same host plant may be
induced by unrelated phytoplasmas (Harrison et al., 2003). The
ability to accurately identify phytoplasmas by using DNA-based
methods has shown that these organisms are more genetically
diverse than was once thought (Davis and Sinclair, 1998). The
geographic occurrence of phytoplasmas is determined largely
by geographic ranges and feeding behavior (mono-, oligo-, or
polyphagous) of the vector species, the relative susceptibility of
the preferred host plant species, and the native host ranges of
plant and insect hosts (Lee et al., 1998a). Phytoplasmas can be
introduced into new geographic regions by long-distance dispersal of infectious vectors (Lee et al., 2003) and by movement
of infected plants or vegetative plant parts. Most recently, phytoplasma DNA has been detected in embryos of aborted seed
from diseased plants (Cordova et al., 2003; Nipah et al., 2007)
and seed transmission of phytoplasmas infecting alfalfa (Medicago sativa L.) has been demonstrated (Khan et al., 2002).
An array of characteristic symptoms is associated with phytoplasma infection of several hundred plant species worldwide. Symptoms vary according to the particular host species,
stage of host infection and the associated phytoplasma strain
(reviewed by Davis and Lee, 1992; Hogenhout et al., 2008;
Kirkpatrick, 1989, 1992; Lee et al., 2000; McCoy et al., 1989;
Seemüller et al., 2002; Sinclair et al., 1994). Some symptoms
indicative of profound disturbances in the normal balance of
growth regulators in plants include virescence (greening of petals), phyllody (conversion of floral organs into leafy structures),
big bud, floral proliferation, sterility of flowers, proliferation
of adventitious or axillary shoots, internode elongation and
etiolation, generalized stunting (small flowers, leaves and fruits
or shortened internodes), unseasonal discoloration of leaves or
shoots (yellow to purple discoloration), leaf curling, cupping
or crinkling, witches’-brooms (bunchy growth at stem apices),
vein clearing, vein enlargement, phloem discoloration, and general plant decline such as die-back of twigs, branches and trunks
(Lee and Davis, 1992; Lee et al., 2000; McCoy et al., 1989).
Infection of herbaceous host plants is followed by rapid
intraphloemic spread of phytoplasma from leaves to roots, often
accompanied by six-fold increases in phytoplasma populations
in these tissues between 14 and 28 d post-inoculation (Kuske
and Kirkpatrick, 1992a; Wei et al., 2004b). Phytoplasma concentrations ranging from 2.2 × 108 to 1.5 × 109 cells per gram of
tissue have been measured in high titer herbaceous hosts such
as periwinkle (Catharanthus roseus) and in certain woody perennial hosts such as alder (Alnus) and most poplar (Populus) species. Lowest phytoplasma concentrations, from 370 to 34,000
cells per gram of tissue, were detected in apple trees that were
grafted on resistant rootstocks and in oak (Quercus robur) or
hornbeam (Carpinus betulus) trees exhibiting nonspecific leaf
yellowing symptoms (Berg and Seemüller, 1999).
Colonization is usually marked by phloem dysfunction and
a reduction in photosynthetic capacity. Alterations in phloem
function have been correlated with structural degeneration of
sieve elements due possibly to physical blockage by colonizing
phytoplasma or the action of a phytotoxin (Guthrie et al., 2001;
Siddique et al., 1998). The onset of symptoms may be accompanied by substantial impairment of the photosynthetic rate of
mature leaves and by fluctuations in carbohydrate and amino
acid levels in source versus sink leaves (Lepka et al., 1999; Tan
and Whitlow, 2001). Leaf yellowing is associated with: decreases
in chlorophyll content, carotenoids, and soluble proteins (Bertamini and Nedunchezhian, 2001); abnormal stomatal function (Martinez et al., 2000); histopathological changes such the
amount of total polyphenols; loss of cellular integrity (Musetti
et al., 2000); fluctuations in hydrogen peroxide; peroxidase
activity and glutathione content in diseased versus healthy plant
tissues (Musetti et al., 2004); and increases in calcium (Ca2+)
ions in cells (Musetti and Favali, 2003; Rudzinska-Langwald
and Kaminska, 2003). Such adverse changes are accompanied
by differential regulation of genes encoding proteins involved
in floral development (Pracros et al., 2006), photosynthesis,
sugar transport, and response to stress or in pathways of lipid
and phenylpropanoid or phytosterol synthesis (Albertazzi
et al., 2009; Carginale et al., 2004; Hren et al., 2009; JagoueixEveillard et al., 2001).
The organisms degenerate and lose their cellular contents
following treatment of infected plants with tetracycline antibiotics (Kamińska and Śliwa, 2003; Sinha and Peterson, 1972).
Tetracycline sensitivity and the lack of sensitivity to cell wallinhibiting antibiotics such as penicillin (Davis and Whitcomb,
1970; Ishii et al., 1967) also support their inclusion in the Mollicutes. Protective or therapeutic treatments with tetracycline
antibiotics for phytoplasma disease control have been extended
to a few high-value crop plants such as coconut for control of
palm lethal yellowing, and to cherries and peaches for control
of X-disease (McCoy, 1982; Nyland, 1971; Raju and Nyland,
1988). Administered by trunk injection, treatment of each tree
with 1.0 g (protective dose) or 3.0 g (therapeutic dose) three
times per year was sufficient for control of coconut lethal yellowing disease (McCoy, 1982).
Isolation of phytoplasma-enriched fractions from plant and
insect host tissues is possible by differential centrifugation and
filtration after first disrupting tissues in osmotically-augmented
buffers (Kirkpatrick et al., 1995; Lee et al., 1988; Sinha, 1979;
Thomas and Balasundaran, 2001). Further purification of phytoplasmas is possible by centrifugation of enriched preparations
in discontinuous Percoll density gradients (Davis et al., 1988;
Gomez et al., 1996; Jiang and Chen, 1987) or by affinity chromatography using phytoplasma-specific antibodies coupled to
Protein A-Sepharose columns (Jiang et al., 1988; Seddas et al.,
1995). Viability of these enriched preparations may be assessed
by infectivity tests in which aliquots of the phytoplasma preparations are micro-injected into vector insects, which are then
fed on healthy indicator plants (Nasu et al., 1974; Sinha, 1979;
Whitcomb et al., 1966a, b). Separation of enriched phytoplasma
DNA from mixtures with host DNA is also possible by use of
cesium chloride-bisbenzimide buoyant density gradient centrifugation (Kollar and Seemüller, 1989). Present as an uppermost
band in final gradients, phytoplasma DNA fractionated by this
means was suitable for endonuclease digestion and cloning for
DNA probe development (Harrison et al., 1992, 1991; Kollar
and Seemüller, 1990).
Viable phytoplasmas have been maintained for at least 6 years
in intact vector insects frozen at −70°C (Chiykowski, 1983).
Viable X-disease phytoplasmas have been maintained for 2
weeks in salivary glands suspended in a tissue culture medium
(Nasu et al., 1974). Extracts of phytoplasma-infected insects
prepared in a MgCl2/glycine buffer, osmotically adjusted to
800 milliosmoles/kg with sucrose, retained their infectivity for
up to 3 d (Smith et al., 1981). Phytoplasma strains have been
routinely maintained in diseased plants kept in an insect-proof
greenhouse or in plantlets grown in tissue culture (Bertaccini
et al., 1992; Davis and Lee, 1992; Jarausch et al., 1996; Sears
and Klomparens, 1989; Wongkaew and Fletcher, 2004). While
plant to plant transmission is accomplished naturally by vector
insects and, in some cases, through grafts, experimental transmissions commonly include the use of plant parasitic dodders
(Cuscuta sp.) (Marcone et al., 1999a). Although phytoplasma
strains are commonly maintained in plants by periodic graft
inoculation, maintenance of phytoplasmas exclusively in plants
can result in strain attenuation over time and an associated loss
of transmissibility by vector insects (Chiykowski, 1988; Denes
and Sinha, 1992).
Differentiation of the genus “Candidatus Phytoplasma”
from other genera
Phytoplasma-specific nucleic acid probes and PCR technology have largely supplanted traditional methods of electron
microscopy and biological criteria for sensitive detection, identification, and genetic characterization of phytoplasmas. Molecular-based analyses have shown phytoplasma genomes to be
A+T rich (Kollar and Seemuller, 1989; Oshima et al., 2004) and
to range from 530 to 1350 kbp in size (Marcone et al., 1999b,
2001; Neimark and Kirkpatrick, 1993). Before any phytoplasma
genomes were sequenced, phytoplasmas were shown to contain two rRNA operons (Davis, 2003a; Harrison et al., 2002; Ho
et al., 2001; Jomantiene et al., 2002; Jung et al., 2003a; Lee et al.,
1998b; Liefting et al., 1996; Marcone et al., 2000; Schneider
and Seemüller, 1994a). Other genes that have been identified
include ribosomal protein genes (Gundersen et al., 1994; Lee
et al., 1998b; Lim and Sears, 1992; Martini et al., 2007; Miyata
et al., 2002a; Toth et al., 1994) of the S10-spc operon (Miyata
et al., 2002a), a nitroreductase gene (Jarausch et al., 1994),
DNA gyrase genes (Chuang and Lin, 2000), genes encoding
elongation factors G and Tu (An et al., 2006; Berg and Seemüller, 1999; Koui et al., 2003; Marcone et al., 2000; Miyata et al.,
2002b; Schneider et al., 1997), secA, secY, and secE genes of a
functional Sec protein translocation system (Kakizawa et al.,
2001, 2004), gidA, potB, potC, and potD (Mounsey et al., 2006),
701
a gene encoding an RNase P ribozyme (Wagner et al., 2001),
recA (Chu et al., 2006), rpoC (Lin et al. 2006), polC (Chi and
Lin., 2005), and insertion sequence (IS)-like elements (Lee
et al., 2005). Numerous other putative genes or pseudogenes
have been identified recently after partially or fully sequencing
random fragments of genomic DNA cloned from phytoplasmas
by various methods (Bai et al., 2004; Cimerman et al., 2006,
2009; Davis et al., 2003b, 2005; Garcia-Chapa et al., 2004; Liefting and Kirkpatrick, 2003; Melamed et al., 2003; Miyata et al.,
2003; Streten and Gibb, 2003).
Development of phytoplasma-specific rRNA gene primers
has permitted PCR-mediated amplification of various regions
of the rRNA operons (Ahrens and Seemüller, 1992; Baric and
Dalla-Via, 2004; Davis and Lee, 1993; Deng and Hiruki, 1991;
Gundersen and Lee, 1996; Lee et al., 1993b) (Namba et al.,
1993; Smart et al., 1996). RFLP analysis of PCR-amplified rDNA
provided a practical solution to the problem of phytoplasma
identification and classification (Lee et al., 2000, 1998b). Pairwise comparisons of disparate strains were marked by considerable differences in RFLP patterns, whereas strains that were
considered closely related on the basis of similar biological
properties were often, although not always, indistinguishable on
the basis of RFLP patterns. Alternatively, heteroduplex mobility
analysis has demonstrated greater sensitivity than RFLP analysis for detecting minor variability in 16S rRNA genes of closely
related phytoplasma strains (Cousin et al., 1998; Wang and
Hiruki, 2000), since RFLP analysis is limited to detection of recognition sites for restriction endonucleases. Cluster analysis of
rDNA RFLP patterns provided the first means to differentiate
between known and unknown phytoplasmas from a wide range
of plant hosts and geographic locations, and to resolve phytoplasmas into well-defined phylogenetic groups and subgroups
(Ahrens and Seemüller, 1992; Lee et al., 1993b; Schneider
et al., 1993, 1995).
Taxonomic comments
The inability to cultivate phytoplasmas outside of their plant
and insect hosts has thus far rendered traditional methods
impractical as aids for taxonomy of these organisms. Unlike
their culturable Mollicute relatives, which were originally classified based only upon biological and phenotypic properties
in pure culture, phytoplasmas cannot be classified by these
criteria. Through application of DNA-based methods, it is now
possible to accurately identify and characterize phytoplasmas
and to assess their genetic interrelationships. These capabilities
have assisted development of classification systems, first based
on hybridization data, later based on 16S rDNA RFLPs, and ultimately on phylogenetic analysis of 16S rRNA genes and other
conserved gene sequences. Classification schemes founded
upon these molecular criteria have been refined and expanded
upon over time, with the goal of defining a taxonomy for these
unique organisms. In a phytoplasma classification scheme proposed by Lee et al. (1993b), based on analyses of rDNA RFLPs,
a total of nine primary 16S rDNA groups (termed 16Sr groups)
and 14 subgroups were initially recognized. Phytoplasma groups
delineated by these analyses were consistent with genomic
strain clusters previously identified by DNA hybridization analysis (Lee et al., 1992), although a greater diversity among strains
comprising group 16SrI (aster yellows and related strains) was
indicated by the earlier hybridization data. Subgroups within a
702
given 16Sr group were distinguished by the presence of one or
more restriction sites in a phytoplasma strain that differed from
those in all existing members of a given subgroup. For those
strains in which intra-rRNA operon heterogeneity was detected,
subgroup designations were assigned according to the combined patterns of both 16S rRNA genes.
RFLP analysis of more variable ribosomal protein genes
(Gundersen et al., 1994; Lee et al., 2004a, b) or tuf genes
(Marcone et al., 2000; Schneider et al., 1997) has provided a
means for more detailed subdivision of phytoplasma primary
groups delineated by 16S rDNA RFLP data. This strategy for
finer subgroup differentiation has been used to modify and
expand upon earlier classifications and to incorporate many
newly identified phytoplasma strains. Based on RFLP analysis of nearly full-length 16S rRNAs, at least 15 primary 16Sr
groups have been recognized (Lee et al., 1998b; Montano et al.,
2001): 16SrI, Aster yellows; 16SrII, Peanut witches’-broom;
16SrIII, X-disease; 16SrIV, Coconut lethal yellows; 16SrV, Elm
yellows; 16SrVI, Clover proliferation; 16SrVII, Ash yellows;
16SrVIII, Loofah witches’-broom; 16SrIX, Pigeonpea witches’broom; 16SrX, Apple proliferation; 16SrXI, Rice yellow dwarf;
16SrXII, Stolbur; 16SrXIII, Mexican periwinkle virescence;
16SrXIV, Bermuda grass white leaf; and 16SrXV Hibiscus witches’broom. A total of 45 subgroups were identified when ribosomal
protein gene RFLP data was also considered in the analyses.
Sequencing of 30 nearly full-length amplified 16S rRNA
genes was undertaken by Namba et al. (1993), Gundersen
et al. (1994), and Seemüller et al. (1994) from a diversity of
strains previously characterized by rDNA RFLP analysis. These
collective efforts, combined with earlier studies (Kuske and
Kirkpatrick, 1992b; Lim and Sears, 1989), provided the first
comprehensive phytoplasma phylogeny. In recognition of their
unique phylogenetic status, the trivial name “phytoplasma” was
initially proposed (Sears and Kirkpatrick, 1994) and has since
been adopted formally (IRPCM Phytoplasma/Spiroplasma
Working Team – Phytoplasma Taxonomy Group, 2004) to collectively name these fastidious, phytopathogenic mollicutes
previously known as mycoplasma-like organisms. Within the
phytoplasma clade, major subclades (primary groups representing “Candidatus” species) include: (1) Stolbur; (2) Aster
yellows; (3) Apple proliferation; (4) Coconut lethal yellowing;
(5) Pigeonpea witches’-broom; (6) X-disease; (7) Rice yellow
dwarf; (8) Elm yellows; (9) Ash yellows; (10) Sunnhemp witches’-broom; (11) Loofah witches’-broom; (12) Clover proliferation; and (13) Peanut witches’-broom (Kirkpatrick et al., 1995;
Schneider et al., 1995; White et al., 1998). Primary phytoplasma
groups including 19 novel groups, namely Australian grapevine
yellows (AUSGY), Italian bindweed stolbur (IBS), Buckthorn
witches’-broom (BWB), Spartium witches’-broom (SpaWB),
Galactia little leaf (GaLL), Vigna little leaf (ViLL), Clover yellow
edge (CYE), Hibiscus witches’-broom (HibWB), Pear decline
(PD), European stone fruit yellows (ESFY), Japanese hydrangea phyllody (JHP), Psammotettix cephalotes-borne (BVK), Italian
alfalfa witches’-broom (IAWB), Cirsium phyllody (CirP),
Bermuda grass white leaf (BGWL), Sugarcane white leaf
(SCWL), Tanzanian lethal decline (TLD), Stylosanthes little leaf
(StLL), and Pinus sylvestris yellows (PinP), that were absent from
previous classification schemes have been subsequently defined.
These new taxonomic entities were delineated on the basis of
phylogenetic tree branching patterns, differences in 16S rRNA
gene sequence similarities that were 1.2–2.3% or greater and,
in some instances, by additional considerations such as plant
host and vector specificity, primer specificity, and RFLP comparisons of ribosomal and nonribosomal DNA, as well as serological comparisons (Seemüller et al., 2002, 1998).
Most recently, Wei et al. (2007) applied computer-simulated
RFLP analysis for classification of phytoplasma strains. Through
comparisons of virtual RFLP patterns of 16S rRNA genes and
calculations of coefficients of RFLP similarity, the authors
classified all available 16S rRNA gene sequences, including sequences from 250 previously unclassified phytoplasma
strains, into a total of 28 16Sr RFLP groups. These included ten
new groups and dozens of new subgroup lineages (Cai et al.,
2008; Wei et al., 2008b). Each new group represents a potential
“Candidatus Phytoplasma” species level taxon. This information was used to augment the 16Sr RFLP classification system
(Lee et al., 2000, 1998b, 1993b) with the following additional
groups: 16SrXVI, Sugarcane yellow leaf syndrome; 16SrXVII,
Papaya bunchy top group; 16SrXVIII, American potato purple
top wilt group; 16SrXIX, Japanese chestnut witches’-broom
group; 16SrXX, Buckthorn witches’-broom group; 16SrXXI,
Pine shoot proliferation group; 16SrXXI, Nigerian coconut
lethal decline (LDN) group; 16SrXXIII, Buckland valley grapevine yellows group; 16SrXXIV, Sorghum bunchy shoot group;
16SrXXV, Weeping tea witches’-broom group; 16SrXXVI, Mauritius sugarcane yellow D3T1 group; 16SrXXVII, Mauritius sugarcane yellow D3T2 group; and 16SrXXVIII, Havana derbid
phytoplasma group. The virtual RFLP patterns are available
for online use as reference patterns at http://www.ba.ars.usda.
gov/data/mppl/virtualgel.html.
