Molecular epidemiology and AMR of avian pathogenic and commensal E. coli from low-antibiotic use broiler production
Antibiotic susceptibility and resistance genes in Escherichia coli from broilers reared in a low-antibiotic-use production system   Antimicrobial resistance (AMR) is a major global concern for animal and human health. This study investigated the occurrence and patterns of AMR in Escherichia coli (E. coli) isolated from Swedish broiler flocks reared under low-antibiotic-use conditions. During routine necropsy examinations of 80 broilers from 40 flocks with increased mortality associated with colibacillosis, liver samples were collected for bacteriological analysis. E. coli isolated from the liver were classified as clinical E. coli. In addition, boot sock samples were taken to collect feces from the litter of 60 broiler flocks with no signs of disease or increased mortality. E. coli isolates (n = 109) obtained from boot sock samples were classified as non-clinical E. coli. Susceptibility to 15 antibiotics was assessed using broth microdilution, and resistance-associated genes and mutations were identified through whole-genome sequencing (WGS). Overall resistance was low, with all isolates susceptible to 9 of the 15 tested antibiotics: meropenem, azithromycin, amikacin, gentamicin, tigecycline, ceftazidime, cefotaxime, chloramphenicol, and colistin. Resistance was significantly more frequent in non-clinical than clinical isolates for the six antibiotics with detected resistance (P < 0.05) and was strongly correlated with the presence of known AMR genes or mutations. Among clinical isolates, 93.7% were fully susceptible to all tested antibiotics, compared with 49.5% of non-clinical isolates. The highest resistance rates were observed in non-clinical isolates against ampicillin (34%), sulfamethoxazole (32.1%), and trimethoprim (28.4%). The results of this study indicate that in low-antibiotic-use production systems, factors beyond direct antibiotic use—such as horizontal gene transfer, vertical transmission, and environmental contamination—may contribute to AMR dissemination. Higher AMR rates in non-clinical isolates suggest that these isolates may serve as reservoirs of resistance genes. This highlights the importance of monitoring commensal E. coli and farm environments to support AMR mitigation and sustainable broiler production.   Molecular epidemiology reveals distinct lineages and genomic profiles of avian pathogenic and commensal Escherichia coli in broiler flocks   Escherichia coli is predominantly an intestinal commensal; however, avian pathogenic E. coli (APEC) causes colibacillosis in poultry. The APEC pathotype lacks a clear genetic definition, further complicated by the opportunistic nature of E. coli. To compare the genomic characteristics of avian pathogenic and commensal E. coli, isolates from diseased and healthy broiler flocks in Sweden collected between 2022 and 2024 were analysed. Clinical isolates (n = 202) were collected at necropsy from 40 flocks during colibacillosis outbreaks, and non-clinical isolates (n = 109) were obtained from litter using sock sampling in 60 unaffected flocks. Whole-genome sequencing was performed to determine sequence types (STs), serotypes, phylogroups, virulence-associated genes (VAGs), and to identify ColV plasmids. A five-gene APEC marker panel targeting plasmid-associated virulence genes (iutA, hlyF, iss, iroN, ompT) was used to classify isolates as APEC or non-APEC, and high-risk clones were identified according to the APECtyper scheme. Clinical isolates comprised 22 STs and 25 serotypes and were dominated by the ST23 O78:H4 clone within phylogroup C (59%). Non-clinical isolates were more diverse (44 STs, 67 serotypes), primarily within phylogroups A (48%) and B1 (33%), with no clone predominating. Clinical isolates carried significantly more VAGs (p < 0.001). Overall, 97% of clinical isolates were identified as APEC, all of which carried a ColV plasmid. Among non-clinical isolates, 28% were APEC, of which 80% were ColV-positive. However, clinical APEC isolates carried significantly more ColV-associated virulence gene clusters than non-clinical APEC isolates (p < 0.001). Only 5% of non-APEC isolates were ColV-positive. High-risk clones were restricted to clinical APEC isolates (63%). These findings indicate that colibacillosis in Swedish broilers was largely driven by a dominant APEC clone during the study period, highlighting the need for coordinated surveillance and targeted control of high-risk clones. The presence of VAG reservoirs among isolates from unaffected flocks, together with the limitations of marker-based APEC typing, supports integrated frameworks combining lineage, VAG profiles, and plasmid content for more reliable APEC identification and pathogenicity assessment.