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2. Diversity and Resistance Profiles of ESBL-Producing Gram-Negative Bacteria from Dairy Farms in Southern Türkiye.

Author

Kürekci, Cemil, Yüksel, Murat, Celil Ozaslan, Büşra Gülay, Tan, Sait, Jäckel, Claudia, Grobbel, Mirjam, and Hammerl, Jens Andre

Subjects
ESCHERICHIA coli, GRAM-negative bacteria, DAIRY farms, CITROBACTER freundii, SHEEP ranches, GOAT milk, KLEBSIELLA pneumoniae
Abstract

Background/Objectives: The increasing occurrence of extended-spectrum β-lactamase (ESBL)–producing Enterobacterales, most commonly Escherichia coli, has become a serious problem. The aim of this study was to determine the presence of ESBL-producing Gram-negative bacteria in dairy cattle, goat and sheep farms located in southern Türkiye. Methods: Samples (409 quarter milk samples and 110 fresh faecal samples from cattle, 75 bulk tank milk samples and 225 rectal swab samples from goats and sheep) were subjected to selective isolation on MacConkey agar with ceftazidime (2 µg/mL). Isolates were identified by MALDI-ToF MS. The antimicrobial susceptibility profile of the isolates was determined by the broth microdilution method. To obtain a deeper insight into the genetic diversity of isolates substantially contributing to an efficient spread of their ESBL-determinants (23-MO00001: an E. coli from mastitis and 23-MO00002 Citrobacter freundii), the transmission potential and the genetic background of the plasmid carrying the blaCTX-M determinant was studied with whole genome analysis using Illumina sequencing. Results: Of the samples tested, 47 from the bovine faecal samples, 1 from the subclinical mastitis milk sample, 9 from the goat/sheep rectal swab samples and 5 from the goat/sheep bulk tank milk samples had ceftazidime-resistant Gram-negative strains with the ESBL phenotype. Of the 33 ESBL-producing E. coli isolates, 66.6% were resistant to tetracycline, 57.6% to sulfamethoxazole, 48.9% to nalidixic acid, 42.4% to ciprofloxacin and 33.3% to trimethoprim. Pulsed field gel electrophoresis (PFGE) results showed that the majority of E. coli isolates (16/33) and all Enterobacter spp. isolates (n = 5) were not clonally related (80% similarity cut value). The sequenced strains were observed to efficiently transfer their ceftazidime resistance to the recipient strain E. coli J53 at 37 °C (transfer rates: 101–102 transconjugants per donor cell). S1-PFGE showed that the transconjugants J53(p23MO01-T1) and J53(p23MO02-T1) had acquired plasmids of about 82 kb and 55 kb plasmids, respectively. According to WGS results, the E. coli isolate was assigned to ST162, while the C. freundii isolate was assigned to ST95. Conclusions: This study demonstrates that dairy animals are reservoirs of ESBL-producing bacteria. [ABSTRACT FROM AUTHOR]

Published
2024
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3. Elimination of Methicillin-Resistant Staphylococcus aureus from Mammary Glands of Dairy Cows by an Additional Antibiotic Treatment Prior to Dry Cow Treatment.

Author

Tenhagen, Bernd-Alois, Wörmann, Mirka Elisabeth, Gretzschel, Anja, Grobbel, Mirjam, Maurischat, Sven, and Lienen, Tobias

Subjects
METHICILLIN-resistant staphylococcus aureus, LICENSED products, RAW milk, DAIRY cattle, COWS, MAMMARY glands
Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) have been isolated from quarter milk samples of dairy cows, raising concerns over transmission to consumers of raw milk. This study investigates whether pre-treatment before dry-off can increase the success rate of dry cow treatment against MRSA. MRSA positive cows were assigned to two treatment groups. Both groups received dry cow treatment with a licensed product. The test group was additionally treated intramammarily with pirlimycin over seven days prior to the dry-off treatment. The use of pirlimycin increased the elimination of MRSA from previously MRSA positive udder quarters significantly (96.0 vs. 53.3%). However, MRSA were still present in noses and udder clefts of cows in MRSA negative quarter milk samples. New infections were observed in some quarters in both groups. Quarters that remained positive carried the same strain as prior to treatment. All MRSA isolates were associated with clonal complex CC398. Resistance to pirlimycin associated with the genes erm(C) or lnu(B) was observed in one isolate each from new infections after calving. Pretreatment supported the elimination of MRSA from the udder but did not eliminate MRSA from other body sites. Using the treatment will not eliminate the bacteria from the herd. [ABSTRACT FROM AUTHOR]

