Chryseomonas luteola - International Journal of Systematic and

INTERNATIONAL
JOURNAL
OF SYSTEMATIC
BACTERIOLOGY,
July 1987, p. 245-250
0020-7713/87/030245-06$02.00/0
Copyright 0 1987, International Union of Microbiological Societies
Vol. 37, No. 3
Chryseomonas luteola comb. nov. and Flavimonas oryzihabitans
gen. nov. , comb. nov. , Pseudomonas-Like Species from Human
Clinical Specimens and Formerly Known, Respectively, as Groups
Ve-1 and Ve-2
B. HOLMES,’ A. G. STEIGERWALT,2 R. E. WEAVER,2* AND DON J. BRENNER2
National Collection of Type Cultures, Central Public Health Laboratory, London NW9 5HT, England,’ and Division of
Bacterial Diseases, Centers for Disease Control, Atlanta, Georgia 303332
Pseudomonas luteoh Kodama et al. 1985 is a senior subjective synonym of Chryseomonas polytricha Holmes
et al. 1986 (formerly group Ve-1). The type strain of P . luteola (JCM 3352) was 78% related to the type strain
of C . polytricha (NCTC 11843). We propose the new combination Chryseomonas luteola for this organism. For
Pseudomonas oryzihabitans Kodama et al. 1985 (formerly group Ve-2) we propose the new genus and new
combination Flavimonas oryzihabitans. We were in the process of proposing a scientific name for group Ve-2
on the basis of an examination of 19 strains when the description of P . oryzihabitans was published. We have
included the type strain of P . oryzihabitans for comparison and here present our results for all 20 strains. Each
of the 20 strains was examined for 129 characteristics, including reactions in 58 enzyme tests (API ZYM
system). These bacteria are rod shaped, aerobic, gram negative, yellow pigmented, motile by a single polar
flagellum, and saccharolytic, but they do not produce cytochrome oxidase. The mean guanine-plus-cytosine
content of the deoxyribonucleicacid of three representative strains was 63.6 f 1.6 mol%. The deoxyribonucleic
acid relatedness of 11strains of group Ve-2 to the strain we would have proposed as the type strain, CL162/81
(= NCTC 11850), averaged 93% (hydroxyapatite method, 70°C). The type strain of P . oryzihabitans (JCM
2952) was 93% related to CL162/81. The relatedness of CL162/81to the respective type strains of the five major
ribosomal ribonucleic acid hybridization groups of the genus Pseudomonas and to the type strain of C .
polytricha (formerly group Ve-1) was 5 % or less. All strains, except the type strain, of F. oryzihabitans were
isolated from human clinical specimens.
Group Ve was divided into two biogroups by Tatum et al.
(19), who first described the taxon. They considered that the
differences between the two biogroups were sufficient to
warrant their acceptance as separate species. Biogroup 1
strains possess multitrichous polar flagella, whereas
biogroup 2 strains have a single polar flagellum, in addition
to which there are several phenotypic differences. Gilardi et
al, (8) reported that strains of group Ve-1 have a guanineplus-cytosine (G+C) content of 56.8 mol%, whereas for
strains of group Ve-2 the corresponding value is 68.9%.
These findings also support the view that these two taxa
should be regarded as separate species. The cellular fatty
acid composition of group Ve strains is most similar to that
of Pseudomonas species (4), despite the fact that group Ve
strains are oxidase negative.
Pedersen et al. (14) used the name “Chromobacteriurn
typhijlavum” to describe nonfermentative strains with the
characteristics of group Ve. However, it was unfortunate
that they did so, as the name “Bacterium typhgavum” was
first applied to yellow-pigmented fermentative bacteria,
which were subsequently demonstrated to be members of
the genus Erwinia. Of the strains reported by Pedersen et al.