The spacer region (SR) separating the 16S from the 23S
rRNA gene of phytoplasmas was also shown to be a reliable
phylogenetic marker. Phylogenetic trees derived from the
entire 16S–23S SR (Gibb et al., 1998; Kenyon et al., 1998) or
variable regions flanking the tRNAile gene (Kirkpatrick et al.,
1995; Schneider et al., 1995) differentiated phytoplasmas into
groups that were concordant with the major groups established
previously from analyses of 16S rRNA genes. Phytoplasmas collectively differ in their 16S rRNA gene sequence by no more
than 14%, whereas their respective 16S–23S SR sequences differ by as much as 22%. This added variation has contributed
to improved accuracy of phytoplasma classification at the subgroup level. Similarly, phylogenetic analysis of ribosomal protein genes, secY, secA, or 23S rRNA genes has been employed
to differentiate closely related phytoplasma strains, as well as to
aid the group and subgroup classification of diverse phytoplasmas (Daire, 1993; Hodgetts et al., 2008; Lee et al., 1998b, 2004a,
2006b; Martini et al., 2007; Reinert, 1999). Such studies have
led to finer differentiation among phytoplasma subgroups and
to enriched descriptions of “Candidatus Phytoplasma” species
(Lee et al., 2004a, 2006a, b).
A polyphasic system for taxonomy based on integration of
genotypic, phenotypic, and phylogenetic information employed
for bacterial classification (Murray et al., 1990; Stackebrandt
and Goebel, 1994) has proved problematic for nonculturable
phytoplasmas. In response to a rapidly growing database of
phylogenetic markers, even in the absence of species-defining
biological or phenotypic characters, the Working Team on Phytoplasmas of the International Research Programme of Comparative Mycoplasmology (IRPCM Phytoplasma/Spiroplasma
Working Team – Phytoplasma Taxonomy Group, 2004) proposed that taxonomy of phytoplasmas be based primarily upon
phylogenetic analyses. This proposal was agreed to and adopted
as policy by the ICSB Subcommittee on the Taxonomy of
Mollicutes (1993, 1997), which also recommended that the provisional taxonomic status of “Candidatus”, originally proposed
by Murray and Schleifer (1994), be used for assigning genera
names as follows: “Candidatus Phytoplasma” (from phytos, Greek
for plant; plasma, Greek for thing molded) [(Mollicutes) NC;
NA; O; NAS (GenBank no. M30790); oligonucleotide sequence
of unique region of the 16S rRNA gene is CAA GAY BAT KAT
GTK TAG CYG GDC T; P (Plant, phloem; Insect, salivary gland);
M]. (IRPCM Phytoplasma/Spiroplasma Working Team –
Phytoplasma Taxonomy Group, 2004). By this same approach,
major groups within the genus also delineated by phylogenetic
analysis of near full-length 16S rRNA gene sequences were considered to represent one or more distinct species.
Current guidelines for “Candidatus Phytoplasma” species
descriptions (Anonymous, 2000; Firrao et al., 2005; IRPCM
Phytoplasma/Spiroplasma Working Team – Phytoplasma Taxonomy Group, 2004) are based upon identification of a significantly unique 16S rRNA gene sequence >1200 bp in length.
The strain from which the sequence is obtained should be
designated as the reference strain. Strains with minimal differences in the 16S rRNA sequence, relative to the reference strain,
should be referred to as related strains. In general, a strain can
be described as a new “Candidatus Phytoplasma” species if its
16S rRNA gene sequence has less than 97% identity to any previously described “Candidatus Phytoplasma” species (ICSB Subcommittee on the Taxonomy of Mollicutes, 2001). There are
cases in which phytoplasmas may share more than 97% of their
16S rRNA gene sequence, but clearly represent ecologically
distinct populations and, thus, they may warrant description as
703
separate species. In such cases, the description of two different
species is recommended when all of the following conditions
apply: (1) the two phytoplasmas are transmitted by different
vector species; (2) the two phytoplasmas have a different natural plant host, or at least their symptomatology is significantly
different in the same plant host; (3) there is evidence of significant molecular diversity between phytoplasmas as determined
by DNA hybridization assays with cloned nonribosomal DNA
markers, serological reactions, or by PCR-based assays. The
taxonomic rank of subspecies should not be used. Reference
strains should be available to the scientific community in graftinoculated or in vitro micropropagated host plants or as DNA
if perpetuation of strains in infected host plants is not feasible.
Descriptions of “Candidatus Phytoplasma” species should be
preferably submitted to the International Journal of Systematic
and Evolutionary Microbiology (http://ijs.sgmjournals.org/).
Recent phylogenetic investigations, including the present analyses (Figure 116), suggest 97.5% 16S rRNA gene
sequence similarity may represent a more suitable upper
threshold for “Candidatus Phytoplasma” species separation,
in that taxonomic subgroups designated based on 16S rRNA
gene sequence similarities of £97.5% more consistently define
species that are phylogenetically distinct from nearest related
species. Regardless of homology criteria, a taxonomy is emerging for the phytoplasmas in the absence of cultivability where
species and related strains of a species are clearly recognized
with due consideration of the genetic, ecological, and environmental constraints unique to this group of plant- and insectassociated Mollicutes. To a large extent, the present taxonomy
employs vernacular names based on associated diseases, but
is constantly shifting towards a traditional taxonomy as more
and more “Candidatus Phytoplasma” species continue to be
recognized and proposed.
List of species of the genus “Candidatus Phytoplasma”
In accordance with the current guidelines for “Candidatus
Phytoplasma” species descriptions, the following species have
been designated. Proposed assignments to the class Mollicutes
are based on nucleic acid sequences. None of these species have
been cultivated independently of their host, and their metabolism and growth temperatures are unknown.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-GTTTCTTCGGAAA-3¢
(68–80), 5¢-GTTAGAAATGACT-3¢ (142–153), 5¢-GCTGGTGGCTT-3¢ (1438–1448).
Habitat, association, or host: Solanum tuberosum phloem.
1. “Candidatus Phytoplasma allocasuarinae” Marcone, Gibb,
Streten and Schneider 2004a, 1028
Vernacular epithet: Allocasuarina yellows phytoplasma,
strain AlloYR.
Gram reaction: not applicable.
Morphology: other.
Unique region of 16S rRNA gene: 5¢-TTTATTCGAGAGGGCG-3¢.
Habitat, association, or host: phloem of Allocasuarina muelleriana (Slaty she-oak).
3. “Candidatus Phytoplasma asteris” Lee, Gundersen-Rindal,
Davis, Bottner, Marcone and Seemüller 2004a, 1046
Vernacular epithet: Aster yellows (AY) phytoplasma, strain
OAY R.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-GGGAGGA-3¢,
5¢-CTGACGGTACC-3¢, and 5¢-CACAGTGGAGGTTATCAGTTG-3¢.
Habitat, association, or host: phloem of Oenothera hookeri
(Evening primrose).
2. “Candidatus Phytoplasma americanum” Lee, Bottner, Secor
and Rivera-Varas 2006a, 1596
Vernacular epithet: Potato purple top, strain APPTW12NER.
4. “Candidatus Phytoplasma aurantifolia” Zreik, Carle, Bové
and Garnier 1995, 452
Vernacular epithet: Witches’-broom disease of lime phytoplasma, strain WBDLR.
704
P.trifoli (CP)
77
P. fraxini (AshY1)
P. ulmi (EY1)
79
100
100
P. ziziphi (JWB-G1)
LfWB
StLL
93
56
LDG
LDT
LY-c2
SBS
P. castaneae (CnWB)
72
88
b
P. pini (PinP)
CIRP
GaLL
98
BVK
100
P. oryzae (RYD)
SCWL
P. cynodontis (BGWL)
P. phoenecium (AlmWB-A4)
98
100
ViLL
WX
GLL-eth
59
P. brasiliense (HibWB)
IAWB
SPLL
100
P. australasia (PpYC)
P. aurantifolia (WBDL)
91
WTWB
P. mali (AP15)
P. prunorum (ESFY-G1)
a
P. pyri (PD1)
100
P. spartii (SPAR)
P. allocasuarinae (AlloY)
P. rhamni (BWB)
STOL
P. australiense (AusGY)
100
P. japonicum (JHP)
IBS
100
P. asteris (MIAY)
MPV
A. palmae
A. laidlawii
10 changes
FIGURE 116. Phylogenetic analysis of the phytoplasmas. Phylogenetic trees were constructed by parsimony analy-
ses of phytoplasma 16S rRNA gene sequences using the computer program PAUP (Swofford, 1998). The closely
related culturable Acholeplasma palmae was employed as the outgroup. Because phytoplasma taxa are too numerous
to present in a single inclusive tree, a global phylogeny of representative phytoplasmas is first presented. The global
tree is divided into lower (a) and upper (b) regions. Each region of the global phylogeny is then expanded into
inclusive trees, a and b, which collectively include 145 phytoplasmas from diverse geographic origins. Taxonomic
subgroups, representing phytoplasmas sharing at least 97.5% 16S rRNA gene sequence similarity, are identified on
each inclusive tree. Each phylogenetically distinct subgroup is equivalent to a subclade (or putative species) within
the genus “Candidatus Phytoplasma”. In all trees, branch lengths are proportional to the number of inferred character state transformations. Bootstrap (confidence) values greater than or equal to 50 are shown on the branches.
Phytoplasmas for which 16S rRNA gene sequences of at least 1200 bp in length have been determined (312 total)
are listed by subgroup in Table 144 along with their sequence accession numbers.
705
a
84
100
100
52
100
87
SUNHP
SPWB
PnWB
AlWB
GPh
P. australasia (PpYC)
CoAWB
IAWB
PEP
SPLL
GLL-eth
P. brasiliense (HibWB)
CaM
CaWB1
FBP
P. aurantifolia (WBDL)
BoLL
WTWB
P. mali (AP15)
AP1/93
AP2
AT
P. pyri (PD1)
PD
100
PPER
P. prunorum (ESFY-G1)
82
ESFY-142
ESFY4
100
ESFY5
P. spartii (Spar)
P. allocasuarinae (AlloY)
100
P. rhamni (BWB)
P. asteris (MIAY)
PPT
MBS
PaWB
MD
Bstv2M.f12
HyPH
APWB
IOWB
PRIVC
RPh
ApSL
AAY
SAY
ValY
WcWB
OY-M
CabD3
AY-WB
100
BB
HYDP
CPh
STRAWB2
BBS
ACLR
CWL
SY
PY
100 VK
STOL
STOL2
80
PpDB
PYL
65
SLY
SV3101
95
P. australiense (AusGY)
P. japonicum (JHP)
51
IBS
MPV
100
STRAWB1
CbY1
A. palmae
P. australasia
IAWB
SPLL
GLL-eth
P. brasiliense
P. aurantifolia
BoLL
WTWB
P. mali
P. pyri
P. prunorum
P. spartii
P. allocasuarinae
P. rhamni
P. asteris
STOL
P. australiense
P. japonicum
IBS
MPV
5 changes
FIGURE 116. (Continued)
Morphology: other.
Unique region of 16S rRNA gene: 5¢-GCAAGTGGTGAACCATTTGTTT-3¢.
Habitat, association, or host: phloem of Citrus; hemolymph
and salivary glands of Hishimonus phycitis (Cicadellidae).
5. “Candidatus Phytoplasma australasia” White, Blackall, Scott
and Walsh 1998, 949
Vernacular epithet: Papaya yellow crinkle phytoplasma,
strain PpYCR.
Morphology: other.
Sequence accession no. (16S rRNA gene): Y10097.
Unique regions of 16S rRNA gene: 5¢-TAAAAGGCATCTTTTATC-3¢ and 5¢-CAAGGAAGAAAAGCAAATGGCGAACCATTTGTTT-3¢.
Habitat, association, or host: phloem of Carica papaya and
Lycopersicon esculentum.
6. “Candidatus Phytoplasma australiense” Davis, Dally, Gundersen, Lee and Habili 1997, 268
Vernacular epithet: Australian grapevine yellows phytoplasma, strain AUSGYR.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-CGGTAGAAATAT
CGT-3¢ and 5¢-TTTATCTTTAAAAGACCTCGCAAGA-3¢.
Habitat, association, or host: Vitis phloem.
7. “Candidatus Phytoplasma brasiliense” Montano, Davis,
Dally, Hogenhout, Pimentel and Brioso 2001, 1117
Vernacular epithet: Hibiscus witches’-broom (HibWB)
phytoplasma, strain HibWB26R.
706
b
56
51
97
88
100
100
P. trifolii (CP)
PWB
BLL-bd
BLL
100
CSV
BLTVA
88
VR
ArAWB
EriWB
100
P. fraxini (AshY1)
ASHY
LWB3
AshY3
ALY
86
FD
HD1
SpaWB229
VC
RuS
P. ulmi (EY1)
ULW
100
100
JWB-ch
P. ziziphi (JWB-G1)
CLY-5
NecY-In1
JWB-Ka
LfWB
100
LfWB-t
StLL
CY
LY-c2
PanD
ScY
LDY
81
LfY1
LDG
72
LDN
LDT
SBS
SGS-v1
SCWL
BVK
100
CIRP
GaLL
P. oryzae (RYD-J)
RYD-Th
BGWL-2
CWL
P. cynodontis (BGWL-C1)
BGWL
P. casteneae (CnWB)
P. pini (Pin127)
PinG
KAP
PPWB-f
P. phoenecium (AlmWB-A4)
ViLL
BBP
TWB
VAC
CYE-C
DanVir-a
GDIII
CbY18
WWB-a
BLWB
PoiBI
VGYIII
CX
ScYPI-Afr
WX
LP
A. palmae
P. trifolii
EriWB
P. fraxini
P. ulmi
P. ziziphi
LfWB
StLL
LY
LDG
LDT
SBS
SCWL
BVK
CIRP
GaLL
P. oryzae
P. cynodontis
P. casteneae
P. pini
P. phoenecium
ViLL
WX
5 changes
FIGURE 116. (Continued)
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-GAAAAAGAAAG-3¢,
5¢-TCTTTCTTT-3¢, 5¢-CAG-3¢, 5¢-ACTTTG-3¢, and 5¢-GTCA
AAAC-3¢.
Habitat, association, or host: Hibiscus phloem.
8. “Candidatus Phytoplasma caricae” Arocha, López, Piñol,
Fernández, Picornell, Almeida, Palenzuela, Wilson and
Jones 2005, 2462
Vernacular epithet: Cuban papaya phytoplasma, strain PAYR.
Morphology: other.
