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, Jaroslav.Hrabak@lfp.cuni.cz. 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. 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