Isolation from a Nonclinical Sample of VIM-1 Leclercia adecarboxylata Producing a -Lactamase

Isolation from a Nonclinical Sample of
Leclercia adecarboxylata Producing a VIM-1
Metallo- β-Lactamase
Costas C. Papagiannitsis, Vendula Studentová, Jaroslav
Hrabák, Jan Kubele, Vlastimil Jindrák and Helena
Zemlicková
Antimicrob. Agents Chemother. 2013, 57(6):2896. DOI:
10.1128/AAC.00052-13.
Published Ahead of Print 25 March 2013.
These include:
REFERENCES
CONTENT ALERTS
This article cites 14 articles, 10 of which can be accessed free
at: http://aac.asm.org/content/57/6/2896#ref-list-1
Receive: RSS Feeds, eTOCs, free email alerts (when new
articles cite this article), more»
Information about commercial reprint orders: http://journals.asm.org/site/misc/reprints.xhtml
To subscribe to to another ASM Journal go to: http://journals.asm.org/site/subscriptions/
Downloaded from http://aac.asm.org/ on October 6, 2014 by guest
Updated information and services can be found at:
http://aac.asm.org/content/57/6/2896
LETTER TO THE EDITOR
Isolation from a Nonclinical Sample of Leclercia adecarboxylata
Producing a VIM-1 Metallo-␤-Lactamase
Costas C. Papagiannitsis,a Vendula Študentová,a Jaroslav Hrabák,a Jan Kubele,b Vlastimil Jindrák,b Helena Žemlicˇkovác
Department of Microbiology, Faculty of Medicine and University Hospital, Charles University, Plzen, Czech Republica; Department of Clinical Microbiology, Na Homolce
Hospital, Prague, Czech Republicb; National Reference Laboratory for Antibiotics, National Institute of Public Health, Prague, Czech Republicc
L
(50 ␮g/ml) as selective agents (Table 1) (7). Plasmid content analysis showed a single type of transconjugant (Trc Lec-476) with a
plasmid (pLec-476) of approximately 290 kb that hybridized
strongly with a blaVIM-specific probe (8; not shown). pLec-476
was nontypeable by the replicon typing method (9).
Characterization of the region flanking the blaVIM-1 gene was
carried out by PCR mapping and sequencing (10). blaVIM-1 was
the first gene cassette of a class 1 integron similar to In110 from
Pseudomonas putida isolates from Italy (11). In110 has also been
found in Pseudomonas aeruginosa from Italy (12) and Enterobacteriaceae strains from Spain and Germany (10, 13). Unlike In110
from Enterobacteriaceae in Spain, the integron was not located
within a Tn21-like element, and a defective Tn402tni module was
not identified beyond the 3= conserved segment (3=CS). However,
the 5=CS was truncated in the noncoding region by an IS26 element, as in In-e541 from pNL194 (14). A second copy of IS26 that
may be implicated in integron mobilization was not found in the
vicinity of the 3=CS.
This study, along with previous reports of antibiotic-resistant
L. adecarboxylata (1, 2), shows that this rarely isolated and inherently susceptible species is capable of acquiring and maintaining
resistance plasmids. To our knowledge, this is the first report of an
M␤L-producing L. adecarboxylata strain. It is of note that Lec-476
was recovered from a nonclinical sample from a hospital that was
considered free of M␤L- or other carbapenemase-producing isolates. The origin of this isolate is not known. Such a finding indicates the spreading potential of carbapenemase genes via routes
that remain largely unknown. Acquisition of a self-transferable,
VIM-1-encoding plasmid by the clinically insignificant species L.
adecarboxylata is disquieting, since such bacteria can act as hidden
sources of clinically important resistance determinants.
Published ahead of print 25 March 2013
Address correspondence to Jaroslav Hrabák, [email protected].
