Genotype, serotype, and antibiotic resistance of sorbitol-negative Escherichia coli isolates from feedlot cattle.

Rectal fecal samples from 80 steers receiving Rumensin, Revalor-S, and Liquamycin alone or in combination for growth promotion and disease prevention were examined for the presence of non-O157:H7 Shiga toxin-producing Escherichia coli. All isolates were identified with the API 20E test, virulence genes were detected with a PCR assay, and antibiotic susceptibilities were determined with the Sensititre system. Of the 153 E. coli isolates recovered 126 (82.3%) were sorbitol negative. Isolates were classified into 14 biochemical E. coli groups; 51.6% were negative for arginine dihydrolase, ornithine decarboxylase, sorbitol, and saccharose reactions but positive for lysine decarboxylase, indole production, and rhamnose reactions. Twenty-one O:H serotypes were detected in the 153 E. coli isolates. The most frequent serotypes were O2:H42 (49.7% of isolates), O49:NM (13.7%), O?:H25 (9.2%), and O10:NM (7.2%). One isolate of E. coli O172:H25 and one of E. coli O157: H39 were found. The stx1 gene was found in the two E. coli O98:H25 isolates. The eaeA and e-hlyA genes were detected in 21, 14, and 10 isolates of serotypes O49:NM, O?:H25, and O10:NM, respectively, and in each isolate of serotype O156:H25 and O172:H25. Four E. coli O132:H18 isolates were multiresistant to ampicillin, chloramphenicol, kanamycin, streptomycin, and sulfisoxazole. Tetracycline resistance due to the tet(B) gene was observed in 74 of the 76 E. coli O2:H42 isolates. Except for one isolate, all tetracycline-resistant isolates were negative for the virulence genes eaeA and e-hlyA or stx1. Pulsed-field gel electrophoresis typing revealed that the tetracycline-resistant serotypes were genetically diverse. Our data illustrate that cattle are a potential source of some atypical antibiotic-resistant E. coli isolates that harbor virulence genes.

Growth performance and shedding of some pathogenic bacteria in feedlot cattle treated with different growth-promoting agents.

Eighty steers with a mean body weight of 319 kg were used in a study to evaluate the effect of a growth-promoting implant (trenbolone acetate plus estradiol benzoate), monensin, and oxytetracycline on the steer performance and shedding of some foodborne pathogens. The steers were allotted to one of eight treatment combinations according to a randomized complete block design with 16 pens of five animals. Rectal fecal samples were collected before treatment commenced and over a period of more than 24 weeks to study the influence of treatments on the intestinal microbiology of the animals. Results supported the beneficial effect of the hormonal implant on the performance of feedlot steers (average daily gain, feed efficiency, and fat thickness), on carcass characteristics (hot carcass weight, lean yield), and economic value of the carcasses (P < 0.01). The levels of Escherichia coli in feces were not affected by treatments but remained high throughout the study period. Antibiotic-resistant isolates of E. coli were more frequently found as the study progressed but were not associated with any specific treatment. Also independently of treatment, we observed a reduction over time in the shedding of Campylobacter and Yersinia during the feeding period, whereas the shedding of Enterococcus was increased. The results of this study confirmed the beneficial economic effect of growth-promoting agents in beef production and showed that the agents tested did not specifically affect the overall microbial evolution of the animal gut. However, the study also showed, independently of the growth promoter used, the shedding of Campylobacter, Yersinia, and antibiotic-resistant E. coli in the feedlot environment. These bacteria also may be found in the colonic tissue of steers at slaughter and might be a source of carcasses contamination.

Antibiotic resistance and hypermutability of Escherichia coli O157 from feedlot cattle treated with growth-promoting agents.

In a longitudinal study (165 days), we investigated the effect of growth-promoting agents (monensin and trenbolone acetate-estradiol) and an antibiotic (oxytetracycline) on the incidence in feedlot steers of Escherichia coli O157, including antibiotic-resistant and hypermutable isolates. Eighty steers in 16 pens were treated with eight combinations of promoters, and each treatment was duplicated. Fecal samples were collected at nine different sampling times for detection of E. coli O157. Overall, 50 E. coli O157 isolates were detected in treated animals, and none were found in untreated animals. Compared with untreated controls, there was a significant association between the utilization of growth-promoting agents or antibiotics and the shedding of E. coli O157 at day 137 (P = 0.03), when a prevalence peak was observed and 50% of the isolates were detected. Multiplex PCR assays were conducted for some virulence genes. PCR results indicated that all except one isolate possessed at least the Shiga toxin gene stx2. MICs for 12 antibiotics were determined, and eight oxytetracycline-resistant E. coli O157 strains were identified. Antibiotic-resistant strains were considered a distinct subpopulation of E. coli O157 by pulsed-field gel electrophoresis typing. Seven of these antibiotic-resistant strains were isolated early in the study (on or before day 25), and among them two were also hypermutable as determined by rifampin mutation frequencies. The proportion of hypermutable strains among E. coli O157 isolates remained relatively constant throughout the study period. These results indicate that the use of growth-promoting agents and antibiotics in beef production may increase the risk of environmental contamination by E. coli O157.