(14), four were recovered in small numbers and in mixed
cultures from sputum, another was found with Pseudornonas
aeruginosa in a urine specimen, two were from blood
cultures, one was from inhalation therapy equipment, one
was from an eye swab, and one was recovered with Neisseria gonorrhoeae from a cervical discharge. None of the 10
strains was considered to be of clinical significance. The
~~
* Corresponding author
group Ve strains received by Tatum et al. (19) were from a
variety of human anatomical sites, most frequently from
wounds and abscesses. Gilardi et al. (8) reported group Ve
strains primarily from wounds (five strains) and sputum (two
strains), but there was one strain from each of the following:
ear, inhalation therapy equipment, cervical swab, urine, eye,
and throat. Group Ve-2 strains have been recovered from
cultures of evacuated blood collection tubes (20) and have
been reported as a cause of bacteremia (15) and as a cause of
peritonitis in a patient on continuous ambulatory peritoneal
dialysis (17).
Our proposal of a scientific name for group Ve-1 was
already in press (10) and that for group Ve-2 was submitted
when the similar proposals by Kodama et al. (11) came to
our attention. Our manuscript on group Ve-2 was withdrawn
and modified as presented here. In the present study the type
strain of Pseudomonas luteola was examined for phenotypic
characteristics and by deoxyribonucleic acid (DNA)-DNA
hybridization to see how it compared with our strains of
Chryseomonas polytricha (10). Nineteen strains of group
Ve-2 and the type strain of Pseudornonas oryzihabitans were
examined for phenotypic characteristics, DNA base composition, and DNA relatedness to determine whether they
constituted a homogeneous taxon. For these 20 strains the
new genus and new combination Flavirnonas oryzihabitans
is proposed, with JCM 2952 (= KS0036) as the type strain.
MATERIALS AND METHODS
Bacterial strains. The 20 bacterial strains studied and the
sources from which they were isolated are given in Table 1.
245
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INT. J. SYST.BACTERIOL.
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2146
TABLE 1. Strains of F . oryzihabituns gen. nov., comb. nov., studied
~~~~
Straina
Source
CL236/74 .............................................................................
.Hip arthroplasty; Glasgow, Scotland
CL458/75 ............................................................................. .Pus; Zurich, Switzerland
(3L326/77 .............................................................................
.Conjunctiva; Munich, Federal Republic of Germany
CL595/77 ............................................................................. .Pleural fluid; Bangkok, Thailand
(3L623/78 ............................................................................. .Maxillary sinus washing; London, England
CL806/79 ............................................................................. .Pleural aspirate; Wrexham, Wales
CL365/80 .............................................................................
.Central venous lineb; London, England
CL162181 ( = NCTC 11850). .......................................................... .Pus, appendicular abscess; Kuala Lumpur, Malaysia
CL327/81 ( = NCTC 11851). ..........................................................
.Blood; Chichester, England
CL234/82 .............................................................................
.Central venous pressure line; London, England
CL87/83 ..............................................................................
.Blood"; Kuala Lumpur, Malaysia
CL320183 ............................................................................. .Blood; Ballymena, Northern Ireland
N993.. ................................................................................
.Ear; Indiana
132524. ................................................................................
.Unknown; Colorado
135066. ................................................................................ .Finger exudate; California
135142................................................................................ ..Blood; New York City
135398. ................................................................................
.Eye; Alaska
13726 ( = NCTC 11852) ..............................................................
.Blood; Virginia
136980. .................................................................................Breast drainage; Pennsylvania
JCM 2952= ( = KS0036T = L-lT = AJ 2197T = IAM 1568T)......................... .Rice paddy; Japan
~
~~
~~
~~
Key to sources: strains prefixed by CL are strains referred for identification to the National Collection of Type Cultures (NCTC), Central Public Health
Laboratory, London, England; the strain prefixed by JCM was received from the Japan Collection of Microorganisms, Saitama, Japan; all other strains are strains
referred for identification to the Special Bacteriology Reference Laboratory, Centers for Disease Control, Atlanta, Ga.
From a patient with infective endocarditis.
From a patient with systemic lupus erythematosus.
a
The strains were isolated over a period of several years and
from diverse geographical locations.
