Molecular characterization of plasmid-mediated AmpC beta-lactamase- and extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae among corvids (Corvus brachyrhynchos and Corvus corax) roosting in Canada

Abstract

This study evaluated the carriage of AmpC and extended-spectrum beta-lactamase (ESBL) genes and associated plasmids in faecal bacteria of Canadian corvids. Faecal samples from 449 birds in five roosting sites across Canada were analyzed using selective media, screening for AmpC and ESBL genes by PCR, and sequencing. Genomic relatedness was determined by PFGE and MLST. Plasmid mobility was studied by conjugation and transformation experiments, followed by plasmid typing. In total, 96 (21%, n = 449) cefotaxime-resistant Escherichia coli and three (0.7%) Klebsiella pneumoniae isolates were identified. ESBL genes blaCTX-M-1 (n = 3), blaCTX-M-14 (n = 2), blaCTX-M-32 (n = 2) and blaCTX-M-124 (n = 1) were detected in eight E. coli isolates, whereas blaSHV-2 (2) was found in two K. pneumoniae. E. coli isolates contained blaCMY-2 (n = 83) and blaCMY-42 (n = 1). The high genetic diversity of the isolates and presence of clinically important E. coli ST69 (n = 1), ST117 (n = 7) and ST131 (n = 1) was revealed. AmpC genes were predominantly carried by plasmids of incompatibility groups I1 (45 plasmids), A/C (10) and K (7). The plasmid IncI1/ST12 was most common and found in diverse E. coli STs in all sites. Highly diverse E. coli isolates containing AmpC and ESBL genes, including clinically important clones and emerging plasmids, are in circulation throughout Canadian wildlife.

Antibiotic Resistance of E. coli Isolated From a Constructed Wetland Dominated by a Crow Roost, With Emphasis on ESBL and AmpC Containing E. coli

“…We found a predominance of bla cmy-2 gene in the AmpC phenotype in the crow (16.8%) and water (18.36%) isolates. bla cmy-2 has been shown to be the most common plasmid borne beta lactamase in human, animal, and environmental bacterial isolates, and that includes large corvids in United States and Canada (Pitout et al, 2007; Mataseje et al, 2010; Folster et al, 2011; Martin et al, 2012; Jamborova et al, 2017, 2018). In a recent report 18.7% of Corvids from Canada were shown to carry the bla cmy-2 , which was substantially more than that reported from Corvids from European countries (4.4%).…”

“…In addition, crows can acquire AR bacteria by foraging on a variety of wastes such as garbage dumps, hospital and animal wastes, and animal feed lots (Verbeek and Caffrey, 2002; Guenther et al, 2011). Several recent studies have reported crows and rooks shedding bacteria that were resistant to one or more antibiotics (Literak et al, 2007; Hasan et al, 2015; Jamborova et al, 2015, 2018). E. coli , which lives as a harmless commensal in the gut of all animal and birds, has proved to be not only an indicator of fecal coliform but also of antibiotic resistance present in the environment (van Den Bogaard et al, 2000; Dolejská et al, 2009; Guenther et al, 2011; Jamborova et al, 2015, 2018).…”

“…Several recent studies have reported crows and rooks shedding bacteria that were resistant to one or more antibiotics (Literak et al, 2007; Hasan et al, 2015; Jamborova et al, 2015, 2018). E. coli , which lives as a harmless commensal in the gut of all animal and birds, has proved to be not only an indicator of fecal coliform but also of antibiotic resistance present in the environment (van Den Bogaard et al, 2000; Dolejská et al, 2009; Guenther et al, 2011; Jamborova et al, 2015, 2018). From the United States, only one study investigating antibiotic resistance in E. coli in crows has been reported (Jamborova et al, 2017).…”

Wide Spread of blaCTX–M–9/mcr-9 IncHI2/ST1 Plasmids and CTX-M-9-Producing Escherichia coli and Enterobacter cloacae in Rescued Wild Animals

“…Numerous studies were also based on fecal dejections collected in various places where birds live or transit, such as landfills, beaches, urban parks, nests, and other habitats, but associating fecal samples to the right bird species may be challenging. In some situations however, these approaches valuably allowed studying AMR in large cohorts of individuals of the same bird species in their natural environment, as for instance illustrated for black kites ( Milvus milvus ), corvids ( Corvus brachyrhynchos , Corvus corax ), gulls ( Larus glaucescens, Larus ridibundus ), pigeons ( Columba livia ) or storks ( Ciconia ciconia ) ( Bonnedahl et al, 2010 , 2014 ; Jamborova et al, 2018 ; Tarabai et al, 2019 ; Zendri et al, 2020 ). Here, we adopted a different strategy by systematically sampling all incoming wild birds at a French rescue center over a 6-month period.…”

Using whole-genome sequence data to examine the epidemiology of antimicrobial resistance in Escherichia coli from wild meso-mammals and environmental sources on swine farms, conservation areas, and the Grand River watershed in southern Ontario, Canada

“…Exploratory work, primarily in the form of cross-sectional surveys, has demonstrated that avian and mammalian wildlife may carry a variety of zoonotic agents (e.g., Salmonella, Campylobacter), some of which have been shown to be resistant to antimicrobials considered critical to human health [6][7][8]. In addition, bacterial clones of international importance (e.g., E. coli ST131), extended spectrum beta-lactamase-producers (ESBLs), and organisms resistant to last-resort antimicrobials (e.g., colistin, vancomycin, carbepenems) mediated by mobile resistance genes have all been isolated from and documented [9][10][11][12][13][14][15][16]. Much of the work examining the epidemiology of AMR in wildlife has been focused on wild birds [4,17,18].…”