707
Table 144. Provisional groupings, strain designations, associated plant disease, geographic origin and accession numbers of 16S rRNA gene
sequences derived from phytoplasmasa
Subgroup
Strain
AlloY
AP
ESFY
AshY
EriWB
AlloY
AP15R
AT
AP2
AP1/93
D365/04
APSb
ESFY-G1R
ESFY5
ESFY4
PPER
ESFY-142
ESFY-173
ESFY-215
AshY1
AshY3
ASHY
LWB3
EriWB
ArAWB
AusGY
AY
AusGY
PpDB
PYL
PYLb
SLY
SV3101
MIAY
OY-M
MBS
APWB
IOWB
HyPH
HYPh
RPh
MD
GDS
AYWB_ro4
PaWB
PY1
BVGY
AAY
CabD4
AY-BW
ApSL
SAY
ValY
PRIVC
WcWB
CabD3
AY-sb
Bstv2Mf12
ACLR-AY
ACLR
BBS3
STRAWB2
PoY
KVG
Associated plant disease
Allocasuarina yellows
Apple proliferation
Apple proliferation
Apple proliferation
Apple proliferation
Apple proliferation
Apple proliferation
European stone fruit yellows
Ash yellows
Ash yellows
Ash yellows
Lilac witches’-broom
Erigeron witches’-broom
Argentinian alfalfa witches’broom
Australian grapevine yellows
Papaya die-back
Phormium yellow leaf, rrnA
Phormium yellow leaf
Strawberry lethal yellows
Strawberry virescence
Oenothera virescence
Onion yellows
Maize bushy stunt
Aphanamixis polystachya
witches’-broom
Ipomoea obscura witches’broom
Hydrangea phyllody
Hydrangea phyllody
Oilseed rape phyllody
Mulberry dwarf
Aster yellows witches’broom
Paulownia witches’-broom
Periwinkle yellows
American aster yellows
Cabbage proliferation
Aster yellows
Apple sessile leaf
Severe aster yellows
Valeriana yellows, rrnA
Primrose virescence
Watercress witches’-broom
Cabbage proliferation
Sugar beet aster yellows
Apricot chlorotic leafroll
Apricot chlorotic leafroll
Blueberry stunt
Strawberry green petal
Populus yellows
Clover phyllody
“Candidatus
Phytoplasma species”
Geographic origin
16S accession no.a
P. allocasuarinae
P. mali
P. prunorum
P. fraxini
Australia
Italy
Germany
Germany
France
Slovenia
Italy
Germany
Austria
Czech Republic
Germany
Spain
Spain
Spain
USA, New York
USA, Utah
Germany
USA, Massachusetts
Brazil
Argentina
AY135523*
AJ542541*
X68375*
AF248958*
AJ542542*
EF025917
EF193361
AJ542544*
AY029540*
Y11933*
X68374
AJ575108*
AJ575106
AJ575105
AF092209*
AF105315*
X68339*
AF105317*
AY034608
AY147038
P. australiense
P. asteris
Australia
Australia
New Zealand
New Zealand
Australia
Tonga
USA, Michigan
Japan
Mexico
Bangladesh
L76865
Y10095*
U43569
U43570*
AJ243045*
AY377868*
M30790*
NC005303*
AY265208*
AY495702*
Taiwan
AY265205*
Italy
France
Czech Republic
South Korea
Ohio, USA
AY265207*
AY265219
U89378*
AY075038*
DQ112021
NC007716
Korea
China
Southern USA
USA, Texas
USA, Ohio
Lithuania
USA, California
Lithuania
Germany
USA, Hawaii
USA, Texas
Hungary
Spain
Europe
USA, Michigan
USA, Florida
Croatia
Germany
AF279271*
AF453328
AY083605
X68373*
AY180932
AY389820
AY734454
M86340*
AY102274*
AY265210*
AY665676*
AY180947*
AF245439
AY180951
AY265211
X68338*
AY265213
U96616*
AF503568
X83870
(continued)
708
Table 144. (continued)
Subgroup
Strain
THP
BWB
BGWL
BVK
CPh
HYDP
AY-WB
BB
PPT
THP
Derbid
BWB
BGWL-C1
BGWL
BGWL-2
CWL
BVK
CIRP
CnWB
CP
CIRP
CnWB
CPR
BLL
BLTVA
VR
PWB
CSV
EY
EY1
ULW
FD
RuS
HD1
VC
SpaWB
JWB
SpaWB229
JWB-G1T
JWB-Ka
JWB-ch
FBP
NecY-In1
CLY-5
WBDL
BoLL
GaLL
GLL-eth
HibWB
IAWB
IBS
StrawY
JHP
CaWB-YNO1
FBP
PPLL
BoLL
GaLL
GLL-eth
HibWB
IAWB
PEP
IBS
StrawY
JHP
LDG
LDT
LfWB
LY
LDG
LDN
LDT
LfWB
LfWB-t
CPY
LDY
Clover phyllody
Hydrangea phyllody
Aster yellows
Tomato big bud
Potato purple top
Tomato ‘hoja de perejil’
Derbid phytoplasma
Buckthorn witches’-broom
Bermudagrass white leaf
Bermudagrass white leaf
Bermudagrass white leaf Cynodon white leaf
Psammotettic cephalotesborne
Cirsium phyllody
Chestnut witches’- broom
Clover proliferation
Brinjal little leaf
Columbia basin potato
purple top
Vinca virescence
Potato witches’-broom
Centauria stolstitialis
virescence
Elm yellows
Ulmus witches’-broom
Flavescence doree
Rubus stunt
Hemp dogbane yellows
Asymptomatic Virginia
creeper
Spartium witches’-broom
Jujube witches’-broom Gifu
isolate 1
Jujube witches’-broom
Korea isolate 1
Ziziphus jujube witches’broom
Nectarine yellows
Cherry lethal yellows
Witches’-broom disease of
lime
Cactus witches’-broom
Faba bean phyllody
Pigeon pea little leaf
Bonamia little leaf
Galactia little leaf
Gliricidia little leaf
Hibiscus witches’-broom
Alfalfa witches’-broom
Pichris echioides phyllody
Italian bindweed yellows
Strawberry lethal yellows
Japanese hydrangea
phyllody
Cape St Paul wilt
Awka disease of coconut
Coconut lethal disease
Loofah WB
Loofah WB
Carludovica palmata yellows
Yucatan coconut decline
“Candidatus
Geographic origin
16S accession no.a
P. lycopersici
P. rhamni
P. cynodontis
Canada
Belgium
USA, Ohio
USA, Arkansas
Mexico
Bolivia
Cuba
Germany
Italy
Italy
Thailand
Australia
Germany
AF222066*
AY265215*
AY389827*
AY180955*
AF217247*
AY787136
AY744945
X76431*
AJ550984*
Y16388*
AF248961*
AF509321*
X76429*
P. castaneae
P. trifolii
Germany
Korea
Canada
India
USA, Washington
X83438*
AB054986*
AY390261*
X83431*
AY692280*
USA, California
Canada
Italy
AY500817*
AY500818*
AY270156*
P. ulmi
USA, New York
Italy
Italy
Italy
USA, New York
USA, Florida
AY197655*
X68376*
X76560*
AY197648*
AY197654*
AF305198*
P. spartii
P. ziziphi
Italy
Japan
AY197652*
AB052876*
Korea
AB052879*
China
AF305240
P. aurantifolia
India
China
Oman
AY332659*
AY197659*
U15442*
P. brasiliense
P. fragariae
P. japonicum
China
Sudan
Australia
Australia
Australia
Ethiopia
Brazil
Italy
Italy
Southern Italy
Lithuania
Japan
AJ293216
X83432*
AJ289191
Y15863*
Y15865*
AF361018*
AF147708*
Y16390*
Y16393*
Y16391*
DQ086423
AB010425
Ghana
Nigeria
Tanzania
Taiwan
Taiwan
Mexico
Mexico
Y13912*
Y14175*
X80177*
L33764*
AF086621*
AF237615
U18753*
(continued)
709
Table 144. (continued)
Subgroup
Strain
ScY
MPV
LfY1
LfY5(PE65)
LY-c2
LY-JC8
PanD
ScY
SCD3T2
SCD3T1
MPV
PD
PinP
PPWB
RYD
SBS
SCWL
PerWB-FL
CbY1
STRAWB1
PD1
PD
PYLR
EPC
Pin127R
PinG
AlmWB-A4
KAP
PPWB-f
RYD-J
RYD-Th
SBS
SCWL
SGS-v1
SpaWB
SPLL
SPWB
StLL
STOL
ViLL
CIWB
WTWB
WX
Spar
SPLL
PpYC
GPh
PnWB
CoAWB
SPLL
SUNHP
AlWB
TBB
StLL
STOL
VK
2642BN
ViLL
IM-3
WTWB
BBP
BLWB
CbY18
CX
CYE
DanVir-a
LP
PoiBI
ScYP I-Afr
TWB
VAC
VGYIII
WWB-a
WX
Coconut leaf yellowing
Coconut leaf yellowing
Coconut lethal yellows
Coconut lethal yellows
Pandanus decline
Sugarcane yellows, group 4
Mexican periwinkle
virescence
Periwinkle witches’-broom
Chinaberry yellows
Strawberry green petal
Pear decline
Pear decline
Peach yellow leafroll
Pear decline
Pinus halepensis yellows
Pinus sylvestris yellows
Almond witches’-broom
Knautia arvensis phyllody
Pigeonpea witches’-broom
Rice yellow dwarf
Rice yellow dwarf
Sorghum bunchy shoot
Sugarcane white leaf
Sorghum grassy shoot,
variant 1
Spartium witches’-broom
Sweet potato little leaf
Papaya yellow crinkle
Gerbera phyllody
Peanut witches’ broom
Cocky apple witches’-broom
Sweet potato little leaf
Sunnhemp phyllody
Alfalfa witches’-broom
Australian tomato big bud
Stylosanthes little leaf
Stolbur of Capsicum annum
Grapevine yellows
Grapevine yellows
Vigna little leaf
Cassia italica witches’-broom
Weeping tea witches’-broom
Blueberry proliferation
Black locust witches’-broom
Chinaberry yellows
Canadian peach X
Clover yellow edge
Dandelion virescence, rrnA
Little peach
Poinsettia branch-inducing
Sugarcane yellows
Tsuwabuki WB
Vaccinium witches’-broom
Virginia grapevine yellows
Walnut witches’-broom,
rrnA
Western X
“Candidatus
Geographic origin
16S accession no.a
Mexico
Mexico
USA, Florida
Jamaica
USA, Florida
Mauritius
Mauritius
Mauritius
Mexico
AF500329*
AF500334
AF498309*
AF498307
AF361020*
AJ539178*
AJ539180
AJ539179
AF248960*
P. pyri
P. pini
P. phoenecium
P. oryzae
USA, Florida
Bolivia
USA, Florida
Italy
Germany
USA, California
Iran
Spain
Germany
Lebanon
Italy
USA, Florida
Japan
Thailand
Australia
Thailand
Australia
AY204549
AF495882*
U96614*
AJ542543*
X76425*
Y16394
DQ471321
AJ632155*
AJ310849*
AF515636*
Y18052*
AF248957*
D12581
AB052873*
AF509322*
X76432*
AF509324*
P. spartii
P. australasia
P. solani
P. omaniense
Italy
Australia
Australia
Japan?
Taiwan
Australia
Australia
Thailand
Oman
Australia
Australia
Europe
Europe
France
Australia
Oman
Australia
Lithuania
USA, Maryland
Bolivia
Canada
Canada
Lithuania
USA, S. Carolina
Southern USA
Africa
Japan
Germany
USA, Virginia
USA, Georgia
X92869*
X90591*
Y10097*
AB026155*
L33765*
AJ295330*
AJ289193
X76433*
AY169322*
Y08173
AJ289192*
X76427*
X76428*
AJ964960
Y15866*
EF666051
AF521672*
AY034090*
AF244363*
AF495657*
L33733*
AF175304*
AF370119*
AF236122*
AF190223*
AF056095*
D12580*
X76430*
AF060875*
AF190226*
USA, California
L04682*
Accession numbers denoted by an asterisk were used as sources of 16S rRNA gene sequences for comprehensive phylogenetic analysis of subgroup phytoplasmas from
diverse geographic origins.
a
710
Unique regions of 16S rRNA gene: 5¢-AAA-3¢ (196–198), 5¢ATT-3¢ (600–603), 5¢-AGGCGCC-3¢ (1089–1095), 5¢-GCGGATTTAGTCACTTTTCAGGC-3¢ (1379–1401).
Habitat, association, or host: Carica papaya phloem.
9. “Candidatus Phytoplasma castaneae” Jung, Sawayanagi,
Kakizawa, Nishigawa, Miyata, Oshima, Ugaki, Lee, Hibi and
Namba 2002, 1548
Vernacular epithet: Chestnut witches’ broom phytoplasma,
strain CnWBR.
Morphology: other.
Sequence accession no. (16S rRNA gene): AB054986.
Unique regions of 16S rRNA gene: 5¢-CTAGTTTAAAAACAATGCTC-3¢ and 5¢-CTCATCTTCCTCCAATTC-3¢.
Habitat, association, or host: Castanea crenata phloem.
10. “Candidatus Phytoplasma cynodontis” Marcone, Schneider
and Seemüller 2004b, 1081
Vernacular epithet: Bermuda grass white leaf (BGWL) phytoplasma, strain BGWl-C1R.
Morphology: other.
Unique region of 16S rRNA gene: 5¢-AATTAGAAGGCAT
CTTTTAAT-3¢.
Habitat, association, or host: phloem of Cynodon dactylon
(Bermuda grass).
11. “Candidatus Phytoplasma fragariae” Valiunas, Staniulis and
Davis 2006, 280
Vernacular epithet: Strawberry yellows phytoplasma, strain
StrawY R.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-GTGCAATGCTCAACGTTGTGAT-3¢, 5¢-AATTGCA-3¢, and 5¢-TGAGTAATCAAGAGGGAG-3¢.
Habitat, association, or host: phloem of Fragaria x ananassa.
12. “Candidatus Phytoplasma fraxini” Griffiths, Sinclair, Smart
and Davis 1999, 1613
Vernacular epithet: Ash yellows phytoplasma, strain AshYR
and lilac witches’-broom (LWB) phytoplasma.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-CGGAAACCCCTCAAAAGGTTT-3¢ and 5¢-AGGAAAGTC-3¢.
Habitat, association, or host: phloem of Fraxinus and
Syringa.
13. “Candidatus Phytoplasma graminis” Arocha, López, Piñol,
Fernández, Picornell, Almeida, Palenzuela, Wilson and
Jones 2005, 2462
Vernacular epithet: Sugarcane yellow leaf phytoplasma,
strain SCYPR.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-TTTG-3¢ (465–468),
5¢-TTG-3¢ (478–480), 5¢-GGG-3¢ (1552–1554), 5¢-TAA-3¢
(1381–1383), and 5¢-ATTTACGTTTCTG-3¢ (1392–1404).
Habitat, association, or host: Saccharum officinarum phloem.
14. “Candidatus
Phytoplasma
japonicum”
Sawayanagi,
Horikoshi, Kanehira, Shinohara, Bertaccini, Cousin, Hiruki
and Namba 1999, 1284
Vernacular epithet: Japanese Hydrangea phyllody phytoplasma, strain JHPR.
Morphology: other.
Sequence accession no. (16S rRNA gene): AB010425.
Unique regions of 16S rRNA gene: 5¢-GTGTAGCCGGGCTGAGAGGTCA-3¢ and 5¢-TCCAACTCTAGCTAAACAGTTTCTG-3¢.
Habitat, association, or host: Hydrangea phloem.
15. “Candidatus Phytoplasma lycopersici” Arocha, Antesana,
Montellano, Franco, Plata and Jones 2007, 1709
Vernacular epithet: Tomato “hoja de perejil” phytoplasma,
strain THPR.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-CTTA-3¢ (positions
175–178), 5¢-AATGGT-3¢ (198–203), 5¢-ATA-3¢ (229–231),
5¢-TGGAGGAA-3¢ (234–242), 5¢-CACG-3¢ (302–305),
5¢-TCT-3¢ (315–317), 5¢-GCT-3¢ (334–336), 5¢-TAT-3¢
(336–338), 5¢-TAC-3¢ (413–415), and 5¢-AGC-3¢ (434–436).
Habitat, association, or host: Lycopersicon esculentum
phloem.
16. “Candidatus Phytoplasma mali” Seemüller and Schneider
2004, 1224
Vernacular epithet: Apple proliferation (AP) phytoplasma,
strain AP15R.
Morphology: other.
Unique region of 16S rRNA gene: 5¢-AATACTCGAAACCAGTA-3¢.
Habitat, association, or host: Malus phloem.
17. “Candidatus Phytoplasma omanense” Al-Saady, Khan,
Calari, Al-Subhi and Bertaccini 2008, 464
Vernacular epithet: Cassia witches’-broom (CWB) phytoplasma, strain IM-1R.
Morphology: other.
Sequence accession no. (16S rRNA gene): EF666051.
Unique regions of 16S rRNA gene: 5¢-AAAAAACAGT-3¢ (467–
474), 5¢-TTGC-3¢ (642–645), 5¢-GTTAAAG-3¢ (853–861),
5¢-TAATT-3¢ (1010–1014), and 5¢-AAATT-3¢ (1052–1056).
Habitat, association, or host: Cassia italica phloem.
18. “Candidatus Phytoplasma oryzae” Jung, Sawayanagi, Wongkaew, Kakizawa, Nishigawa, Wei, Oshima, Miyata, Ugaki,
Hibi and Namba 2003c, 1928
Vernacular epithet: Rice yellow dwarf (RYD) phytoplasma,
strain RYD-ThR.
Morphology: other.
Sequence accession nos (16S rRNA gene): D12581, AB052873
(RYD-Th).
Unique regions of 16S rRNA gene: 5¢-AACTGGATAGGAAATTAAAAGGT-3¢ and 5¢-ATGAGACTGCCAATA-3¢.
Habitat, association, or host: Oryza sativa phloem.
19. “Candidatus Phytoplasma phoenicium” Verdin, Salar, Danet,
Choueiri, Jreijiri, El Zammar, Gélie, Bové and Garnier 2003, 837
Vernacular epithet: Almond witches’-broom (AlmWB)
phytoplasma, strain AlmWB-A4R.
Morphology: other.
Unique region of 16S rRNA gene: 5¢-CCTTTTTCGGAAGGTATG-3¢.
Habitat, association, or host: Prunus amygdalus phloem.
20. “Candidatus Phytoplasma pini” Schneider, Torres, Martín,
Schröder, Behnke and Seemüller 2005, 306
Vernacular epithet: Pinus halepensis yellows (Pin) phytoplasma, strain Pin127SR.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-GGAAATCTTTCGGGATTTTAGT-3¢ and 5¢-TCTCAGTGCTTAACGCTGTTCT-3¢.
Habitat, association, or host: Pinus phloem.
21. “Candidatus Phytoplasma prunorum” Seemüller and Schneider 2004, 1224
Vernacular epithet: European stone fruit yellows (ESFY)
phytoplasma, strain ESFY-G1R.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-AATACCCGAAACCAGTA-3¢ and 5¢-TGAAGTTTTGAGGCATCTCGAA-3¢.
Habitat, association, or host: Prunus phloem.
22. “Candidatus Phytoplasma pyri” Seemüller and Schneider
2004, 1224
Vernacular epithet: Pear decline (PD) phytoplasma, strain
PD1R.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-AATACTCAAAACCAGTA-3¢ and 5¢-ATACGGCCCAAACTCATACGGA-3¢.
Habitat, association, or host: Pyrus phloem.
23. “Candidatus Phytoplasma rhamni” Marcone, Gibb, Streten
and Schneider 2004a, 1028
Vernacular epithet: Buckthorn witches’-broom phytoplasma, strain BWBR.
Morphology: other.
Sequence accession nos (16S rRNA gene): X76431, AJ583009.
Unique regions of 16S rRNA gene: 5¢-CGAAGTATTTCGATAC-3¢.
Habitat, association, or host: phloem of Rhamnus catharticus
(buckthorn).
24. “Candidatus Phytoplasma solani” Firrao, Gibb and Streton
2005, 251
711
Vernacular epithet: Stolbur phytoplasma; subgroup A reference type of the stolbur phytoplasma taxonomic group
16SrXII (Lee et al., 2000).
Morphology: other.
Sequence accession no. (16S rRNA gene): AJ970609 (strain
PO; Cimerman et al., 2006).
Unique region of 16S rRNA gene: not reported.
Habitat, association, or host: many species of Solanaceae plus
several species in other plant families, and Fulguromorpha
spp. planthopper vectors.
25. “Candidatus Phytoplasma spartii” Marcone, Gibb, Streten
and Schneider 2004a, 1028
Vernacular epithet: Spartium witches’-broom phytoplasma,
strain SpaWBR.
Morphology: other.
Sequence accession no. (16S rRNA gene): X92869.
Unique region of 16S rRNA gene: 5¢-TTATCCGCGTTAC-3¢.
Habitat, association, or host: phloem of Spartium junceum
(Spanish broom).
26. “Candidatus Phytoplasma tamaricis” Zhao, Sun, Wei, Davis,
Wu and Liu 2009, 2496
Vernacular epithet: Salt cedar witches’-broom phytoplasma,
strain SCWB1R.
Morphology: other.
Sequence accession no. (16S rRNA gene): FJ432664.
Unique regions of 16S rRNA gene: 5¢-ATTAGGCATCTAGTAACTTTG-3¢, 5¢-TGCTCAACATTGTTGC-3¢, 5¢-AGCTTTGCAAAGTTG-3¢, and 5¢-TAACAGAGGTTATCAGAGTT-3¢.
Habitat, association, or host: phloem of Tamarix chinensis
(salt cedar).
27. “Candidatus Phytoplasma trifolii” Hiruki and Wang 2004, 1352
Vernacular epithet: Clover proliferation phytoplasma,
strain CPR.
Morphology: other.
Unique regions of 16S rRNA gene: 5¢-TTCTTACGA-3¢ and
5¢-TAGAGTTAAAAGCC-3¢.
Habitat, association, or host: Trifolium phloem.
28. “Candidatus Phytoplasma ulmi” Lee, Martini, Marcone and
Zhu 2004b, 345
Vernacular epithet: Elm yellows phytoplasma (EY) phytoplasma, strain EY1R.
Morphology: other.
Sequence accession nos (16S rRNA gene): AY197655,
AY197675, and AY197690.
Unique regions of 16S rRNA gene: 5¢-GGAAA-3¢ and 5¢-CGTTAGTTGCC-3¢.
Habitat, association, or host: Ulmus americana phloem.
29. “Candidatus Phytoplasma vitis” Firrao, Gibb and Streton
2005, 251
Vernacular epithet: Flavescence dorée phytoplasma; strains
are genetically heterogenous and vary in degree of virulence,
but all are referable to subgroups C or D of the elm yellows
712
phytoplasma taxonomic group 16SrV (Lee et al., 2000).
Morphology: other.
Sequence accession nos (16S rRNA gene): AY197645 (16SrV
subgroup C), AY197644 (16SrV subgroup D1).
Unique regions of 16S rRNA gene: not reported.
Habitat, association, or host: grapevines (Vitis vinifera) and
the leafhopper vector Scaphoideus titanus.
30. “Candidatus Phytoplasma ziziphi” Jung, Sawayanagi, Kakizawa, Nishigawa, Wei, Oshima, Miyata, Ugaki, Hibi and
Namba 2003b, 1041
References
Ahrens, U. and E. Seemüller. 1992. Detection of DNA of plant pathogenic mycoplasma-like organisms by a polymerase chain reaction
that amplifies a sequence of the 16S rRNA gene. Phytopathology 82:
828–832.
Al-Saady, N.A., A.J. Khan, A. Calari, A.M. Al-Subhi and A. Bertaccini.
