Progress 02/17/01 to 02/16/06
Outputs
Progress Report 1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? Food animals are the primary source of bacterial human foodborne pathogens. Following USDA-Food Safety and Inspection Service (FSIS)- Hazard Analysis and Critical Control Points (HACCP) regulation and consumer concerns, livestock producers are attempting to reduce the on- farm prevalence of these pathogens. The project has three goals: (1) Develop rapid and sensitive molecular-based techniques, such as enyzyme- linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) assays, DNA fingerprinting methods, pulsed field gel electrophoresis (PFGE), oligonucleotide fingerprinting of ribosomal RNA genes (OFRG) and randomly amplified polymorphic DNA (RAPD) to detect, enumerate, and characterize intestinal microbial communities and specific bacterial foodborne pathogens in livestock;
(2) Delineate the dynamics of foodborne pathogens in livestock and their environment and develop intervention strategies to reduce transmission on-farm; (3) Determine pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the dynamics of the normal intestinal microbial flora and the presence of foodborne pathogens. Pathogens to be studied include: Campylobacter, Salmonella, Listeria, and emerging bacterial agents (e.g., Yersinia, Arcobacter). Molecular methods, such as PCR-based protocols, combined with the ecological perspective will identify management practices to reduce the prevalence of foodborne pathogens in hogs and turkeys. Sampling methods for isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). Management practices include feed withdrawal, transport,
holding, biosecurity, housing, etc., (Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1) as well as stress, commingling, and environmental contamination (Action Plan Priority Objective 1.2.1.2). This research is central to achieving effective intervention strategies to reduce on-farm transmission (Action Plan Priority Objective 1.4.1). In-house efforts are complemented by external support (USDA-CSREES-NRI-CGP, USDA Program 406 and the Food Safety Consortium. In the U.S., approximately 5 million cases of bacterial foodborne illness occur annually at a cost of approximately $3.2 billion. In the U. S., over 300,000 swine per day are slaughtered. Approximately 3% to 50% of those animals harbor Salmonella when they reach the kill floor. Hogs are also a major animal reservoir for Yersinia enterocolitica, a bacterial agent, which is transmitted to humans by pork. The U.S. turkey industry is valued at $7.8 billion with per capita consumption ~ 18 pounds annually. Of the 458,000
pounds of whole or cut-up turkey parts exported, nearly 10% is destined for Russia. Thus, the Russian embargo against U.S. poultry attributed to Salmonella contamination, impacted the industry. Turkeys, which represent the second commodity addressed in this project, are a natural reservoir for C. jejuni and Salmonella. Reducing the prevalence of these bacteria in hogs and turkeys may lower human foodborne illnesses and deaths. 2. List by year the currently approved milestones (indicators of research progress) Objective 1 Year 1 (FY 2001): Optimize methods for Salmonella. Year 2 (FY 2002): Optimize methods for Campylobacter. Year 3 (FY 2003): Opimize protocols for Yersinia. Year 4 (FY 2004): Optimize protocols for Listeria monocytogenes. Year 5 (FY 2005): Optimize protocols for Arcobacter. Objective 2 Year 1 (FY 2001): On-farm environmental studies: planning and data collection. Year 2 (FY 2002): On-farm studies: inputs (feed and water) and outputs (hog and runoff
contamination): analysis and reporting. Year 3 (FY 2003): On-farm studies: data collection, analysis. Year 4 (FY 2004): On-farm studies: data collection, analysis. Year 5 (FY 2005): On-farm studies: reporting. Objective 3 Year 1 (FY 2001): Lairage/transportation study: planning. Year 2 (FY 2002): Lairage/transportation study: data collection, analysis, reporting. Year 3 (FY 2003): Case control studies: planning, data collection. Year 4 (FY 2004): Case control studies: planning, data collection, analysis. Abattoir: planning data collection analysis; Manure management: planning. Year 5 (FY 2005): Manure management: data collection, analysis. 4a List the single most significant research accomplishment during FY 2006. This accomplishment focuses on understanding the ecology and epidemiology of food borne pathogens in swine and turkey production processes under Section 1.1.2 (Epidemiology); 1.1.3 (Ecology, host pathogens and chemical residue relationships); and 1.2.3
(Production and processing ecology). Maturation of the microbiota of the turkey ceca: A period of community transition in the ceca of domestic turkeys was detected between 9 and 12 weeks of age. This transition was marked by a conversion from Clostridial to Bacteroides predominance. Sequence analysis indicated the transition included a period in which the communities were dominated by multiple strains of Bacteroides uniformis (84% and 79% for two trials, respectively) indicating a possible role in maturation of the intestinal community. The transition may be a result of decreasing levels of growth hormone or a decreased dietary need for protein during this time frame. A significant increase of Campylobacter coli was also detected at week 12 in one trial, suggesting a correlation between microbiota destabilization and pathogen prevalence. 4b List other significant research accomplishment(s), if any. Sampling isolation, identification, characterization, and quantification of foodborne
pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). The in-house research and extramural projects will utilize statistically-based methods to identify pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the prevalence of foodborne pathogens. Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., as summarized in Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Effects caused by stress, co-mingling, and environmental contamination (Action Plan Priority Objective 1.2.1.2). Salmonella populations in the turkey are not altered during perimarketing events: We compared the prevalence of Salmonella enterica in turkeys on six commercial flocks pre- (30 birds/flock) and post- (30 birds/flock) transport in collaboration with scientists at the Iowa State University and partially funded by the Iowa
Turkey Federation. No statistical difference was found between the overall prevalence found on- farm and at slaughter, based on any sample type analyzed (crop, ceca, liver/gall bladder, and spleen) with cecal contents having the highest relative sensitivity. This demonstrates that the perimarketing practices of feed withdrawal, catching, loading, transportation, and pre-slaughter holding do not significantly affect the prevalence of S. enterica in market-age turkeys in contrast to what we have previously reported for hogs. Comparison of DNA extraction protocols: We compared 14 protocols to determine the best method for DNA extraction to be used in analysis of the microbial ecology of the turkey cecum. Electrophoretic and spectrophotometric analysis showed a wide range of DNA yield, shearing and contamination resulted from the methods. In addition, bacterial ribosomal small subunit quantification of the extraction products indicated a wide range of PCR inhibition resulting from
template contaminants. This indicates that the DNA extraction protocol must be carefully selected prior to use in analysis of microbial communities. PCR assay for C. fetus: Bovine semen contaminated with C. fetus subsp venerealis cannot be exported. To develop highly specific assays to identify this pathogen, we screened 65 field strains of C. fetus with applied amplified fragment length polymorphism (AFLP) analysis in collaboration with University of Utrecht, College of Veterinary Medicine. Based on AFLP comparisons of these isolates obtained from Europe, Africa, North America and the Middle East, PCR specific primers were designed which amplified only C. fetus subspecies venerealis. Multilocus Sequence Typing (MLST) of Campylobacter coli: C. coli is a food-borne pathogen associated increasingly with human illness. We examined 488 C. coli strains from 4 different food animals (cattle, chickens, swine and turkeys), collected over a 6-year period throughout the United States in
collaboration with ARS Albany, California, Athens, Georgia, and North Carolina State University. The majority of the sequence types (STs) could be linked to a specific host origin. The presence of host-associated C. coli MLST alleles may track potential animal sources during sporadic or outbreaks of human clinical C. coli. Pathogenic Yersinia enterocolitica in pigs: Y. enterocolitica a zoonotic pathogen impacting US pork production and is included in the FoodNet surveillance for human food-borne pathogens. To determine the prevalence of pathogenic strains, 2,793 fecal samples from hog 77 farms were screened for pathogenic Y. enterocolitica over a period of 27 weeks in collaboration with APHIS-CEAH. Of the 77 farms samples 53% (41 sites) contained at least one fecal sample positive for the ail sequence indicating that the US swine are contaminated and thus are reservoirs for human Y.NBenterocolitica infections. Intestinal microbiota transition in the pre-adolescent turkey: A period
