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Applied and Environmental Microbiology, September 2001, p. 3810-3818, Vol. 67, No. 9
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.9.3810-3818.2001

Genotypic Analyses of Escherichia coli O157:H7 and O157 Nonmotile Isolates Recovered from Beef Cattle and Carcasses at Processing Plants in the Midwestern States of the United States

Genevieve A. Barkocy-Gallagher,* Terrance M. Arthur, Gregory R. Siragusa,dagger James E. Keen, Robert O. Elder,Dagger William W. Laegreid, and Mohammad Koohmaraie

Roman L. Hruska U.S. Meat Animal Research Center, Agricultural Research Service, United States Department of Agriculture, Clay Center, Nebraska 68933-0166

Received 5 January 2001/Accepted 8 June 2001

Escherichia coli O157:H7 and O157 nonmotile isolates (E. coli O157) previously were recovered from feces, hides, and carcasses at four large Midwestern beef processing plants (R. O. Elder, J. E. Keen, G. R. Siragusa, G. A. Barkocy-Gallagher, M. Koohmaraie, and W. W. Laegreid, Proc. Natl. Acad. Sci. USA 97:2999-3003, 2000). The study implied relationships between cattle infection and carcass contamination within single-source lots as well as between preevisceration and postprocessing carcass contamination, based on prevalence. These relationships now have been verified based on identification of isolates by genomic fingerprinting. E. coli O157 isolates from all positive samples were analyzed by pulsed-field gel electrophoresis of genomic DNA after digestion with XbaI. Seventy-seven individual subtypes (fingerprint patterns) grouping into 47 types were discerned among 343 isolates. Comparison of the fingerprint patterns revealed three clusters of isolates, two of which were closely related to each other. Remarkably, isolates carrying both Shiga toxin genes and nonmotile isolates largely fell into specific clusters. Within lots analyzed, 68.2% of the postharvest (carcass) isolates matched preharvest (animal) isolates. For individual carcasses, 65.3 and 66.7% of the isolates recovered postevisceration and in the cooler, respectively, matched those recovered preevisceration. Multiple isolates were analyzed from some carcass samples and were found to include strains with different genotypes. This study suggests that most E. coli O157 carcass contamination originates from animals within the same lot and not from cross-contamination between lots. In addition, the data demonstrate that most carcass contamination occurs very early during processing.

* Corresponding author. Mailing address: USDA, ARS, Roman L. Hruska U.S. Meat Animal Research Center, P.O. Box 166, Spur 18D, Clay Center, NE 68933-0166. Phone: (402) 762-4228. Fax: (402) 762-4149. E-mail: gallagher{at}emailmarc.usda.gov.

dagger Present address: USDA, ARS, Russell Research Center, Athens, GA 30604-5677.

Dagger Present address: USDA, ARS, South Plains Agricultural Research Center, College Station, TX 77845.

Applied and Environmental Microbiology, September 2001, p. 3810-3818, Vol. 67, No. 9
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.9.3810-3818.2001


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