2008. ‘Candidatus Phytoplasma omanense’, associated with witches’broom of Cassia italica (Mill.) Spreng. in Oman. Int. J. Syst. Evol.
Microbiol. 58: 461–466.
Albertazzi, G., A.C. J. Milc, E. Francia, E. Roncaglia, F. Ferrari, E. Tagliafico, E. Stefani and N. Pecchioni. 2009. Gene expression in grapevine
cultivars in response to Bois Noir phytoplasma infection. Plant Science 176: 792–804.
Alma, A., D. Bosco, A. Danielli, A. Bertaccini, M. Vibio and A. Arzone.
1997. Identification of phytoplasmas in eggs, nymphs and adults of
Scaphoideus titanus Ball reared on healthy plants. Insect. Mol. Biol. 6:
115–121.
An, F.-Q., Y.-F. Wu, X.-Q. Sun, P.-W. Gu and Y. Yang. 2006. Homologic
analysis of tuf gene for elongation factor Tu of phytoplasma from
wheat blue dwarf. Sci. Agric. Sinica 39: 74–80.
Anonymous. 2000. Report of consultations: Phytoplasmas, spiroplasmas, mesoplasmas and entomoplasmas working team. Presented at
the International Research Programme on Comparative Mycoplasmology (IRPCM) of the International Organization for Mycoplasmology (IOM) Fukuoka, Japan.
Arashida, R., S. Kakizawa, Y. Ishii, A. Hoshi, H.Y. Jung, S. Kagiwada,
Y. Yamaji, K. Oshima and S. Namba. 2008. Cloning and characterization of the antigenic membrane protein (Amp) gene and in situ
detection of Amp from malformed flowers infected with Japanese
hydrangea phyllody phytoplasma. Phytopathology 98: 769–775.
Arocha, Y., M. Lopez, B. Pinol, M. Fernandez, B. Picornell, R. Almeida,
I. Palenzuela, M.R. Wilson and P. Jones. 2005. ‘Candidatus Phytoplasma graminis’ and ‘Candidatus Phytoplasma caricae’, two novel
phytoplasmas associated with diseases of sugarcane, weeds and
papaya in Cuba. Int. J. Syst. Evol. Microbiol. 55: 2451–2463.
Arocha, Y., O. Antesana, E. Montellano, P. Franco, G. Plata and P. Jones.
2007. ‘Candidatus Phytoplasma lycopersici’, a phytoplasma associated
with ‘hoja de perejil’ disease in Bolivia. Int. J. Syst. Evol. Microbiol.
57: 1704–1710.
Bai, X., J. Zhang, I.R. Holford and S.A. Hogenhout. 2004. Comparative genomics identifies genes shared by distantly related insecttransmitted plant pathogenic mollicutes. FEMS Microbiol. Lett. 235:
249–258.
Bai, X., J. Zhang, A. Ewing, S.A. Miller, A. Jancso Radek, D.V.
Shevchenko, K. Tsukerman, T. Walunas, A. Lapidus, J.W. Campbell
and S.A. Hogenhout. 2006. Living with genome instability: the adaptation of phytoplasmas to diverse environments of their insect and
plant hosts. J. Bacteriol. 188: 3682–3696.
Barbara, D.J., A. Morton, M.F. Clark and D.L. Davies. 2002. Immunodominant membrane proteins from two phytoplasmas in the
aster yellows clade (chlorante aster yellows and clover phyllody) are
Vernacular epithet: Jujube witches’-broom phytoplasma,
strain JWBR.
Morphology: other.
Sequence accession nos (16S rRNA gene):AB052875–
AB052879.
Unique regions of 16S rRNA gene: 5¢-TAAAAAGGCATCTT
TTTGTT-3¢ and 5¢-AATCCGGACTAAGACTGT-3¢.
Habitat, association, or host: Ziziphyus jujube phloem.
highly divergent in the major hydrophilic region. Microbiology 148:
157–167.
Baric, S. and J. Dalla-Via. 2004. A new approach to apple proliferation
detection: a highly sensitive real-time PCR assay. J. Microbiol. Methods 57: 135–145.
Berg, M., D.L. Davies, M.F. Clark, H.J. Vetten, G. Maier, C. Marcone and
E. Seemüller. 1999. Isolation of the gene encoding an immunodominant membrane protein of the apple proliferation phytoplasma and
expression and characterization of the gene product. Microbiology
145: 1937–1943.
Berg, M. and E. Seemüller. 1999. Chromosomal organization and nucleotide sequence of the genes coding for the elongation factors G and
Tu of the apple proliferation phytoplasma. Gene 226: 103–109.
Bertaccini, A., R.E. Davis, R.W. Hammond, M. Vibio, M.G. Bellardi and
I.-M. Lee. 1992. Sensitive detection of mycoplasmalike organisms in
field-collected and in vitro propagated plants of Brassica, Hydrangea
and Chrysanthemum by polymerase chain reaction. Ann. Appl. Biol.
121: 593–599.
Bertaccini, A., J. Fráova, S. Paltrinieri, M. Martini, M. Navrátil, C. Lugaresi, Nebesárová and M. Simkova. 1999. Leek proliferation: a new
phytoplasma disease in the Czech Republic and Italy. Eur. J. Plant
Pathol. 105: 487–493.
Bertamini, M. and N. Nedunchezhian. 2001. Effects of phytoplasma
[stolbur-subgroup (Bois noir-BN)] on photosynthetic pigments, saccharides, ribulose 1,5-biphosphate carboxylase, nitrate and nitrite
reductases, and photosynthetic activities in field-grown grapevine (Vitis
vinifera L. cv. Chardonnay) leaves. Photosynthetica 39: 119–122.
Bianco, P.A., R.E. Davis, J.P. Prince, I.-M. Lee, D.E. Gundersen, A. Fortusini and G. Belli. 1993. Double and single infections by aster yellows
and elm yellows MLOs in grapevines and symptoms characteristic of
Flavescence dorée. Rev. Patol. Veg. 3: 69–82.
Blomquist, C.L., D.J. Barbara, D.L. Davies, M.F. Clark and B.C. Kirkpatrick. 2001. An immunodominant membrane protein gene from
the Western X-disease phytoplasma is distinct from those of other
phytoplasmas. Microbiology 147: 571–580.
Cai, H., W. Wei, R.E. Davis, H. Chen and Y. Zhao. 2008. Genetic diversity
among phytoplasmas infecting Opuntia species: virtual RFLP analysis
identifies new subgroups in the peanut witches’-broom phytoplasma
group. Int. J. Syst. Evol. Microbiol. 58: 1448–1457.
Carginale, V., G. Maria, C. Capasso, E. Ionata, F. La Cara, M. Pastore,
A. Bertaccini and A. Capasso. 2004. Identification of genes expressed
in response to phytoplasma infection in leaves of Prunus armeniaca by
messenger RNA differential display. Gene 332: 29–34.
Carraro, L., N. Loi and P. Ermacora. 2001. Transmission characteristics
of the European stone fruit yellows phytoplasma and its vector Cacopsylla pruni. Eur. J. Plant Pathol. 107: 695–700.
Chang, F.L., C.C. Chen and C.P. Lin. 1995. Monoclonal antibody for the
detection and identification of a phytoplasma associated with rice
yellow dwarf. Eur. J. Plant Pathol. 101: 511–518.
Chen, M.H. and C. Hiruki. 1978. The preservation of membranes of
tubular bodies associated with mycoplasma-like organisms by tannic
acid. Can. J. Bot. 56: 2878–2882.
Chen, T.A., D.A. Lei and C.P. Lin. 1989. Detection and identification
of plant and insect mollicutes. In The Mycoplasmas, vol. 5 (edited by
Whitcomb and Tully). Academic Press, New York, pp. 393–424.
Chi, K.L. and C.P. Lin. 2005. Cloning and analysis of polC gene of phytoplasma associated with peanut witches’ broom. Plant Pathol. Bull.
14: 51–58.
Chiykowski, L.N. 1983. Frozen leafhoppers as a vehicle for long-term
storage of different isolates of the aster yellows agents. Can. J. Plant
Pathol. 5: 101–106.
Chiykowski, L.N. 1988. Maintenance of yellows-type mycoplasmalike
organisms. In Tree Mycoplasmas and Mycoplasma Diseases (edited
by Hiruki). The University of Alberta Press, Edmonton, Alberta,
Canada, pp. 123–134.
Chiykowski, L.N. and R.C. Sinha. 1990. Differentiation of MLO diseases
by means of symptomatology and vector transmission. Rec. Adv.
Mycoplasmol. Suppl. 20: 280–287.
Christensen, N.M., M. Nicolaisen, M. Hansen and A. Schulz. 2004. Distribution of phytoplasmas in infected plants as revealed by real-time
PCR and bioimaging. Mol. Plant Microbe. Interact. 17: 1175–1184.
Chu, Y.R., W. Y. Chen and C.P. Lin. 2006. Cloning and sequence analses
of recA gene of phytoplasma associated with peanut witches’ broom.
Plant Pathol. Bull. 15: 211–218.
Chuang, J.G. and C.P. Lin. 2000. Cloning of gyrB and gyrA genes of phytoplasma associated with peanut witches’ broom. Plant Pathol. Bull.
9: 157–166.
Cimerman, A., G. Arnaud and X. Foissac. 2006. Stolbur phytoplasma
genome survey achieved using a suppression subtractive hybridization approach with high specificity. Appl. Environ. Microbiol. 72:
3274–3283.
Cimerman, A., D. Pacifico, P. Salar, C. Marzachi and X. Foissac. 2009.
Striking diversity of vmp1, a variable gene encoding a putative membrane protein of the stolbur phytoplasma. Appl. Environ. Microbiol.
75: 2951–2957.
Clark, M.F. 1992. Immunodiagnostic techniques for plant mycoplasmalike organisms. In Techniques for the Rapid Detection of Plant
Pathogens (edited by Duncan and Torrance). Blackwell Scientific
Publications, Oxford, pp. 34–45.
Cordova, I., P. Jones, N.A. Harrison and C. Oropeza. 2003. In situ PCR
detection of phytoplasma DNA in embryos from coconut palms with
lethal yellowing disease. Mol. Plant Pathol. 4: 99–108.
Cousin, M.T., J. Roux, E. Boudon-Padieu, R. Berges, E. Seemüller and
C. Hiruki. 1998. Use of heteroduplex mobility analysis (HMA) for differentiating phytoplasma isolates causing witches’ broom disease of
Populus nigra vc Italica and stolbur or big bud symptoms on tomato.
J. Phytopathol. 146: 97–102.
D’Arcy, C.J. and L.R. Nault. 1982. Insect transmission of plant viruses
and mycoplasmalike and rickettsialike organisms. Plant Dis. 66:
99–104.
Daire, X., D. Clair, J. Larrue, E. Boudon-Padieu and A. Caudwell. 1993.
Diversity among mycoplasma-like organisms inducing grapevine yellows in France. Vitis 32: 159–163.
Davis, M.J., J.H. Tsai, R.L. Cox, L.L. McDaniel and N.A. Harrison. 1988.
Cloning of chromosomal and extrachromosomal DNA of the mycoplasma-like organism that causes maize bushy stunt disease. Mol.
Davis, R.E. and R.F. Whitcomb. 1970. Evidence on possible mycoplasma
etiology of aster yellows disease. I. Suppression of symptom development in plants by antibiotics. Infect. Immun. 2: 201–208.
Davis, R.E. and I.-M. Lee. 1992. Mycoplasmalike organisms as plant disease agents. ATCC Quart. Newsl. 4: 8–11.
Davis, R.E. and I.-M. Lee. 1993. Cluster-specific polymerase chain reaction amplification of 16S rDNA sequences for detection and identification of mycoplasmalike organisms. Phytopathology 63: 1008–1011.
Davis, R.E., E.L. Dally, D.E. Gundersen, I.M. Lee and N. Habili. 1997.
“Candidatus Phytoplasma australiense,” a new phytoplasma taxon
associated with Australian grapevine yellows. Int. J. Syst. Bacteriol.
47: 262–269.
713
Davis, R.E. and W.A. Sinclair. 1998. Phytoplasma identity and disease
etiology. Phytopathology 88: 1372–1376.
Davis, R.E., R. Jomantiene, A. Kalvelyte and E.L. Dally. 2003a. Differential amplification of sequence heterogeneous ribosomal RNA genes
and classification of the ‘Fragaria multicipita’ phytoplasma. Microbiol.
Res. 158: 229–236.
Davis, R.E., R. Jomantiene, Y. Zhao and E.L. Dally. 2003b. Folate biosynthesis pseudogenes, PsifolP and PsifolK, and an O-sialoglycoprotein
endopeptidase gene homolog in the phytoplasma genome. DNA
Cell Biol. 22: 697–706.
Davis, R.E., R. Jomantiene and Y. Zhao. 2005. Lineage-specific decay of
folate biosynthesis genes suggests ongoing host adaptation in phytoplasmas. DNA Cell Biol. 24: 832–840.
Davis, R.E., R. Jomantiene, E. L. Dally and T. K. Wolf. 1998. Phytoplasmas associated with grapevine yellows in Virginia belong to group
16SrI, subgroup A (tomato big bud phytoplasma subgroup), and
group 16SrIII, new subgroup I. Vitis 37: 131–137.
Denes, A.S. and R.C. Sinha. 1991. Extrachromosomal DNA elements of
plant-pathogenic mycoplasma-like organisms. Can. J. Plant Pathol.
13: 26–32.
Denes, A.S. and R.C. Sinha. 1992. Alteration of clover phyllody mycoplasma DNA after in vitro culturing of phyllody-diseased clover. Can. J.
Plant Pathol. 14: 189–196.
Deng, S. and C. Hiruki. 1991. Amplification of 16S rRNA genes from
culturable and non-culturable mollicutes. J. Microbiol. Meth. 14:
53–61.
Doi, Y., M. Teranaka, K. Yora and H. Asuyama. 1967. Mycoplasma or
PLT-group-like organisms found in the phloem elements of plants
infected with mulberry dwarf, potato witches-broom, aster yellows,
or Paulownia witches broom. Ann. Phytopathol. Soc. Jpn. 33: 256–
266.
Errampelli, D. and J. Fletcher. 1993. Production of monospecific polyclonal antibodies made against aster yellows MLO-associated antigen. Phytopathology 83: 1279–1282.
Esau, K., A.C. Magyarosy and V. Breazeale. 1976. Studies of the mycoplasma-like organism (MLO) in spinach leaves affected by the aster
yellows disease. Protoplasma 90: 189–203.
Evert, R.F. 1977. Phloem structure and histochemistry. Annu. Rev. Plant
Physiol. 28: 199–222.
Firrao, G., C.D. Smart and B.C. Kirkpatrick. 1996. Physical map of
the western X-disease phytoplasma chromosome. J. Bacteriol. 178:
3985–3988.
Firrao, G., K. Gibb and C. Streten. 2005. Short taxonomic guide to the
genus ‘Candidatus Phytoplasma’. J. Plant Pathol. 87: 249–263.
Florance, E.R. and H.T. Cameron, 1978. Three-dimensional structure
and morphology of mycoplasma-like bodies associated with albino
disease of Prunus avium. Phytopathology 68: 75–80.
Flores, H.E., J. M. Vivanco and V.M. Loyola-Vargas. 1999. ‘Radicle’ biochemistry: the biology of root-specific metabolism. Trends Plant Sci.
4: 220–226.
Galetto, L., J. Fletcher, D. Bosco, M. Turina, A. Wayadande and
C. Marzachi. 2008. Characterization of putative membrane protein
genes of the ‘Candidatus Phytoplasma asteris’, chrysanthemum yellows isolate. Can. J. Microbiol. 54: 341–351.
Garcia-Chapa, M., A. Batlle, D. Rekab, M.R. Rosquete and G. Firrao.
2004. PCR-mediated whole genome amplification of phytoplasmas.
J. Microbiol. Methods 56: 231–242.
Garnier, M., X. Foissac, P. Gaurivaud, F. Laigret, J. Renaudin, C. Saillard
and J.M. Bové. 2001. Mycoplasmas, plants, insect vectors: a matrimonial triangle. C. R. Acad. Sci. III 324: 923–928.
Gibb, K.S., B. Schneider and A.C. Padovan. 1998. Differential detection
and genetic relatedness of phytoplasmas in papaya. Plant Pathol. 47:
325–332.
Gomez, G.G., L.R. Conci, D.A. Ducasse and S.F. Nome. 1996. Purification of the phytoplasma associated with China-tree (Melia azedarach L.)
decline and the production of a polyclonal antoserum for its detection. J. Phytopathol. 144: 473–477.
714
Griffiths, H.M., W.A. Sinclair, C.D. Smart and R.E. Davis. 1999. The phytoplasma associated with ash yellows and lilac witches’-broom: ‘Candidatus Phytoplasma fraxini’. Int. J. Syst. Bacteriol. 49: 1605–1614.
Gundersen, D.E., I.M. Lee, S.A. Rehner, R.E. Davis and D.T. Kingsbury.
1994. Phylogeny of mycoplasmalike organisms (phytoplasmas): a
basis for their classification. J. Bacteriol. 176: 5244–5254.
Gundersen, D.E. and I.M. Lee. 1996. Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathol. Mediterr. 35: 144–151.
Guo, Y.H., Z.M. Cheng, J.A. Walla and Z. Zhang. 1998. Diagnosis of
X-disease phytoplasma in stone fruits by a monoclonal antibody developed directly from a woody plant. J. Environ. Hortic. 16: 33–37.
Guthrie, J.N., K.B. Walsh, P.T. Scott and T.S. Rasmussen. 2001. The phytopathology of Australian papaya dieback: a proposed role for the
phytoplasma. Physiol. Mol. Plant Pathol. 58: 23–30.
Haggis, G.H. and R.C. Sinha. 1978. Scanning electron microscopy of
mycoplasmalike organisms after freeze fracture of plant tissues affected
with clover phyllody and aster yellows. Phytopathology 68: 677–680.
Hanboonsong, Y., C. Choosai, S. Panyim and S. Damak. 2002. Transovarial transmission of sugarcane white leaf phytoplasma in the insect
vector Matsumuratettix hiroglyphicus (Matsumura). Insect. Mol. Biol.
11: 97–103.
Harrison, N.A., J.H. Tsai, C.M. Bourne and P.A. Richardson. 1991.
Molecular cloning and detection of chromosomal and extrachromosomal DNA of mycoplasma-like organisms associated with witches’
broom disease of pigeon pea in Florida. Mol. Plant Microbe Interact.
4: 300–307.
Harrison, N.A., C.M. Bourne, R.L. Cox, J.H. Tsai and P.A. Richardson. 1992. DNA probes for detection of mycoplasma-like organisms
associated with lethal yellowing disease of palms in Florida. Phyto
pathology 82: 216–224.
Harrison, N.A., W. Myrie, P. Jones, M.L. Carpio, M. Castillo, M.M. Doyle
and C. Oropeza. 2002. 16S rRNA interoperon sequence heterogeneity distinguishes strain populations of palm lethal yellowing phytoplasma in the Caribbean region. Ann. Appl. Biol. 141: 183–193.
