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Open Access Highly Accessed Research article

Longitudinal characterization of antimicrobial resistance genes in feces shed from cattle fed different subtherapeutic antibiotics

Trevor W Alexander1, Jay L Yanke2, Tim Reuter2, Ed Topp3, Ronald R Read4, Brent L Selinger5 and Tim A McAllister2*

Author Affiliations

1 Department of Animal Science, University of Vermont, Burlington, Vermont, 05405, USA

2 Agriculture and Agri-Food Canada Research Centre, Lethbridge, Alberta, T1J 4B1, Canada

3 Agriculture and Agri-Food Canada Research Centre, London Ontario, N5V 4T3, Canada

4 Faculty of Medicine, University of Calgary, Calgary, Alberta, T2N 4N1, Canada

5 Department of Biological Sciences, University of Lethbridge, Lethbridge, Alberta, T1K 3M4, Canada

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BMC Microbiology 2011, 11:19  doi:10.1186/1471-2180-11-19

Published: 24 January 2011

Abstract

Background

Environmental transmission of antimicrobial-resistant bacteria and resistance gene determinants originating from livestock is affected by their persistence in agricultural-related matrices. This study investigated the effects of administering subtherapeutic concentrations of antimicrobials to beef cattle on the abundance and persistence of resistance genes within the microbial community of fecal deposits. Cattle (three pens per treatment, 10 steers per pen) were administered chlortetracycline, chlortetracycline plus sulfamethazine, tylosin, or no antimicrobials (control). Model fecal deposits (n = 3) were prepared by mixing fresh feces from each pen into a single composite sample. Real-time PCR was used to measure concentrations of tet, sul and erm resistance genes in DNA extracted from composites over 175 days of environmental exposure in the field. The microbial communities were analyzed by quantification and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S-rRNA.

Results

The concentrations of 16S-rRNA in feces were similar across treatments and increased by day 56, declining thereafter. DGGE profiles of 16S-rRNA differed amongst treatments and with time, illustrating temporal shifts in microbial communities. All measured resistance gene determinants were quantifiable in feces after 175 days. Antimicrobial treatment differentially affected the abundance of certain resistance genes but generally not their persistence. In the first 56 days, concentrations of tet(B), tet(C), sul1, sul2, erm(A) tended to increase, and decline thereafter, whereas tet(M) and tet(W) gradually declined over 175 days. At day 7, the concentration of erm(X) was greatest in feces from cattle fed tylosin, compared to all other treatments.

Conclusion

The abundance of genes coding for antimicrobial resistance in bovine feces can be affected by inclusion of antibiotics in the feed. Resistance genes can persist in feces from cattle beyond 175 days with concentrations of some genes increasing with time. Management practices that accelerate DNA degradation such as frequent land application or composting of manure may reduce the extent to which bovine feces serves as a reservoir of antimicrobial resistance.