Copyright © 2013, American Society for Microbiology. All Rights Reserved.
doi:10.1128/AAC.00052-13
TABLE 1 Antibiotic susceptibilities of strains harboring VIM-1-encoding plasmids
MIC (mg/liter) of a:
Isolate
Pip
Ptz
Ctx
Caz
Fep
Atm
Mem
Gen
Amk
Col
Tmp
Cip
L. adecarboxylata Lec-476
E. coli trc Lec-476b
E. coli A15 (recipient)
⬎64
64
ⱕ0.5
64
64
1
⬎8
⬎8
ⱕ0.0625
⬎32
⬎32
ⱕ0.25
⬎16
4
ⱕ0.125
1
ⱕ0.25
ⱕ0.25
4
0.25
ⱕ0.125
16
0.125
0.125
1
0.5
0.5
ⱕ0.125
ⱕ0.125
ⱕ0.125
⬎32
⬎32
1
2
ⱕ0.0625
ⱕ0.0625
a
Pip, piperacillin; Ptz, piperacillin-tazobactam (inhibitor fixed at 4 mg/liter); Ctx, cefotaxime; Caz, ceftazidime; Fep, cefepime; Atm, aztreonam; Mem, meropenem; Gen,
gentamicin; Amk, amikacin; Col, colistin; Tmp, trimethoprim; Cip, ciprofloxacin. MICs were interpreted using guidelines from the European Committee on Antimicrobial
Susceptibility Testing (EUCAST) from 2011 (http://www.eucast.org/).
b
Transconjugant of L. adecarboxylata Lec-476.
2896
aac.asm.org
Antimicrobial Agents and Chemotherapy
p. 2896 –2897
June 2013 Volume 57 Number 6
Downloaded from http://aac.asm.org/ on October 6, 2014 by guest
eclercia adecarboxylata belongs to the family Enterobacteriaceae
and is rarely isolated from clinical material, especially from
immunocompromised patients. This bacterium is usually susceptible to most commonly used antibiotics, including beta-lactams.
However, a few cases of antibiotic-resistant L. adecarboxylata
strains have been reported (1, 2). Here, we report a case of a VIM1-producing L. adecarboxylata strain.
A survey study focused on compliance of hand hygiene among
the staff was performed in Na Homolce Hospital, Prague, Czech
Republic, in May 2011. Both hands were pressed onto the surface
of blood agar, which was then incubated overnight at 35°C. The
bacteria grown on blood agar were preliminarily identified by matrix-assisted laser desorption–ionization time of flight mass spectrometry (MALDI-TOF MS) (Bruker Daltonik, GmBH, Bremen,
Germany), and only potentially pathogenic bacteria (e.g., Enterobacteriaceae, Staphylococcus aureus) were subjected to susceptibility testing (3). Non-carbapenem-susceptible strains were further
investigated.
The only detected non-carbapenem-susceptible Enterobacteriaceae strain was L. adecarboxylata Lec-476. Identification of L.
adecarboxylata was, additionally, confirmed by its 16S rRNA gene
sequence (1). Lec-476 was resistant or nonsusceptible to piperacillin, piperacillin-tazobactam, cefotaxime, ceftazidime, cefepime,
and meropenem but susceptible to aztreonam (Table 1), as determined by the broth dilution method (3) and interpreted according to the criteria of the European Committee on Antimicrobial
Susceptibility Testing (EUCAST). The isolate was also resistant to
various non-␤-lactam antibiotics. Carbapenemase production
was hypothesized by a positive result in the MALDI-TOF MS
meropenem hydrolysis assay (4). Lec-476 tested Klebsiella pneumoniae carbapenemase (KPC) negative by the boronic acid-meropenem-combined disk test (5). The respective EDTA-meropenem
test appeared to be positive, indicating metallo-␤-lactamase
(M␤L) production (5). blaVIM-specific PCR (6) followed by amplicon sequencing identified the gene to be blaVIM-1.
Conjugal transfer of blaVIM-1 to the rifampin-resistant Escherichia coli A15 strain was achieved by mating experiments in
mixed-broth cultures, using rifampin (150 ␮g/ml) and ampicillin
Letter to the Editor
Nucleotide sequence accession number. The blaVIM-1-carrying sequence from L. adecarboxylata Lec-476 has been assigned
GenBank accession number KC430094.
ACKNOWLEDGMENTS
This work was supported by research project grant NT11032-6/2010 from
the Ministry of Health of the Czech Republic and by the Charles University Research Fund (project number P36). C. C. Papagiannitsis was supported by the project Support of Establishment, Development, and Mobility of Quality Research Teams at Charles University (registration
number CZ.1.07/2.3.00/30.0022), financed by The Education for Competitiveness Operational Programme (ECOP), funded by the ESF and the
government budget of the Czech Republic.