Harrison, N.A., E. Boa and M.L. Carpio. 2003. Characterization of phytoplasmas detected in Chinaberry trees with symptoms of leaf yellowing and decline in Bolivia. Plant Pathol. 52: 147–157.
Hearon, S.S., R.H. Lawson, F.F. Smith, J.T. Mckenzie and J. Rosen. 1976.
Morphology of filamentous forms of a mycoplasmalike organism
associated with hydrangea virescence. Phytopathology 66: 608–616.
Hiruki, C. and K. Wang. 2004. Clover proliferation phytoplasma:
‘Candidatus Phytoplasma trifolii’. Int. J. Syst. Evol. Microbiol. 54:
1349–1353.
Ho, K.C., C.C. Tsai and T.L. Chung. 2001. Organization of ribosomal
RNA genes from a Loofah witches’ broom phytoplasma. DNA Cell
Biol. 20: 115–122.
Hodgetts, J., N. Boonham, R. Mumford, N. Harrison and M. Dickinson. 2008. Phytoplasma phylogenetics based on analysis of secA and
23S rRNA gene sequences for improved resolution of candidate
species of ‘Candidatus Phytoplasma’. Int. J. Syst. Evol. Microbiol. 58:
1826–1837.
Hogenhout, S.A., K. Oshima, D. Ammar el, S. Kakizawa, H.N. Kingdom
and S. Namba. 2008. Phytoplasmas: bacteria that manipulate plants
and insects. Mol. Plant. Pathol. 9: 403–423.
Hren, M., M. Ravnikar, J. Brzin, P. Ermacora, L. Carraro, P. A. Bianco,
P. Casati, M. Borgo, E. Angelini, A. Rotter and K. Gruden. 2009.
Induced expression of sucrose synthase and alcohol dehydrogenase
I genes in phytoplasma-infected grapevine plants grown in the field.
Hsu, H.T., I.M. Lee, R.E. Davis and Y.C. Wang. 1990. Immunization for
generation of hybridoma antibodies specifically reacting with plants
infected with a mycoplasmalike organism (MLO) and their use in
detection of MLO antigens. Phytopathology 80: 946–950.
ICSB Subcommittee on the Taxonomy of Mollicutes. 1993. Minutes of
the Interim meetings, 1 and 2 August, 1992, Ames, Iowa. Int. J. Syst.
Bacteriol. 43: 394–397.
the interim meetings, 12 and 18 August, 1996, Orlando, Florida, USA
the interim meetings, 13 and 19 July 2000, Fukuoka, Japan. Int. J.
IRPCM Phytoplasma/Spiroplasma Working Team – Phytoplasma Taxonomy Group. 2004. Description of the genus ‘Candidatus Phytoplasma’,
a taxon for the wall-less non-helical prokaryotes that colonize plant
phloem and insects. Int. J. Syst. Evol. Microbiol. 54: 1243–1255.
Ishii, T., Y. Doi, K. Yora and H. Asuyama. 1967. Suppressive effects
of antibiotics of tetracycline group on symptom development
of mulberry dwarf disease. Ann. Phytopathol. Soc. Jpn. 33: 267–275.
Jagoueix-Eveillard, S., F. Tarendeau, K. Guolter, J.L. Danet, J.M.
Bové and M. Garnier. 2001. Catharanthus roseus genes regulated
differentially by mollicute infections. Mol. Plant Microbe Interact.
14: 225–233.
Jarausch, W., C. Saillard, F. Dosba and J.M. Bové. 1994. Differentiation
of mycoplasmalike organisms (MLOs) in European fruit trees by PCR
using specific primers derived from the sequence of a chromosomal
fragment of the apple proliferation MLO. Appl. Environ. Microbiol.
60: 2916–2923.
Jarausch, W., C. Saillard and F. Dosba. 1996. Long-term maintenance
of nonculturable apple proliferation phytoplasmas in their micropropagated natural host plant. Plant Pathol. 45: 778–786.
Jiang, Y.P. and T.A. Chen. 1987. Purification of mycoplasma-like organisms
from lettuce with aster yellows disease. Phytopathology 77: 949–953.
Jiang, Y.P., J.D. Lei and T.A. Chen. 1988. Purification of aster yellows
agent from diseased lettuce using affinity chromatography. Phytopathology 78: 828–831.
Jiang, Y.P., T.A. Chen, L.N. Chiykowski and R.C. Sinha. 1989. Production of monoclonal antibodies to peach eastern-X disease and their
use in disease detection. Can. J. Plant Pathol. 11: 325–331.
Jomantiene, R., R.E. Davis, D. Valiunas and A. Alminaite. 2002. New group
16SrIII phytoplasma lineages in Lithuania exhibit rRNA interoperon
sequence heterogeneity. Eur. J. Plant Pathol. 108: 507–517.
Jomantiene, R. and R.E. Davis. 2006. Clusters of diverse genes existing
as multiple, sequence-variable mosaics in a phytoplasma genome.
Jomantiene, R., Y. Zhao and R.E. Davis. 2007. Sequence-variable mosaics:
composites of recurrent transposition characterizing the genomes of
phylogenetically diverse phytoplasmas. DNA Cell Biol. 26: 557–564.
Jones, P. 2002. Phytoplasma plant pathogens. In Plant Pathologists
Pocketbook (edited by Waller). CAB International, Wallingford, UK,
pp. 126–139.
Jung, H.Y., T. Sawayanagi, S. Kakizawa, H. Nishigawa, S. Miyata,
K. Oshima, M. Ugaki, J.T. Lee, T. Hibi and S. Namba. 2002. ‘Candidatus Phytoplasma castaneae’, a novel phytoplasma taxon associated
with chestnut witches’ broom disease. Int. J. Syst. Evol. Microbiol. 52:
1543–1549.
Jung, H.Y., S. Miyata, K. Oshima, S. Kakizawa, H. Nishigawa, W. Wei, S.
Suzuki, M. Ugaki, T. Hibi and S. Namba. 2003a. First complete nucleotide sequence and heterologous gene organization of the two rRNA
operons in the phytoplasma genome. DNA Cell Biol. 22: 209–215.
Jung, H.Y., T. Sawayanagi, S. Kakizawa, H. Nishigawa, W. Wei,
K. Oshima, S. Miyata, M. Ugaki, T. Hibi and S. Namba. 2003b. ‘Candidatus Phytoplasma ziziphi’, a novel phytoplasma taxon associated
with jujube witches’-broom disease. Int. J. Syst. Evol. Microbiol. 53:
1037–1041.
Jung, H.Y., T. Sawayanagi, P. Wongkaew, S. Kakizawa, H. Nishigawa,
W. Wei, K. Oshima, S. Miyata, M. Ugaki, T. Hibi and S. Namba. 2003c.
‘Candidatus Phytoplasma oryzae’, a novel phytoplasma taxon associated
with rice yellow dwarf disease. Int. J. Syst. Evol. Microbiol. 53: 1925–1929.
Kakizawa, S., K. Oshima, T. Kuboyama, H. Nishigawa, H. Jung,
T. Sawayanagi, T. Tsuchizaki, S. Miyata, M. Ugaki and S. Namba. 2001.
Cloning and expression analysis of Phytoplasma protein translocation
genes. Mol. Plant Microbe Interact. 14: 1043–1050.
Kakizawa, S., K. Oshima, H. Nighigawa, H.Y. Jung, W. Wei, S. Suzuki,
M. Tanaka, S. Miyata, M. Ugaki and S. Namba. 2004. Secretion of
immunodominant membrane protein from onion yellows phytoplasma through the Sec protein-translocation system in Escherichia
coli. Microbiology 150: 135–142.
Kakizawa, S., K. Oshima, Y. Ishii, A. Hoshi, K. Maejima, H.Y. Jung, Y.
Yamaji and S. Namba. 2009. Cloning of immunodominant membrane protein genes of phytoplasmas and their in planta expression.
Kamińska, M. and H. liwa. 2003. Effect of antibiotics on symptoms
of stunting disease of Magnolia lilliflora plants. J. Phytopathol. 151:
59–63.
Kawakita, H., T. Saiki, W. Wei, W. Mitsuhashi, K. Watanabe and M. Sato.
2000. Identification of mulberry dwarf phytoplasmas in the genital
organs and eggs of leafhopper Hishimonoides sellatiformis. Phytopathology 90: 909–914.
Kenyon, L., N.A. Harrison, G.R. Ashburner, E.R. Boa and P.A. Richardson. 1998. Detection of a pigeon pea witches’ broom-related phytoplasma in trees of Gliricidia sepium affected by little-leaf disease in
Central America. Plant Pathol. 47: 671–680.
Khan, A.J., S. Botti, S. Paltrinieri, A.M. Al-Subhi and A.F. Bertaccina. 2002.
Phytoplasmas in alfalfa seedlings: infected or contaminated seed?
Proceedings of the 14th International Congress of the International
Organization for Mycoplasmology, Vienna, Austria, p. 148.
Kirkpatrick, B.C., D.C. Stenger, T.J. Morris and A.H. Purcell. 1987.
Cloning and detection of DNA from a nonculturable plant pathogenic Mycoplasma-like organism. Science 238: 197–200.
Kirkpatrick, B.C. 1989. Strategies for characterizing plant pathogenic
mycoplasma-like organisms and their effects on plants. In PlantMicrobe Interactions, Molecular and Genetic Perspectives (edited by
Kosuge and Nester). McGraw-Hill, New York, pp. 241–293.
Kirkpatrick, B.C. 1992. Mycoplasma-like organisms: plant and invertebrate pathogens. In The Prokaryotes: a Handbook on the Biology of
Bacteria: Ecophysiology, Isolation, Identification, Applications, 2nd
edn, vol. 4 (edited by Balows, Trüper, Dworkin, Harder and Schleifer). Springer, New York, pp. 4050–4067.
Kirkpatrick, B.C., N.A. Harrison, I.-M. Lee, H. Neimark and B.B. Sears.
1995. Isolation of Mycoplasma-like organism DNA from plant and
insect hosts. In Molecular and Diagnostic Procedures in Mycoplasmology, vol. 2 (edited by Razin and Tully). Academic Press, New
York, pp. 105–117.
Kison, H., B. Schneider and E. Seemüller. 1994. Restriction fragment
length polymorphisms within the apple proliferation mycoplasmalike organism. J. Phytopathol. 141: 395–401.
Kison, H., B.C. Kirkpatrick and E. Seemüller. 1997. Genetic comparison
of the peach yellow leaf roll agent with European fruit tree phytoplasmas of the apple proliferation group. Plant Pathol. Bull. 46: 538–544.
Kollar, A. and E. Seemüller. 1989. Base composition of the DNA of
mycoplasmalike organisms associated with various plant diseases.
J. Phytopathol. 127: 177–186.
Kollar, A. and E. Seemüller. 1990. Chemical composition of the phloem
exudate of Mycoplasma-infected trees. J. Phytopathol. 128: 99–111.
Koui, T., T. Natsuaki and S. Okuda. 2002. Antiserum raised against
gyrase A of Acholeplasma laidlawii reacts with phytoplasma proteins.
Koui, T., N. Tomohide and S. Okuda. 2003. Phylogenetic analysis of
elongation factor Tu gene of phytoplasmas from Japan. J. Gen. Plant
Pathol. 69: 316–319.
Kube, M., B. Schneider, H. Kuhl, T. Dandekar, K. Heitmann, A. M. Migdoll, R. Reinhardt and E. Seemüller. 2008. The linear chromosome
of the plant-pathogenic Mycoplasma ‘Candidatus Phytoplasma mali’.
BMC Genomics 9: 306.
Kuboyama, T., C.C. Huang, X. Lu, T. Sawayanagi, T. Kanazawa,
T. Kagami, I. Matsuda, T. Tsuchizaki and S. Namba. 1998. A plasmid isolated from phytopathogenic onion yellows phytoplasma and
its heterogeneity in the pathogenic phytoplasma mutant. Mol. Plant
Microbe Interact. 11: 1031–1037.
715
Kunkel, L.O. 1926. Studies on aster yellows. Am. J. Bot. 13: 646–705.
Kuske, C.R. and B.C. Kirkpatrick. 1990. Identification and characterization of plasmids from the western aster yellows mycoplasmalike
organism. J. Bacteriol. 172: 1628–1633.
Kuske, C.R., B.C. Kirkpatrick and E. Seemüller. 1991. Differentiation of
virescence MLOS using western aster yellows mycoplasma-like organism chromosomal DNA probes and restriction fragment length polymorphism analysis. J. Gen. Microbiol. 137: 153–159.
Kuske, C.R. and B.C. Kirkpatrick. 1992a. Distribution and multiplication
of western aster yellows mycoplasmalike organisms in Catharanthus
roseus as determined by DNA hybridization analysis. Phytopathology
82: 457–462.
Kuske, C.R. and B.C. Kirkpatrick. 1992b. Phylogenetic relationships
between the western aster yellows mycoplasmalike organism and
other prokaryotes established by 16S rRNA gene sequence. Int. J.
Lauer, U. and E. Seemüller. 2000. Physical map of the chromosome of
the apple proliferation phytoplasma. J. Bacteriol. 182: 1415–1418.
Lee, I.-M. and R.E. Davis. 1992. Mycoplasmas which infect plants and
insects. In Mycoplasmas: Molecular Biology and Pathogenesis (edited
Lee, I.-M., A. Bertaccini, M. Vibio and D.E. Gundersen. 1988. Detection and investigation of genetic relatedness among aster yellows
and other mycoplasmalike organisms by using cloned DNA and RNA
probes. Mol. Plant Microbe Interact. 1: 303–310.
Lee, I.-M., R.E. Davis, T.A. Chen, L.N. Chiykowski, J. Fletcher, C. Hiruki
and D.A. Schaff. 1992. A genotype-based system for identification
and classification of mycoplasmalike organisms (MLOs) in the aster
yellows MLO strain cluster. Phytopathology 82: 977–986.
Lee, I.-M., R.E. Davis and H.T. Hsu. 1993a. Differentiation of strains in
the aster yellows mycoplasmalike organism strain cluster by serological assay with monoclonal antibodies. Plant Dis. 77: 815–817.
Lee, I.-M., R.W. Hammond, R.E. Davis and D.E. Gundersen. 1993b.
Universal amplification and analysis of pathogen 16S rDNA for classification and identification of mycoplasmalike organisms. Phytopathology 83: 834–842.
Lee, I.-M., A. Bertaccini, M. Vibio and D.E. Gundersen. 1995. Detection
of multiple phytoplasmas in perennial fruit trees with decline symptoms in Italy. Phytopathology 85: 728–735.
Lee, I.-M., D.E. Gundersen-Rindal and A. Bertaccini. 1998a. Phytoplasma:
ecology and genomic diversity. Phytopathology 88: 1359–1366.
Lee, I.-M., D.E. Gundersen-Rindal, R.E. Davis and I.M. Bartoszyk.
1998b. Revised classification scheme of phytoplasmas based an RFLP
analyses of 16S rRNA and ribosomal protein gene sequences. Int. J.
Lee, I.-M., R.E. Davis and D.E. Gundersen-Rindal. 2000. Phytoplasma:
phytopathogenic Mollicutes. Annu. Rev. Microbiol. 54: 221–255.
Lee, I.-M., M. Martini, K.D. Bottner, R.A. Dane, M.C. Black and N. Troxclair. 2003. Ecological implications from a molecular analysis of phytoplasmas involved in an aster yellows epidemic in various crops in
Texas. Phytopathology 93: 1368–1377.
Lee, I.-M., D.E. Gundersen-Rindal, R.E. Davis, K.D. Bottner, C. Marcone
and E. Seemüller. 2004a. ‘Candidatus Phytoplasma asteris’, a novel
phytoplasma taxon associated with aster yellows and related diseases.
Int. J. Syst. Evol. Microbiol. 54: 1037–1048.
Lee, I.-M., M. Martini, C. Marcone and S.F. Zhu. 2004b. Classification of
phytoplasma strains in the elm yellows group (16SrV) and proposal
of ‘Candidatus Phytoplasma ulmi’ for the phytoplasma associated
with elm yellows. Int. J. Syst. Evol. Microbiol. 54: 337–347.
Lee, I.-M., Y. Zhao and K.D. Bottner. 2005. Novel insertion sequence-like
elements in phytoplasma strains of the aster yellows group are putative
new members of the IS3 family. FEMS Microbiol. Lett. 242: 353–360.
Lee, I.-M., K.D. Bottner, G. Secor and V. Rivera-Varas. 2006a. “Candidatus Phytoplasma americanum”, a phytoplasma associated with a
potato purple top wilt disease complex. Int. J. Syst. Evol. Microbiol.
56: 1593–1597.
716
Lee, I.-M., Y. Zhao and K.D. Bottner. 2006b. SecY gene sequence analysis for finer differentiation of diverse strains in the aster yellows phytoplasma group. Mol. Cell. Probes 20: 87–91.
Lefol, C., J. Lherminier, E. Boudon-Padieu, J. Larrue, C. Louis and A.
Caudwell. 1994. Propagation of Flavescence Dorèe MLO (mycoplasma-like organisms) in the leafhopper vector Euscelidius variegatus. Kbm. J. Invertebr. Pathol. 63: 285–293.
Lepka, P., M. Stitt, E. Moll and E. Seemüller. 1999. Effect of phytoplasmal infection on concentration and translocation of carbohydrates
and amino acids in periwinkle and tobacco. Physiol. Mol. Plant
Pathol. 55: 59–68.
Liefting, L.W., M.T. Andersen, R.E. Beever, R.C. Gardner and R.L.S.
Forster. 1996. Sequence heterogeneity in the two 16S rRNA genes
of Phormium yellow leaf phytoplasma. Appl. Environ. Microbiol. 62:
3133–3139.
Liefting, L.W. and B.C. Kirkpatrick. 2003. Cosmid cloning and sample
sequencing of the genome of the uncultivable mollicute, western
X-disease phytoplasma, using DNA purified by pulsed-field gel electrophoresis. FEMS Microbiol. Lett. 221: 203–211.
Liefting, L.W., M.E. Shaw and B.C. Kirkpatrick. 2004. Sequence analysis
of two plasmids from the phytoplasma beet leafhopper-transmitted
virescence agent. Microbiology 150: 1809–1817.
Liefting, L.W., M.T. Andersen, T.J. Lough and R.E. Beever. 2006. Comparative analysis of the plasmids from two isolates of “Candidatus Phytoplasma australiense”. Plasmid 56: 138–144.
Lim, P.O. and B.B. Sears. 1989. 16S rRNA sequence indicates that plantpathogenic mycoplasmalike organisms are evolutionarily distinct
from animal mycoplasmas. J. Bacteriol. 171: 5901–5906.
Lim, P.O. and B.B. Sears. 1992. Evolutionary relationships of a plantpathogenic mycoplasmalike organism and Acholeplasma laidlawii
deduced from two ribosomal protein gene sequences. J. Bacteriol.
174: 2606–2611.
