Methods for recovering Escherichia coli O157:H7 from cattle fecal, hide, and carcass samples: Sensitivity and improvements

Abstract

The Meats Research Unit (MRU) methods, developed by MRU scientists of the U.S. Meat Animal Research Center, have been used to study the prevalence of Escherichia coli O157:H7 in cattle carcass, hide, and fecal samples. The sensitivity of these methods for recovery of injured E. coli O157:H7 cells from inoculated and uninoculated samples was determined, and potential improvements to these methods were evaluated. When using the conventional MRU methods, 91% of the preevisceration carcass samples tested positive for E. coli O157:H7 when inoculated with 5 to 10 CFU, 100% of hide samples tested positive for E. coli O157:H7 when inoculated with 30 to 50 CFU, and 96% of the fecal samples produced positive results when inoculated with 300 to 400 CFU per 10 g. The addition of a phosphate buffer to the tryptic soy broth enrichment improved recovery of E. coli O157:H7 from feces. Using the modified enrichment, 92% of the samples were identified as positive when inoculated with 10 to 30 CFU per 10 g. Substituting a commercially available wash buffer for the phosphate-buffered saline (PBS) plus Tween 20 wash buffer during immunomagnetic separation of hide samples improved recovery of the target organism at lower inoculum concentrations. When comparing uninoculated samples, substituting a PBS buffer plus a zwitterionic detergent for PBS plus Tween 20 also had a positive effect on recovery of E. coli O157:H7 from hide samples. Data presented here indicate that the MRU methods are highly effective at recovering injured E. coli O157:H7 from fecal, hide, and beef carcass samples; however, modifications can be added to increase the sensitivity.