Evaluation and validation of laboratory procedures for the surveillance of ESBL-, AmpC-, and carbapenemase-producing Escherichia coli from fresh meat and caecal samples

Frontiers in Microbiology

Microbiology

Henrik Hasman¹  Valeria Bortolaia¹  Lina M. Cavaco¹  Yvonne Agersø²  Beatriz Guerra³  Rene S. Hendriksen⁴  Christina Aaby Svendsen⁴  Hanne Nørgaard Nielsen⁴  Jette Sejer Kjeldgaard⁴  Susanne Karlsmose Pedersen⁴  Mette Fertner⁴ 

[1] Department for Bacteria, Parasites and Fungi, Statens Serum Institut, Reference Laboratory for Antimicrobial Resistance, Copenhagen, Denmark;Department of Veterinary and Animal Sciences, University of Copenhagen University, Copenhagen, Denmark;European Food Safety Authority, Parma, Italy;National Food Institute, Technical University of Denmark (DTU Food), European Union Reference Laboratory for Antimicrobial Resistance, Research Group for Global Capacity Building, Kgs. Lyngby, Denmark;

关键词: extended-spectrum beta-lactamase;    carbapenemase;    isolation method;    Escherichia coli;    protocol;    surveillance;    European Union;   

DOI  :  10.3389/fmicb.2023.1229542

 received in 2023-05-26, accepted in 2023-07-12,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionExtended-spectrum β-lactamase- (ESBL) and AmpC- β-lactamase-producing Enterobacterales are widely distributed and emerging in both human and animal reservoirs worldwide. A growing concern has emerged in Europe following the appearance of carbapenemase-producing Escherichia coli (E. coli) in the primary production of food animals. In 2013, the European Commission (EC) issued the Implementing Decision on the monitoring and reporting of antimicrobial resistance in zoonotic and commensal bacteria. The European Union Reference Laboratory for Antimicrobial Resistance (EURL-AR) was tasked with providing two laboratory protocols for samples derived from meat and caecal content, respectively, for the isolation of ESBL- and AmpC-producing E. coli (part 1) and carbapenemase-producing (CP) E. coli (part 2). In this study, we describe the current protocols, including the preparatory work for the development.MethodsUp to nine laboratory procedures were tested using minced meat as the matrix from beef, pork, and chicken as well as six procedures for the caecal content of cattle, pigs, and chicken. Variables included sample volume, pre-enrichment volume, pre-enrichment broth with and without antimicrobial supplementation, and incubation time/temperature. The procedures were evaluated against up to nine E. coli strains harboring different AMR genes and belonging to the three β-lactamase groups.Results and discussionThe laboratory procedures tested revealed that the most sensitive and specific methodologies were based on a Buffered Peptone Water pre-enrichment of 225 ml to 25 g or 9 ml to 1 g for minced meat and caecal content, respectively, incubated at 37°C overnight, followed by inoculation onto MacConkey agar supplemented with 1 mg/L cefotaxime for detecting ESBL- and AmpC-producing E. coli and Chrom ID SMART (Chrom ID CARBA and OXA) for CP E. coli, incubated overnight at 37 and 44°C, respectively. We provided two isolation protocols for the EU-specific monitoring of ESBL- and AmpC- producing E. coli (part 1) and CP E. coli (part 2) from fresh meat (protocol 1) and caecal (protocol 2) samples, which have been successfully implemented by all EU Member States for the monitoring period 2014–2027 (EU 2020/1729).

【 授权许可】

Unknown   
Copyright © 2023 Hendriksen, Cavaco, Guerra, Bortolaia, Agersø, Svendsen, Nielsen, Kjeldgaard, Pedersen, Fertner and Hasman.