Published
2024
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4. Azithromycin resistance in Escherichia coli and Salmonella from food-producing animals and meat in Europe

Author

Ivanova, Mirena, primary, Ovsepian, Armen, additional, Leekitcharoenphon, Pimlapas, additional, Seyfarth, Anne Mette, additional, Mordhorst, Hanne, additional, Otani, Saria, additional, Koeberl-Jelovcan, Sandra, additional, Milanov, Mihail, additional, Kompes, Gordan, additional, Liapi, Maria, additional, Černý, Tomáš, additional, Vester, Camilla Thougaard, additional, Perrin-Guyomard, Agnès, additional, Hammerl, Jens A, additional, Grobbel, Mirjam, additional, Valkanou, Eleni, additional, Jánosi, Szilárd, additional, Slowey, Rosemarie, additional, Alba, Patricia, additional, Carfora, Virginia, additional, Avsejenko, Jelena, additional, Pereckiene, Asta, additional, Claude, Dominique, additional, Zerafa, Renato, additional, Veldman, Kees T, additional, Boland, Cécile, additional, Garcia-Graells, Cristina, additional, Wattiau, Pierre, additional, Butaye, Patrick, additional, Zając, Magdalena, additional, Amaro, Ana, additional, Clemente, Lurdes, additional, Vaduva, Angela M, additional, Romascu, Luminita-Maria, additional, Milita, Nicoleta-Manuela, additional, Mojžišová, Andrea, additional, Zdovc, Irena, additional, Escribano, Maria Jesús Zamora, additional, De Frutos Escobar, Cristina, additional, Overesch, Gudrun, additional, Teale, Christopher, additional, Loneragan, Guy H, additional, Guerra, Beatriz, additional, Beloeil, Pierre Alexandre, additional, Brown, Amanda M V, additional, Hendriksen, Rene S, additional, Bortolaia, Valeria, additional, and Kjeldgaard, Jette Sejer, additional

Published
2024
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5. Azithromycin resistance in Escherichia coli and Salmonella from food-producing animals and meat in Europe

Author

Ivanova, Mirena, Ovsepian, Armen, Leekitcharoenphon, Pimlapas, Seyfarth, Anne Mette, Mordhorst, Hanne, Otani, Saria, Koeberl-Jelovcan, Sandra, Milanov, Mihail, Kompes, Gordan, Liapi, Maria, Černý, Tomáš, Vester, Camilla T., Perrin-Guyomard, Agnès, Hammerl, Jens A., Grobbel, Mirjam, Valkanou, Eleni, Jánosi, Szilárd, Slowey, Rosemarie, Alba, Patricia, Carfora, Virginia, Avsejenko, Jelena, Pereckiene, Asta, Claude, Dominique, Zerafa, Renato, Veldman, Kees, Boland, Cécile, Garcia-Graells, Cristina, Wattiau, Pierre, Butaye, Patrick, Zając, Magdalena, Amaro, Ana, Clemente, Lurdes, Vaduva, Angela, Romascu, Luminita-Maria, Milita, Nicoleta-Manuela, Mojžišová, Andrea, Zdovc, Irena, Escribano, Maria, De Frutos Escobar, Cristina, Overesch, Gudrun, Teale, Christopher, Loneragan, Guy, Guerra, Beatriz, Beloeil, Pierre A., Brown, Amanda, Hendriksen, Rene, Bortolaia, Valeria, Kjeldgaard, Jette S., Ivanova, Mirena, Ovsepian, Armen, Leekitcharoenphon, Pimlapas, Seyfarth, Anne Mette, Mordhorst, Hanne, Otani, Saria, Koeberl-Jelovcan, Sandra, Milanov, Mihail, Kompes, Gordan, Liapi, Maria, Černý, Tomáš, Vester, Camilla T., Perrin-Guyomard, Agnès, Hammerl, Jens A., Grobbel, Mirjam, Valkanou, Eleni, Jánosi, Szilárd, Slowey, Rosemarie, Alba, Patricia, Carfora, Virginia, Avsejenko, Jelena, Pereckiene, Asta, Claude, Dominique, Zerafa, Renato, Veldman, Kees, Boland, Cécile, Garcia-Graells, Cristina, Wattiau, Pierre, Butaye, Patrick, Zając, Magdalena, Amaro, Ana, Clemente, Lurdes, Vaduva, Angela, Romascu, Luminita-Maria, Milita, Nicoleta-Manuela, Mojžišová, Andrea, Zdovc, Irena, Escribano, Maria, De Frutos Escobar, Cristina, Overesch, Gudrun, Teale, Christopher, Loneragan, Guy, Guerra, Beatriz, Beloeil, Pierre A., Brown, Amanda, Hendriksen, Rene, Bortolaia, Valeria, and Kjeldgaard, Jette S.