Lim, P.O., B.B. Sears and K.L. Klomparens. 1992. Membrane properties
of a plant-pathogenic mycoplasmalike organism. J. Bacteriol. 174:
682–686.
Lin, C.-L., T. Zhou, H.-F. Li, Z.-F. Fan, Y. Li, C.-G. Piao and G.-Z. Tian.
2009. Molecular characterisation of two plasmids from paulownia
witches’-broom phytoplasma and detection of a plasmid encoded
protein in infected plants. Eur. J. Plant Pathol. 123: 321–330.
Lin, C.-Y., Chen, W.-Y., and C.P. Lin. 2006. Cloning and analysis of rpoC
gene of phytoplasma associated with peanut witches’ broom. Plant
Pathol. Bull. 15: 129–138.
Loi, N., P. Ermacora, T.A. Chen, L. Carraro and R. Osler. 1998. Monoclonal antibodies for the detection of tagetes witches’ broom agent.
J. Plant Pathol. 80: 171–174.
Loi, N., P. Ermacora, L. Carraro, R. Osler and T.A. Chen. 2002. Production of monoclonal antibodies against apple proliferation phytoplasma
and their use in serological detection. Eur. J. Plant Pathol. 108: 81–86.
Marcone, C., A. Ragozzino, B. Schneider, U. Lauer, C.D. Smart and
E. Seemüller. 1996. Genetic characterization and classification of two
phytoplasmas associated with spartium witches’ broom disease. Plant
Dis. 80: 365–371.
Marcone, C., F. Hergenhahn, A. Ragozzino and E. Seemüller. 1999a.
Dodder transmission of pear decline, European stone fruit yellows,
rubus stunt, Picris echioides yellows and cotton phyllody phytoplasmas
to periwinkle. J. Phytopathol. 147: 187–192.
Marcone, C., H. Neimark, A. Ragozzino, U. Lauer and E. Seemüller.
1999b. Chromosome sizes of phytoplasmas composing major phylogenetic groups and subgroups. Phytopathology 89: 805–810.
Marcone, C., I.-M. Lee, R.E. Davis, A. Ragozzino and E. Seemüller.
2000. Classification of aster yellows-group phytoplasmas based on
combined analyses of rRNA and tuf gene sequences. Int. J. Syst. Evol.
Microbiol. 50: 1703–1713.
Marcone, C., A. Ragozzino, I. Camele, G.L. Rana and E. Seemüller.
2001. Updating and extending genetic characterization and classification of phytoplasmas from wild and cultivated plants in southern
Italy. J. Plant Pathol. 83: 133–138.
Marcone, C. and E. Seemüller. 2001. A chromosome map of the
European stone fruit yellows phytoplasma. Microbiology 147:
1213–1221.
Marcone, C., K.S. Gibb, C. Streten and B. Schneider. 2004a. ‘Candidatus
Phytoplasma spartii’, ‘Candidatus Phytoplasma rhamni’ and ‘Candidatus Phytoplasma allocasuarinae’, respectively associated with spartium witches’-broom, buckthorn witches’-broom and allocasuarina
yellows diseases. Int. J. Syst. Evol. Microbiol. 54: 1025–1029.
Marcone, C., B. Schneider and E. Seemüller. 2004b. ‘Candidatus Phytoplasma cynodontis’, the phytoplasma associated with Bermuda grass
white leaf disease. Int. J. Syst. Evol. Microbiol. 54: 1077–1082.
Martinez, S., I. Cordova, B.E. Maust, C. Oropeza and J.M. Santamaria.
2000. Is abscisic acid responsible for abnormal stomatal closure in coconut palms showing lethal yellowing? J. Plant Physiol. 156: 319–322.
Martini, M., I.M. Lee, K.D. Bottner, Y. Zhao, S. Botti, A. Bertaccini,
N.A. Harrison, L. Carraro, C. Marcone, A.J. Khan and R. Osler. 2007.
Ribosomal protein gene-based phylogeny for finer differentiation
and classification of phytoplasmas. Int. J. Syst. Evol. Microbiol. 57:
2037–2051.
Marzachi, C., R.G. Milne and D. Bosco. 2004. Phytoplasma-plantvector relationships. In Recent Research Developments in Plant
Pathology, vol. 3 (edited by Pandalai). Research Signpost, Trivandrum, India.
McCoy, R.E. 1979. Mycoplasmas and yellows diseases. In The Mycoplasmas, vol. III, Plant and Insect Mycoplasmas (edited by Whitcomb and
McCoy, R.E. 1982. Use of tetracycline antibiotics to control yellows diseases. Plant Dis. 66: 539–542.
McCoy, R.E., A. Caudwell, C.J. Chang, T.A. Chen, L.N. Chiykowski,
M.T. Cousin, J.L. Dale, G.T.N. deLeeuw, D.A. Golino, K.J. Hackett,
B.C. Kirkpatrick, R. Marwitz, H. Petzold, R.C. Sinha, M. Suguira,
R.F. Whitcomb, I.L. Yang, B.M. Zhu and E. Seemüller. 1989. Plant
diseases associated with mycoplasma-like organisms. In The Mycoplasmas, vol. V (edited by Whitcomb and Tully). Academic Press, San
Diego, pp. 545–640.
Melamed, S., E. Tanne, R. Ben-Haim, O. Edelbaum, D. Yogev and
I. Sela. 2003. Identification and characterization of phytoplasmal
genes, employing a novel method of isolating phytoplasmal genomic
DNA. J. Bacteriol. 185: 6513–6521.
Miyata, S., K. Furuki, K. Oshima, T. Sawayanagi, H. Nishigawa, S. Kakizawa, H.Y. Jung, M. Ugaki and S. Namba. 2002a. Complete nucleotide
sequence of the S10-spc operon of phytoplasma: Gene organization
and genetic code resemble those of Bacillus subtilis. DNA Cell Biol.
21: 527–534.
Miyata, S., K. Furuki, T. Sawayanagi, K. Oshima, T. Kuboyama,
T. Tsuchizaki, M. Ugaki and S. Namba. 2002b. The gene arrangement
and sequence of str operon of phytoplasma resemble those of Bacillus
more than those of Mycoplasma. J. Gen. Plant Pathol. 68: 62–67.
Miyata, S., K. Oshima, S. Kakizawa, H. Nishigawa, H.Y. Jung,
T. Kuboyama, M. Ugaki and S. Namba. 2003. Two different thymidylate kinase gene homologues, including one that has catalytic activity,
are encoded in the onion yellows phytoplasma genome. Microbiology 149: 2243–2250.
Montano, H.G., R.E. Davis, E.L. Dally, S. Hogenhout, J.P. Pimentel and
P.S. Brioso. 2001. ‘Candidatus Phytoplasma brasiliense’, a new phytoplasma taxon associated with hibiscus witches’ broom disease. Int. J.
Moriwaki, N., K. Matsuchita, M. Nishina and Y. Kono. 2003. High concentrations of trehalose in aphid hemolymph Appl. Entomol. Zool.
38: 241–248.
Morton, A., D.L. Davies, C.L. Blomquist and D.J. Barbara. 2003. Characterization of homologues of the apple proliferation immunodominant membrane protein gene from three related phytoplasmas. Mol.
Mounsey, K.E., C. Streten and K.S. Gibb. 2006. Sequence characterization of four putative membrane-associated proteins from sweet
potato little strain V4 phytoplasma. Plant Pathol. 55: 29–35.
Murray, R.G.E., D.J. Brenner, R.R. Colwell, P. de Vos, P. Goodfellow,
P.A.D. Grimont, N. Pfennig, E. Stackebrandt and G.A. Zavarin. 1990.
Report of the ad hoc committee on approaches to taxonomy within
the proteobacteria. Int. J. Syst. Bacteriol. 40: 213–215.
Murray, R.G.E. and K.H. Schleifer. 1994. Taxonomic notes: a proposal
for recording the properties of putative taxa of procaryotes. Int. J.
Musetti, R., M.A. Favali and L. Pressacco. 2000. Histopathology and
polyphenol content in plants infected by phytoplasmas. Cytobios
102: 133–147.
Musetti, R. and M.A. Favali. 2003. Cytochemical localization of calcium
and X-ray microanalysis of Catharanthus roseus L. infected with phytoplasmas. Micron 34: 387–393.
Musetti, R., L.S. Di Toppi, P. Ermacora and M.A. Favali. 2004. Recovery in apple trees infected with the apple proliferation phytoplasma: an ultrastructural and biochemical study. Phytopathology
94: 203–208.
Nakashima, K. and T. Hayashi. 1997. Sequence analysis of extrachromosomal DNA of sugarcane white leaf phytoplasma. Ann. Phytopathol.
Soc. Jpn. 63: 21–25.
Namba, S., H. Oyaizu, S. Kato, S. Iwanami and T. Tsuchizaki. 1993. Phylogenetic diversity of phytopathogenic mycoplasmalike organisms.
Namba, S. 2002. Molecular biological studies on phytoplasmas. J. Gen.
Nasu, S., D.D. Jensen and J. Richardson. 1970. Electron microscopy
of mycoplasma-like bodies associated with insect and plant hosts of
peach western X-disease. Virology 41: 583–595.
Nasu, S., D. D. Jensen and J. Richardson. 1974. Primary culture of the
western X-disease mycoplasma-like organism from Colladonus montanus leafhopper vectors. Appl. Entomol. Zool. 9: 115–126.
Neimark, H. and B.C. Kirkpatrick. 1993. Isolation and characterization
of full-length chromosomes from non-culturable plant-pathogenic
mycoplasma-like organisms. Mol. Microbiol. 7: 21–28.
Nielson, M.W. 1979. Taxonomic relationships of leafhopper vectors of plant
pathogens. In Leafhopper Vectors and Plant Disease Agents (edited by
Maromorosch and Harris). Academic Press, New York, pp. 3–27.
Nipah, J.O., P. Jones and M.J. Dickinson. 2007. Detection of lethal yellowing phytoplasmas in embryos from coconuts infected with Cape
St. Paul wilt disease in Ghana. Plant Pathol. 56: 777–784.
Nishigawa, H., S. Miyata, K. Oshima, T. Sawayanagi, A. Komoto,
T. Kuboyama, I. Matsuda, T. Tsuchizaki and S. Namba. 2001. In planta
expression of a protein encoded by the extrachromosomal DNA of a
phytoplasma and related to geminivirus replication proteins. Microbiology 147: 507–513.
Nishigawa, H., K. Oshima, S. Kakizawa, H.Y. Jung, T. Kuboyama,
S. Miyata, M. Ugaki and S. Namba. 2002a. A plasmid from a non-insecttransmissible line of a phytoplasma lacks two open reading frames that
exist in the plasmid from the wild-type line. Gene 298: 195–201.
Nishigawa, H., K. Oshima, S. Kakizawa, H.Y. Jung, T. Kuboyama, S.
Miyata, M. Ugaki and S. Namba. 2002b. Evidence of intermolecular
recombination between extrachromosomal DNAs in phytoplasma: a
trigger for the biological diversity of phytoplasma? Microbiology 148:
1389–1396.
Nishigawa, H., K. Oshima, S. Miyata, M. Ugaki and S. Namba. 2003.
Complete set of extrachromosomal DNAs from three pathogenic
lines of onion yellows phytoplasma and use of PCR to differentiate
each line. J. Gen. Plant Pathol. 69: 194–198.
Nyland, G. 1971. Remission of symptoms of pear decline in pear and
peach X-disease in peach after treatment with a tetracycline. Phytopathology 61: 904–905.
Oparka, K.J. and R. Turgeon. 1999. Sieve elements and companion
cells-traffic control centers of the phloem. Plant Cell. 11: 739–750.
Oshima, K., S. Kakizawa, H. Nishigawa, T. Kuboyama, S. Miyata,
M. Ugaki and S. Namba. 2001a. A plasmid of phytoplasma encodes
a unique replication protein having both plasmid- and virus-like
domains: clue to viral ancestry or result of virus/plasmid recombination? Virology 285: 270–277.
717
Oshima, K., T. Shiomi, T. Kuboyama, T. Sawayanagi, H. Nishigawa,
S. Kakizawa, S. Miyata, M. Ugaki and S. Namba. 2001b. Isolation and
characterization of derivative lines of the onion yellows phytoplasma
that do not cause stunting or phloem hyperplasia. Phytopathology
91: 1024–1029.
Oshima, K., S. Miyata, T. Sawayanagi, S. Kakizawa, H. Nishigawa, H. Jung,
K. Furuki, M. Yanazaki, S. Suzuki, W. Wei, T. Kuboyama, M. Ugaki and
S. Namba. 2002. Minimal set of metabolic pathways suggested from the
genome of onion yellow phytoplasma. J. Plant Pathol. 68: 225–236.
Oshima, K., S. Kakizawa, H. Nishigawa, H.Y. Jung, W. Wei, S. Suzuki,
R. Arashida, D. Nakata, S. Miyata, M. Ugaki and S. Namba. 2004.
Reductive evolution suggested from the complete genome sequence
of a plant-pathogenic phytoplasma. Nat. Genet. 36: 27–29.
Oshima, K., S. Kakizawa, R. Arashida, Y. Ishii, A. Hoshi, Y. Hayashi,
S. Kakiwada and S. Namba. 2007. Presence of two glycolytic gene
clusters in a severe pathogenic line of Candidatus Phytoplasma asteris.
Mol. Plant Pathol. 8: 481–489.
Padovan, A.C., G. Firrao, B. Schneider and K.S. Gibb. 2000. Chromosome
mapping of the sweet potato little leaf phytoplasma reveals genome
heterogeneity within the phytoplasmas. Microbiology 146: 893–902.
Pracros, P., J. Renaudin, S. Eveillard, A. Mouras and M. Hernould. 2006.
Tomato flower abnormalities induced by stolbur infection are associated with changes of expression of floral development genes. Mol.
Rajan, J., M.F. Clark, M. Barba and A. Hadidi. 1995. Detection of apple
proliferation and other MLOs by immuno-capture PCR (IC-PCR).
Acta Hortic. 386: 511–514.
Raju, B.C. and G. Nyland. 1988. Chemotherapy of mycoplasma diseases
of fruit trees. In Tree Mycoplasmas and Mycoplasma Diseases (edited
by Hiruki). University of Alberta Press, Edmonton, Alberta, Canada,
pp. 207–216.
Reinert, W. 1999. Detection and further differentiation of plant pathogenic phytoplasmas (Mollicutes, Eubacteria) in Germany regarding
phytopathological aspects. PhD Dissertation. Dem Fachbereich Biologie der Technischen Universität Darmstadt (in German), p. 148.
Rekab, D., L. Carraro, B. Schneider, E. Seemüller, J. Chen, C.J. Chang,
R. Locci and G. Firrao. 1999. Geminivirus-related extrachromosomal
DNAs of the X-clade phytoplasmas share high sequence similarity.
Microbiology 145: 1453–1459.
Rudzinska-Langwald, A. and M. Kaminska. 1999. Cytopathological evidence
for transport of phytoplasma in infected plants. Bot. Pol. 68: 261–266.
Rudzinska-Langwald, A. and M. Kaminska. 2003. Changes in the ultrastructure and cytoplasmic free calcium in Gladiolus x hybridus Van
Houtte roots infected by aster yellows phytoplasma. Acta Soc. Bot.
Pol. 72: 269–282.
Saglio, P.H.M. and R.F. Whitcomb. 1979. Diversity of wall-less prokaryotes in plant vascular tissue, fungi and invertebrate animals. In The
Sawayanagi, T., N. Horikoshi, T. Kanehira, M. Shinohara, A. Bertaccini,
M.T. Cousin, C. Hiruki and S. Namba. 1999. ‘Candidatus Phytoplasma
japonicum’, a new phytoplasma taxon associated with Japanese
Hydrangea phyllody. Int. J. Syst. Bacteriol. 49: 1275–1285.
Schneider, B., U. Ahrens, B.C. Kirkpatrick and E. Seemüller. 1993.
Classification of plant-pathogenic mycoplasma-like organisms using
restriction-site analysis of PCR-amplified 16S rDNA. J. Gen. Microbiol. 139: 519–527.
Schneider, B. and E. Seemüller. 1994a. Presence of two sets of ribosomal genes in phytopathogenic mollicutes. Appl. Environ. Microbiol. 60: 3409–3412.
Schneider, B. and E. Seemüller. 1994b. Studies on taxonomic relationships of mycoplasma-like organisms by Southern blot analysis. J. Phytopathol. 141: 173–185.
Schneider, B., E. Seemüller, C.D. Smart and B.C. Kirkpatrick. 1995. Phylogenetic classification of plant pathogenic mycoplasmalike organisms or phytoplasmas. In Molecular and Diagnostic Procedures in
Mycoplasmology: Molecular Characterization, vol. 1 (edited by Razin
and Tully). Academic Press, San Diego, pp. 369–380.
718
Schneider, B., K.S. Gibb and E. Seemüller. 1997. Sequence and RFLP
analysis of the elongation factor Tu gene used in differentiation and
classification of phytoplasmas. Microbiology 143: 3381–3389.
Schneider, B., E. Torres, M.P. Martin, M. Schroder, H.D. Behnke and
E. Seemuller. 2005. ‘Candidatus Phytoplasma pini’, a novel taxon
from Pinus silvestris and Pinus halepensis. Int. J. Syst. Evol. Microbiol.
55: 303–307.
Sears, B.B. and K.L. Klomparens. 1989. Leaf tip cultures of the evening
primrose allow stable, aspectic culture of mycoplasma-like organism.
Can. J. Plant Pathol. 11: 343–348.
Sears, B.B. and B.C. Kirkpatrick. 1994. Unveiling the evolutionary relationships of plant pathogenic mycoplasmalike organisms. ASM News
60: 307–312.
Seddas, A., R. Meignoz, C. Kuszala and E. Boudon-Padieu. 1995. Evidence for the physical integrity of flavescence dorée phytoplasmas
purified by affinity chromatography by immunoaffinity from infected
plants or leafhoppers and the plant pathogenicity of phytoplasmas
from leafhoppers. Plant Pathol. 44: 971–978.
Seemüller, E. and B. Schneider. 2007. Differences in virulence and
genomic features of strains of ‘Candidatus Phytoplasma mali’, the
Apple Proliferation agent. Phytopathology 97: 964–970.
Seemüller, E., B. Schneider, R. Mäurer, U. Ahrens, X. Daire, H. Kison, K.H.
Lorenz, G. Firrao, L. Avinent, B.B. Sears and E. Stackebrandt. 1994.
Phylogenetic classification of phytopathogenic mollicutes by sequence
analysis of 16S ribosomal DNA. Int. J. Syst. Bacteriol. 44: 440–446.
Seemüller, E., C. Marcone, U. Lauer, A. Ragozzino and M. Göschl. 1998.
Current status of molecuar classification of the phytoplasmas. J. Plant
Pathol. 80: 3–26.
Seemüller, E., M. Garnier and B. Schneider. 2002. Mycoplasmas of
plants and insects. In Molecular Biology and Pathogenicity of Mycoplasmas (edited by Razin and Hermann). Kluwer Academic/Plenum
Publishers, Dordrecht, The Netherlands, pp. 91–116.
