Masters Theses

Date of Award

5-2001

Degree Type

Thesis

Degree Name

Master of Science

Major

Animal Science

Major Professor

Alan G. Mathew

Committee Members

Frank Masincupp, David Golden

Abstract

This study was an attempt to locate the apramycin resistance gene in E.coli isolated from cold-stressed swine. Six pigs were administered apramycin for 14 days during a 150-day study and were exposed to cold stress in the form of an 8 °C reduction in recommended daily temperature. Fecal swabs were taken on eight different days for the recovery of Escherichia coli, and isolates were tested for sensitivity to apramycin. Plasmids were isolated from selected E. coli from each day of sampling and were electroporated into receptive bacteria to see whether those bacteria in turn would become receptive so that a resistant plasmid could be identified. Electroporated plasmids could not be recovered so AP PCR was examined instead to identify resistant plasmids. Fingerprints were made of all plasmids using two different primers. PFGE of DNA, using two different restriction endonucleases, was also performed. Profiles from AP PCR and PFGE were compared separately for similarity by eye and also by using a molecular analyst software program that creates dendrograms. No resistant plasmids were identified, the source of resistance could not be linked to a plasmid or plasmids, and the apramycin resistance gene was not discovered.

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