Published
2024

8. Rare Plasmid-Mediated AmpC Beta-Lactamase DHA-1 Located on Easy Mobilized IS26-Related Genetic Element Detected in Escherichia coli from Livestock and Food in Germany.

Author

Manfreda, Chiara, Kaesbohrer, Annemarie, Schmoger, Silvia, Skladnikiewicz-Ziemer, Tanja, Grobbel, Mirjam, and Irrgang, Alexandra

Subjects
ESCHERICHIA coli, BETA-lactamase inhibitors, THIRD generation cephalosporins, LIVESTOCK, FOOD chains, TRANSPOSONS, FOSFOMYCIN
Abstract

AmpC beta-lactamases cause resistance to third-generation cephalosporins, including beta-lactamase inhibitors. In Escherichia coli from the German food production chain, the majority of AmpC beta-lactamase activity can be attributed to plasmid-mediated CMY-2 or overproduction of chromosomal AmpC beta-lactamase, but occasionally other enzymes like DHA-1 are involved. This study investigated the prevalence of the AmpC beta-lactamase DHA-1 in ESBL/AmpC-producing E. coli (n = 4706) collected between 2016 and 2021 as part of a German antimicrobial resistance monitoring program along the food chain. Eight isolates (prevalence < 0.2%) were detected and further characterized by PFGE, transformation and conjugation experiments as well as short-read and long-read sequencing. All eight strains harbored blaDHA-1 together with qnrB4, sul1 and mph(A) resistance genes on an IS26 composite transposon on self-transferable IncFII or IncFIA/FIB/II plasmids. During laboratory experiments, activation of the translocatable unit of IS26-bound structures was observed. This was shown by the variability of plasmid sizes in original isolates, transconjugants or transferred plasmids, and correspondingly, duplications of resistance fragments were found in long-read sequencing. This activation could be artificial due to laboratory handling or naturally occurring. Nevertheless, DHA-1 is a rare AmpC beta-lactamase in livestock and food in Germany, and its dissemination will be monitored in the future. [ABSTRACT FROM AUTHOR]

Published
2024
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18. HIGH DIVERSITY OF PLASMIDS CARRYING QNR RESISTANCE GENES IN FLUOROQUINOLONE RESISTANT ESCHERICHIA COLI ISOLATED IN GERMANY IN 2017

Author

Juraschek, Katharina, Schmoger, Silvia, Grobbel, Mirjam, Malorny, Burkhard, Käsbohrer, Annemarie, Schwarz, Stefan, Meemken, Diana, and Hammerl, Jens Andre