of community transition in the ceca of domestic turkeys was detected between 9 and12 weeks of age. This transition was marked by a conversion from Clostridial to Bacteroides predominance. Sequence analysis indicated the transition included a period in which the communities were dominated by multiple strains of Bacteroides uniformis (84% and 79% for two trials, respectively) indicating a possible role in maturation of the intestinal community. The transition may be a result of decreasing levels of growth hormone or a decreased dietary need for protein during this time frame. A significant increase of Campylobacter coli was also detected at week 12 in one trial, suggesting a correlation between microbiota destabilization and pathogen prevalence. Intestinal bacterial communities differ between domestic and wild turkeys: We compared 2990 16S rRNA clones from 13 wild and 13 domesticated turkeys in collaboration with University of California- Riverside. Bacterial library composition was
determined to include Bacteroidetes (28% domestic, 29% wild), Clostridiales clusters XIV, IV and IX (16% domestic, 12% wild), proteobacteria (2% domestic, 0.4% wild) and unknown organisms (6% domestic, 0.9% wild). Fingerprint cluster analysis identified domestic- and wild-turkey specific clusters of Bacteroidetes, Clostridiales. Deferribacterales were found almost exclusively in the domestic animals, as were the proteobacteria and an unidentified Lower levels of Campylobacter in the intestines of the wild birds may indicate the presence of competing microbes or increased immunity to the food borne pathogen. Thus understanding the ecology of the intestinal tract is vital to development of intervention strategies to remove Campylobacter from the food supply. Transport may impact cecal microbiota: Library comparisons indicated significant community differences between on-farm birds at Farm A and birds from Farm A that were transported and killed at the abattoir. In addition, total gut
communities from Farm A were significantly different from those from Farm B. Sustained exposure to growth-promoting antibiotics and intermittent whole-flock treatment with therapeutic antibiotics greatly alters the intestinal microbiota of poultry, and thus immune status and health. Description of rumen protozoal diversity: Rumen-associated protozoa include species for which 18S rRNA sequences do not yet exist. Fingerprint analysis detected no correlation between protozoa species and their associated bacterial communities. We developed a set of protozoa- specific primers to amplify the 18S rRNA genes from individually isolated protozoa, which allowed sequence-based identification of 14 isolates as Dasytricha ruminantium (8 isolates, 96-100% sequence similarity) Isotricha prostoma (1 isolate, 96% sequence similarity), Epidinium caudatum (1 isolate, 98% sequence similarity) and four without similarity to known protozoa sequences. Rumen acidosis results from the activities of unknown
protozoa, however complete defaunation of the animals threatens their well-being. A description of natural protozoal communities in collaboration with PHFSED, NADC and protozoal communities correlated with acidosis will help identify the culprits and allow selective defaunation. Antimicrobial drug resistant Campylobacter in market weight turkeys: We screened Campylobacter jejuni (n=31 isolates) and C. coli (n=33 isolates) from commercially raised turkeys for resistance to nine antimicrobials included in the NARMS panel. All of the isolates were resistant to tetracycline whereas the majority of C. jejuni (84%) and C. coli (734%) were resistant to fluoroquinoles although the flocks had no history of fluoroquinolone use. This suggests the importance of collecting on-farm drug histories and that naturally occurring fluoroquinolone-resistance may be unrelated to drug use but may be related to Campylobacters ability to replicate in bile. 5. Describe the major accomplishments to date and
their predicted or actual impact. These major accomplishments are consistent with the National Program Action Plan addressing sampling, isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment (Action Plan Priority Objective 1.1.1). The in- house research and extramural projects utilized statistically-based methods to identify pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the prevalence of foodborne pathogens. Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., as summarized in Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Effects caused by stress, co- mingling, and environmental contamination (Action Plan Priority Objective 1.2.1.2). Our eight studies indicate that the holding pen at the abattoir poses a significant risk factor for acute antemortem infection of hogs with Salmonella. This led to field trials
including disinfection of pens and bypassing the holding pen by directly unloading hogs into the slaughter line. A major program shift occurred in April 2003 when more emphasis was placed on elucidating the sources of Campylobacter and Salmonella contamination in turkeys. In response, we conducted field studies in 2003 and 2004 to determine the role of transportation and holding in turkeys immediately prior to slaughter. Whereas population shifts are seen in Campylobacter, no such changes were documented for Salmonella in turkeys. This is in contrast to perimarketing events in swine which are associated with a statistically significant increase in Salmonella prevalence. In addition, we have adapted molecular methods, such as real- time PCR formats to identify and speciate and now quantitate foodborne pathogens in livestock, indicating the ease of application of molecular methods. Ability to differentiate C. jejuni from C. coli is fundamental to understanding dynamics of
Campylobacter in the avian gut and in evaluating on-farm pathogen reduction interventions. By PFGE profiles, C. coli strains obtained from turkeys were genetically more diverse than C. jejuni, indicating that C. coli may be better adapted to survive environmental stress. Ability to monitor the microbial communities in the turkey ceca with molecular techniques will elucidate the dynamics of the interaction of commensal microbes with frank pathogens and will indicate the optimal time for administration of pro- and prebiotics on- farm. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Details of the effect of feed withdrawal, transport, and holding on the intestinal populations of Salmonella, Campylobacter jejuni and C. coli in the market weight turkey
have been published and presented to scientific and commodity group meetings, including the annual meeting of the National Turkey Federation (February 10, 2006), APHIS Salmonella Summit (August 2006). Similarly, the lack of effect of perimarketing events on Salmonella shedding in turkeys in contrast to what occurs in market weight hogs is being presented to regulatory, scientific, and commodity industry representatives. Despite the FSIS emphasis on Salmonella reduction, although specific on-farm interventions are well known, the economics of such undertaking may be cost prohibitive. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Presented, Identification of host-specific alleles and detection of potential lateral transfer events using novel Campylobacter and Arcobacter MLST methods. 7th International Meeting on Microbial Epidemiological
Markers. May 11-14, 2005. Presented, Host specificity of Campylobacter coli multilocus sequence typing (MLST) alleles. International Union of Microbiological Societies meeting, July 23-28, 2005. Presented, Prevalence of Salmonella enterica in market-age turkeys. Research Works in Animal Diseases Conference. 2005 Presented, Prevalence of Salmonella enterica in turkeys on-farm and at slaughter. FSIS Salmonella in Poultry Symposium, Athens, GA August 25- 27, 2005. Presented, Salmonella and Campylobacter colonization in the context of microbial community development in the ceca of the domestic turkey. FSIS Salmonella in Poultry Symposium, Athens, GA August 25-27, 2005. Presented, The effect of transport and holding on Salmonella and Campylobacter in commercial turkeys. FSIS Salmonella in Poultry Symposium, Athens, GA August 25-27, 2005. Presented, Detection of heat-injured but viable L. monocytogenes by reverse transcriptase real-time PCR. Winter meeting of the Iowa Turkey
Federation, Des Moines, Iowa. 2005 Presented a seminar to faculty, students, and staff, highlighting molecular methods for characterizing foodborne pathogens, with an emphasis on Campylobacter in market-weight birds at the Autonomous University of Nuevo Leon, Monterrey, Mexico, November 2005. Presented, Prevalence of pathogenic Yersinia enterocolitica strains in pigs in the United States. 2005. Presented, Identification of host-associated alleles by multilocus sequence typing of Campylobacter coli strains from food animals. 2006.
Impacts
(N/A)
Publications
- Andersen, M.M., Wesley, I.V., Muraoka, W.T., Nestor, E.J., Bouchard, C.T. 2005. Prevalence of Arcobacter in commercial turkey production [abstract]. International Workshop on Campylobacter, Helicobacter, and Related Organisms. p. 101.
- Dassanayake, R.P., Stryker, C.J., Johnson, R.K., Muraoka, W.T., Wesley, I. V., Duhamel, G.E. 2005. Characterization of a novel Campylobacter cytolethal distending toxin from Campylobacter hyointestinalis subsp. hyointestinalis isolated from humans and pigs [abstract]. Rushmore Conference on Mechanisms in Pathogenesis of Enteric Diseases. p. 33.
- Dassanayake, R.P., Stryker, C.J., Johnson, R.K., Gebhart, C.J., Post, K.W., Hinkley, S., Muraoka, W.T., Wesley, I.V., Duhamel, G.E. 2005. The U.S. Porcine Campylobacter coli are negative for cytolethal distending toxin activity [abstract]. Research Workers in Animal Diseases Conference Proceedings. p. 88.
- Lechtenberg, T., Muraoka, W.T., Wesley, I.V. 2005. Quantification of Salmonella enterica via the revised RX tube method [abstract]. North Central Branch of the American Society for Microbiology. p.78.
- Rostagno, M.H., Wesley, I.V., Trampel, D.W., Hurd, H.S. 2005. Prevalence of Salmonella enterica in turkeys on-farm and at slaughter [abstract]. North Central Branch-American Society for Microbiology Annual Meeting. p. 67.