Bacteriological investigations. The methods used for this
study have been described previously (9).
G C contents. The G +C contents of DNAs were determined optically in triplicate by thermal denaturation (5).
DNA from Serratia marcescens SM-6 (G+C content, 62.0
mol%) was used as the control.
DNA relatedness. The preparation and purification of DNA
and the conditions used to determine DNA relatedness by
the hydroxyapatite method have been described previously
(2). DNA from group Ve-2 strain CL162/81 was labeled with
"*PO4in vitro (3). Owing to the high G+C contents of group
Ve-2 DNAs, hybridization reactions were done at 70°C
(optimal reassociation) and 85°C (stringent reassociation).
Median organism. The median organism was determined
by the method of Lapage and Willcox (12).
+
RESULTS AND DISCUSSION
Labeled DNA from the type strain of C . polytricha (NCTC
111843) was 78% related (at 70°C) to unlabeled DNA from the
type strain of P.luteola (Table 2). Labeled DNA from group
Ve-2 strain CL162/81 was 83 to 99% related (average, 93%)
to unlabeled DNAs from 11other group Ve-2 strains and the
type strain of P . oryzihabitans (JCM 2952) in hybridization
reactions done at 70"C, a temperature that is close to the
optimum for DNA reassociation, and 72 to 99% related
(average, 93%) to the same 12 DNAs in reactions done at
M"C, at which only very closely related DNA sequences can
reassociate (Table 2). The divergence (percent unpaired
bases within related sequences) was 0.0 to 2.0%.
The DNA-DNA relatedness of group Ve-2 strains to
strains representing the major ribosomal ribonucleic acid
(rRNA) hybridization groups of the genus Pseudomonas and
C. polytricha (formerly group Ve-1 [lo]) was 1to 5% (Table
2).
Our phenotypic data (B. Holmes and R. E. Weaver,
unpublished results) and the DNA relatedness data pre-
sented here show that P . luteola and C . polytricha are
synonymous. The former species has priority, and for it we
here propose the new combination Chryseomonas luteola.
Our DNA relatedness (Table 2) and phenotypic (Table 3)
data also show that the 19 strains of group Ve-2 bacteria
studied here and the type strain of P . oryzihabitans constitute a homogeneous taxon. Other workers have failed to find
an existing named species to which their strains of group
Ve-2 correspond, so the recognition of this taxon as a new
species has not been questioned. It is generic assignment
that is not clear, although group Ve-2 is generally regarded
as Pseudomonas-like in phenotypic characteristics (7) and in
DNA base composition (8). The cellular fatty acids of group
Ve-2 strains are also most similar to those of Pseudomonas
strains (4).
Brenner (1) considers that, at least until a good genetic
means is found, the primary consideration for a genus is that
it contains biochemically similar species that are covenient
or important to consider as a group and that must be
separated from one another "at the bench." If one accepts
this view, then group Ve-2 could be recognized as an
additional species io the genus Pseudomonas (a view chosen
by Kodama et al. [ll]). However, strains of group Ve-2 fail
to produce oxidase, while those of Pseudomonas species
generally do. Also, the genus Pseudomonus as presently
defined is known by rRNA-DNA hybridization studies and
other techniques to contain five major groups that are not
closely related phylogenetically despite a close phenotypic
similarity (13). As attempts are made to classify Pseudomonus species to better represent their phylogenetic relationships, many existing species may be transferred to other
genera and perhaps to families other than the family
Pseudomonadaceae. It has already been proposed that
Pseudomonas maltophilia be transferred to the genus
Xanthomonas as Xanthomonas maltophilia in recognition of
the close phylagenetic relationship of this species to
Xanthomonas species (18). Such nomenclatural changes are,
however, unlikely to be accepted by many clinical microbiologists and others while the determination of phenotypic
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F. ORYZIHABITANS GEN. NOV., COMB. NOV.
