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Applied and Environmental Microbiology, January 2007, p. 156-163, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.01551-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Antimicrobial-Resistant Enteric Bacteria from Dairy Cattle{triangledown}

Ashish A. Sawant,1 Narasimha V. Hegde,1 Beth A. Straley,1 Sarah C. Donaldson,1 Brenda C. Love,1 Stephen J. Knabel,2 and Bhushan M. Jayarao1*

Department of Veterinary and Biomedical Sciences,1 Department of Food Science, Pennsylvania State University, University Park, Pennsylvania2

Received 5 July 2006/ Accepted 27 October 2006

A study was conducted to understand the descriptive and molecular epidemiology of antimicrobial-resistant gram-negative enteric bacteria in the feces of healthy lactating dairy cattle. Gram-negative enteric bacteria resistant to ampicillin, florfenicol, spectinomycin, and tetracycline were isolated from the feces of 35, 8, 5, and 42% of 213 lactating cattle on 74, 39, 9, 26, and 82% of 23 farms surveyed, respectively. Antimicrobial-resistant gram-negative bacteria accounted for 5 (florfenicol) to 14% (tetracycline) of total gram-negative enteric microflora. Nine bacterial species were isolated, of which Escherichia coli (87%) was the most predominant species. MICs showing reduced susceptibility to ampicillin, ceftiofur, chloramphenicol, florfenicol, spectinomycin, streptomycin, and tetracycline were observed in E. coli isolates. Isolates exhibited resistance to ampicillin (48%), ceftiofur (11%), chloramphenicol (20%), florfenicol (78%), spectinomycin (18%), and tetracycline (93%). Multidrug resistance (≥3 to 6 antimicrobials) was seen in 40% of E. coli isolates from healthy lactating cattle. Of 113 tetracycline-resistant E. coli isolates, tet(B) was the predominant resistance determinant and was detected in 93% of isolates, while the remaining 7% isolates carried the tet(A) determinant. DNA-DNA hybridization assays revealed that tet determinants were located on the chromosome. Pulsed-field gel electrophoresis revealed that tetracycline-resistant E. coli isolates (n = 99 isolates) belonged to 60 subtypes, which is suggestive of a highly diverse population of tetracycline-resistant organisms. On most occasions, E. coli subtypes, although shared between cows within the herd, were confined mostly to a dairy herd. The findings of this study suggest that commensal enteric E. coli from healthy lactating cattle can be an important reservoir for tetracycline and perhaps other antimicrobial resistance determinants.

* Corresponding author. Mailing address: Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park, PA 16802. Phone: (814) 863-5939. Fax: (814) 863-6140. E-mail: bmj3{at}psu.edu.

{triangledown} Published ahead of print on 10 November 2006.

Applied and Environmental Microbiology, January 2007, p. 156-163, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.01551-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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