- Scupham, A.J., Baldwin, J., Rasmussen, M.A. 2005. Fingerprint analysis of bacterial communities associated with single protozoa [abstract]. North Central Branch-American Society for Microbiology. p. 61.
- Scupham, A.J., Jones, J., Bent, E., Borneman, J. 2005. Intestinal bacterial communities of domestic and wild turkeys [abstract]. North Central Branch-American Society for Microbiology. p. 62.
- Wesley, I.V., Muraoka, W.T., Scupham, A.J., Rostagno, M.H., Trampel, D. 2005. Prevalence of Campylobacter jejuni and Campylobacter coli in market weight turkeys on-farm and at slaughter [abstract]. International Workshop on Campylobacter, Helicobacter, and Related Organisms. p. 109.
- Van Bergen, M.A., Simons, G., Van Der Graaf, L., Van Putten, J.M., Rombout, J., Wesley, I.V., Wagenaar, J.A. 2005. Amplified fragment length polymorphism based identification of genetic markers and novel PCR assay for differentiation of campylobacter fetus subspecies. Journal of Medical Microbiology. 54(12):1217-1224.
- Wesley, I.V., Hurd, H.S., Muraoka, W.T., Harbaugh, E., Trampel, D. 2005. The effect of transport and holding on Salmonella and Campylobacter. World Poultry. 21(9):28-30.
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Progress 10/01/04 to 09/30/05
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Food animals are the primary source of bacterial human foodborne pathogens. Following USDA-Food Safety and Inspection Service (FSIS)- Hazard Analysis and Critical Control Points (HACCP) regulation and consumer concerns, livestock producers are attempting to reduce the on- farm prevalence of these pathogens. The project has three goals: (1) Develop rapid and sensitive molecular-based techniques, such as enyzyme- linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) assays, DNA fingerprinting methods, pulsed field gel electrophoresis (PFGE), oligonucleotide fingerprinting of ribosomal RNA genes (OFRG) and randomly amplified polymorphic DNA (RAPD) to detect, enumerate, and characterize intestinal microbial communities and specific bacterial foodborne pathogens in livestock; (2) Delineate
the dynamics of foodborne pathogens in livestock and their environment and develop intervention strategies to reduce transmission on-farm; (3) Determine pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the dynamics of the normal intestinal microbial flora and the presence of foodborne pathogens. Pathogens to be studied include: Campylobacter, Salmonella, Listeria, and emerging bacterial agents (e.g., Yersinia, Arcobacter). Molecular methods, such as PCR-based protocols, combined with the ecological perspective will identify management practices to reduce the prevalence of foodborne pathogens in hogs and turkeys. Sampling methods for isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). Management practices include feed withdrawal, transport, holding,
biosecurity, housing, etc., (Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1) as well as stress, commingling, and environmental contamination (Action Plan Priority Objective 1.2.1.2). This research is central to achieving effective intervention strategies to reduce on-farm transmission (Action Plan Priority Objective 1.4.1). In-house efforts are complemented by external support (USDA-CSREES-NRI-CGP, USDA Program 406, Food Safety Consortium, Iowa Turkey Federation, Midwest Poultry Federation, National Pork Board, and National Alliance for Food Safety funding). In the U.S., approximately 5 million cases of bacterial foodborne illness occur annually at a cost of approximately $3.2 billion. In the U. S., over 300,000 swine per day are slaughtered. Approximately 3% to 50% of those animals harbor Salmonella when they reach the kill floor. Hogs are also a major animal reservoir for Yersinia enterocolitica, a bacterial agent, which is transmitted to humans by pork. The U.S. turkey
industry is valued at $7.8 billion with per capita consumption 18 pounds annually. Of the 458,000 pounds of whole or cut-up turkey parts exported, nearly 10% is destined for Russia. Thus, the Russian embargo against U.S. poultry attributed to Salmonella contamination, impacted the industry. Turkeys, which represent the second commodity addressed in this project, are a natural reservoir for C. jejuni and Salmonella. Reducing the prevalence of these bacteria in hogs and turkeys may lower human foodborne illnesses and deaths. 2. List the milestones (indicators of progress) from your Project Plan. Objective 1 Year 1 (FY 2001): Optimize methods for Salmonella. Year 2 (FY 2002): Optimize methods for Campylobacter. Year 3 (FY 2003): Opimize protocols for Yersinia. Year 4 (FY 2004): Optimize protocols for Listeria monocytogenes. Year 5 (FY 2005): Optimize protocols for Arcobacter. Objective 2 Year 1 (FY 2001): On-farm environmental studies: planning and data collection. Year 2 (FY
2002): On-farm studies: inputs (feed and water) and outputs (hog and runoff contamination): analysis and reporting. Year 3 (FY 2003): On-farm studies: data collection, analysis. Year 4 (FY 2004): On-farm studies: data collection, analysis. Year 5 (FY 2005): On-farm studies: reporting. Objective 3 Year 1 (FY 2001): Lairage/transportation study: planning. Year 2 (FY 2002): Lairage/transportation study: data collection, analysis, reporting. Year 3 (FY 2003): Case control studies: planning, data collection. Year 4 (FY 2004): Case control studies: planning, data collection, analysis. Abattoir: planning data collection analysis; Manure management: planning. Year 5 (FY 2005): Manure management: data collection, analysis. 3a List the milestones that were scheduled to be addressed in FY 2005. For each milestone, indicate the status: fully met, substantially met, or not met. If not met, why. 1. Identification protocols for the bacterium Arcobacter will be optimized.
Milestone Fully Met 2. Identify system inputs and outputs from on-farm studies which influence food safety pathogens. Milestone Substantially Met 3. Correlate swine production practices with microbial contamination on hog carcasses. Milestone Not Met Redirection of Research focus due to change in priorities 3b List the milestones that you expect to address over the next 3 years (FY 2006, 2007, and 2008). What do you expect to accomplish, year by year, over the next 3 years under each milestone? The project is scheduled to be included in the NP 108 Food Safety Panel Review, conducted by the Office of Scientific Quality Review, to begin December 2004 with the new project plan expected to be certified and implemented October 2005. Thus, the year's milestones listed below reflect those of the pending, but yet to be approved CRIS project plan and accompanying milestones. Year 1 (FY 2006) Objective 1 Subobjective 1.1 - Track entry of Campylobacter and Salmonella into the brooder house;
Begin flock sampling. Subobjective 1.2 - Document flock management practices; with FSIS design management questionnaire. Objective 2 Subobjective 2.1 - Map distribution of Campylobacter in intestine. Subobjective 2.4 - Recruit 0.5 SY. Objective 3 Subobjective 3.1 - Description of bacterial and fungal communities; begin bacterial fungal OFRG. Subobjective 3.2 - Total community flux; complete on-farm flock data collection. Objective 4 Subobjective 4.1 - Optimize detection and enumeration of Salmonella; evaluate single Salmonella MPN method. Year 2 (FY 2007) Objective 1 Subobjective 1.1 - Evaluate data; redesign sampling strategy, if needed. Subobjective 1.2 - With risk assessors analyze questionnaire. Objective 2 Subobjective 2.2 - Begin in vivo virulence assays with C. coli. Subobjective 2.2 - Begin in vitro virulence assays with C. coli. Objective 3 Subobjective 3.1 - Complete bacterial, fungal OFRG. Subobjective 3.2 - Complete analysis. Subobjective 3.3 - Begin antibiotic dissection
of gut community. Objective 4 Subobjective 4.1 - Evaluate DNA extraction protocols. Subobjective 4.2 - Evaluate DNA and real-time PCR forms with various organs. Subobjective 4.3 - Plan studies with FSIS; secure plant access. Year 3 FY 2008 Objective 1 Subobjective 1.1 - Data analysis; retest if needed. Subobjective 1.2 - Present data and survey responses to risk assessors. Objective 2 Subobjective 2.2 - Continue screening in vivo models. Subobjective 2.3 - Continue screening cell lines. Subobjective 2.4 - Begin C. coli microarray analysis. Objective 3 Subobjective 3.1 - Present and publish results. Subobjective 3.3 - OFRG and validation. Objective 4 Subobjective 4.1 - Evaluate DNA extraction protocols. Sobobjective 4.3 - Begin in-plant sampling. 4a What was the single most significant accomplishment this past year? Distribution of Campylobacter associated with perimarketing events in turkeys: Viscera of birds (crops, duodenum, jejunum, ileum, colon, ceca) from six commercial flocks
in the Midwest were examined for Campylobacter before (n = 30/flock) and after (n = 30/flock) transport to the abattoir in collaboration with scientists at the USDA-ARS-National Animal Disease Center (NADC), Ames, IA, and Iowa State University (D. Trampel), and partially funded by the Iowa Turkey Federation. Overall, when data for the six farms are combined, no difference was seen in Campylobacter spp. recovered from ceca, duodenum, ileum, spleen, and large intestine, whereas after transport, recovery was significantly higher from the gall bladder and crop when compared to on-farm levels (P < 0.05). Overall, C. coli was isolated more often from the crop and cecum, whereas C. jejuni predominates in the intestine (duodenum, ileum, colon) resulting in its more frequent isolation from cloacal swabs. Ability to differentiate C. jejuni from C. coli is fundamental to understanding dynamics of Campylobacter in the avian gut and in evaluating on-farm pathogen reduction interventions. 4b