VOL.37, 1987
characteristics remains the major method of identifying
these orgahisms at the bench. We consider it inappropriate
to make the genus Pseudomonas more heterogeneous by
including group Ve-2 within it. Group Ve-2 strains fail to
produce oxidase. They show no appreciable DNA relatedness to strains representing the major Pseudomonas rRNA
hybridization groups (Table 2). Studies of rRNA relatedness
or rRNA sequencing are necessary to determine the rRNA
group to which group Ve-2 belongs at the level of phylum or
superfamily. There are no guidelines for the use of rRNA
data for classification at the family or genus level, although
some workers have used rRNA data for this purpose. We
therefore leave this topic for future reconsideration by
experts in the rRNA field. We would place group Ve-2 in a
separate, new genus but as a compromise would choose a
generic name reflecting the close phenotypic similarity of
group Ve-2 to species presently placed in the genus Pseudomonas.
For this genus we propose the name Flavimonas
(fla.vi'mo.nas. L. adj. flavus, yellow; Gr. n. monas, a unit;
TABLE 2. DNA relatedness among F. oryzihabitans and
C. polytricha strains
Source of unlabeled DNA
32P04-labeledDNA from
F. oryzihubitans CL162/81
( = NCTC 11850)
RBR" at
70°C
F. oryzihabitans CL162/81
F. oryzihabitans CL236/74
F. oryzihabitans CL320/83
F. oryzihabitans CL327/81
F. oryzihabitans B2524
F. oryzihabitans B5066
F. oryzihabitans B5142
F. oryzihabitans CL234/82
F. oryzihabitans B993
F. oryzihabitans JCM 2952T
F. oryzihabitans CL365/80
F. oryzihabitans E5726
F. oryzihabitans CL806/79
P. aeruginosa (rRNA homology
group I) NCTC 10332T
P. cepacia (rRNA homology
group 11) NCTC 10743T
P. acidovorans (rRNA homology
group 111) NCTC 10683T
P. diminuta (rRNA homology
group IV) NCTC 8545T
P. rnaltophilia (rRNA homology
group V) NCTC 10257T
C. polytricha NCTC 11843T
100
99
97
96
95
95
95
94
94
93
93
85
83
1
C. polytricha NCTC 11843T
P. luteola JCM 3352T
load
Db
RBR at
85°C
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.5
2.0
0.0
0.5
0.0
NT'
100
98
94
94
72
94
95
99
91
90
97
92
99
NT
3
NT
NT
1
NT
NT
1
NT
NT
5
NT
NT
16
10.0
NT
7gd
O.Od
lSd
lood
76d
RBR, Relative binding index = percent heterologous DNA bound to
hydroxyapatitelpercent homologous DNA bound to hydroxyapatite x 100.
D, Divergence. D was calculated on the assumption that each 1°C
decrease in the thermal stability of a heterologous DNA duplex as compared
with that of the homologous DNA duplex was caused by 1% unpaired bases
within the duplex. D was calculated to the nearest 0.5%. All reactions were
done at least twice. The values shown are averages. Before normalization to
loo%, the percentage of DNA bound to hydroxyapatite in homologous
reactions was approximately 5 5 . The amount of labeled DNA that bound to
hydroxyapatite in control reactions that did not contain unlabeled DNA was
0.7 to 3.9%. These control values were subtracted from all reassociation
reactions before normalization.
NT, Not tested.
Hybridization with C. polytricha NCTC 11843TDNA.
a
247
N. L. fem. n. flavimonas, a yellow unit or little thing). We
further propose the new combination Flavimonas oryzihabitans for strains of group Ve-2. We consider it appropriate to include the new genus in the family Pseudomonadaceae.
Fkvimonas gen. nov. Rods with parallel sides and rounded
ends. Intracellular granules of poly-P-hydroxybutyrate are
generally absent. Do not produce prosthecae and are not
surrounded by sheaths. No resting stages are known. Gramnegative. Motile by a single polar flagellum. Aerobic, having
a strictly respiratory type of metabolism. Grow at temperatures from 18 to 42°C. Growth on solid media is pigmented
pale yellow to deeply yellow. Colonies are typically circular
(diameter, 1 mm), low convex, smooth (occasional strains
produce wrinkled colonies), and shiny, with entire edges.