Abstract

Aim: The prevalence and diversity of qnr-carrying isolates among (fluoro-)quinolone (FQ)-resistant, commensal E. coli from the German annual antimicrobial resistance monitoring in livestock and food in 2017 was determined. The data will be used to understand mechanisms involved in FQ-resistance development and the diversity of mobile genetic elements associated with their spread. Methods: A total of 3,409 E. coli isolates from the German National Reference Laboratory for Antimicrobial Resistance was investigated. Antimicrobial resistance testing was conducted by broth microdilution according to CLSI guidelines. MIC values for ciprofloxacin and nalidixic acid were evaluated using EUCAST epidemiological cut-off values. Isolates exhibiting resistances to FQ were subjected to qnr-PCR, XbaI-/S1-PFGE, whole genome sequencing (WGS) and bioinformatics analysis. Results: Overall, 504 isolates were classified as quinolone-resistant, while only 107 of them harbored a qnr gene. XbaI-macrorestriction demonstrated a high heterogeneity, letting us assume that no predominant E. coli lineages are associated with FQ resistance development in Germany. S1-PFGE analysis showed a variety of extrachromosomal elements of various sizes. Short-read WGS revealed qnr to be associated with bla as well as with the disinfectant qacEΔ1. The most abundant plasmid replicon type among our identified qnr reference plasmids were IncN, IncY and IncX1-IncX3. Further, we frequently detected chromosomal point mutations leading to an altered susceptibility against FQs. Conclusions: qnr genes were shown to be carried by multiple plasmid types and in frequent association with other resistance genes. This diversity of qnr plasmids in combination with their multi-resistance demonstrates the risk associated with qnr positive E. coli.

Published
2021
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19. Comparison of antibiotic resistance in Escherichia coli from clinical diagnostic submissions and isolates of healthy broilers, turkeys and calves from surveillance and monitoring systems in Germany and France

Author

Mesa-Varona, Octavio, Mader, Rodolphe, Granier, Sophie.A, Perrin, Jouy, Eric, Madec, Jean-Yves, Kaspar, Heike, Anjum, Muna, Grobbel, Mirjam, Velasova, Martina, and Tenhagen, Bernd-Alois

Abstract

The effectiveness of antibiotics has been reduced since their discovery. Surveillance and monitoring systems are key elements to collect and assess the trends of antibiotic resistance (ABR). Livestock data on ABR are traditionally collected from bacterial populations of clinical and non-clinical isolates. Resistance data on non-clinical isolates are based on the Decision 2013/652/EU in Europe. For clinical isolates, different standards, laboratory methods and methodologies are applied to collect ABR data. Further, a clinical interpretation is preferred in clinical isolates, while an epidemiological approach is frequently used in non-clinical isolates. Lack of harmonization between data types (clinical vs. non-clinical isolates) prevents the data comparison. The Normalized Resistance Interpretation (NRI) method was applied to circumvent the lack of ABR data harmonization between and within countries. Analyses were performed to identify associations between resistance to four antibiotics (ampicillin, gentamicin, nalidixic acid and tetracycline) and the data type variable per animal category (broiler, turkey and calf) within countries. Within countries, higher resistance proportions were found in clinical isolates vs commensal isolates to: gentamicin in broilers from France and in calves from Germany and France, nalidixic acid in calves from France and Germany and tetracycline for calves from France and Germany. In contrast, a higher probability of resistance in non-clinical isolates was encountered for tetracycline in broilers and turkeys from Germany and France and to gentamicin in turkeys from Germany. It seems that the higher presence of resistance in one data type (i.e. clinical or non-clinical isolates) is strongly associated with the relationship between the animal species and the antibiotic. The NRI identifies the wild-type distribution providing approximate epidemiological cut-offs that allow comparing quantitative results from different ABR systems with a different level of harmonization. This method might be regularly used in veterinary medicine and in One Health studies to compare not harmonized ABR systems until international harmonization of ABR is achieved.

Published
2021
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25. Clostridioides difficile antimicrobial susceptibility testing using disc diffusion

Author

Maurischat, Sven, Witt, P, Maneck, C, and Grobbel, Mirjam

Subjects
disk diffusion, Clostridioides difficile
Abstract

Routine antimicrobial susceptibility testing (AST) is a prerequisite for understanding and treating Clostridioides (formerly Clostridium, C.) difficile infections (CDI) which cause thousands of fatal cases in the EU each year. The disc diffusion (DD) methodology proved to bea suitable and convenient alternative to the current gold standard, the agar dilution (AD) for moxifloxacin, metronidazole and vancomycin [1]. Yet, DD has major drawbacks regarding reproducibility and the unknown influence of anaerobic conditions on the testresult. The aim of this study within the One Health European Joint Programme project IMPART (IMproving Phenotypic AntimicrobialResistance Testing) project is to optimize and propose a protocol for a robust DD method for C. difficile for a broader range ofantimicrobials and to find out, under which conditions and cut-off values a resistance determination is reliable.