We declare that we have no conflict of interest.
REFERENCES
June 2013 Volume 57 Number 6
aac.asm.org 2897
Downloaded from http://aac.asm.org/ on October 6, 2014 by guest
1. Mazzariol A, Zuliani J, Fontana R, Cornaglia G. 2003. Isolation from
blood culture of a Leclercia adecarboxylata strain producing an SHV-12
extended-spectrum beta-lactamase. J. Clin. Microbiol. 41:1738 –1739.
2. Shin GW, You MJ, Lee HS, Lee CS. 2012. Catheter-related bacteremia
caused by multidrug-resistant Leclercia adecarboxylata in a patient with
breast cancer. J. Clin. Microbiol. 50:3129 –3132.
3. European Committee on Antimicrobial Susceptibility Testing. 2003.
Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by broth dilution. Clin. Microbiol. Infect. 9:1–7.
4. Hrabak J, Studentova V, Walkova R, Zemlickova H, Jakubu V, Chudackova E, Gniadkowski M, Pfeifer Y, Perry JD, Wilkinson K,
Bergerova T. 2012. Detection of NDM-1, VIM-1, KPC, OXA-48, and
OXA-162 carbapenemases by MALDI-TOF mass spectrometry. J. Clin.
Microbiol. 50:2441–2443.
5. Nordmann P, Gniadkowski M, Giske CG, Poirel L, Woodford N,
Miriagou V, European Network on Carbapenemases. 2012. Identification and screening of carbapenemase producing Enterobacteriaceae. Clin.
Microbiol. Infect. 18:432– 438.
6. Fiett J, Baraniak A, Mrówka A, Fleischer M, Drulis-Kawa Z, Naumiuk
Ł, Samet A, Hryniewicz W, Gniadkowski M. 2006. Molecular epidemiology of the acquired-metallo-␤-lactamase-producing bacteria in Poland.
Antimicrob. Agents Chemother. 50:880 – 886.
7. Giakkoupi P, Tzouvelekis LS, Tsakris A, Loukova V, Sofianou D,
Tzelepi E. 2000. IBC-1, a novel integron-associated class A ␤-lactamase
with extended-spectrum properties produced by an Enterobacter cloacae
clinical strain. Antimicrob. Agents Chemother. 44:2247–2253.
8. Barton BM, Harding GP, Zuccarelli AJ. 1995. A general method for
detecting and sizing large plasmids. Anal. Biochem. 226:235–240.
9. Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. 2005.
Identification of plasmids by PCR-based replicon typing. J. Microbiol.
Methods 63:219 –228.
10. Tato M, Coque TM, Baquero F, Canton R. 2010. Dispersal of carbapenemase blaVIM-1 gene associated with different Tn402 variants,
mercury transposons, and conjugative plasmids in Enterobacteriaceae
and Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 54:
320 –327.
11. Lombardi G, Luzzaro F, Docquier JD, Riccio ML, Perilli M, Coli A,
Amicosante G, Rossolini GM, Toniolo A. 2002. Nosocomial infections
caused by multidrug-resistant isolates of Pseudomonas putida producing
VIM-1 metallo-beta-lactamase. J. Clin. Microbiol. 40:4051– 4055.
12. Riccio ML, Pallechi L, Docquier JD, Cresti S, Catania MR, Pagani L,
Lagatolla C, Cornaglia G, Fontana R, Rossolini GM. 2005. Clonal
relatedness and conserved integron structures in epidemiologically unrelated Pseudomonas aeruginosa strains producing the VIM-1 metallo-␤lactamase from different Italian hospitals. Antimicrob. Agents Chemother. 49:104 –110.
13. Fischer J, Rodriguez I, Schmoger S, Friese A, Roesler U, Helmuth R,
Guerra B. 2012. Escherichia coli producing VIM-1 carbapenemase isolated
on a pig farm. J. Antimicrob. Chemother. 67:1793–1795.
14. Miriagou V, Papagiannitsis CC, Kotsakis SD, Loli A, Tzelepi E, Legakis
NJ, Tzouvelekis LS. 2010. Sequence of pNL194, a 79.3-kilobase IncN
plasmid carrying the blaVIM-1 metallo-␤-lactamase gene in Klebsiella
pneumoniae. Antimicrob. Agents Chemother. 54:4497–5502.