Seemüller, E. and B. Schneider. 2004. ‘Candidatus Phytoplasma mali’,
‘Candidatus Phytoplasma pyri’ and ‘Candidatus Phytoplasma prunorum’, the causal agents of apple proliferation, pear decline and
European stone fruit yellows, respectively. Int. J. Syst. Evol. Microbiol.
54: 1217–1226.
Shen, W.C. and C.P. Lin. 1993. Production of monoclonal antibodies against a mycoplasmalike organism associated with sweetpotato
witches’ broom. Phytopathology 83: 671–675.
Shen, W.C. and C.P. Lin. 1994. Application of immunofluorescent staining,
tissue blotting techniques against a mycoplasmalike organism assoicated
with sweetpotato witches’ broom. Plant Pathol. Bull. 3: 79–83.
Siddique, A.B.M., J.N. Giuthrie, K.B. Walsh, D.T. White and P.T. Scott.
1998. Histopathology and within-plant distribution of the phytoplasma associated with Australian papaya dieback. Plant Dis. 82:
1112–1120.
Siller, W., B. Kuhbandner, R. Marwitz, H. Petzold and E. Seemüller.
1987. Occurrence of mycoplasma-like organisms in parenchyma cells
of Cuscuta odorata (Ruiz et Pav.). J. Phytopathol. 119: 147–159.
Sinclair, W.A., H.M. Griffiths and I.M. Lee. 1994. Mycoplasmalike
organisms as causes of slow growth and decline of trees and shrubs.
J. Arboric. 20: 176–189.
Sinha, R.C. and E.A. Peterson. 1972. Uptake and persistence of oxytetracycline in aster plants and vector leafhoppers in relation to inhibition of clover phyllody agent. Phytopathology 62: 377–383.
Sinha, R.C. 1979. Purification and serology of mycoplasma-like organisms from aster yellows-infected plants. Can. J. Plant Pathol. 1: 65–70.
Sjölund, R.D. 1997. The phloem sieve element: a river runs through it.
Plant Cell 9: 1137–1146.
Smart, C.D., B. Schneider, C.L. Blomquist, L.J. Guerra, N.A. Harrison,
U. Ahrens, K.H. Lorenz, E. Seemuller and B.C. Kirkpatrick. 1996.
Phytoplasma-specific PCR primers based on sequences of the 16S–
23S rRNA spacer region. Appl. Environ. Microbiol. 62: 2988–2993.
Smith, A.J., R.E. McCoy and J.H. Tsai. 1981. Maintenance in vitro of
the aster yellows mycoplasmalike organism. Phytopathology 71:
819–822.
Stackebrandt, E. and B.M. Goebel. 1994. Taxonomic note: a place for
DNA–DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst. Bacteriol. 44: 846–849.
Streten, C. and K.S. Gibb. 2003. Identification of genes in the tomato
big bud phytoplasma and comparison to those in sweet potato little
leaf-V4 phytoplasma. Microbiology 149: 1797–1805.
Suzuki, S., K. Oshima, S. Kakizawa, R. Arashida, H.Y. Jung, Y. Yamaji, H. Nishigawa, M. Ugaki and S. Namba. 2006. Interaction between the membrane
protein of a pathogen and insect microfilament complex determines
insect-vector specificity. Proc. Natl. Acad. Sci. U. S. A. 103: 4252–4257.
Swofford, D.L. 1998. PAUP: Phylogenetic analysis using parsimony and
other methods, 4th edn. Sinauer Associates, Sunderland, MA.
Tan, P.K. and T. Whitlow. 2001. Physiological responses of Catharanthus
roseus (periwinkle) to ash yellows phytoplasmal infection. New Phytol. 150: 759–769.
Tanne, E., E. Boudon-Padieu, D. Clair, M. Davidovich, S. Melamed and
M. Klein. 2001. Detection of phytoplasma by polymerase chain reaction of insect feeding medium and its use in determining vectoring
ability. Phytopathology 91: 741–746.
Tedeschi, R., V. Ferrato, J. Rossi and A. Alma. 2006. Possible phytoplasma transovarial transmission in the psyllids Cacopsylla melanoneura and Cacopsylla pruni. Plant Pathol. 55: 18–24.
Thomas, D.L. 1979. Mycoplasmalike bodies associated with lethal
declines of palms in Florida. Phytopathology 69: 928–934.
Thomas, S. and M. Balasundaran. 2001. Purification of sandal spike
phytoplasma for the production of polyclonal antibody. Curr. Sci. 80:
1489–1494.
Toth, K.F., N. Harrison and B.B. Sears. 1994. Phylogenetic relationships among members of the class Mollicutes deduced from rps3 gene
sequences. Int. J. Syst. Bacteriol. 44: 119–124.
Tran-Nguyen, L.T. and K.S. Gibb. 2006. Extrachromosomal DNA isolated from tomato big bud and Candidatus Phytoplasma australiense
phytoplasma strains. Plasmid 56: 153–166.
Tran-Nguyen, L.T., M. Kube, B. Schneider, R. Reinhardt and K.S. Gibb.
2008. Comparative genome analysis of “Candidatus Phytoplasma australiense” (subgroup tuf-Australia I; rp-A) and “Ca. Phytoplasma asteris” strains OY-M and AY-WB. J. Bacteriol. 190: 3979–3991.
Tsai, J.H. 1979. Vector transmission of mycoplasmal agents of plant diseases. In The Mycoplasmas, vol. III, Plant and Insect Mycoplasmas
(edited by Whitcomb and Tully). Academic Press, New York, pp.
266–307.
Valiunas, D., J. Staniulis and R.E. Davis. 2006. ‘Candidatus Phytoplasma fragariae’, a novel phytoplasma taxon discovered in yellows
diseased strawberry, Fragaria x ananassa. Int. J. Syst. Evol. Microbiol.
56: 277–281.
Van Helden, M., W.F. Tjallinghii and T.A. Van Beek. 1994. Phloem collection from lettuce (Lactuca sativa L.): Chemical comparison among
collection methods. J. Chem. Ecol. 20: 3191–3206.
Verdin, E., P. Salar, J.L. Danet, E. Choueiri, F. Jreijiri, S. El Zammar,
B. Gelie, J.M. Bové and M. Garnier. 2003. ‘Candidatus Phytoplasma
phoenicium’ sp. nov., a novel phytoplasma associated with an emerging lethal disease of almond trees in Lebanon and Iran. Int. J. Syst.
Wagner, M., C. Fingerhut, H.J. Gross and A. Schön. 2001. The
first phytoplasma RNase P RNA provides new insights into the
sequence requirements of this ribozyme. Nucleic Acids Res. 29:
2661–2665.
Wang, K. and C. Hiruki. 2000. Heteroduplex mobility assay detects DNA
mutations for differentiation of closely related phytoplasma strains.
J. Microbiol. Methods 41: 59–68.
Waters, H. and P. Hunt. 1980. The in vivo three-dimensional form of
a plant mycoplasma-like organism revealed by the analysis of serial
ultra-thin sections. J. Gen. Microbiol. 116: 111–131.
Webb, D.R., R.G. Bonfiglioli, L. Carraro, R. Osler and R.H. Symons.
1999. Oligonucleotides as hybridization probes to localize phytoplasmas in host plants and insect vectors. Phytopathology 89:
894–901.
Order IV. Anaeroplasmatales
Wei, W., S. Kakizawa, H.Y. Jung, S. Suzuki, M. Tanaka, H. Nishigawa, S.
Miyata, K. Oshima, M. Ugaki, T. Hibi and S. Namba. 2004a. An antibody against the SecA membrane protein of one phytoplasma reacts
with those of phylogenetically different phytoplasmas. Phytopatho
logy 94: 683–686.
Wei, W., S. Kakizawa, S. Suzuki, H.Y. Jung, H. Nishigawa, S. Miyata,
K. Oshima, M. Ugaki, T. Hibi and S. Namba. 2004b. In planta dynamic
analysis of onion yellows phytoplasma using localized inoculation by
insect transmission. Phytopathology 94: 244–250.
Wei, W., R.E. Davis, I.M. Lee and Y. Zhao. 2007. Computer-simulated
RFLP analysis of 16S rRNA genes: identification of ten new phytoplasma groups. Int. J. Syst. Evol. Microbiol. 57: 1855–1867.
Wei, W., R.E. Davis, R. Jomantiene and Y. Zhao. 2008a. Ancient, recurrent phage attacks and recombination shaped dynamic sequencevariable mosaics at the root of phytoplasma genome evolution. Proc.
Natl. Acad. Sci. U. S. A. 105: 11827–11832.
Wei, W., I.M. Lee, R.E. Davis, X. Suo and Y. Zhao. 2008b. Automated
RFLP pattern comparison and similarity coefficient calculation for
rapid delineation of new and distinct phytoplasma 16Sr subgroup
lineages. Int. J. Syst. Evol. Microbiol. 58: 2368–2377.
Weintraub, P.G. and L. Beanland. 2006. Insect vectors of phytoplasmas.
Annu. Rev. Entomol. 51: 91–111.
Weisburg, W., J. Tully, D. Rose, J. Petzel, H. Oyaizu, D. Yang, L. Mandelco, J. Sechrest, T. Lawrence and J. Van Etten. 1989. A phylogenetic
analysis of the mycoplasmas: basis for their classification. J. Bacteriol.
171: 6455–6467.
719
Whitcomb, R.F., D.D. Jensen and J. Richardson. 1966a. The infection of
leafhoppers by western X-disease. virus: II. Fluctuation of virus concentration in the hemolymph after injection. Virology 28: 454–458.
Whitcomb, R.F., D.D. Jensen and J. Richardson. 1966b. The infection of
leafhoppers by the western X-disease virus: I. Frequency of transmission after injection or acquisition feeding. Virology 28: 448–453.
White, D.T., L.L. Blackall, P.T. Scott and K.B. Walsh. 1998. Phylogenetic
positions of phytoplasmas associated with dieback, yellow crinkle and
mosaic diseases of papaya, and their proposed inclusion in ‘Candidatus Phytoplasma australiense’ and a new taxon, ‘Candidatus Phytoplasma australasia’. Int. J. Syst. Bacteriol. 48: 941–951.
Wongkaew, P. and J. Fletcher. 2004. Sugarcane white leaf phytoplasma
in tissue culture: long-term maintenance, transmission, and oxytetracycline remission. Plant Cell Rep. 23: 426–434.
Yu, Y.L., K.W. Yeh and C.P. Lin. 1998. An antigenic protein gene of a
phytoplasma associated with sweet potato witches’ broom. Microbiology 144: 1257–1262.
Zhao, Y., Q. Sun, W. Wei, R.E. Davis, W. Wu and Q. Liu. 2009. ‘Candidatus Phytoplasma tamaricis’, a novel taxon discovered in witches’broom-diseased salt cedar (Tamarix chinensis Lour.). Int. J. Syst. Evol.
Microbiol. 59: 2496–2504.
Zreik, L., P. Carle, J.M. Bové and M. Garnier. 1995. Characterization
of the mycoplasmalike organism associated with witches’ broom disease of lime and proposition of a Candidatus taxon for the organism, “Candidatus Phytoplasma aurantifolia”. Int. J. Syst. Bacteriol. 45:
449–453.
Order IV. Anaeroplasmatales Robinson and Freundt 1987, 81VP
A.na.e.ro.plas.ma.ta¢les. N.L. neut. n. Anaeroplasma, -atos type genus of the order; -ales
ending to denote an order; N.L. fem. pl. n. Anaeroplasmatales the Anaeroplasma order.
This order in the class Mollicutes represents a unique group of
strictly anaerobic, wall-less prokaryotes (trivial name, anaeroplasmas) first isolated from the bovine and ovine rumen.
Other than their anaerobiosis, the description of organisms
in the order is essentially the same as for the class. A single
family, Anaeroplasmataceae, with two genera, was proposed
to recognize the two most prominent characteristics of the
organisms: a requirement of sterol supplements for growth by
those strictly anaerobic organisms now assigned to the genus
Anaeroplasma; and strictly anaerobic growth in the absence of
sterol supplements by those now assigned to the genus Asteroleplasma. Genome sizes range from 1542 to 1794 kbp as estimated by renaturation kinetics. The DNA G+C content ranges
from 29 to 40 mol%. All species examined utilize the universal genetic code in which UGA is a stop codon. Phylogenetic
studies indicate that members of the Anaeroplasmatales are
much more closely related to the Acholeplasmatales than to the
Mycoplasmatales or Entomoplasmatales (Weisburg et al., 1989).
Type genus: Anaeroplasma Robinson, Allison and Hartman
1975, 179AL.
The initial proposal for elevation of the anaeroplasmas to an
order of the class Mollicutes (Robinson and Freundt, 1987)
was based upon the description of three novel species and the
observation that some anaeroplasmas did not have a sterol
requirement for growth.
The obligate requirement for anaerobic growth conditions is the single most important property in distinguishing
members of the Anaeroplasmatales from other mollicutes. The
anaeroplasmas exist in a natural environment where the oxidation potential is maintained at a low level by the metabolism of associated micro-organisms. Anaerobic methods for
preparing media and culture techniques for the organisms
are essentially those described by Hungate (1969), with
media and inocula maintained in closed vessels and exposure to air avoided during inoculation and incubation. A
primary isolation medium and clarified rumen fluid broth
have been described (Bryant and Robinson, 1961; Robinson,
1983; Robinson et al., 1975).
References
Bryant, M.P. and I.M. Robinson. 1961. An improved nonselective culture medium for ruminal bacteria and its use in determining diurnal variation in numbers of bacteria in the rumen. J. Dairy Sci. 44:
1446–1456.
Hungate, R.E. 1969. A roll tube method for cultivation of strict anaerobes. In Methods in Microbiology, vol. 3B (edited by Norris and
Ribbons). Academic Press, London, pp. 117–132.
Robinson, I.M. 1983. Culture media for anaeroplasmas. In Methods in
Mycoplasmology, vol. 1, (edited by Razin and Tully). Academic Press,
Robinson, I.M. and E.A. Freundt. 1987. Proposal for an amended
classification of anaerobic mollicutes. Int. J. Syst. Bacteriol. 37:
78–81.
Weisburg, W., J. Tully, D. Rose, J. Petzel, H. Oyaizu, D. Yang, L. Mandelco,
J. Sechrest, T. Lawrence and J. Van Etten. 1989. A phylogenetic analysis of the mycoplasmas: basis for their classification. J. Bacteriol. 171:
6455–6467.
720
Family I. Anaeroplasmataceae
Family I. Anaeroplasmataceae Robinson and Freundt 1987, 80VP
A.na.e.ro.plas.ma.ta.ce¢ae. N.L. neut. n. Anaeroplasma, -atos type genus of the family; -aceae ending
to denote a family; N.L. fem. pl. n. Anaeroplasmataceae the Anaeroplasma family.
All members have an obligate requirement for anaerobiosis.
Organisms assigned to the genus Anaeroplasma require sterol
supplements for growth. Organisms assigned to the genus Asteroleplasma grow in the absence of sterol supplements. Other
characteristics are as described for the type genus.
Type genus: Anaeroplasma Robinson, Allison and Hartman
1975, 179AL.
The obligate requirement for anaerobic growth conditions and
for growth only in media containing cholesterol is established
with the methods described by Hungate (1969), with media
and inocula maintained in closed vessels and exposure to air
avoided during inoculation and incubation. The primary isolation medium and the clarified rumen fluid broth supplemented
with cholesterol have been described (Robinson, 1983).
Genus I. Anaeroplasma Robinson, Allison and Hartman 1975, 179AL
A.na.e.ro.plas¢ma. Gr. prefix an without; Gr. masc. n. aer air; Gr. neut. n. plasma a form; N.L. neut. n. Anaeroplasma intended to denote “anaerobic mycoplasma”.
Cells are predominantly coccoid, about 500 nm in diameter;
clusters of up to ten coccoid cells may be joined by short filaments. Older cells have a variety of pleomorphic forms. Cells
lack a cell wall and are bound by a single plasma membrane.
Gram-stain-negative due to absence of cell wall. Obligately
anaerobic; the inhibitory effect of oxygen on growth is not
alleviated during repeated subcultures. Require sterol supplements for growth. Nonmotile. Optimal temperature, 37°C; no
growth at 26 or 47°C. Optimal pH, 6.5–7.0. Surface colonies
have a dense center with a translucent periphery, or “fried-egg”
appearance. Subsurface colonies are golden, irregular, and
often multilobed. Strains vary in their ability to ferment various carbohydrates. The products of carbohydrate fermentation
include acids (generally acetic, formic, propionic, lactic, and
succinic), ethanol, and gases (primarily CO2, but some strains
also produce H2). Bacteriolytic and nonbacteriolytic strains have
been described. Commensals in the bovine and ovine rumen.
Type species: Anaeroplasma abactoclasticum Robinson, Allison
and Hartman 1975, 179AL.
Cells of Anaeroplasma examined by phase-contrast microscopy
appear as single cells, clumps, dumbbell forms, and clusters
of coccoid forms joined by short filaments. In electron micrographs of negatively stained preparations, pleomorphic forms
are observed; these include filamentous cells, budding cells,
and cells with bleb-like structures.
All species examined have similar fermentation products of
acetate, formate, lactate, ethanol, and carbon dioxide (Robinson et al., 1975). Anaeroplasma abactoclasticum is the only species
known not to digest casein. Anaeroplasma abactoclasticum strains
are the only ones known to produce succinate through fermentation. Anaeroplasma bactoclasticum, Anaeroplasma intermedium,
and Anaeroplasma varium are the only species known to produce
hydrogen and propionate during their fermentation.
The roll-tube anaerobic culture technique (Hungate,
1969), with pre-reduced medium maintained in a system for
exclusion of oxygen, is used to culture the organisms (Robinson, 1983; Robinson and Allison, 1975; Robinson et al., 1975;
Robinson and Hungate, 1973). Anaerobic mollicutes in a sewage sludge digester were cultured in an anaerobic cabinet
(Rose and Pirt, 1981). Although it is possible that other types
of anaerobic culture techniques might be acceptable (anaerobic culture jar or GasPak system), the effective use of such
equipment has not been demonstrated (Robinson, 1983).
Strains with bacteriolytic activity are detected with the addition of autoclaved Escherichia coli cells to the Primary Isolation
Medium (PIM) described below. Clear zones around colonies
of anaeroplasmas, when viewed by a stereoscopic microscope,
are suggestive of bacteriolytic anaeroplasmas. Colonies can be
subcultured to clarified rumen fluid broth (CRFB) medium
described below.
A slide agglutination test was first used to show that the antigens of anaerobic mollicutes were not related to established
Mycoplasma or Acholeplasma species found in cattle (Robinson
and Hungate, 1973). Later, the agglutination test was adapted
to either a plate or tube test and combined with an agar gel
diffusion test and a modified growth inhibition procedure to
examine the antigenic interrelationships among the anaerobic mollicutes (Robinson and Rhoades, 1977). On the basis
of these tests, a serological grouping of anaerobic mollicutes
appeared compatible with the group separations based upon
cultural, biochemical, and biophysical properties of the organisms (Robinson, 1979; Robinson and Rhoades, 1977).
