Extended-Spectrum-Cephalosporin Resistance Genes in Escherichia coli from Beef Cattle

Abstract

e R esistance to extended-spectrum cephalosporins (ESC) has rarely been detected in Enterobacteriaceae from beef cattle in Canada (1); in contrast, in the United States, it has repeatedly been found in bacteria from cattle (see, for instance, references 2 to 5). To assess whether similar bacteria are present in beef cattle in Alberta, Canada, we used enrichment cultures and PCR to increase the sensitivities of our detection methods.Four hundred thirty-two composite fecal samples were collected from 73 pens in four major feedlots in Alberta from May 2014 to April 2015. The samples were grown in EC broth (BD Difco) containing cefotaxime (2 g/ml), and aliquots were plated on MacConkey agar with ceftriaxone (1 g/ml) and then on Trypticase soy agar with ampicillin (32 g/ml). Lactose-fermenting, indole-positive isolates were investigated further. The remaining enrichment cultures were centrifuged, and the sediment was used for DNA extractions using a QIAamp DNA minikit. Real-time PCR with custom TaqMan primer/probe combinations for the bla CTX-M-1 , bla CTX-M-2 , bla CTX-M-8 , bla CTM-X-9 , bla CTX-M-25 , bla CMY , bla SHV , and bla TEM gene groups were performed using these extracts. Mock extractions and no-template controls were run with every batch to control for nonspecific background signals. A multivariate probit analysis was conducted to assess dependencies among the gene groups under consideration and their associations with season, feedlot, and pen. Sequencing of PCR products was used to identify gene variants in a subset of samples and isolates (http://www.lahey.org/Studies/).The PCR results from the enrichment cultures (Table 1) demonstrate that ESC resistance genes are widespread in feces from beef cattle in Alberta. Testing isolates from a subsample of these cultures confirmed the accuracy of these results (53/59 isolates were positive for one or two of these gene groups). Genes of the TEM group were almost exclusively (n ϭ 234; 96%) found together with genes of the other groups (Table 1) and were likely not extended-spectrum ␤-lactamases. The following ␤-lactamase genes (and numbers of samples [n ϭ 29]) were found in the indicated groups: TEM group, bla TEM-1 (4 [all 4 were from samples positive for one or several of the other ␤-lactamase groups]); SHV group, bla SHV-2a (2); CMY group, bla CMY-2 (2); CTX-M-1 group, bla CTX-M-15 (2), bla (1), bla CTX-M-55 (8), bla CTX-M-61 (1), and bla (1 [GenBank accession number KT251041]); CTX-M-2 group, bla CTX-M-2 (1) and bla CTX-M-115 (2); and CTX-M-9 group, bla CTX-M-24 (1), bla CTX-M-27 (2), and bla CTX-M-65 (2). Although more positive samples were detected in the summer, the

Extended‐Spectrum β‐lactam Resistance in the Enteric Flora of Patients at a Tertiary Care Medical Centre

“…CTX‐M‐15 is the most frequently identified CTX‐M enzyme in both human clinical isolates (Castanheira et al., ) and companion animals in the US (Canton et al., ). This sequence type has been rarely reported in food animals, found in the enteric flora of dairy cows on 4% of Ohio herds (Mollenkopf et al., ) and swine flora from 2% of US finishing barns (Mollenkopf et al., ); however, a more recent study reported a higher prevalence of CTX‐M‐15 (7%) compared to CTX‐M‐1(0) in isolates from North American beef feedlot cattle, suggesting a transmission from humans to animals (Cormier et al., ). The 2010 SENTRY Antimicrobial Surveillance Program reported that 6 of the 85 CTX‐positive Enterobacteriaceae from 26 US hospitals carried CTX‐M‐27(Castanheira et al., ), similar to the rate in beef feedlot cattle (7% of CTX isolates) (Cormier et al., ), and it was the predominant sequence type found recently on one Ohio dairy (Mathys et al., ).…”

Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in cattle production – a threat around the world

“…Faecal (Velasova et al, 2019) Infection (Hunter et al, 2010) ND (Stokes, 2014) Czech Republic Faecal (Dolejska et al, 2011b) Sick (Dolejska et al, 2013) R eunion ND (Gay et al, 2018) Slovakia Faecal (Kme t and Buj n akov a, 2018) Sweden Faecal (Duse et al, 2015) Switzerland Faecal (Endimiani et al, 2012;Geser et al, 2012a;Zurfluh et al, 2015) Turkey Faecal (AslantaŞ et al, 2017;Pehlivanoglu et al, 2016) North America Canada ND (Cormier et al, 2019) USA Faecal (Mir et al, 2016;Mollenkopf et al, 2012;Wittum et al, 2010) Asia China Mastitis (Ali et al, 2016(Ali et al, , 2017 South Korea Mastitis (Tark et al, 2017) Indonesia Faecal (Sudarwanto et al, 2016) Japan Mastitis (Ohnishi et al, 2013b) Klebsiella pneumoniaeEurope Italy Mastitis (Locatelli et al, 2010) Klebsiella ozaenae Italy Faecal (Stefani et al, 2014) Salmonella enterica Germany ND (Rodríguez et al, 2009) CTX-M-1/61 E. coli Europe Germany Faecal (Dahms et al, 2015) CTX-M-2 E. coli South America Brazil Faecal (Palmeira et al, 2018) Europe Germany Mastitis (Eisenberger et al, 2017;Freitag et al, 2017;Michael et al, 2017) Sick (Michael et al, 2017) Netherlands Faecal (Ceccarelli et al, 2019;Heuvelink et al, 2019) North America Canada Faecal (Cormier et al, 2016) Asia Japan Faecal (Shiraki et al, 2004) Diarrheic (Ohnishi et al, 2013a) Infection (Asai et al, 2011) K. pneumoniae Japan Mastitis (Ohnishi et...…”

“…In North America only 2 countries registered E-ESBL description in cattle, Canada and the United States, the last one being the world's largest cattle producer (USDA et al, 2017). CTX-M-15, CTX-M-32 and CTX-M-65 were the only beta-lactamases described in both countries, the Figure 3 shows the complete described ESBL from cattle in the North America (Cormier et al, 2016;Mir et al, 2016;Poole et al, 2017;Tate et al, 2017).…”

“…CTX ND-ESBL were described in E-ESBL in cattle in Canada, which was the country where the largest variety of CTX-M's types were described (Awosile et al, 2018;Cormier et al, 2016;Lussier, 2010). All E-ESBLs were E. coli of faecal origin.…”

Establishing Statistical Equivalence of Data from Different Sampling Approaches for Assessment of Bacterial Phenotypic Antimicrobial Resistance

“…Tetracyclines, penicillins, and aminoglycosides were introduced in the 1940s and macrolides in the 1970s ( 40 ). Consequently, multiple genes encoding various degrees of susceptibility to these drug classes have been observed in fecal E. coli and S. enterica isolates from farm animals ( 41 – 45 ). The tetracycline resistance gene pool is especially diverse, with several tens of tet genes described to date for different animal and human hosts ( 41 , 46 ).…”