2007 Annual Report 1a.Objectives (from AD-416) (1) Using optimized sampling strategies, enumeration, and molecular diagnostic, identify management practices resulting in high and low Salmonella and Campylobacter prevalence turkey farms and monitor the efficacy of on-farm intervention strategies targeting specific risk factors. (2) Identify key virulence attributes to differentiate Salmonella and Campylobacter avirulent commensals from those pathogenic strains that pose a public health threat in humans. (3) Develop molecular methods to assess the dynamics of the microbial intestinal flora throughout hog and turkey production. Identify microbes associated with gut colonization by, and population shifts of, foodborne pathogens. (4) Determine prevalence and quantities of recognized foodborne pathogens, principally Salmonella but also Campylobacter and Yersinia, in hog carcasses and organs. 1b.Approach (from AD-416) Time of entry of Salmonella and Campylobacter will be monitored in turkeys. The study will document flock management practices which affect the prevalence of these foodborne pathogens initially in the brooder period and ultimately throughout live production. Key virulence attributes of C. jejuni and C. coli strains recovered from turkeys will be characterized in vitro (cell invasion assays and in vivo day of hatch poult model). Ultimately, differential gene expression formats will be used to differentiate a virulent from virulent isolates of C. coli and C. jejuni. Bacterial and fungal communities of the ceca of domestic and wild turkeys will be described. This initial survey will provide a measurement of diversity between wild and domestic birds. Analysis of total community flux over time will focus on kinetics of Campylobacter community development and stability in the turkey ceca. Organisms that correlate with Campylobacter colonization or exclusion will be identified. Second generation PCR assays will be developed to detect and quantify Salmonella and Campylobacter on hog and turkey carcasses. Monitoring viscera will serve as an indicator of on-farm versus in-plant sources of contamination. These studies will assist in determining the critical control points of contamination during slaughter. 4.Accomplishments Development of a method for identifying functional microbial species in vivo: It is important for studies of food pathogen ecology to understand how the microbes respond to changes in the intestinal environment, however to date no method was available to perform these experiments. We demonstrated that the thymidine analog bromodeoxyuridine (BrdU), when supplied in the drinking water of turkey poults, will be incorporated into the DNA of actively dividing intestinal microbes. Thus BrdU can be used for identification of turkey intestinal microbes that grow in response to in vivo environmental changes resulting from feed withdrawal or other stressors experienced by the fowl host. This method will allow examination of microbial and host-microbe interactions affecting animal health, nutrition and food safety. This work aligns with Component 1.1 of the ARS NP 108 Food Safety Action Plan and addresses Problem Statement: 1.1.3 (Ecology, Host Pathogen and Chemical Contaminants Relationships). Showed how commercial bird housing systems impact the prevalence of bacterial foodborne pathogens. Ceca derived from hens raised under three different commercial systems were screened for Campylobacter and Salmonella. When the three housing types were compared during the winter, the prevalence of Campylobacter was highest in the non-caged birds. In contrast, for the summer sampling, there was no difference in either Campylobacter or Salmonella prevalence for the three housing types. This indicates that commercial bird housing systems evaluations should consider animal welfare issues as well as microbial food safety impact. This work aligns with Component 1.1 of the ARS NP 108 Food Safety Action Plan and addresses Problem Statement: 1.1.2 (Epidemiology). 5.Significant Activities that Support Special Target Populations Presented a seminar (“Foodborne Pathogens of Public Health Significance”) to faculty and veterinary students and reviewed USDA Capacity Building Grants at Tuskegee University, Tuskegee, Alabama, February 14, 2007. 6.Technology Transfer Number of non-peer reviewed presentations and proceedings 18 Number of newspaper articles and other presentations for non-science audiences 2 Review Publications Harbaugh, E., Trampel, D., Wesley, I.V., Hoff, S., Griffith, R., Hurd, H.S. 2006. Rapid aerosol transmission of Salmonella among turkeys in a simulated holding-shed environment. Poultry Science. 85(10):1693-1699. Scupham, A.J. 2007. Examination of the microbial ecology of the avian intestine in vivo using bromodeoxyuridine. Environmental Microbiology. 9(7):1801:1809. Rostagno, M.H., Wesley, I.V., Trampel, D.W., Hurd, H.S. 2006. Salmonella prevalence in market-age turkeys on-farm and at slaughter. Poultry Science. 85(10):1838-1842. Scupham, A.J. 2007. Succession in the intestinal microbiota of pre-adolescent turkeys. Federation of European Microbiological Societies Microbiology Ecology. 60(1):136-147. Bhaduri, S., Wesley, I.V. 2006 Isolation and characterization of yersinia enterocolitica from swine feces recovered during the National Animal Health Monitoring System Swine 2000 Study. Journal of Food Protection. Vol.69:1552-1560. Borneman, J., Becker, J.O., Bent, E., Lanoil, B., Gardener, B.M., Olatinwo, R., Presley, L., Scupham, A.J., Valinsky, L., Yin, B. 2007. Identifying microorganisms involved in specific in situ functions: experimental design considerations for rRNA gene-based population studies and sequence-selective PCR assays. In: Hurst, C.J. editor. Manual of Environmental Microbiology. 3rd edition. Washington, D.C.: ASM Press. p.748-757. Wesley, I.V. 2007. Listeriosis in Animals. In: Listeria, Listeriosis, and Food Safety, 3rd edition. Boca Raton, FL:CRC Press. p.55-84. Andersen, M.E., Wesley, I.V., Nestor, E., Trampel, D.W. 2007. Prevalence of Arcobacter species in market-weight commercial turkeys. Antonie Van Leeuwenhoek. 92:309-317.