Whole genome sequence analysis of antimicrobial resistance genes, multilocus sequence types and plasmid sequences in ESBL/AmpC Escherichia coli isolated from broiler caecum and meat.

Plasmid-encoded extended-spectrum β-lactamase and AmpC gene-carrying Escherichia coli (ESBL/AmpC E. coli) is an increasing cause of human infections worldwide. Increasing carbapenem and colistin resistance further complicate treatment of these infections. The aim of this study was to assess the occurrence of ESBL/AmpC E. coli in different broiler flocks and farms, as well as in broiler meat, in a country with no antimicrobial usage in broiler production. An additional goal was to assess the genetic characteristics of ESBL/AmpC E. coli isolates by using whole genome sequencing (WGS). Altogether 520 caecal swabs and 85 vacuum-packed broiler meat samples were investigated at the slaughterhouse level. WGS of the bacterial isolates revealed acquired antimicrobial resistance (AMR) genes, multilocus sequence types (MLST) and plasmid sequences. ESBL/AmpC E. coli was identified in 92 (18%) of the caecum and 27 (32%) of the meat samples. ESBL/AmpC E. coli-carrying birds derived from six (33%) out of 18 farms. Of the two blaESBL/AmpC genes detected by PCR, blaCMY-2 (96%) was predominant over blaCTX-M-1 (4%). Furthermore, WGS revealed an additional AMR gene sul2. Carbapenemase, colistin, and other AMR genes were not detected from the isolates of either the caecal or meat samples. Altogether seven MLSTs (ST101, ST117, ST212, ST351, ST373, ST1594 and an unknown ST) and a variety of different plasmid sequences (IncB/O/K/Z, IncI1, IncFII, IncII, IncFIB, IncFIC, IncX1 and an additional set of Col-plasmids) were detected. This is the first study on genomic epidemiology of ESBL/AmpC E. coli on broiler farms and flocks with no antimicrobial usage, by using WGS analysis. Results show that ESBL/AmpC E. coli occurrence is common both in the caecum and in the packaged meat. However, compared to other European countries, the occurrence is low and the presence of AMR genes other than blaCMY-2 and blaCTX-M-1 is rare. More studies are needed to understand the ESBL/AmpC E. coli occurrence in broiler production to prevent the meat from contamination during slaughter and processing, thereby also preventing zoonotic transmission of ESBL/AmpC E. coli. Additionally, more studies are needed to understand the ecology and fitness cost of Enterobacteriaceae plasmids in animal production in order to prevent their acquisition of plasmid-encoded antimicrobial resistance genes such as carbapenem and colistin resistance genes, as this would pose a great hazard to food safety.

Päivärinta M ，Latvio S ，Fredriksson-Ahomaa M ，Heikinheimo A ... -  《-》

Occurrence of the Colistin Resistance Gene mcr-1 and Additional Antibiotic Resistance Genes in ESBL/AmpC-Producing Escherichia coli from Poultry in Lebanon: A Nationwide Survey.

Mikhayel M ，Leclercq SO ，Sarkis DK ，Doublet B ... -  《Microbiology Spectrum》

High Heterogeneity of Escherichia coli Sequence Types Harbouring ESBL/AmpC Genes on IncI1 Plasmids in the Colombian Poultry Chain.

Castellanos LR ，Donado-Godoy P ，León M ，Clavijo V ，Arevalo A ，Bernal JF ，Timmerman AJ ，Mevius DJ ，Wagenaar JA ，Hordijk J ... -  《PLoS One》

Contamination of chicken meat with extended-spectrum beta-lactamase producing- Klebsiella pneumoniae and Escherichia coli during scalding and defeathering of broiler carcasses.

Extended-spectrum beta-lactamase- (ESBL-) producing Klebsiella (K.) pneumoniae and Escherichia (E.) coli are of critical importance in human and veterinary medicine. Animal food products, especially broiler chickens, are discussed as a possible source for the exposure of humans with antibiotic resistant bacteria. Although the occurrence and vertical transmission of ESBL-/AmpC-producing Enterobacteriaceae in the broiler production has been reported before, detailed investigations concerning the dissemination along the slaughter processing line are missing. In this study, we investigated cross-contamination with ESBL-producing Enterobacteriaceae during the processing of two different broiler flocks in one slaughterhouse. The ESBL-status during the fattening period of the flocks was determined and environmental samples from the slaughterhouse were taken before processing of the respective flocks. These isolates were compared to those found in samples from the carcasses after processing using whole genome sequencing. Phylogenetic analyses of seven ESBL-producing K. pneumoniae and 14 E. coli revealed close relationships between isolates from scalding water and the defeathering machine, respectively, which were collected before the processing of the broiler flocks, to those isolates found in samples from skin and filet of the respective flock carcasses. In conclusion, using high resolution molecular data we found evidence for the cross-contamination of carcasses with ESBL-producing Enterobacteriaceae during scalding and defeathering in the slaughterhouse.

Projahn M ，von Tippelskirch P ，Semmler T ，Guenther S ，Alter T ，Roesler U ... -  《-》

Low Occurrence of Extended-Spectrum β-lactamase-Producing Escherichia coli in Finnish Food-Producing Animals.

ESBL/AmpC-producing Escherichia coli is increasingly isolated from humans and animals worldwide. The occurrence of ESBL/AmpC-producing E. coli was studied in food-producing animals in Finland, a country with a low and controlled use of antimicrobials in meat production chain. A total of 648 cattle, 531 pig, 495 broiler and 35 turkey faecal samples were collected from four Finnish slaughterhouses to determine the presence of extended-spectrum β-lactamase (ESBL/AmpC)-producing E. coli. In addition, 260 broiler and 15 turkey samples were screened for carbapenemase-producing E. coli. Susceptibility to different class of cephalosporins and meropenem was determined with disc diffusion tests according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Determination of ESBL/AmpC production was performed with a combination disc diffusion test according to the recommendations of the European Food Safety Authority (EFSA). Plasmidic blaESBL/AmpC  genes were characterized by polymerase chain reaction and sequencing. A collection of isolates producing AmpC enzyme but not carrying plasmidic blaAmpC  was analysed by PCR and sequencing for possible chromosomal ampC promoter area mutations. Altogether ESBL/AmpC-producing E. coli was recovered from five cattle (0.8%), eight pig (1.5%) and 40 broiler samples (8.1%). No ESBL/AmpC-producing E. coli was found in turkey samples. Carbapenem resistance was not detected. Altogether ESBL/AmpC-producing E. coli was found on 4 (2.0%), 3 (4.5%) and 14 (25%) cattle, pig and broiler farms, respectively. From cattle samples 3 (27%) blaCTX-M-1  and from broiler samples 13 (33%) blaCTX-M-1  and 22 (55%) blaCMY-2 gene-carrying isolates were detected. In pigs, no plasmidic blaESBL/AmpC gene-carrying isolates were found. In all analysed isolates, the same mutations in the promoter region of chromosomal ampC were detected. The results showed low occurrence of ESBL/AmpC-producing E. coli in Finnish food-producing animals. In pigs, plasmidic blaESBL/AmpC -carrying E. coli was not detected at all.

Päivärinta M ，Pohjola L ，Fredriksson-Ahomaa M ，Heikinheimo A ... -  《-》