List other significant accomplishments, if any. Salmonella populations are not altered during perimarketing events: We compared the prevalence of Salmonella enterica in turkeys on six commercial flocks pre- (30 birds/flock) and post- (30 birds/flock) transport in collaboration with scientists at the Iowa State University (D. Trampel), and partially funded by the Iowa Turkey Federation. No statistical difference was found between the overall prevalence found on- farm and at slaughter, based on any sample type analyzed (crop, ceca, liver/gall bladder, and spleen) with cecal contents having the highest relative sensitivity. This demonstrates that the perimarketing practices of feed withdrawal, catching, loading, transportation, and pre-slaughter holding do not significantly affect the prevalence of S. enterica in market-age turkeys in contrast to what we have previously reported for hogs. Therefore, it may be possible to estimate on-farm prevalence based on samples collected at
slaughter. Vitamin E does not facilitate survival of L. monocytogenes in irradiated turkey meat: We monitored survival of Listeria monocytogenes (LM) following irradiation in ground meat from turkeys fed diets supplemented with vitamin E. These collaborative efforts between scientists at the USDA-ARS-NADC and Iowa State University were partially funded by USDA- CSREES-Special Program 406. Dietary vitamin E, which we have previously shown augments the immune response of turkeys, did not diminish the survival of LM in meat following irradiation. This suggests that vitamin E while boosting the immune response of birds does not compromise the bactericidal effects of irradiation on turkey meat. Culture methods bias the recovery of Salmonella from swine feces: Four culture methods (A, B, C, and D) were evaluated for their ability to recover Salmonella enterica from pooled swine fecal samples (n = 100). None of the methods was able to isolate the Salmonella at the same frequency and of
the same serotype from all positive samples. We concluded that culture methods differ in efficiency of recovering S. enterica serotypes from naturally contaminated swine fecal samples. Therefore, depending on the objective(s) of investigations on the ecology and epidemiology of S. enterica populations in swine, a method or a combination of methods should be considered for more reliable results. Comparison of DNA extraction protocols: We performed preliminary experiments for analysis of the microbial ecology of the turkey cecum, including comparison of 14 DNA extraction methods. Electrophoretic and spectrophotometric analysis showed a wide range of DNA yield, shearing and contamination resulted from the methods. In addition, bacterial ribosomal small subunit quantification of the extraction products indicated a wide range of PCR inhibition resulting from template contaminants. This indicates that the DNA extraction protocol must be carefully selected prior to use in analysis of
microbial communities. Seasonal distribution of Arcobacter: The prevalence of Arcobacter detected by cloacal swab (6/298) and cecal contents (3/145) suggests that Arcobacter colonizes the intestinal tract at very low levels. The overall prevalence of Arcobacter in the drinker water decreased from 63. 04% (29/46) in the summer of 2003 to 24.66% (18/73) in the spring of 2004 with a concommitant shift in the species of Arcobacter during this interval. The prevalence of Arcobacter in the water appears to be related to the chlorination level present in the drinker water. Antibiotic resistance profiles of Campylobacter isolated from swine: Human Campylobacter infections are commonly treated with quinolone or macrolide antibiotics, such as erythromycin. The observed rise in the number of Campylobacter resistant to quinolone and macrolide antibiotics may be related to the use of growth promotants such as the macarolide tylosin in animal agriculture. C. coli strains (n = 152) isolated
from swine farms in the Midwest were resistant to ciprofloxacin (7.9), to doxycycline (57.8%), erythromycin (65.1%), but not to gentamicin (0.0%), and minimally to meropenem (1.3%). Together these findings suggest that the swine C. coli isolates are highly resistant to erythromycin, which may be due to the use of macrolides as growth promoters during swine production. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. We have adapted molecular methods to identify and speciate foodborne pathogens in livestock, indicating the ease of application of molecular methods. In addition, the real-time PCR format is being used to monitor the levels of Salmonella in experimentally infected and naturally infected turkeys indicating the feasibility of this approach. By PFGE profiles, C. coli strains obtained from turkeys were genetically more diverse than C. jejuni, indicating that C. coli may be better adapted to survive environmental
stress. Our studies indicate that the holding pen at the abattoir poses a significant risk factor for acute antemortem infection of hogs with Salmonella. This led to field trials including disinfection of pens and bypassing the holding pen by directly unloading hogs into the slaughter line. A major program shift occurred in April 2003 when more emphasis was placed on elucidating the sources of Campylobacter and Salmonella contamination in turkeys. In response, we conducted field studies in 2003 and 2004 to determine the role of transportation and holding in turkeys immediately prior to slaughter. Whereas population shifts are seen in Campylobacter, no such changes were documented for Salmonella. This is in contrast to perimarketing events in swine which are associated with a statistically significant increase in Salmonella prevalence. These major accomplishments are consistent with the National Program Action Plan and milestones in the project plan. Sampling isolation,
identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). The in-house research and extramural projects will utilize statistically-based methods to identify pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the prevalence of foodborne pathogens. Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., as summarized in Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Effects caused by stress, co-mingling, and environmental contamination (Action Plan Priority Objective 1.2.1.2). 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of
the technology products? Details of the effect of feed withdrawal, transport, and holding on the intestinal populations of Campylobacter jejuni and C. coli in the market weight turkey have been published and presented to various user groups. Similarly, the lack of effect of perimarketing events on Salmonella shedding in turkeys in contrast to what occurs in market weight hogs is being presented to regulatory, scientific, and commodity groups. Despite the FSIS emphasis on Salmonella reduction, although specific on-farm interventions are well known, the economics of such undertaking may be cost prohibitive. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Reviewed research findings on the effect of transport on the dynamics of Campylobacter and Salmonella in turkeys to the Salmonella Summit hosted at corporate headquarters of Jennie-O Turkey, the
world's largest turkey processor, March 31, 2005, Willmar, Minnesota. Presented a seminar to faculty, students, and staff, highlighting molecular methods for characterizing foodborne pathogens, with an emphasis on Campylobacter in market-weight birds at the Autonomous University of Nuevo Leon, Monterrey, Mexico, November 2004. Interviewed by the Poultry World to summarize role of transport on shedding of Campylobacter and Salmonella in market weight turkeys.
Impacts
(N/A)
Publications
- Romero, M.G., Mendonca, A.F., Ahn, D.U., Wesley, I.V. 2005. Influence of dietary Vitamin E on behavior of Listeria monocytogenes and color stability in ground turkey meat following electron beam irradiation. Journal of Food Protection. 68(6):1159-1164.
- Rostagno, M.H., Gailey, J.K., Hurd, H.S., McKean, J.D., Leite, R.C. 2005. Culture methods differ on the isolation of Salmonella enterica serotypes from naturally contaminated swine fecal samples. Journal of Veterinary Diagnostic Investigation. 17(1):80-83.
- Wesley, I.V., Muraoka, W.T., Trampel, D., Hurd, H.S. 2005. Effect of pre- slaughter events on prevalence of Campylobacter jejuni and Campylobacter coli in market-weight turkeys. Applied and Environmental Microbiology. 71(6):2824-2831.
- Wesley, I.V., Muraoka, W.T., Bouchard, C.T., Scupham, A.J., Trampel, D. 2004. Prevalence of Campylobacter jejuni (CJ) and Campylobacter coli (CC) in market weight turkeys pre- and post-transport [abstract]. Conference of Research Workers in Animal Diseases. p. 114.
- Behm, S., Pereira, S., Wesley, I.V., Bouchard, C.T., Zhang, Q. 2004. Antibiotic resistance profiles of Campylobacter strains isolated from swine [abstract]. Conference of Research Workers in Animal Diseases. p. 91.
