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Volume 9
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10.3390/microorganisms9051105
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Article

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

by
Natalie Pauly
1,
Yvonne Klaar
1,
Tanja Skladnikiewicz-Ziemer
1,
Katharina Juraschek
1,
Mirjam Grobbel
1,
Jens André Hammerl
1,
Lukas Hemmers
2,
Annemarie Käsbohrer
1,3,
Stefan Schwarz
4,5,
Diana Meemken
6,
Bernd-Alois Tenhagen
1 and
Alexandra Irrgang
1,*
1
German Federal Institute for Risk Assessment (Bundesinstitut für Risikobewertung), D-10589 Berlin, Germany
2
State Office for Health and Social Affairs Berlin, D-10639 Berlin, Germany
3
Unit for Veterinary Public Health and Epidemiology, University of Veterinary Medicine, AT-1210 Vienna, Austria
4
Institute of Microbiology and Epizootics, Freie Universität Berlin, D-14163 Berlin, Germany
5
FAO Reference Centre for Antimicrobial Resistance, Freie Universität Berlin, D-14163 Berlin, Germany
6
Institute of Food Safety and Food Hygiene, Freie Universität Berlin, D-14163 Berlin, Germany
*
Author to whom correspondence should be addressed.
Microorganisms 2021, 9(5), 1105; https://doi.org/10.3390/microorganisms9051105
Submission received: 30 March 2021 / Revised: 13 May 2021 / Accepted: 14 May 2021 / Published: 20 May 2021
(This article belongs to the Special Issue Farm Animal and Wildlife Zoonotic Microorganisms)
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Abstract

Due to the increasing reports of carbapenemase-producing Enterobacteriaceae (CPE) from livestock in recent years, the European Reference Laboratory for Antimicrobial Resistances (EURL-AR) provided a protocol for their recovery from caecum and meat samples. This procedure exhibited limitations for the detection of CPE with low carbapenem MIC values. Therefore, it was modified by a second, selective enrichment in lysogeny broth with cefotaxime (CTX 1 mg/L) and with meropenem (MEM 0.125 mg/L) at 37 °C under microaerophilic conditions. By Real-time PCR, these enrichments are pre-screened for the most common carbapenemase genes. Another adaptation was the use of in-house prepared MacConkey agar with MEM and MEM+CTX instead of commercial selective agar. According to the EURL-method, we achieved 100% sensitivity and specificity using the in-house media instead of commercial agar, which decreased the sensitivity to ~75%. Comparing the method with and without the second enrichment, no substantial influence on sensitivity and specificity was detected. Nevertheless, this enrichment has simplified the CPE-isolation regarding the accompanying microbiota and the separation of putative colonies. In conclusion, the sensitivity of the method can be increased with slight modifications.
Keywords: Isolation; carbapenemase; CPE detection; selective media; specificity; sensitivity Isolation; carbapenemase; CPE detection; selective media; specificity; sensitivity

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MDPI and ACS Style

Pauly, N.; Klaar, Y.; Skladnikiewicz-Ziemer, T.; Juraschek, K.; Grobbel, M.; Hammerl, J.A.; Hemmers, L.; Käsbohrer, A.; Schwarz, S.; Meemken, D.; et al. Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity. Microorganisms 2021, 9, 1105. https://doi.org/10.3390/microorganisms9051105

AMA Style

Pauly N, Klaar Y, Skladnikiewicz-Ziemer T, Juraschek K, Grobbel M, Hammerl JA, Hemmers L, Käsbohrer A, Schwarz S, Meemken D, et al. Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity. Microorganisms. 2021; 9(5):1105. https://doi.org/10.3390/microorganisms9051105

Chicago/Turabian Style

Pauly, Natalie, Yvonne Klaar, Tanja Skladnikiewicz-Ziemer, Katharina Juraschek, Mirjam Grobbel, Jens André Hammerl, Lukas Hemmers, Annemarie Käsbohrer, Stefan Schwarz, Diana Meemken, and et al. 2021. "Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity" Microorganisms 9, no. 5: 1105. https://doi.org/10.3390/microorganisms9051105

APA Style

Pauly, N., Klaar, Y., Skladnikiewicz-Ziemer, T., Juraschek, K., Grobbel, M., Hammerl, J. A., Hemmers, L., Käsbohrer, A., Schwarz, S., Meemken, D., Tenhagen, B.-A., & Irrgang, A. (2021). Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity. Microorganisms, 9(5), 1105. https://doi.org/10.3390/microorganisms9051105

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