Published
2020
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27. Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity

Author

Pauly, Natalie, primary, Klaar, Yvonne, additional, Skladnikiewicz-Ziemer, Tanja, additional, Juraschek, Katharina, additional, Grobbel, Mirjam, additional, Hammerl, Jens André, additional, Hemmers, Lukas, additional, Käsbohrer, Annemarie, additional, Schwarz, Stefan, additional, Meemken, Diana, additional, Tenhagen, Bernd-Alois, additional, and Irrgang, Alexandra, additional

Published
2021
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29. Comparison of Phenotypical Antimicrobial Resistance between Clinical and Non-Clinical E. coli Isolates from Broilers, Turkeys and Calves in Four European Countries

Author

Mesa-Varona, Octavio, primary, Mader, Rodolphe, additional, Velasova, Martina, additional, Madec, Jean-Yves, additional, Granier, Sophie A., additional, Perrin-Guyomard, Agnes, additional, Norstrom, Madelaine, additional, Kaspar, Heike, additional, Grobbel, Mirjam, additional, Jouy, Eric, additional, Anjum, Muna F., additional, and Tenhagen, Bernd-Alois, additional

Published
2021
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30. Wildlife as Sentinels of Antimicrobial Resistance in Germany?

Author

Plaza-Rodríguez, Carolina, primary, Alt, Katja, additional, Grobbel, Mirjam, additional, Hammerl, Jens Andre, additional, Irrgang, Alexandra, additional, Szabo, Istvan, additional, Stingl, Kerstin, additional, Schuh, Elisabeth, additional, Wiehle, Lars, additional, Pfefferkorn, Beatrice, additional, Naumann, Steffen, additional, Kaesbohrer, Annemarie, additional, and Tenhagen, Bernd-Alois, additional

Published
2021
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35. ChromID® CARBA Agar Fails to Detect Carbapenem-Resistant Enterobacteriaceae With Slightly Reduced Susceptibility to Carbapenems

Author

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

Published
2020
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41. High heterogeneity of plasmid-mediated quinolone resistance in Escherichia coli isolates recovered from livestock and food in Germany

Author

Juraschek, Katharina, Grobbel, Mirjam, Käsbohrer, Annemarie, Tenhagen, Bernd-Alois, and Hammerl, Jens Andre

Abstract

Introduction: Resistance to quinolones can be chromosomally encoded or plasmid-mediated (PMQR). One PMQR mechanism is mediated by Qnr proteins. The horizontal gene transfer of this plasmid-mediated quinolone resistance increases the threat of fallible treatment with quinolones. Objectives: To better understand the distribution of PMQR, in particular qnr genes, Escherichia (E.) coli isolates recovered in 2017 from livestock and food were phenotypically and genotypically characterized. Materials & Methods: 3,409 E. coli isolates from the German National Reference Laboratory for Antimicrobial Resistance were investigated. The isolates were received in the German national monitoring program for antimicrobial resistance. Antimicrobial resistance was determined by broth microdilution according to CLSI guidelines. MIC values for ciprofloxacin and nalidixic acid were evaluated using EUCAST epidemiological cut-off values (MICNAL ≥16 mg/L, MICCIP ≥0.06 mg/L). E.coli resistant to quinolones were subjected to qnr-PCR, XbaI-PFGE, S1-PFGE, WGS and bioinformatic analysis. Six different qnr-PCRs were conducted to identify the respective qnr-variants. Results: Overall, 504 isolates were classified as quinolone-resistant. Of those, 107 were found to harbor a qnr gene. The most abundant qnr-variant was qnrS. PFGE profiling for the 107 qnr positive isolates demonstrated a high heterogeneity, indicating that they are not associated to a predominant E. coli clone spreading via vertical transmission. S1-PFGE plasmid profiling showed a variety of extrachromosomal elements of various sizes. 43 Isolates, selected according their XbaI- and S1-PFGE pattern were further screened for their genetic setting through short read whole genome sequencing (WGS). Sequencing confirmed the high genetic diversity of the quinolone-resistant E. coli strains. Conclusion: Quinolone-resistance could not be attributed to a specific lineage of E. coli. Further analysis is needed for a better understanding of the plasmid diversity within qnr-harboring E. coli and the prerequisites of their spread.&nbsp