- Wesley, I.V., Muraoka, W.T., Bouchard, C.T., Scupham, A.J., Trampel, D. 2004. Prevalence of Campylobacter jejuni (CJ) and Campylobacter coli (CC) in market weight turkeys pre- and post-transport [abstract]. U.S.-Japan Cooperative Program in Natural Resources Panel of Animal and Avian Health Meeting. 39:5.
- Koski, J., Bouchard, C.T., Wesley, I.V., Muraoka, W.T., Trampel, D. 2004. Prevalence of Campylobacter spp. in the crop and ceca of market turkeys pre- and post-transport [abstract]. In: The Food Safety Consortium Annual Meeting, October 3-5, 2004, Ames, Iowa. 2004 CDROM.
- Andersen, M., Wesley, I.V., Muraoka, W.T., Nestor, E.J., Bouchard, C.T., Trampel, D. 2004. On-farm prevalence of Arcobacter species in market- weight commercial turkeys as determined by two isolation protocols [abstract]. In: The Food Safety Consortium Annual Meeting, October 3-5, 2004, Ames, Iowa. 2004 CDROM.
- Wesley, I.V., Muraoka, W.T., Bouchard, C.T., Scupham, A.J., Trampel, D. 2004. Distribution of Campylobacter jejuni (CJ) and Campylobacter coli (CC) in market weight turkeys pre- and post-transport: comparative prevalence in cloacal swabs, crop and ceca [abstract]. In: The Food Safety Consortium Annual Meeting, October 3-5, 2004, Ames, Iowa. 2004 CDROM.
- Wesley, I.V., Hurd, H.S., Muraoka, W.T., Harbaugh, E., Trampel, D. 2004. The effect of transport and holding on Salmonella and Campylobacter in commercial turkeys [abstract]. In: Proceedings of the North Central Avian Disease Conference. The Food Safety Consortium Annual Meeting, October 3-5, 2004, Ames, Iowa. 2004 CDROM.
- Scupham, A.J., Rardin, J.A., Wesley, I.V. 2004. Importance of DNA isolation method on representation of microbial diversity [abstract]. In: Proceedings of the North Central Avian Disease Conference. The Food Safety Consortium Annual Meeting, October 3-5, 2004, Ames, Iowa. 2004 CDROM.
- Scupham, A.J., Wesley, I.V. 2004. Extraction of total microbial DNA from the cecal contents of turkeys: comparison of methods [abstract]. North Central Branch of the American Society for Microbiology. Paper No. 67.
- Scupham, A.J., Wesley, I.V. 2004. Extraction of total microbial DNA from the ceceal contents of turkeys: comparison of methods [abstract]. Conference of Research Workers in Animal Diseases. p. 91.
- Scupham, A.J. 2005. Microbial community development in the ceca of the domestic turkey (Meleagris gallopavo) [abstract]. American Society for Microbiology. p. 397.
- Rostagno, M.H., Hurd, H.S., McKean, J.D. 2004. Bacteriological and serological Salmonella prevalence in finishing pigs [abstract]. Conference of Research Workers in Animal Diseases. p. 121.
- Gailey, J., O'Connor, A., McKean, J., Hurd, H.S. 2004. Enumeration of Salmonella in postmortem porcine samples [abstract]. In: Proceedings of the Food Safety Consortium Annual Meeting. The Food Safety Consortium Annual Meeting, October 3-5, 2004, Ames, Iowa. 2004 CDROM.
- Hurd, H.S., Gailey, J.K., McKean, J.K., Griffith, R.W. 2005. Variable abattoir conditions affect Salmonella enterica prevalence and meat quality in swine and pork. Foodborne Pathogens and Disease. 2(1):77-81.
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Progress 10/01/03 to 09/30/04
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Food animals are the primary source of bacterial human foodborne pathogens. Following USDA-Food Safety and Inspection Service (FSIS)- Hazard Analysis and Critical Control Points (HACCP) regulation and consumer concerns, livestock producers are attempting to reduce the on- farm prevalence of these pathogens. The project has three goals: (1) Develop rapid and sensitive molecular-based techniques, such as enyzyme- linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) assays, DNA fingerprinting methods, pulsed field gel electrophoresis (PFGE), and randomly amplified polymorphic DNA (RAPD) to detect, enumerate, and characterize bacterial foodborne pathogens in livestock and their environment; (2) Delineate the dynamics of foodborne pathogens in livestock and their environment and develop
intervention strategies to reduce transmission on-farm; (3) Determine pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the presence of foodborne pathogens. Pathogens to be studied include: Campylobacter, Salmonella, Listeria, and emerging bacterial agents (e.g., Yersinia). Molecular methods, such as PCR-based protocols, combined with the ecological perspective will identify management practices to reduce the prevalence of foodborne pathogens in hogs and turkeys. Sampling isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., (Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1) as well as stress, commingling, and environmental contamination (Action Plan Priority
Objective 1.2.1.2). This research is central to achieving effective intervention strategies to reduce on-farm transmission (Action Plan Priority Objective 1.4.1). In-house efforts are complemented by external support (USDA-CSREES-NRI- CGP, USDA Program 406, Food Safety Consortium, Iowa Turkey Federation, Midwest Poultry Federation, National Pork Board, and National Alliance for Food Safety funding). In the U.S., approximately 5 million cases of bacterial foodborne illness occur annually at a cost of approximately $3.2 billion. In the U. S., over 300,000 swine per day are slaughtered. Approximately 3% to 50% of those animals harbor Salmonella when they reach the kill floor. Hogs are also a major animal reservoir for Yersinia enterocolitica, a bacterial agent, which is transmitted to humans by pork. The U.S. turkey industry is valued at $7.8 billion with per capita consumption 18 pounds annually. Of the 458,000 pounds of whole or cut-up turkey parts exported, nearly 10% is
destined for Russia. Thus, the Russian embargo against U.S. poultry attributed to Salmonella contamination, impacted the industry. Turkeys, which represent the second commodity addressed in this project, are a natural reservoir for Salmonella and C. jejuni. Reducing the prevalence of these bacteria in hogs and turkeys may lower human foodborne illnesses and deaths. 2. List the milestones (indicators of progress) from your Project Plan. Objective 1 Year 1 (FY 2001): Optimize methods for Salmonella Year 2 (FY 2002): Optimize methods for Campylobacter Year 3 (FY 2003): Opimize protocols for Yersinia Year 4 (FY 2004): Optimize protocols for Listeria monocytogenes Year 5 (FY 2005): Optimize for Arcobacter Objective 2 Year 1 (FY 2001): On-farm environmental studies: planning and data collection Year 2 (FY 2002): On-farm studies: inputs (feed and water) and outputs (hog and runoff contamination): analysis and reporting Year 3 (FY 2003): On-farm studies: data collection, analysis
Year 4 (FY 2004): On-farm studies: data collection, analysis Year 5 (FY 2005): On-farm studies: reporting Objective 3 Year 1 (FY 2001): Lairage/transportation study: planning Year 2 (FY 2002): Lairage/transportation study: data collection, analysis, reporting Year 3 (FY 2003): Case control studies: planning, data collection Year 4 (FY 2004): Case control studies: planning, data collection, analysis. Abattoir: planning data collection analysis; Manure management: planning Year 5 (FY 2005): Manure management: data collection, analysis 3. Milestones: A. List the milestones (from the list in Question #2) that were scheduled to be addressed in FY 2004. How many milestones did you fully or substantially meet in FY 2004 and indicate which ones were not fully or substantially met, briefly explain why not, and your plans to do so. Objective 1. Optimize protocols for Listeria monocytogenes - The PCR assay to detect Listeria monocytogenes in real time (5' nuclease, TaqMan) is now being
adapted in a reverse transcriptase format to detect only viable L. monocytogenes. Methods to genotype this foodborne pathogen by PFGE were completed during FY 2002 and are continuing for Campylobacter, Arcobacter, and Salmonella. Thus this milestone is substantially met. Objective 2. On-farm studies: data collection, analysis - Milestones for conducting on-farm studies as prelude for determining on-farm risk factors were partially met and will continue for Salmonella and Campylobacter in turkeys and hogs. Objective 3. Case control studies: planning, data collection, analysis. Abattoir: planning data collection analysis; Manure management: planning - Interventions, such as no holding, holding (2-3 hours) on the transport as well as floor type (slated versus solid concrete), disinfection, and other pen treatments were evaluated. Thus, milestones for this ambitious objective were partially accomplished and raised additional research questions. Research involving manure management
originally incorporated into the project was refocused to address the issue of pen contamination and disinfection. B. List the milestones (from the list in Question #2) that you expect to address over the next 3 years (FY 2005, 2006, & 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? The project is scheduled to be included in the NP 108 Food Safety Panel Review, conducted by the Office of Scientific Quality Review, to begin December 2004 with the new project plan expected to be certified and implemented October 2005. The Year 5 (FY 2005) milestones are listed below with a description of the anticipated outcomes. Year 5 (FY 2005) Construct new project plan to focus on delineating the dynamics of foodborne pathogens and the interaction with the resident host intestinal flora throughout the livestock production as a basis for developing intervention strategies to reduce pathogen transmission. Develop novel molecular methods to monitor
host intestinal communities. Identify on-farm risk factors leading to the introduction of Campylobacter and Salmonella into turkey hatchery. Assuming that the subsequent project plan continues the research along its present course, we anticipate accomplishing in: Year 6 (FY 2006) Analyze microbial community changes during the lifetime of domestic turkeys and identify intestinal microbes whose populations change concordant with the colonization by Salmonella. Identify on-farm risk factors leading to the introduction of Campylobacter and Salmonella into brooder flocks by monitoring poults at the time of placement and weekly thereafter. Identify on-farm risk factors by correlating management practices (feeding, live haul, holding, biosecurity, housing, etc.) with the presence of foodborne agents in hogs and turkeys. Year 7 (2007) Based on progress realized in Year 6 (FY 2006), continue analysis of microbial community dynamics during the lifetime of domestic turkeys and initiate parallel