Catalase positive but oxidase negative. Agar is not digested.
Chemoorganotrophic. Saccharolytic. Found in the general
environment, they are apparently saprophytes or commensals of humans and other warm-blooded animals, in which
they may occasionally prove pathogenic. The G+C content
of the DNA is 62 to 65 mol% as determined by the thermal
denaturation method.
Type species: Flavimonas oryzihabitans.
Flavimonas oryzihabitans gen. nov., comb. nov. The 20
strains which we studied were strictly aerobic, gramnegative, nonsporeforming, uniformly stained rods with parallel sides and rounded ends. Except for the type strain, JCM
2952, the cells were motile, by means of a single polar
flagellum (Fig. l),in hanging drop preparations after overnight growth in nutrient broth incubated at room temperature (18 to 22°C). Colonies on nutrient agar after 24 h were
typically circular, low convex, about 1 mm in diameter,
smooth (strains CL458/75, CL326/77, CL623/78, CL806/79,
CL365/80, CL327/81, CL320/83, and E5726 produced deepyellow, wrinkled colonies similar to those produced by
certain strains of Pseudomonas stutzeri), shiny, entire, and
pigmented pale yellow to deeply yellow. Strains CL806/79,
CL327/81, CL320/83, and E5726 caused concern that the
cultures might be mixed. In each of these strains, after
prolonged incubation, bright yellow colonies developed on
top of the initial growth of paler yellow colonies. However,
upon subculturing, single colonies of both types eventually
reverted to colonies identical to the other type. No evidence
could be found that these cultures were mixed, and it is
possible that the bright yellow colonies grew by utilizing
nutrients released by the dead, paler yellow, original growth.
Only 10 of the 20 strains produced a dark brown, diffusible,
melaninlike pigment on tyrosine agar. Hemolysis was not
present after aerobic growth for 24 h on 5% (voVvo1) horse
blood agar. The strains were not fluorescent on King medium B.
A total of 95 characteristics were common to all strains
(either all positive or all negative), and there were 34
characteristics in which one or more of the strains differed
(Table 3). No strain conformed to the median organism, but
the strain we would have proposed as the type strain,
CL162/81, was the centrotype strain. Strain JCM 2952T was
the least typical member of the group, differing from the
other 19 strains in six biochemical characteristics. The
atypical nature of the type strain is probably a result of the
fact that it was the only strain of the 20 studied here to have
come from a nonclinical environment. All strains had the
following characteristics in common: growth at room temperature and at 37°C but not at 5 or 42°C; production of
catalase but not of cytochrome oxidase; oxidative reaction in
glucose oxidation-fermentation medium; hydrolysis of
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INT.J. SYST.BACTERIOL.
HOLMES ET AL.
