Genomic diversity, virulence and source of Campylobacter jejuni contamination in Irish poultry slaughterhouses by whole genome sequencing.

AIMS: The aim was to exploit whole genome sequencing (WGS) to assess genomic diversity, identify virulence genes and deduce the proportion of Campylobacter colonized broilers that directly contaminate their carcasses. METHODS AND RESULTS: Campylobacter jejuni isolates (107) from caeca and carcass neck skin samples (50 pairs from the same batch plus 7 individual caeca) sampled at three poultry slaughterhouses over a one-year period were selected for sequencing (MiSeq; Illumina). FastQ files were submitted to BioNumerics for analysis using the wgMLST scheme for allele calling. Campylobacter cgMLST and hierarchical clustering was performed by applying the single linkage algorithm. Sequence types (STs) were determined in silico from the WGS data and isolates were assigned into clonal complexes (CCs) using the Campylobacter PubMLST.org database. Virulence genes were determined by downloading core sequences from the virulence factor database (VFDB) and the National Center for Biotechnology Information (NCBI). A high degree of diversity was observed with 23 different STs identified. ST257 and CC-21 were the most common STs and CCs, respectively. cgMLST analysis suggested that 56% of carcass contamination was a direct result of contamination from caeca from the same batch. Virulence genes known to play a role in human C. jejuni infection were identified such as the wlaN gene and the genes associated with lipooligosaccharide synthesis, which were identified in 30% of isolates. CONCLUSIONS: Caecal colonization was the more plausible occurring source of C. jejuni contamination of broiler carcasses, compared with cross-contamination from another batch or the environment. The high rate of genetic diversity observed amongst caecal isolates is consistent with a wide variety of Campylobacter strains circulating in poultry flocks in Ireland. SIGNIFICANCE AND IMPACT OF STUDY: The results will further inform broiler processors and regulators about the influence and importance of on-farm colonization versus slaughterhouse cross-contamination and the relationship between C. jejuni in caeca and carcasses during processing.

Detection of bla OXA-1, bla TEM-1, and Virulence Factors in E. coli Isolated From Seals.

Marine mammals are frequently considered good sentinels for human, animal and environmental health due to their long lifespan, coastal habitat, and characteristics as top chain predators. Using a One Health approach, marine mammals can provide information that helps to enhance the understanding of the health of the marine and coastal environment. Antimicrobial resistance (AMR) is the quintessential One Health problem that poses a well-recognised threat to human, animal, and ecosystem health worldwide. Treated and untreated sewage, hospital waste and agricultural run-off are often responsible for the spread of AMR in marine and freshwater ecosystems. Rescued seals (n = 25) were used as sentinels to investigate the levels of AMR in the Irish coastal ecosystem. Faecal swabs were collected from these animals and bacterial isolates (E. coli and cefotaxime-resistant non-E. coli) from each swab were selected for further investigation. E. coli isolates were characterised in terms of phylogenetic group typing, AMR, and virulence factors. All E. coli isolates investigated in this study (n = 39) were ampicillin resistant while 26 (66.6%) were multi-drug resistant (MDR). Resistance genes bla OXA-1 and bla TEM-1 were detected in 16/39 and 6/39 isolates, respectively. Additionally, virulence factors associated with adhesion (sfa, papA, and papC) and siderophores (fyuA and iutA) were identified. An additional 19 faecal cefotaxime-resistant non-E. coli isolates were investigated for the presence of ß-lactamase encoding genes. These isolates were identified as presumptive Leclercia, Pantoea and Enterobacter, however, none were positive for the presence of the genes investigated. To the authors knowledge this is the first study reporting the detection of bla OXA-1 and bla TEM-1 in phocid faecal E. coli in Europe. These results highlight the importance of marine mammals as sentinels for the presence and spread of AMR in the marine and coastal environment.