studies in pigs. Identify microbes whose populations change concordant with the colonization by Campylobacter. Identify on-farm risk factors leading to the introduction of Campylobacter and Salmonella into turkey flocks by monitoring birds at weeks 0, 3, 12, and 16 after placement in the growout house. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004 (one per Research (OOD) Project): In field studies of six commercial turkey farms, perimarketing events which occur immediately prior to slaughter, such as feed withdrawal, transport and holding at the slaughterhouse, were monitored for their impact on Campylobacter and Salmonella prevalence. We documented a shift in the distribution of C. jejuni and C. coli (increased C. coli) in cloacal swabs of market-weight turkeys (n = 1,200) when compared to the on-farm baseline prevalence. In contrast, Salmonella prevalence in turkeys, unlike previous reports in swine,
did not change during transport and holding. In some flocks, a statistically significant increase in the overall levels of Campylobacter spp., as well as C. coli, was observed following transport. By genotype comparison of isolates by PFGE, C. coli was found to be more diverse than C. jejuni at both pre- and post-transport. These data indicate that evaluating the effects of transport stress will require speciation and genotyping in addition to simple prevalence estimates. B. Other significant accomplishment(s), if any We demonstrated that large fans, like those used in holding sheds at commercial slaughterhouses, can spread Salmonella to uninfected birds. Through collaborative efforts between scientists at the USDA-ARS-National Animal Disease Center (NADC), Ames, IA, and Iowa State University (D. Trampel), and partially funded by the Iowa Turkey Federation, we simulated the holding shed environment to determine if Salmonella could be transmitted through aerosols. In two trials,
Salmonella was recovered from the trachea, air sacs, lung and spleen of "clean" market-weight turkeys (n = 30 birds/trial) following aerosol exposure to infected birds (n = 10 birds/trial). This indicates the possibility of aerosol transmission of Salmonella in the holding shed of commercial abattoirs within hours of slaughter. Scientists at the USDA-ARS-NADC in Ames, IA, in collaboration with Iowa State University (J. McKean), completed intervention trials comparing Salmonella prevalence in pigs held in disinfected pens and finalized spatial analysis to determine the best sampling strategies to quantify Salmonella levels on pen floors. The studies indicate that although cleaning and disinfection reduce the amount of Salmonella in the holding pen, the impact on the overall prevalence of Salmonella in market-weight pigs remains inconclusive. We validated the most probable number (MPN) method to enumerate Salmonella in tissues of 200 hogs at three commercial abattoirs. These
collaborative efforts between scientists at the USDA-ARS-NADC and Iowa State University (J. McKean) were partially funded by Food Safety Consortium. Of the 450 samples collected, Salmonella levels were highest in ileocecal lymph nodes but were also readily isolated from cecal and fecal samples. This study provides useful information for a quantitative risk assessment of the sources and levels of Salmonella potentially entering the food chain during slaughter. We monitored survival of Listeria monocytogenes (LM) following irradiation in ground meat from turkey fed diets supplemented with vitamin E. These collaborative efforts between scientists at the USDA- ARS-NADC and Iowa State University were partially funded by USDA-CSREES- Special Program 406. Dietary vitamin E, which we have previously shown augments the immune response of turkeys, did not diminish the survival of LM in meat following irradiation. In addition, survival of LM was greater in vacuum-packaged samples compared
to samples packaged aerobically for both meat from birds fed control and vitamin E diets. We estimated the prevalence of Arcobacter on six Midwestern commercial turkey farms and recorded a shift in the species of Arcobacter during this time. This is a collaborative effort between scientists at the USDA- ARS-NADC and Iowa State University (D. Trampel), Midwest turkey producers, and a central abattoir, and with partial funding by the Midwest Poultry Federation, Food Safety Consortium, and the Iowa Turkey Federation. In contrast to C. jejuni, to which it is closely related, Arcobacter spp. were infrequently detected in cloacal swab (2%) and cecal contents (2%), which suggests that low level of intestinal colonization. The overall prevalence of Arcobacter in the drinker water decreased from summer (63. 04%) to spring (24.66%) and may be related to the chlorination level present in the drinker water. The low Arcobacter prevalence in live birds in contrast to its high prevalence on
carcasses and suggests extensive cross contamination at slaughter. We developed a model of acute Salmonella colonization of the crop of the domestic turkey and investigated the effect of probiotic and prebiotic treatment with lactobaccilli and lactose. This is a collaborative effort between scientists at the USDA-ARS-NADC and Iowa State University (C. Scanes) with partial funding from the Midwest Poultry Federation, the Iowa Turkey Marketing Council, Iowa Livestock Health Advisory Council, and a USDA special grant. Although by immunohistochemical staining Salmonella was juxtaposed with the crop epithelium, lactose and L. acidophilus supplementation did not reduce Salmonella colonization. This crop colonization model may prove critical in screening candidate probiotic and prebiotic interventions for reducing Salmonella in birds. C. Significant activities that support special target populations. None. 5. Describe the major accomplishments over the life of the project, including
their predicted or actual impact. Our molecular-based detection methods are being used in field studies to detect Y. enterocolitica in hogs. We have adapted molecular methods to identify and speciate Campylobacter in turkeys, indicating the ease of application of molecular methods. In addition, the real-time PCR format is being used to monitor the levels of Salmonella in experimentally infected turkeys indicating the feasibility of this approach. By PFGE profiles C. coli strains obtained from turkeys were genetically more diverse than C. jejuni, indicating that C. coli may be better adapted to survive environmental stress. Our studies indicate that the holding pen at the abattoir poses a significant risk factor for acute antemortem infection of hogs with Salmonella. This led to field trials including disinfection of pens and bypassing the holding pen by directly unloading hogs into the slaughter line. A major program shift occurred in April 2003 when more emphasis was placed on
elucidating the sources of Campylobacter and Salmonella contamination in turkeys. In response, we initiated field studies in 2003 to determine the role of transportation and holding in turkeys immediately prior to slaughter. These major accomplishments are consistent with the National Program Action Plan and milestones in the project plan. Sampling isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). The in-house research and extramural projects will utilize statistically-based methods to identify pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the prevalence of foodborne pathogens. Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., as summarized in Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Effects
caused by stress, co- mingling, and environmental contamination (Action Plan Priority Objective 1.2.1.2). 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Findings about the risk of abattoir holding pens have been transferred via meetings and conferences to abattoir management, veterinary practitioners, producers, and other researchers. In spring 2004, at the request of the International Epidemiology Laboratory of the Danish Institute for Food and Veterinary Research, a sabbatical was awarded to Dr. Scott Hurd to evaluate the current and alternative herd classification schemes with regard to their ability to reduce the public health risk of salmonellosis attributable to pork. Specific management practices to minimize cross contamination in holding
pens have yet to be developed although research continues. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. Summarized Salmonella and Campylobacter research and discussed future turkey research with the Executive Board of the Iowa Turkey Federation, October 2003, Ames, Iowa. Unit organized an ARS/Industry workshop entitled "Salmonella in swine: the transport and lairage dilemma," to discuss research progress and direction with industry, November 4, 2003. Highlighted research findings on the effect of transport on the dynamics of Campylobacter and Salmonella in turkeys to the Annual Turkey Day sponsored by the Iowa Turkey Federation, Des Moines, Iowa, December 2003. Presented research findings on the effect of transport on the dynamics of and Salmonella and Campylobacter in turkeys to the annual meeting of the National Turkey Federation, Savannah, Georgia, January 2004. Reviewed research findings on
the effect of transport on the dynamics of and Campylobacter and Salmonella in turkeys to the annual meeting of the Midwest Poultry Federation, Minneapolis, Minnesota, March 2004. Met with Executive Board of the Iowa Turkey Federation to discuss progress in food safety turkey research as a prelude to obtaining formal Specific Cooperative Agreements, Ames, Iowa, April 2004. Summarized Campylobacter and Salmonella research, discussed future turkey research and introduced the potential significance of understanding microbial gut communities of the adult bird with West Liberty Foods, West Liberty, Iowa, April 2004. Met with key turkey industry commodity representatives and described efforts in molecular analysis of the avian microbial gut communities as well as the dynamics of Salmonella and Campylobacter in market-weight birds at the Turkey Research Update, National Animal Disease Center, Ames, Iowa, May 2004. Reviewed the role of holding pens as a risk factor for acute salmonellosis
transmission immediately prior to slaughter to the George A. Young Swine Health and Management Conference, South Sioux City, Nebraska, August 2004. Presented a seminar to faculty, students, and staff, highlighting dynamics of Campylobacter in market-weight birds pre- and post-transport at the Autonomous University of Nuevo Leon, Monterrey, Mexico, November 2003. Interviewed by the Food Safety Consortium to summarize role of cull sows as potential carriers of L. monocytogenes and was published in the 2003 winter quarter newsletter. Interviewed by the ARS magazine staff on the role of vitamin E as a means of reducing foodborne pathogens in turkeys and was published in January 2004 and August 2004. Interviewed by the ARS magazine staff on the role of holding pens as a fomite in the transmission of Salmonella to hogs immediately prior to slaughter and was published in February 2004.