TABLE 3. Characteristics in which the 20 strains of F. oryzihabitans differed from each other
No. of
Characteristic
strains
positive
Acid production froma:
meso-Inositol
Maltose
Acid production from 10% (wt/vol) lactose
Hydrolysis of gelatin (plate method)
Motility at room temperature
Hydrolysis of b:
L-Seryl-L-tyrosyl-p-naphthylamide
2-Naphthyl caprylate'
L-Phenylalanyl- P-naphthylamide
L-Omithyl-P-naphthylamide
L-Tryptophan yl-P-naphth ylamide
Motility at 37°C
Hydrolysis of b:
N-Carbobenzoxy-glycyl-glycyl-L-arginineP-naphth ylamide
N-Benzoyl-~~-arginine-2-naphythylamide'
L-Pyrrolidonyl- P-naphth ylamide
Result for
type strain
(JCM 2952)
Accession no. of strain(s)
with the less common result
19
19
19
19
19
E5398
CL234/82
JCM 2952
JCM 2952
JCM 2952
19
18
18
18
17
17
CL623/78
CL458/75, JCM 2952
CL623/78, E6980
CL234182, JCM 2952
CL458/75, CL234/82, JCM 2952
CL458/75, CL623/78, JCM 2952
13
CL327/81, B2524, B5066, B5142, E5398, E5726, JCM 2952
12
CL806/79, CL162/81, CL327/81, B2524, B5142, E5398, E5726,
JCM 2952
CL326/77, CL623/78, CL36Y80, CL234/82, B993, B2524,
E6980, JCM 2952
CL236/74, CL458/75, CL326/77, CL806/79, CL234/82,
CL320/83, E5398, JCM 2952
CL806/79d, CL162/81, CL327/81, CL234182, B993, B5142,
E5398, E5726, E6980, JCM 2952
CL236/74d, CL623/78, CL365/80, CL162/81, CL234/82,
CL87/83, CL320/83, B2524, E6980, JCM 2952
CL236/74', CL595/77, CL806/79, CL162/81, CL234/82,
CL87/83, CL320183, B993, E5398, JCM 2952
CL326/77, CL595/77, B993, B2524, B5066, B5142, E6980
12
Malonate utilization
12
Hydrolysis of L-isoleucyl-P-naphthylamideb
10
Urease production
io
Pigment on tyrosine agar
10
Deoxyribonuclease production
Hydrolysis of b:
L-Hydrox yprol yl- p-naphth ylamide
7
L-Tyrosyl-p-naphthylamide
Acid production froma:
D-Cellobiose
Sucrose
Hydrolysis of b:
2-Naphthyl phosphate at pH 5.4"
L-Aspartyl-P-naphthylamide
2-Naphthyl butyrate'
~-Valyl-2-naphthylamide'
$-Benz yl-L-c ysteine- P-naphthylamide
2-Naphthyl phosphate at pH 8.5"
Naphthol- AS-B 1-phosphodiamide'
6-Bromo-2-naphthyl-~-~-glucopyranoside'
y-L-Glutamyl-p-naphthylamide
L-Threonyl-P-naphth ylarnide
KCN tolerance
6
CL236/74, CL458/75, CL595/77, CL623/78, CL87/83, B993,
B5142
CL236/74, CL458/75, CL806/79, CL320/83, B5066, E5398
5
5
CL623178, CL365/80, CL234/82, CL87/83, CL320/83
CL234/82, CL320/83, B993, E5398, E6980
4
4
2
2
2
1
1
1
1
1
1
CL623/78, CL806/79, CL320/83, JCM 2952
CL458175, CL623178, CL87/83, CL320/83
CL326/77, CL87/83
CL595/77, CL623/78
CL595/77, CL320/83
JCM 2952
JCM 2952
JCM 2952
CL320/83
CL326/77
CL595/77
a
7
Tested in ammonium salt medium.
Tested by using various API ZYM galleries.
Enzyme test included in the standard API ZYM gallery.
Strains giving negative results.
Strains giving positive results.
tributyrin and Tween 20 but not of gelatin (stab liquefaction
method); no hydrolysis of Tween 80 or tyrosine; no digestion
of casein; no production of opalescence on lecithovitellin
agar; no reduction of nitrate or nitrite; no production of
imdole; no production of hydrogen sulfide (lead acetate paper
and triple sugar iron agar methods); growth on phydroxybutyrate (without the production of lipid inclusion
granules), cetrimide, and MacConkey agar; no hydrolysis of
esculin or starch; alkali production on Christensen citrate;
utilization of citrate (Simmons medium); no oxidation of
gluconate; no production of arginine desimidase, argihine
dihydrolase, lysine decarboxylase, or ornithine decar-
boxylase; no production of 3-ketolactose; no reduction of
selenite; no deamination of phenylalanine; no production of
P-D-galactosidase (o-nitrophenyl-P-D-galactopyranoside
test) or phosphatase; production of acid in ammonium salt
medium under aerobic conditions from D-glucose, Larabinose, ethanol, D-fructose, glycerol, mannitol, Lrhamnose, sorbitol, trehalose, and D-xylose; no production
of acid in ammonium salt medium under aerobic conditions
from adonitol, dulcitol, lactose, raffinose, and salicin; production of acid from 10% (wthol) D-glucose; and no production of acid or gas from D-glucose in peptone water medium.
