ChromID® CARBA Agar Fails to Detect Carbapenem-Resistant Enterobacteriaceae With Slightly Reduced Susceptibility to Carbapenems

Natalie Pauly; Jens A Hammerl; Mirjam Grobbel; Bernd-Alois Tenhagen; Annemarie Käsbohrer; Sandra Bisenius; Jannika Fuchs; Sabine Horlacher; Holger Lingstädt; Ute Mauermann; Silke Mitro; Margit Müller; Stefan Rohrmann; Arthur P Schiffmann; Birgit Stührenberg; Pia Zimmermann; Stefan Schwarz; Diana Meemken; Alexandra Irrgang

doi:10.3389/fmicb.2020.01678

ChromID^® CARBA Agar Fails to Detect Carbapenem-Resistant Enterobacteriaceae With Slightly Reduced Susceptibility to Carbapenems

Front Microbiol. 2020 Aug 11:11:1678. doi: 10.3389/fmicb.2020.01678. eCollection 2020.

Authors

Natalie Pauly¹, Jens A Hammerl¹, Mirjam Grobbel¹, Bernd-Alois Tenhagen¹, Annemarie Käsbohrer^{1

2}, Sandra Bisenius³, Jannika Fuchs⁴, Sabine Horlacher⁵, Holger Lingstädt⁶, Ute Mauermann⁷, Silke Mitro⁸, Margit Müller⁹, Stefan Rohrmann¹⁰, Arthur P Schiffmann¹¹, Birgit Stührenberg¹², Pia Zimmermann¹³, Stefan Schwarz¹⁴, Diana Meemken¹⁵, Alexandra Irrgang¹

Affiliations

¹ German Federal Institute for Risk Assessment, Berlin, Germany.
² Unit for Veterinary Public Health and Epidemiology, University of Veterinary Medicine, Vienna, Austria.
³ Institute for Fish and Fishery Products (LAVES), Cuxhaven, Germany.
⁴ Chemical and Veterinary Investigation Office, Karlsruhe, Germany.
⁵ Chemical and Veterinary Investigation Office, Fellbach, Germany.
⁶ State Office for Consumer Protection Saxony-Anhalt, Stendal, Germany.
⁷ Food and Veterinary Institute, Oldenburg, Germany.
⁸ State Investigation Institute for Health and Veterinary Services, Chemnitz, Germany.
⁹ Chemical and Veterinary Investigation Office Rhein-Ruhr-Wupper, Krefeld, Germany.
¹⁰ Chemical and Veterinary Investigation Office, Arnsberg, Germany.
¹¹ Hessian State Laboratory, Gießen, Germany.
¹² Chemical and Veterinary Investigation Office, Detmold, Germany.
¹³ Bavarian Health and Food Safety Authority, Oberschleißheim, Germany.
¹⁴ Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.
¹⁵ Institute of Food Safety and Food Hygiene, Freie Universität Berlin, Berlin, Germany.

Abstract

After first detections of carbapenemase-producing Enterobacteriaceae (CPE) in animals, the European Union Reference Laboratory for Antimicrobial Resistance has provided a protocol for the isolation of carbapenemase-producing Escherichia (E.) coli from cecum content and meat. Up to now, only few isolates were recovered using this procedure. In our experience, the choice of the selective agar is important for the efficacy of the method. Currently, the use of the prevailing method fails to detect CPE that exhibit a low resistance against carbapenems. Thus, this study aims to evaluate the suitability of selective media with antibiotic supplements and commercial ChromID^® CARBA agar for a reliable CPE detection. For comparative investigations, detection of freeze-dried carbapenemase-resistant bacteria was studied on different batches of the ChromID^® CARBA agar as well as on MacConkey agar supplemented with 1 mg/L cefotaxime and 0.125 mg/L meropenem (McC+CTX+MEM). The suitability of the different media was assessed within a time of 25 weeks, starting at least six weeks before expiration of the media. Carbapenem-resistant isolates exhibiting a serine-based hydrolytic resistance mechanism (e.g., bla _KPC genes) were consistently detected over 25 weeks on the different media. In contrast, carbapenemase producers with only slightly reduced susceptibility and exhibiting a zinc-catalyzed activity (e.g., bla _VIM, bla _NDM, and bla _IMP) could only be cultivated on long-time expired ChromID^® CARBA, but within the whole test period on McC+CTX+MEM. Thus, ChromID^® CARBA agar appears to be not suitable for the detection of CPE with slightly increased minimum inhibitory concentrations (MIC) against carbapenems, which have been detected in German livestock and thus, are of main interest in the national monitoring programs. Our data are in concordance with the results of eleven state laboratories that had participated in this study with their ChromID^® CARBA batches routinely used for the German CPE monitoring. Based on the determined CPE detection rate, we recommend the use of McC+CTX+MEM for monitoring purposes. This study indicates that the use of ChromID^® CARBA agar might lead to an underestimation of the current CPE occurrence in food and livestock samples.

Keywords: carbapenemase; isolation; media; sensitivity; specificity.

Copyright © 2020 Pauly, Hammerl, Grobbel, Tenhagen, Käsbohrer, Bisenius, Fuchs, Horlacher, Lingstädt, Mauermann, Mitro, Müller, Rohrmann, Schiffmann, Stührenberg, Zimmermann, Schwarz, Meemken and Irrgang.