First Detection of VIM-2 Metallo-beta-Lactamase-Producing Pseudomonas putida in Blattella germanica Cockroaches in an Algerian Hospital

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First Detection of VIM-2

Metallo-beta-Lactamase-Producing Pseudomonas putida

in Blattella germanica Cockroaches in an Algerian

Hospital

Lotfi Loucif, Zineb Cherak, Naima Chamlal, Esma Bendjama, Djamila

Gacemi-Kirane, Nadia Grainat, Jean-Marc Rolain

To cite this version:

Lotfi Loucif, Zineb Cherak, Naima Chamlal, Esma Bendjama, Djamila Gacemi-Kirane, et al.. First

Detection of VIM-2 Metallo-beta-Lactamase-Producing Pseudomonas putida in Blattella germanica

Cockroaches in an Algerian Hospital. Antimicrobial Agents and Chemotherapy, American Society for

Microbiology, 2017, 61 (8), �10.1128/AAC.00357-17�. �hal-01774703�

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First Detection of VIM-2 Metallo-

␤-Lactamase-Producing Pseudomonas

putida in Blattella germanica

Cockroaches in an Algerian Hospital

Lotﬁ Loucif,

a,b

_{Zineb Cherak,}

c

_{Naima Chamlal,}

d

_{Esma Bendjama,}

a,b

Djamila Gacemi-Kirane,

e

_{Nadia Grainat,}

f

_{Jean-Marc Rolain}

b

Laboratoire de Biotechnologie des Molécules Bioactives et de la Physiopathologie Cellulaire (LBMBPC), Faculté des Sciences de la Nature et de la Vie, Université de Batna 2, Batna, Algeriaa_{; Unité de Recherche sur les}

Maladies Infectieuses et Tropicales Émergentes (URMITE), UM 63, CNRS 7278, IRD 198, INSERM 1095, IHU Méditérranée Infection, Faculté de Médecine et de Pharmacie, Aix-Marseille-Université, Marseille, Franceb_;

Laboratoire de Génétique, Biotechnologie, et Valorisation des Bio-ressources (GBVB), Faculté des Sciences Exactes et des Sciences de la Nature et de la Vie, Université Mohamed Khider, Biskra, Algeriac_{; Département de}

Biologie, Faculté des Sciences de la Nature et de la Vie et Sciences de la Terre et de l'Univers, Université 08 Mai 1945, Guelma, Algeriad_{; Département de Biochimie, Faculté des Sciences, Université Badji Mokhtar, Annaba,}

Algeriae_{; Faculté de Médecine, Université de Batna 2, Batna, Algeria}f

KEYWORDS

VIM-2 MBL, Pseudomonas putida, Blattella germanica, Algeria

I

n the last few years, carbapenem resistance has increasingly been reported in

Gram-negative bacilli (GNB), particularly via carbapenemase production (1).

Metallo-␤-lactamases (MBLs) are the most commonly acquired carbapenemases identiﬁed

among Pseudomonas spp. (2). Moreover, the VIM-2 MBL, initially reported in a

Pseu-domonas aeruginosa clinical isolate from France (3), is now endemic in many parts of

the world (4). In our recent investigation of hospital cockroaches of the Blattella

germanica species, we reported the carriage of carbapenemase-producing

Enterobac-teriaceae by these insects, which may present a serious health threat (5). Here we report

the ﬁrst detection of VIM-2-MBL-producing Pseudomonas putida in Algeria through its

ﬁrst isolation from hospital cockroaches of the Blattella germanica species.

