Information on the dissemination of antibiotic resistance mechanisms in the environment as well as wild life is needed in North America. A constructed wetland (where ∼15,000 American crows roost) was sampled on the University of Washington Bothell Campus for the presence of antibiotic resistant E. coli (ARE). Crow droppings from individual birds and grab samples of water were collected in 2014–2015. E. coli were isolated by selective agar plating. The most frequent antibiotic resistance (AR) of the fecal isolates was to ampicillin (AMP) (53%), followed by amoxicillin-clavulanic acid (AMC) (45%), streptomycin (S) (40%), and nalidixic acid (NA) (33%). Water isolates had similar AR pattern and ∼40% were multidrug resistant. Isolates from water samples collected during storm events showed higher resistance than isolates from no rain days to tetracycline, AMP, AMC, NA, and gentamycin. Extended spectrum beta lactamase (ESBL) containing E. coli with the bla ctx-M was found in three water and nine fecal isolates while bla cmy-2 in 19 water and 16 fecal isolates. Multilocus Sequence Typing analysis (MLST) yielded 13 and 12 different sequence types (STs) amongst fecal and water isolates, many of which could be correlated to livestock, bird, and humans. MLST identified ESBL E. coli belonging to the clinically relevant ST131 clone in six fecal and one water isolate. Three STs found in feces could be found in water on the same dates of collection but not subsequently. Thus, the strains do not appear to survive for long in the wetland. Phylogenetic analysis revealed similar distribution of the water and fecal isolates among the different phylo-groups, with the majority belonging to the commensal B1 phylo-group, followed by the pathogenic B2 phylo-group. This study demonstrates that corvids can be reservoirs and vectors of ARE and pathogenic E. coli , posing a significant environmental threat.

The study examines whether crows are carriers of extraintestinal pathogenic E. coli (ExPEC) and avian pathogenic E. coli (APEC)-like strains, and if wetland roost areas contribute to their spread. A total of 10 crow feces (n = 71) and 15 water E. coli isolates (n = 134) from a wetland area could be characterized as potentially ExPEC based on the presence of ≥2 of the five cardinal genes iutA, kpsMT2, papEF, pap A/C, papG, sfa/foc, and afa/dra, while six fecal and 14 water isolates could be characterized as potentially APEC-like based on the presence of plasmid associated genes: iutA, episomal iss, ompT, hlyF and iroN. A total of 32 fecal and 27 water isolates tested carried plasmids based on incompatibility typing. Plasmids from 34 of 38 isolates tested could be transferred to another E. coli strain by conjugation with the antibiotic resistance (AR) profile being transferred, indicating their potential to be transferred to indigenous and non-pathogenic strains in the wetland. APEC-like plasmids could be transferred in six of eight isolates tested. Pathogenic E. coli of importance to the medical community and poultry industry may be detected in high levels in surface water due to corvid activity. Regardless of their role in health or disease, water in wetlands and streams can serve as a media for the dissemination of AR and virulence traits of bacteria, with corvids acting as potential vectors for farther dissemination.