All strains hydrolyzed the following substrates (tests were
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F. ORYZZHABZTANS GEN. NOV., COMB. NOV.
249
done with API ZYM galleries): L-leucyl-2-naphthylamide; phosphate; p-nitrophenyl-a-D-xylopyranoside;
p-nitrophenyl-p-D-fucopyranoside; p-nitrophenyl-P-L-fucopyranoside;
L-lysyl-P-naphthylamide; glycyl-P-naphthylamide; L-arginyl-P-naphthylamide; L-alanyl-P-naphthylamide;DL-methio-nitrophenyl-N-acetyl-a-D-glucosaminide;
p-nitrophenylonyl-P-naphthylamide;glycyl-glycyl-P-naphthylamide
lactopyranoside; p-nitrocatechol-sulfate; 4-methylumbellihydrobromide; glycyl-L-phenylalanyl-P-naphthylamide;glycyl-Lferylarabinopyranoside; 4-methylumbelliferylcellobio pyprolyl-p-naphthylamide ; L-leucyl-glycyl-P-naphthylamide ;
ranoside; L-histidyl-P-naphthylamide;N-benzoyl-L-leucylp-naphthylamide; N-carbobenzoxy-~-arginine-4-methoxy-~L-glutamine-P-naphthylamidehydrochloride; L-prolyl-Pnaphthylamide hydrochloride; a-L-glutamyl-p-naphthylnaphthylamide hydrochloride; and L-seryl-p-naphthylamide.
None of the strains hydrolyzed the following substrates
amide .
(tests were done with API ZYM galleries): 2-naphthyl myThe G + C contents of the DNAs of three phenotypically
ristate ; ~-cystyl-2-naphthylamide;
N-glutaryl-phenylalanine- representative strains of F. oryziha bitans included in this
2-naphthylamide; 6-bromo-2-naphthyl-a-~-galactopyranosid study were between 61.8 and 65.4 mol%, with a mean of 63.6
mol% and a standard deviation of k1.6 mol%; the G+C
e; 2-naphthyl-P-~-galactopyranoside; naphthol-AS-BIcontent of the DNA of the strain we would have chosen as
P-D-glucuronic acid; 2-naphthyl-a-~-glucopyranoside;
1the type strain, CL162/81 (= NCTC 11850), was 61.8 mol%
naphthyl-N-acetyl-P-D-glucosaminide;
6-bromo-2-naphthyl(as estimated by the thermal denaturation method). Kodama
a-D-mannopyranoside; 2-naphthyl-a-~-fucopyranoside;
6et al. (11) obtained similar results, with a range of 63.9 to
bromo-2-naphthyl-~-~-xylopyranoside;
bis-(p-nitropheny1)65.6 mol% for their strains and a value of 65.1 mol% for the
type strain (JCM 2952).
Group Ve strains appear to occur only rarely in clinical
material and then most frequently in wounds or abscesses
(19). Some of the strains included in this study may have
been clinically significant, particularly those from blood and
pus, but the details received were insufficient to make a
valuable judgement. No pus cells were seen in the pleural
aspirate from which strain CL806/79 was grown, so this
particular strain was of doubtful clinical significance. Only
once has a group Ve-2 strain been reported as a cause of
bacteremia and then only in a patient who was severely
traumatized following neurosurgery (15). There has also
been only one report of peritonitis caused by a group Ve-2
strain, in a patient on continuous ambulatory peritoneal
dialysis (17). Strains are, in any case, generally susceptible
to several antimicrobial agents (ampicillin, carbenicillin,
cefotaxime, chloramphenicol, colistin, erythromycin, gentamicin, kanamycin, moxalactam, neomycin, polymyxin,
streptomycin, and tetracycline [6, 8]), so that treatment
should present no special difficulties. Treatment with intravenous ampicillin (15) and intraperitoneal tobramycin (17)
has proved effective. However, while strains of C. luteolu
(formerly group Ve-1 [lo, 111) are inhibited by low concentrations of several cephalosponns, strains of F. oryzihabitans show resistance or moderate resistance to most of the
same cephalosporins (16). F. oryzihabituns appears to be
widely distributed, since the 20 strains were isolated in
England, Scotland, Wales, Northern Ireland, seven states in
the United States, Switzerland, the Federal Republic of
Germany, Japan, Malaysia, and Thailand.