Ten German cockroaches (B. germanica) were randomly caught from the burn unit

of Batna University Hospital, Algeria. Samples were subjected to the

bacterial-suspension preparation and preenrichment as previously described (5). Subsequently,

selective enrichment was performed in a vancomycin- and ertapenem-supplemented

brain heart infusion broth (BD, France) (5). One hundred microliters from each positive

tube was plated onto MacConkey agar plates (BD) supplemented with 64 mg/liter of

vancomycin and 1 mg/liter of imipenem. Representative colonies of nonfermenting

GNB were screened for carbapenemase production using the modiﬁed Carba NP test

(MCNP test). Only one positive MCNP test isolate was obtained from an external surface,

identiﬁed as P. putida by Vitek 2 (bioMérieux), and conﬁrmed by matrix-assisted laser

desorption–ionization time of ﬂight mass spectrometry (6). Susceptibility testing was

performed using a disk diffusion technique, and MICs were determined by Vitek 2.

Interpretations were made according to Comité de l’Antibiogramme de la Société

Française de Microbiologie (CA-SFM) breakpoints (7) (Table 1). The P. putida isolate was

resistant to ticarcillin, ticarcillin-clavulanate, piperacillin, piperacillin-tazobactam,

cefta-zidime, imipenem, meropenem, gentamicin, and tobramycin, intermediate to

aztreo-nam, but susceptible to cefepime, ciproﬂoxacin, amikacin, and colistin. MBL production

was conﬁrmed by the MBL double-sided Etest (imipenem and imipenem plus EDTA)

(bioMérieux). The isolate was screened using real-time PCR for the presence of the

following

␤-lactamase genes: bla

CTX-M

, bla

TEM

, bla

SHV

, bla

KPC

, bla

NDM

, bla

VIM

, and bla

IMP

.

Accepted manuscript posted online 30 May

2017

Citation Loucif L, Cherak Z, Chamlal N,

Bendjama E, Gacemi-Kirane D, Grainat N, Rolain J-M. 2017. First detection of VIM-2 metallo-β-lactamase-producing

Pseudomonas putida in Blattella germanica

cockroaches in an Algerian hospital. Antimicrob Agents Chemother 61:e00357-17.https://doi.org/10.1128/AAC.00357-17.

LETTER TO THE EDITOR

crossm

August 2017 Volume 61 Issue 8 e00357-17 Antimicrobial Agents and Chemotherapy aac.asm.org 1

on May 7, 2018 by Bibliotheque - Univ. de la Mediterranee

http://aac.asm.org/

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It was positive only for the bla

VIM

carbapenemase gene. We searched for the antibiotic

resistance-encoding genes bla

PER

, bla

VEB

, bla

GES

, bla

VIM

, aac(3)-Ia, aac(6=)-Ib, aph(3=)-VI,

and armA by standard PCR and sequencing as previously described (8, 9). Sequencing

of the positive-standard PCR products showed that the P. putida isolate carried the

blaVIM-2

and aac(6=)-Ib genes. The transfer of the detected bla

VIM-2

gene by both

conjugation and transformation experiments was unsuccessful. However, this does not

exclude a possible plasmid location of the bla

VIM-2

gene, and further investigations are

required to determine its location. The presence of the class 1 integron was conﬁrmed

by integrase gene PCR targeting the intI1 and intI2 integrase genes as previously

described (10). VIM-2-MBL-producing P. putida was ﬁrst detected in Taiwan and Korea

(11, 12). Subsequently, nosocomial infections caused by VIM-2-producing P. putida have

occasionally been reported (13–15). In an earlier investigation, Saitou et al. searched for

MBL genes of the VIM-2 and IMP-1 types in 45 P. aeruginosa isolates from hospital

cockroaches in Japan. However, all the studied strains were negative for both MBL

genes (16). Moreover, the isolation of imipenem-resistant Pseudomonas ﬂuorescens/

putida from hospital cockroaches has already been reported (17). In addition, the role

of P. putida as a reservoir of MBL genes that can be transferred to Pseudomonas

aeruginosa has already been demonstrated (18), which makes the presence of such

organisms in hospital cockroaches worrisome in spite of their low clinical importance.

To the best of our knowledge, these data document the ﬁrst detection of

VIM-2-producing P. putida in cockroaches; therefore, these pests play a potential role in the

dissemination of multidrug-resistant bacteria.