Published
2020
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42. Genomic and Functional Analysis of Emerging Virulent and Multidrug-Resistant Escherichia coli Lineage Sequence Type 648

Author

Schaufler, Katharina, primary, Semmler, Torsten, additional, Wieler, Lothar H., additional, Trott, Darren J., additional, Pitout, Johann, additional, Peirano, Gisele, additional, Bonnedahl, Jonas, additional, Dolejska, Monika, additional, Literak, Ivan, additional, Fuchs, Stephan, additional, Ahmed, Niyaz, additional, Grobbel, Mirjam, additional, Torres, Carmen, additional, McNally, Alan, additional, Pickard, Derek, additional, Ewers, Christa, additional, Croucher, Nicholas J., additional, Corander, Jukka, additional, and Guenther, Sebastian, additional

Published
2019
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44. Genomic and Functional Analysis of Emerging Virulent and Multidrug-Resistant Escherichia coli Lineage Sequence Type 648

Author

Schaufler, Katharina, Semmler, Torsten, Wieler, Lothar H., Trott, Darren J., Pitout, Johann, Peirano, Gisele, Bonnedahl, Jonas, Dolejska, Monika, Literak, Ivan, Fuchs, Stephan, Ahmed, Niyaz, Grobbel, Mirjam, Torres, Carmen, McNally, Alan, Pickard, Derek, Ewers, Christa, Croucher, Nicholas J., Corander, Jukka, Guenther, Sebastian, Schaufler, Katharina, Semmler, Torsten, Wieler, Lothar H., Trott, Darren J., Pitout, Johann, Peirano, Gisele, Bonnedahl, Jonas, Dolejska, Monika, Literak, Ivan, Fuchs, Stephan, Ahmed, Niyaz, Grobbel, Mirjam, Torres, Carmen, McNally, Alan, Pickard, Derek, Ewers, Christa, Croucher, Nicholas J., Corander, Jukka, and Guenther, Sebastian

Abstract

The pathogenic extended-spectrum-beta-lactamase (ESBL)-producing Escherichia coli lineage ST648 is increasingly reported from multiple origins. Our study of a large and global ST648 collection from various hosts (87 whole-genome sequences) combining core and accessory genomics with functional analyses and in vivo experiments suggests that ST648 is a nascent and generalist lineage, lacking clear phylogeographic and host association signals. By including large numbers of ST131 (n = 107) and ST10 (n = 96) strains for comparative genomics and phenotypic analysis, we demonstrate that the combination of multidrug resistance and high-level virulence are the hallmarks of ST648, similar to international high-risk clonal lineage ST131. Specifically, our in silico, in vitro, and in vivo results demonstrate that ST648 is well equipped with biofilm-associated features, while ST131 shows sophisticated signatures indicative of adaption to urinary tract infection, potentially conveying individual ecological niche adaptation. In addition, we used a recently developed NFDS (negative frequency-dependent selection) population model suggesting that ST648 will increase significantly in frequency as a cause of bacteremia within the next few years. Also, ESBL plasmids impacting biofilm formation aided in shaping and maintaining ST648 strains to successfully emerge worldwide across different ecologies. Our study contributes to understanding what factors drive the evolution and spread of emerging international high-risk clonal lineages., Funding Agencies|German Research Foundation (DFG) [GU 1283/3-1, EW116/2-1]; Feodor Lynen Research Fellowship by the Alexander von Humboldt Foundation, Germany; Wellcome Trust Sanger Institute (Cambridge, United Kingdom) [2433ILB]; project CEITEC 2020 from the Ministry of Education, Youth, and Sports of the Czech Republic [LQ1601]

Published
2019
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45. ChromID® CARBA Agar Fails to Detect Carbapenem-Resistant Enterobacteriaceae With Slightly Reduced Susceptibility to Carbapenems.

Author

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

Subjects
CARBAPENEM-resistant bacteria, CARBAPENEMS, AGAR, DRUG resistance in microorganisms, NATIONAL interest, CECUM
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. [ABSTRACT FROM AUTHOR]

Published
2020
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