Journal
VETERINARY MICROBIOLOGY
Volume 172, Issue 3-4, Pages 499-504Publisher
ELSEVIER
DOI: 10.1016/j.vetmic.2014.05.032
Keywords
Cephalosporins; Biodegradation; Drug residues; Veterinary drugs; Mass spectrometry
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Funding
- cooperative research and development agreement between Zoetis (formerly Pfizer Animal Health) and U.S. Food and Drug Administration
- National Center for Toxicological Research, U.S. Food and Drug Administration
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Ceftiofur is a highly effective veterinary cephalosporin, yet it is rapidly degraded by bacteria in the gut. The goal of this work was to directly determine the mechanism of ceftiofur degradation by the bovine intestinal isolate Bacillus cereus P41. B. cereus P41 was isolated from the feces of a cow that had not been treated with cephalosporins, and was found to rapidly degrade ceftiofur in culture. Analysis of spent culture media by HPLC/UV and HPLC/MS revealed one major metabolite of ceftiofur, with a negative ion m/z of 127. Comparison of ceftiofur, ceftriaxone, and cefpodoxime degradation suggested that the major stable ceftiofur metabolite was the thiofuroic acid group eliminated from the C-3 position of the drug after hydrolysis by beta-lactamase. Genomic DNA from B. cereus P41 was cloned into Escherichia colt, and the transformants were screened for growth in the presence of ceftiofur. DNA sequencing of the plasmid pHSG299-BC-3 insert revealed the presence of a gene encoding a metallo-beta-lactamase. Incubation of ceftiofur with either the E. coli transformant or a commercial B. cereus metallo-beta-lactamase showed degradation of the drug and formation of the same major metabolite produced by B. cereus P41. These data demonstrate that a metallo-beta-lactamase plays a major role in the degradation of ceftiofur by the bovine intestinal bacterium B. cereus P41. Published by Elsevier B.V.
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