ACKNOWLEDGMENTS

We are very grateful to Mohamed Islam Hatcha for his contribution and to Helis

Yassine (Laboratoire d’Analyses Médicales, LAM IBN ROCHD, Batna, Algeria) for Vitek 2

(bioMérieux) utilization.

This work was partly funded by the Centre National de la Recherche Scientiﬁque

(CNRS 7278) and IHU Méditérranée Infection.

We declare no conﬂicts of interest.

REFERENCES

1. Poirel L, Potron A, Nordmann P. 2012. OXA-48-like carbapenemases: the phantom menace. J Antimicrob Chemother 67:1597–1606.https://doi .org/10.1093/jac/dks121.

2. Gniadek TJ, Carroll KC, Simner PJ. 2016. Carbapenem-resistant non-glucose-fermenting Gram-negative bacilli: the missing piece to the puzzle. J Clin Microbiol 54:1700 –1710.https://doi.org/10.1128/JCM.03264-15.

3. Poirel L, Naas T, Nicolas D, Collet L, Bellais S, Cavallo JD, Nordmann P. 2000. Characterization of VIM-2, a carbapenem-hydrolyzing metallo-beta-lactamase and its plasmid- and integron-borne gene from a

Pseu-domonas aeruginosa clinical isolate in France. Antimicrob Agents

Che-mother 44:891– 897.https://doi.org/10.1128/AAC.44.4.891-897.2000. 4. Diene SM, Rolain J-M. 2014. Carbapenemase genes and genetic

plat-forms in Gram-negative bacilli: Enterobacteriaceae, Pseudomonas and

Acinetobacter species. Clin Microbiol Infect 20:831– 838.https://doi.org/ 10.1111/1469-0691.12655.

5. Loucif L, Gacemi-Kirane D, Cherak Z, Chamlal N, Grainat N, Rolain J-M. 2016. First report of German cockroaches (Blattella germanica) as reser-voirs of CTX-M-15 extended-spectrum-beta-lactamase- and OXA-48 carbapenemase-producing Enterobacteriaceae in Batna University Hos-pital, Algeria. Antimicrob Agents Chemother 60:6377– 6380.https://doi .org/10.1128/AAC.00871-16.

6. Seng P, Drancourt M, Gouriet F, La Scola B, Fournier P-E, Rolain JM, Raoult D. 2009. Ongoing revolution in bacteriology: routine identifica-tion of bacteria by matrix-assisted laser desorpidentifica-tion ionizaidentifica-tion time-of-flight mass spectrometry. Clin Infect Dis 49:543–551.https://doi.org/10 .1086/600885.

TABLE 1 MICs of the P. putida isolate

Antimicrobial agenta _{MIC (}_␮g/ml)

PIP 32 TPZ 32 CAZ 16 FEP 2 IPM ⬎32 AMK 0.5 TOB 8 CIP 0.5 CST 0.25

a_{PIP, piperacillin; TPZ, piperacillin-tazobactam; CAZ, ceftazidime; FEP, cefepime; IPM, imipenem; AMK,} amikacin; TOB, tobramycin; CIP, ciproﬂoxacin; CST, colistin.

Letter to the Editor Antimicrobial Agents and Chemotherapy

August 2017 Volume 61 Issue 8 e00357-17 aac.asm.org 2

on May 7, 2018 by Bibliotheque - Univ. de la Mediterranee

http://aac.asm.org/

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7. EUCAST. 2016. Comité de l’Antibiogramme de la Société Française de Microbiologie. Recommandations 2016, v.1.0 Février. Société Française de Microbiologie, Paris, France.http://www.sfm-microbiologie.org/UserFiles/ ﬁles/casfm/CASFM2016_V1_0_FEVRIER.pdf.