There is no current evidence for the pathogenicity of any of
the Anaeroplasma species described so far. Obligately anaerobic
mollicutes appear to be a heterogeneous group that has been
found so far only in the rumen of cattle and sheep (Robinson,
1979; Robinson et al., 1975). Each new isolated group of these
organisms seems to have different properties, suggesting that
additional undescribed species are likely to exist. The ecological
role of these organisms in the rumen has not been determined.
Although the titer of these organisms in the rumen appears
to be low when compared to titers of other rumen organisms,
the mollicutes probably contribute to the pool of microbial
fermentation products at that site. Growth of anaeroplasmas is
inhibited by thallium acetate (0.2%), bacitracin (1000 mg/ml),
streptomycin (200 mg/ml), and d-cycloserine (500 mg/ml),
but not by benzylpenicillic acid (1000 U/ml).
Genus I. Anaeroplasma
The PIM medium used to grow and detect anaerobic mycoplasmas (Robinson, 1983; Robinson et al., 1975; Robinson and Hungate, 1973) contains: 40% (v/v) rumen fluid strained through
cheesecloth, autoclaved, and clarified by centrifugation; 0.05%
(w/v) glucose; 0.05% (w/v) cellobiose; 0.05% (w/v) starch;
3.75% (v/v) of a mineral solution consisting of 1.7 × 10−3 M
K2HPO4, 1.3 × 10−3 M KH2PO4, 7.6 × 10−4 M NaCl, 3.4 × 10−3 M
(NH4)2SO4, 4.1 × 10−4 M CaCl2, and 3.8 × 10−4 M MgSO4·7H2O;
0.2% (w/v) trypticase; 0.1% (w/v) yeast extract; 0.0001% (w/v)
resazurin; 0.5% (w/v) autoclaved Escherichia coli cells; 0.4% (w/v)
Na2CO3; 0.05% (w/v) cysteine hydrochloride; 1.5% (w/v) agar;
and 0.0006% (w/v) benzylpenicillic acid. Pure cultures are established by picking individual colonies from PIM roll tubes and subculturing into CRFB medium. CRFB medium contains the same
ingredients and concentrations as PIM, except: glucose, cellobiose, and starch concentrations are 0.2% (w/v), and autoclaved
Escherichia coli cells, agar, and benzylpenicillic acid are omitted.
Growth also occurs in a rumen fluid-free medium (Medium D)
in which growth factors supplied in rumen fluid are replaced by
lipopolysaccharide (Boivin; Difco) and cholesterol (Robinson,
1983; Robinson et al., 1975), or in a completely defined medium
in which the trypticase, yeast extract, and lipopolysaccharide of
Medium D are replaced by amino acids, vitamins, and phosphatidylcholine esterified with unsaturated fatty acids (Robinson,
1979, 1983). An agar-overlay plating technique carried out in an
anaerobic hood has also been reported to be an effective isolation procedure (Robinson, 1979). Anaerobic mollicutes grow
only in a prereduced medium maintained in a system for exclusion of oxygen. When resazurin is used in the test medium and
becomes oxidized, mollicutes will fail to grow.
Cultures are viable after storage for as long as 5 years at −40°C in
CRFB medium. They may also be preserved by lyophilization using
standard techniques for other mollicutes (Leach, 1983). However,
the type strains of several species of Anaeroplasma are no longer
available from the American Type Culture Collection because it
was impossible to revive the cultures sent by the depositors.
Differentiation of the genus Anaeroplasma
from other genera
sequences. The obligately anaerobic nature of Anaeroplasma species is a distinctive and stable characteristic among these organisms. Strictly anaerobic growth plus the requirement for sterol
supplements for growth exclude assignment to any other taxon
in the class. Moreover, the bacteriolytic capability possessed by
some of the anaeroplasmas has not been reported for other
mycoplasmas. Plasmalogens (alkenyl-glycerol ethers), which
are found in various anaerobic bacteria but not in aerobic bacteria, are major components of polar lipids from anaeroplasmas
(Langworthy et al., 1975); this further supports the contention
that anaeroplasmas are distinct from other mollicutes.
Taxonomic comments
The first organism in the group to be described was referred to
as Acholeplasma bactoclasticum (type strain JRT = ATCC 27112T;
Robinson and Hungate, 1973) because the organism was
721
thought to lack a sterol requirement for growth. Later, when
other obligately anaerobic mollicutes were isolated, these and
the JRT strain were found to require sterol for growth. A proposal was then made to form the new genus Anaeroplasma to
accommodate strain JRT (as Anaeroplasma bactoclasticum; Robinson and Allison, 1975) and a second anaerobic mollicute
designated Anaeroplasma abactoclasticum (Robinson et al., 1975).
These developments prompted a proposal for an amended classification of anaerobic mollicutes, which included descriptions
of Anaeroplasma varium and Anaeroplasma intermedium, the family
Anaeroplasmataceae, and the order Anaeroplasmatales within the
class Mollicutes (Robinson and Freundt, 1987).
Early serological studies suggested the existence of several
distinct species of anaeroplasmas. Subsequent reports on DNA–
DNA hybridization, DNA base composition, and genome size
comparisons of organisms in the group also indicated the existence of a number of species in two distinct genera of anaerobic mollicutes (Christiansen et al., 1986; Stephens et al., 1985).
Strains initially assigned to Acholeplasma abactoclasticum [serovar
3, type strain 6-1T (=ATCC 27879T)] were found to be a single species with about 80% interstrain DNA–DNA hybridization. However, strains A-2T (serovar 1) and 7LAT (serovar 2), previously
included in the description of Acholeplasma bactoclasticum, are the
type strains of separate species designated Anaeroplasma varium
and Anaeroplasma intermedium, respectively (Robinson and Freundt, 1987). Strains of Anaeroplasma all have DNA G+C contents
in the range 29.3–33.7 mol%, whereas the base composition of
serovar 4 strains 161T, 162, and 163 clustered above 40 mol%.
These were assigned to the new genus Asteroleplasma [type strain
161T (=ATCC 27880T)] whose members are anaerobic, but do
not require sterol for growth. Genome sizes ranged from 1542
to 1715 kbp for Anaeroplasma species, as determined by renaturation kinetics (Christiansen et al., 1986). Although the genome
sizes reported were in the expected range for members of the
class Mollicutes, no data are currently available on genome sizes
estimated by the more accurate pulsed-field gel electrophoresis
technique. A phylogenetic analysis of members of the Anaeroplasmatales, based upon 16S rRNA gene sequence comparison,
was carried out by Weisburg et al. (1989). Anaeroplasma and
Acholeplasma are sister genera basal on the mollicute tree.
Acknowledgements
Further reading
Johansson, K.-E. 2002. Taxonomy of Mollicutes. In Molecular
Biology and Pathogenicity of Mycoplasmas (edited by Razin
and Herrmann). Kluwer Academic/Plenum Publishers, New
York, pp. 1–29.
Differentiation of the species of the genus Anaeroplasma
The technical challenges of cultivating these anaerobic mollicutes have led to a current reliance principally on the combination of 16S rRNA gene sequencing and reciprocal serology for
species differentiation. Serological characterization of anaeroplasmas has been performed with agglutination, modified
metabolism inhibition, and immunodiffusion tests (Robinson
and Rhoades, 1977). Failure to cross-react with antisera against
previously recognized species provides substantial evidence for
species novelty. DNA–DNA hybridization values between species
722
Family I. Anaeroplasmataceae
examined are less than 5%. Bacteriolytic and nonbacteriolytic
organisms occur within the genus. When grown on agar media
containing a suspension of autoclaved Escherichia coli cells,
bacteriolytic strains form colonies surrounded by a clear zone
due to lysis of the suspended cells by a diffusible enzyme(s).
On media lacking suspended cells, bacteriolytic and nonbacteriolytic strains of Anaeroplasma cannot be distinguished from
each other on the basis of colonial or cellular morphology.
List of species of the genus Anaeroplasma
1. Anaeroplasma abactoclasticum Robinson, Allison and Hartman 1975, 179AL
a.bac.to.clas¢ti.cum. Gr. pref. a without; Gr. bakt- (L. transliteration bact-) part of the stem of the Gr. dim. n. bakterion
(L. transliteration bacterium) a small rod; N.L. adj. clasticus,
-a, um (from Gr. adj. klastos, -ê, -on broken in pieces) breaking; N.L. neut. adj. abactoclasticum intended to denote “not
bacteriolytic”.
This is the type species of the genus Anaeroplasma. Cells
are coccoid, about 500 nm in diameter, sometimes joined
into short chains by filaments. Colonies on solid medium
are subsurface, but nevertheless present a typical fried-egg
appearance. Growth is inhibited by 20 mg/ml digitonin. A
major distinguishing factor is the lack of the extracellular
bacterioclastic and proteolytic enzymes that characterize the
lytic species.
No evidence of a role in pathogenicity.
Source: occurs primarily in the bovine and ovine rumen.
Type strain: 6-1, ATCC 27879.
2. Anaeroplasma bactoclasticum (Robinson and Hungate
1973) Robinson and Allison 1975, 186AL (Acholeplasma bactoclasticum Robinson and Hungate 1973, 180)
bac.to.clas¢ti.cum. Gr. bakt- (L. transliteration bact-) part of
the stem of the Gr. dim. n. bakterion (L. transliteration bacterium) a small rod; N.L. adj. clasticus, -a, um (from Gr. adj.
klastos, -ê, -on broken in pieces) breaking; N.L. neut. adj. bactoclasticum bacteria-breaking.
Pleomorphic and coccoid cells ranging in size from 550
to 2000 nm in diameter. Cells cluster and sometimes form
short chains. Colonies on solid medium have a typical friedegg appearance. Optimal temperature for growth is between
30 and 47°C. Growth is inhibited by 20 mg/ml digitonin.
Skim milk is cleared by a proteolytic, extracellular enzyme
and certain bacteria are lysed by an extracellular enzyme
that attacks the peptidoglycan layer of the cell wall. Shares
some serological relationship to other established species in
the genus, but can be distinguished by agglutination, modified metabolism inhibition, and agar gel immunodiffusion
precipitation tests.
Source: occurs in the bovine and ovine rumen.
DNA G+C content (mol%): 32.5 to 33.7 (Tm, Bd).
Type strain: JR, ATCC 27112.
3. Anaeroplasma intermedium Robinson and Freundt 1987, 79VP
in.ter.me¢di.um. L. neut. adj. intermedium intermediate.
Cellular morphology and colonial features are similar to
those of Acholeplasma bactoclasticum. Serologically distinct
from other species in the genus by agglutination, metabolism inhibition, and agar gel immunodiffusion precipitation
tests (Robinson and Rhoades, 1977).
Type strain: 7LA, ATCC 43166.
Sequence accession no. (16S rRNA gene): not available.
4. Anaeroplasma varium Robinson and Freundt 1987, 79VP
va¢ri.um. L. neut. adj. varium diverse, varied, intended to
mean different from Anaeroplasma bactoclasticum.
Cellular morphology and colonial features are similar to
those of Acholeplasma bactoclasticum. Serologically distinct
from other species in the genus by agglutination, metabolism inhibition, and agar gel immunodiffusion precipitation
tests (Robinson and Rhoades, 1977).
Type strain: A-2, ATCC 43167.
Genus II. Asteroleplasma Robinson and Freundt 1987, 79VP
Daniel R. Brown, Janet M. Bradbury and Karl-Erik johansson
A.ste.rol.e.plas¢ma. Gr. pref. a not; N.L. neut. n. sterolum sterol; e combining vowel; Gr. neut. n. plasma
something formed or molded, a form; N.L. neut. n. Asteroleplasma name intended to indicate that sterol is
not required for growth.
Cellular and colonial morphology similar to species of the
genus Anaeroplasma. Nonmotile. The three strains that form
the new genus and species are obligately anaerobic and capable
of growth in the absence of cholesterol or serum supplements.
Temperature optimum for growth is 37°C. No evidence of bacteriolytic activity. The organisms are serologically distinct from
other members in the family Anaeroplasmataceae. Occur in the
ovine rumen.
DNA G+C content (mol%): about 40 (Tm, Bd).
Type species: Asteroleplasma anaerobium Robinson and Freundt 1987, 79VP.
The most prominent characteristics of the organisms are strictly
anaerobic growth and growth in the absence of sterol supplements. The G+C contents of strains analyzed to date are higher
than the values for Anaeroplasma species (Stephens et al., 1985).
DNA–DNA reassociation values clearly show that strains 161T,
Genus II. Asteroleplasma
162, and 163 of Asteroleplasma anaerobium are genetically related
and distinct from established species in the genera Acholeplasma
or Anaeroplasma (Stephens et al., 1985). Tube agglutination
tests and gel diffusion precipitation tests showed that strains
assigned to Asteroleplasma anaerobium are serologically distinct
from Anaeroplasma species (Robinson and Rhoades, 1977). No
data have been reported on antibiotic sensitivity or pathogenicity of asteroleplasmas. Strains have been isolated only from
sheep rumen. Isolation and maintenance techniques are similar to those reported for Anaeroplasma species.
Differentiation of the genus Asteroleplasma
from other genera
sequences. The obligately anaerobic nature of Asteroleplasma is
a distinctive and stable characteristic. Strictly anaerobic growth
plus growth in the absence of sterol supplements exclude assignment to any other taxon in the class. Extracellular bacteriolytic
and proteolytic enzymes are absent. Growth is not inhibited by
20 mg/ml digitonin.
Taxonomic comments
The taxonomic position of strains 161T, 162, and 163 of obligately
anaerobic mollicute serovar 4 was delineated through observations that they did not require sterol for growth (Robinson
723
et al., 1975), were serologically distinct (Robinson and Rhoades,
1977), and had G+C contents that were much higher than those
of Anaeroplasma species (Christiansen et al., 1986). Less than 5%
DNA–DNA relatedness existed between these strains and species assigned to the genus Anaeroplasma (Stephens et al., 1985).
Lastly, the significance of the group and the need to clarify its
taxonomic status was emphasized when it was demonstrated
that a significant proportion of the anaerobic mollicute population in the bovine and ovine rumen does not require sterol for
growth (Robinson and Rhoades, 1982). The phylogenetic analysis of Weisburg et al. (1989) indicated that Asteroleplasma anaerobium had branched from the Firmicutes lineage independently
of Acholeplasma and Anaeroplasma. Further, Asteroleplasma shared
two of three important synapomorphies that united the Mycoplasma and Spiroplasma lineages. Thus, the question of possible
monophyly and the true phylogenetic position of Asteroleplasma
with respect to other mollicutes remains open.
Acknowledgements
Further reading
Johansson, K.-E. 2002. Taxonomy of Mollicutes. In Molecular
Biology and Pathogenicity of Mycoplasmas (edited by Razin
and Herrmann). Kluwer Academic/Plenum Publishers, New
York, pp. 1–29.
List of species of the genus Asteroleplasma
1. Asteroleplasma anaerobium Robinson and Freundt 1987, 79VP
a.na.e.ro¢bi.um. Gr. pref. an not; Gr. n. aer air; Gr. n. bios life;
N.L. neut. adj. anaerobium not living in air.
This is the type species of the genus Asteroleplasma. Cell
morphology and colonial characteristics are similar to those
of other members of the order Anaeroplasmatales. Strains 161T,
162, and 163 form a homogeneous and distinct serological
group, as judged by about 80% DNA–DNA hybridization and
References
Christiansen, C., E.A. Freundt and I.M. Robinson. 1986. Genome size
and deoxyribonucleic acid base composition of Anaeroplasma abactoclasticum, Anaeroplasma bactoclasticum, and a sterol-nonrequiring
anaerobic mollicute. Int. J. Syst. Bacteriol. 36: 483–485.
Hungate, R.E. 1969. A roll tube method for cultivation of strict anaerobes. In Methods in Microbiology, vol. 3B (edited by Norris and
Ribbons). Academic Press, London, pp. 117–132.
Langworthy, T., W. Mayberry, P. Smith and I. Robinson. 1975. Plasmalogen composition of Anaeroplasma. J. Bacteriol. 122: 785–787.
Robinson, I.M. and M.J. Allison. 1975. Transfer of Acholeplasma bactoclasticum Robinson and Hungate to genus Anaeroplasma (Anaeroplasma
bactoclasticum Robinson and Hungate comb. nov.), emended description of species. Int. J. Syst. Bacteriol. 25: 182–186.
serological agglutination, metabolism inhibition, and agar
gel immunodiffusion precipitation tests.
Source: all isolates have been identified from the bovine or
ovine rumen.
DNA G+C content (mol%): 40.2–40.5 (Bd, Tm).
Type strain: 161 (the type strain ATCC 27880 no longer
exists).
Robinson, I.M. and K.R. Rhoades. 1977. Serological relationships between
strains of anaerobic mycoplasmas. Int. J. Syst. Bacteriol. 27: 200–203.
Robinson, I.M. 1979. Special features of anaeroplasmas. In The Mycoplasmas, vol. 1 (edited by Barile and Razin). Academic Press, New
York, pp. 515–528.
Robinson, I.M. and K.R. Rhoades. 1982. Serologic relationships between
strains of anaerobic mycoplasmas. Rev. Infect. Dis. 4: S271.
Robinson, I.M. 1983. Culture media for anaeroplasmas. In Methods in
Robinson, I.M. and E.A. Freundt. 1987. Proposal for an amended
classification of anaerobic mollicutes. Int. J. Syst. Bacteriol. 37:
78–81.
Robinson, J.P. and R.E. Hungate. 1973. Acholeplasma bactoclasticum sp.
n., an anaerobic mycoplasma from the bovine rumen. Int. J. Syst.
Bacteriol. 23: 171–181.
Rose, C. and S. Pirt. 1981. Conversion of glucose to fatty acids and methane: roles of two mycoplasmal agents. J. Bacteriol. 147: 248–254.
Stephens, E., I. Robinson and M. Barile. 1985. Nucleic acid relationships
among the anaerobic mycoplasmas. J. Gen. Microbiol. 131: 1223–1227.
Weisburg, W., J. Tully, D. Rose, J. Petzel, H. Oyaizu, D. Yang, L. Mandelco,
J. Sechrest, T. Lawrence and J. Van Etten. 1989. A phylogenetic analysis of the mycoplasmas: basis for their classification. J. Bacteriol. 171:
6455–6467.

here - The International Organization for Mycoplasmology

Transcription

Similar documents

Mycoplasma Testing – An Overview

Staying on top of new technology Tick

bergman

Spirochetes from Digital Dermatitis Lesions in Cattle Are Closely

55 rRNA sequences`of 12 species of flatworms: implications for the

full service salon

Lisa Tully, PhD.

Elizabeth Diller Diller Scofidio + Renfro

European Guideline on the management of non

PDF (Chapter 1)