Impacts
(N/A)
Publications
- HURD, H.S., MCKEAN, J.D., GRIFFITH, R.D., ROSTAGNO, M.H. ESTIMATION OF THE SALMONELLA ENTERICA PREVALENCE IN FINISHING SWINE. EPIDEMIOLOGY AND INFECTION. 2004. V. 132. P. 127-135.
- Johannsen, S.A., Griffith, R.W., Wesley, I.V., Scanes, C.G. 2004. Salmonella enterica serovar typhimurium colonization of the crop in the domestic turkey: influence of probiotic and prebiotic treatment (lactobacillus acidophilus and lactose). Avian Diseases. 48(2):279-286.
- Larsen, S.T., Hurd, H.S., McKean, J.D., Griffith, R.W., Wesley, I.V. 2004. Effect of short-term lairage on the prevalence of Salmonella enterica in cull sows. Journal of Food Protection. 67(7):1489-1493.
- Schmidt, P.L., O'Connor, A.M., McKean, J.D., Hurd, H.S. 2004. The association between cleaning and disinfection of lairage pens and the prevalence of Salmonella enterica in swine at harvest. Journal of Food Protection. 67(7):1384-1388.
- ZHU, M., WESLEY, I.V., NANNAPANENI, R., COX, M., MENDONCA, A., JOHNSON, M. G., AHN, D. THE ROLE OF DIETARY VITAMIN E IN EXPERIMENTAL LISTERIA MONOCYTOGENES INFECTIONS IN TURKEYS. POULTRY SCIENCE. 2003. v. 82. p. 1559- 1564.
- ANDERSEN, M.M., WESLEY, I.V., TRAMPEL, D.W., MURAOKA, W.T., NESTOR, E.J., COATES, E., HURD, H.S. PREVALENCE OF ARCOBACTER IN IOWA'S COMMERCIAL TURKEYS AS DETERMINED BY TWO DIFFERENT ISOLATION PROTOCOLS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2003. ABSTRACT P. 21P.
- ANDERSEN, M.M., WESLEY, I.V., TRAMPEL, D.W., MURAOKA, W.T., NESTOR, E.J., COATES, E., HURD, H.S. PREVALENCE OF ARCOBACTER IN IOWA'S COMMERCIAL TURKEYS AS DETERMINED BY TWO DIFFERENT ISOLATION PROTOCOLS. US-JAPAN COOP PGM IN NATURAL RESOURCES PANEL OF ANIMAL AND AVIAN HEALTH MEETING. 2003. ABSTRACT P. 20.
- Bush, E., Wesley, I.V., Bhaduri, S. 2003. Risk factors for yersinia enterocolitica on u.s. swine farms in 2000. Meeting Abstract. p.54-56
- GRIFFITH, R.W., HURD, H.S., LEVIS, I., MCKEAN, J.D. PERACUTE INFECTION OF SWINE WITH SALMONELLA. Proceedings of the International Symposium on the Epidemiology and Control of Foodborne Pathogens in Pork. 2003. p. 321-322.
- HAMIR, A.N., SONN, R.J., FRANKLIN, S.L., WESLEY, I.V. SMALL INTESTINAL INTUSSUSCEPTION ASSOCIATED WITH CAMPYLOBACTER JEJUNI INFECTION IN A RACCOON. CAMPYLOBACTER HELICOBACTER AND RELATED ORGANISMS INTERNATIONAL WORKSHOP. 2003. ABSTRACT P. 55.
- HURD, H.S., MCKEAN, J.D., GAILEY, J.K., GRIFFITH, R.W., O'CONNOR, A.C. SLATTED PEN FLOORS REDUCE SALMONELLA IN MARKET SWINE HELD IN ABATTOIRS. Proceedings of the International Symposium on the Epidemiology and Control of Foodborne Pathogens in Pork. 2003. p. 203-204.
- Hurd, H.S., Rostagno, M.H., McKean, J.D. 2004. Effect of pre-slaughter holding in abattoir pen or transport trailer on Salmonella prevalence [abstract]. Proceedings of the International Pig Veterinary Society. p. 684.
- HURD, H.S., TRAMPEL, D.W., WESLEY, I.V., REGEN, P.S., MURAOKA, W.T., RIVERA, F., HARBAUGH, E. EFFECT OF TRANSPORT AND HOLDING ON SALMONELLA PREVALENCE IN MARKET WEIGHT TURKEYS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2003. ABSTRACT P. 72.
- LARSEN, S.T., MCKEAN, J.D., HURD, H.S., WESLEY, I.V. ELIMINATING THE ABATTOIR PEN LAIRAGES TO DECREASE THE PREVALENCE OF SALMONELLA IN CULL SOWS. Proceedings of the International Symposium on the Epidemiology and Control of Foodborne Pathogens in Pork. 2003. p. 42-44.
- MURAOKA, W.T., WESLEY, I.V., TRAMPEL, D.W., NESTOR, E.J., ANDERSEN, M.M., HURD, H.S., MORALES, V.A. EFFECT OF TRANSPORT AND HOLDING ON THE PREVALENCE OF CAMPYLOBACTER IN MARKET WEIGHT TURKEYS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2003. ABSTRACT P. 20P.
- O'CONNOR, A.M., GRAY, J.T., HURD, H.S., MCKEAN, J.D., ROSTAGNO, M.H. GENETIC RELATEDNESS OF SALMONELLA FROM PENS AND SWINE AT SLAUGHTER. Proceedings of the International Symposium on the Epidemiology and Control of Foodborne Pathogens in Pork. 2003. p. 78-80.
- OYARZABAL, O.A., WESLEY, I.V., BARBAREE, J.M., LAUERMAN, L.H., CONNER, D.E. IDENTIFICATION AND CHARACTERIZATION OF ARCOBACTER AND CAMPYLOBACTER FROM RETAIL POULTRY BY PHYSIOLOGICAL TESTING AND POLYMERASE CHAIN REACTION ASSAYS. CAMPYLOBACTER HELICOBACTER AND RELATED ORGANISMS INTERNATIONAL WORKSHOP. 2003. ABSTRACT P. 144.
- ROSTAGNO, M.H., GAILEY, J.K., HURD, H.S., MCKEAN, J.D. EFFECT OF CULTURE METHODS ON INVESTIGATIONS OF SALMONELLA ENTERICA ECOLOGY AND EPIDEMIOLOGY IN SWINE. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2003. ABSTRACT P. 77.