8. Mesli E, Berrazeg M, Drissi M, Bekkhoucha SN, Rolain J-M. 2013. Preva-lence of carbapenemase-encoding genes including New Delhi metallo-beta-lactamase in Acinetobacter species, Algeria. Int J Infect Dis 17: e739 – e743.https://doi.org/10.1016/j.ijid.2013.02.024.

9. Mellouk FZ, Bakour S, Meradji S, Al-Bayssari C, Bentakouk MC, Zouyed F, Djahoudi A, Boutefnouchet N, Rolain JM. 17 June 2016. First detection of VIM-4-producing Pseudomonas aeruginosa and OXA-48-producing

Kleb-siella pneumoniae in northeastern (Annaba, Skikda) Algeria. Microb Drug

Resisthttps://doi.org/10.1089/mdr.2016.0032.

10. Koeleman JG, Stoof J, Van Der Bijl MW, Vandenbroucke-Grauls CM, Savelkoul PH. 2001. Identiﬁcation of epidemic strains of Acinetobacter

baumannii by integrase gene PCR. J Clin Microbiol 39:8 –13.https://doi .org/10.1128/JCM.39.1.8-13.2001.

11. Yan JJ, Hsueh PR, Ko WC, Luh KT, Tsai SH, Wu HM, Wu JJ. 2001. Metallo-beta-lactamases in clinical Pseudomonas isolates in Taiwan and identiﬁcation of VIM-3, a novel variant of the VIM-2 enzyme. Antimicrob Agents Chemother 45:2224 –2228. https://doi.org/10.1128/AAC.45.8 .2224-2228.2001.

12. Lee K, Lim JB, Yum JH, Yong D, Chong Y, Kim JM, Livermore DM. 2002.

bla(VIM-2)cassette-containing novel integrons in

metallo-beta-lactamase-producing Pseudomonas aeruginosa and Pseudomonas putida isolates

disseminated in a Korean hospital. Antimicrob Agents Chemother 46: 1053–1058.https://doi.org/10.1128/AAC.46.4.1053-1058.2002. 13. Trevino M, Moldes L, Hernandez M, Martinez-Lamas L, Garcia-Riestra C,

Regueiro BJ. 2010. Nosocomial infection by VIM-2 metallo-beta-lactamase-producing Pseudomonas putida. J Med Microbiol 59:853– 855.

https://doi.org/10.1099/jmm.0.018036-0.

14. Marchiaro P, Viale AM, Ballerini V, Rossignol G, Vila AJ, Limansky A. 2010. First report of a Tn402-like class 1 integron carrying blaVIM-2 in

Pseu-domonas putida from Argentina. J Infect Dev Ctries 4:412– 416.

15. Almuzara M, Radice M, de Garate N, Kossman A, Cuirolo A, Santella G, Famiglietti A, Gutkind G, Vay V. 2007. VIM-2-producing Pseudomonas

putida, Buenos Aires. Emerg Infect Dis 13:668 – 669.https://doi.org/10 .3201/eid1304.061083.

16. Saitou K, Furuhata K, Kawakami Y, Fukuyama M. 2009. Isolation of

Pseudomonas aeruginosa from cockroaches captured in hospitals in

Japan, and their antibiotic susceptibility. Biocontrol Sci 14:155–159.

https://doi.org/10.4265/bio.14.155.

17. Pai H-H. 2013. Multidrug resistant bacteria isolated from cockroaches in long-term care facilities and nursing homes. Acta Trop 125:18 –22.

https://doi.org/10.1016/j.actatropica.2012.08.016.

18. Juan C, Zamorano L, Mena A, Alberti S, Pérez JL, Oliver A. 2010. Metallo-␤-lactamase-producing Pseudomonas putida as a reservoir of multidrug resistance elements that can be transferred to successful Pseudomonas

aeruginosa clones. J Antimicrob Chemother 65:474 – 478.https://doi.org/ 10.1093/jac/dkp491.

Letter to the Editor Antimicrobial Agents and Chemotherapy

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