title: Inoculation of beef with low concentrations of Escherichia coli O157:H7 and examination of factors that interfere with its detection by culture isolation and rapid methods.
abstract: Currently used industry testing programs require the ability to detect Escherichia coli O157:H7 in samples of beef trim or ground beef at levels as low as 1 CFU/375 g. We present a reliable protocol for generating a control inoculum for verification testing at this low concentration and evaluate its use. Results show that half of all samples received no cells when 1 CFU was the target concentration and that targets greater than 3 CFU were much more reliable. Detection by culture isolation and two commercial assays, Qualicon BAX-MP and BioControl GDS, detected 94% ± 11%, 92% ± 10%, and 92% ± 7% of samples inoculated with 5.4 CFU (range 1 to 9 CFU), respectively. We also examined the effect of background aerobic plate count (APC) bacteria and fat content effects on the detection of E. coli O157:H7. At APC concentrations below 6 log CFU/g, the rapid methods detected all beef trim samples inoculated with 26 CFU of E. coli O157:H7 per 65 g. At an APC of 6.7 log CFU/g, culture, BAX-MP, and GDS detected 100, 75, and 13%, respectively, of inoculated samples. Neither commercial method detected E. coli O157:H7 in the samples when APC was 7.7 log CFU/g, whereas culture was able to detect 63% of E. coli O157:H7 in the samples when APC was at this concentration. Increased fat content correlated with decreasing recovery of immunomagnetic separation beads, but this was not observed to interfere with detection of E. coli O157:H7.
authors.affiliation: U.S. Department of Agriculture, Agricultural Research Service, Roman L. Hruska U.S. Meat Animal Research Center, Clay Center, Nebraska 68933-0166, USA. mick.bosilevac@ars.usda.gov
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Bosilevac
authors.forename: Norasak
authors.initials: N
authors.lastname: Kalchayanand
authors.forename: John W
authors.initials: JW
authors.lastname: Schmidt
authors.forename: Steven D
authors.initials: SD
authors.lastname: Shackelford
authors.forename: Tommy L
authors.initials: TL
authors.lastname: Wheeler
authors.forename: Mohammad
authors.initials: M
authors.lastname: Koohmaraie
mesh_terms: D000818:Animals; D002417:Cattle; D015169:Colony Count, Microbial / Q000379:methods / Q000592:standards*; D003257:Consumer Product Safety; D019453:Escherichia coli O157 / Q000254:growth & development / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D006801:Humans; D008460:Meat / Q000382:microbiology*; D012307:Risk Factors; D013997:Time Factors