title: Prevalence and molecular characterization of Escherichia coli O157:H7 in bulk tank milk and fecal samples from cull cows: a 12-month survey of dairy farms in east Tennessee.
abstract: A study on the prevalence of Escherichia coli O157:H7 was conducted on 30 dairy farms in east Tennessee between May 2000 and April 2001. This pathogen was isolated from 8 of 30 (26.7%) dairy farms at various sampling times. A total of 415 fecal samples from cull dairy cows and 268 bulk tank milk samples were analyzed. Overall, 10 of 683 (1.46%) samples (2 of 268 [0.75%] milk samples and 8 of 415 [1.93%] fecal samples) tested positive for E. coli O157:H7. Food and Drug Administration Bacteriological Analytical Manual protocols were used for the conventional isolation and confirmation of E. coli O157:H7. Samples were shake cultured (150 rpm) at 42 degrees C for 24 h in tryptic soy broth containing 2 mg of novobiocin per liter. White colonies isolated on cefixime-tellurite sorbitol MacConkey agar plates were evaluated for fluorescence on sorbitol MacConkey agar supplemented with 0.025 g of methylumbelliferyl-beta-D-glucuronide per liter. Nonfluorescing white colonies were biochemically typed and serologically confirmed. Multiplex polymerase chain reaction profiles of E. coli O157:H7 isolates indicated the presence of common virulence factors (Shiga toxin, enterohemolysin, and intimin) of Shiga toxin-producing E. coli, suggesting the potential human pathogenicity of bacterial isolates. Pulsed-field gel electrophoresis profiles of SpeI and XbaI restriction enzyme-digested genomic DNA were used to establish relatedness among bacterial isolates. Data from this study indicate that both cull dairy cows and bulk tank milk pose a potential hazard with regard to human foodborne illness. It is therefore imperative to develop on-farm and preharvest pathogen reduction programs to control the carriage of E. coli O157:H7 pathogens.
authors.affiliation: University of Tennessee Food Safety Center of Excellence, Knoxville 37996, USA.
authors.forename: S E
authors.initials: SE
authors.lastname: Murinda
authors.forename: L T
authors.initials: LT
authors.lastname: Nguyen
authors.forename: S J
authors.initials: SJ
authors.lastname: Ivey
authors.forename: B E
authors.initials: BE
authors.lastname: Gillespie
authors.forename: R A
authors.initials: RA
authors.lastname: Almeida
authors.forename: F A
authors.initials: FA
authors.lastname: Draughon
authors.forename: S P
authors.initials: SP
authors.lastname: Oliver
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology*; D004269:DNA, Bacterial / Q000032:analysis*; D003612:Dairying; D016521:Electrophoresis, Gel, Pulsed-Field; D004927:Escherichia coli Infections / Q000453:epidemiology / Q000662:veterinary*; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D005243:Feces / Q000382:microbiology*; D005260:Female; D008892:Milk / Q000382:microbiology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D015995:Prevalence; D013714:Tennessee / Q000453:epidemiology; D014774:Virulence
chemical_list: D004269:DNA, Bacterial