The biochemical characteristics of the field and reference
strains examined agree well with those presented by other
authors, except for some minor discrepancies, as follows.
All the strains studied by Gilardi (7) and Gilardi et al. (8)
failed to produce deoxyribonuclease, whereas two of the
strains studied by Kodama et al. (11) and seven of our group
Ve-2 strains were able to do so. Conversely, none of our
strains or those of Kodama et al. (11) hydrolyzed starch,
whereas several strains produced a positive result in this test
for Gilardi (7). Kodama et al. (11) found that all but one of
their strains produced urease, whereas only half of our
strains did so; they (11) also found that some of their strains
were able to accumulate poly-P-hydroxybutyrate, whereas
none of our stains did so; and although they (11) found that
most of their strains were able to grow at 42"C, their
reference strains of group Ve-2, like ours, failed to grow at
FIG. 1. Electron micrograph of a cell of F . oryzihubitans
this temperature. The G+C content determined for strains of
CL162/81 (= NCTC 11850), showing the monotrichous polar flagF. oryzihabitans in this study (63.6 mol%) is in accordance
ellar arrangement. Magnification, ~25,000.
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250
INT.J. SYST.BACTERIOL.
HOLMES ET AL.
TABLE 4. Characteristicsfor the differentiation of C. luteola
(formerly group Ve-1 [lo, 113) from F. oryzihabitans
(formerly group Ve-2)
Resultufor:
Characteristic
Acid produced in ammonium salt
medium from salicin
Acid produced in ammonium salt
medium from sorbitol
Arginine desimidase production
Arginine dihydrolase production
Esculin hydrolysis
Growth at 42°C
Hydrolysis of 6-bromo-2-naphthylP-D-glucopyranoside'
Hydrolysis of 4-methylumbelliferylarabinopyranoside"
Malonate utilization
Nitrate reduction
Tween 80 hydrolysis
Tyrosine hydrolysis
P-Galactosidaseproduction (o-nitrophenyl-P-D-galactopyranosidetest)
C. luteola
:i~zi
619
-
119
+
819
1/20
+
+
+
819
+
+
5/9
519
+
+
-
12120
-
+ ,All strains positive; -, all strains negative; x/x, no. positiveho. tested.
Tested in the standard API ZYM gallery.
Tested in an experimental API ZYM I1 gallery.
with the values reported by Kodama et al. (11) (range, 63.9
to 65.6 mol%) but is about 5% lower than the value of 68.9
mol% reported by Gilardi et al. (8) for strains of group Ve-2.
The reasons for this discrepancy are unknown.
Characteristics for the differentiation of F. oryzihabitans
from C . Zuteola (formerly group Ve-1 [lo, 111) are given in
Table 4.
The following additional strains of F. oryzihabitans have
been deposited in the National Collection of Type Cultures:
CL327/81 as NCTC 11851and E5726 as NCTC 11852. B2524
is available from the American Type Culture Collection as
ATCC 35564.
ACKNOWLEDGMENTS
We are most grateful to API Systems S.A. for the supply of API
ZYM galleries. We also thank T. 0. MacAdoo for advising us on the
etymology of the new generic epithet proposed here and E. Roe for
secretarial assistance. We thank the staff of the National Collection
of Type Cultures Computer IdentificationLaboratory for their help,
in particular, E. Cain for technical assistance. We thank A. A.
Porter for electron microscopy.
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