- ROSTAGNO, M.H., HURD, H.S., MCKEAN, J.D. ARE THERE HIGH AND LOW SALMONELLA PREVALENCE FARMS?. PROCEEDINGS OF THE AMERICAN ASSOCIATION OF SWINE VETERINARIANS ANNUAL MEETING. 2004. P. 445-446.
- Rostagno, M.H., Hurd, H.S., McKean, J.D. 2004. Bacteriological and serological Salmonella prevalence in finishing pigs [abstract]. Proceedings of the International Pig Veterinary Society. p. 649.
- ROSTAGNO, M.H., HURD, H.S., GAILEY, J.K., MCKEAN, J.D. SAMPLING CECAL CONTENTS OR ILEOCECAL LYMPH NODES: IS IT DIFFERENT?. Proceedings of the International Symposium on the Epidemiology and Control of Foodborne Pathogens in Pork. 2003. p. 103-104.
- ROSTAGNO, M.H., HURD, H.S., MCKEAN, J.D., GAILEY, J.K. EVALUATING CECAL CONTENT AND ILEOCECAL LYMPH NODE SAMPLING TO DETERMINE SALMONELLA ENTERICA PREVALENCE IN SWINE. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2003. ABSTRACT P. 59.
- SCHMIDT, P.L., O'CONNOR, A.M., MCKEAN, J.D., HURD, H.S. THE ROLE OF DISINFECTION IN THE CONTROL OF SALMONELLA IN SWINE LAIRAGE PENS. INTERNATIONAL CONFERENCE ON EMERGING ZOONOSES. 2003. ABSTRACT p. 27.
- Wesley, I.V., Bhaduri, S., Bush, E. 2004. 5' fluorogenic PCR (real time) assays to determine the prevalence of Yersinia enterocolitica in U.S. market weight hogs [abstract]. American Society for Microbiology. p. 671.
- WESLEY, I.V., HURD, H.S., MURAOKA, W.T., HARBAUGH, E., TRAMPEL, D. EFFECT OF TRANSPORTATION AND HOLDING ON THE PREVALENCE OF SALMONELLA AND CAMPYLOBACTER IN COMMERCIAL TURKEYS. PROCEEDINGS OF THE MIDWEST POULTRY FEDERATION. 2004. P. 58-68.
- WESLEY, I.V., MURAOKA, W.T., TRAMPEL, D. CAMPYLOBACTER JEJUNI: A MAJOR FOODBORNE BACTERIAL PATHOGEN. CD-ROM. MONTERREY, MEXICO: FOOD SAFETY CONGRESS. 2003.
- WESLEY, I.V., SCHROEDER-TUCKER, L., FRANKLIN, S.L. RECOVERY OF ARCOBACTER SPP. FROM EXOTIC ANIMAL SPECIES. CAMPYLOBACTER HELICOBACTER AND RELATED ORGANISMS INTERNATIONAL WORKSHOP. 2003. ABSTRACT P. 54.
- Hurd, H.S. 2004. Salmonellosis in livestock. In: McGraw-Hill Yearbook of Science and Technology. New York, NY: McGraw Hill. p. 297-299.
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Progress 10/01/02 to 09/30/03
Outputs
1. What major problem or issue is being resolved and how are you resolving it? Food animals are the primary source of bacterial human foodborne pathogens, such as Campylobacter, Salmonella, Listeria, and Yersinia. Following USDA-Food Safety and Inspection Service (FSIS)-Hazard Analysis and Critical Control Points (HACCP) regulation and consumer concerns, livestock producers are attempting to reduce the on-farm prevalence of these pathogens. We are designing molecular-based strategies to detect, quantitate, and characterize bacterial foodborne pathogens in livestock. We are using these assays to study the epidemiology of human bacterial foodborne pathogens on-farm, during transport, and at the abattoir. This study will examine animal-to-animal transmission, environment-to-animal transmission, as well as environmental sources and reservoir hosts of bacterial foodborne agents. Rapid methods combined with the ecological perspective will identify management practices to
reduce the prevalence of foodborne pathogens in hogs and turkeys. Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., as summarized in Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Stress, commingling, and environmental contamination will also be considered (Action Plan Priority Objective 1.2.1.2). In-house efforts are complemented by external support (USDA-CSREES-NRI-CGP, USDA Program 406, National Pork Board, and National Alliance for Food Safety funding). This research is central to achieving effective intervention strategies to reduce on-farm transmission (Action Plan Priority Objective 1.4.1). 2. How serious is the problem? Why does it matter? In the U.S., approximately 5 million cases of bacterial foodborne illness occur annually at a cost of approximately $3.2 billion. In the U.S., over 300,000 swine per day are slaughtered. Approximately 3% to 50% of those animals harbor Salmonella when they reach the kill floor. Hogs are
also a major animal reservoir for Yersinia enterocolitica, a bacterial agent, which is transmitted to humans by pork. The U.S. turkey industry is valued at $7.8 billion with per capita consumption 18 pounds annually. Of the 458,000 pounds of whole or cut-up turkey parts exported, nearly 10% is destined for Russia. Thus, the Russian embargo against U. S. poultry attributed to Salmonella contamination, impacted the industry. Turkeys, which represent the second commodity addressed in this project, are a natural reservoir for Salmonella and C. jejuni. Reducing the prevalence of these bacteria in hogs and turkeys may lower human foodborne illnesses and deaths. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? National Program 108, Food Safety (animal and plant products) (100%). Sampling isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major
research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). In-house efforts are augmented by specific cooperative agreements (funded through the National Alliance for Food Safety). The in-house research and extramural project (National Pork Board) will utilize statistically-based methods to identify pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the prevalence of foodborne pathogens. Management practices include feed withdrawal, transport, holding, biosecurity, housing, etc., as summarized in Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Stress, commingling, and environmental contamination will also be considered (Action Plan Priority Objective 1.2.1.2). In-house efforts are complemented by external support (USDA-CSREES-NRI-CGP, USDA Program 406, National Pork Board, and National Alliance for Food Safety funding). This research is central to achieving effective intervention
strategies to reduce on-farm transmission (Action Plan Priority Objective 1.4.1). 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2003: Research has continued to find sources of Salmonella infection in market pigs during transport and holding prior to slaughter. Through a collaborative effort between USDA-ARS-National Animal Disease Center (NADC), Ames, IA, and J. McKean and R. Griffith, Iowa State University, various Midwest producers and abattoirs, we tested market swine, culled sows, and holding pen environments for Salmonella and Listeria in studies partially funded by the National Alliance for Food Safety. We have demonstrated that holding pens pose a significant risk for Salmonella but not for Listeria contamination of pork. Efforts are now underway to determine if the transportation and holding of turkeys immediately prior to slaughter similarly impact contamination of birds with Salmonella and
Campylobacter. B. Other Significant Accomplishment(s), if any: We designed a statistically validated method for spatial sampling of the abattoir holding pens. This is a collaboration between NADC and Iowa State University (J. McKean), funded by the USDA-CSREES-National Research Initiative-Competitive Grants Program ($783,400, "Perimarketing Control Strategies to Reduce Salmonella Contamination of Pork Carcasses"). During FY 03, we completed three intervention trails to evaluate methods for reducing Salmonella infection of swine immediately prior to slaughter, including no holding in pens (20-30 minutes from unload), holding on slatted versus concrete floors, and cleaning and disinfection of pens. Samples were collected from market swine in one single commercial packing plant, cultured for Salmonella, and the effect of disinfectants was evaluated. We developed a multiplex polymerase chain reaction (PCR) assay for the simultaneous detection of Listeria and L. monocytogenes. Scientists
at NADC, using this assay, detected L. monocytogenes in 38% of mechanically separated turkey meat samples obtained from a single processing plant. We also used this multiplex PCR assay to identify L. monocytogenes in brains of sheep. Although the sheep had died after consuming contaminated sprouts, DNA profiles of isolates obtained by pulsed field gel electrophoresis (PFGE) did not identify the exact source of contamination. To determine the on-farm prevalence of Yersinia enterocolitica in hogs, we sampled pen fecal and tonsil swab samples collected on 127 farms located in the 17 pork-producing states during the USDA-APHIS-Veterinary Services, Center for Animal Health Monitoring (CAHM), National Animal Health Monitoring System (NAHMS) Swine 2000 survey. This is a collaboration between scientists at NADC, ARS-Eastern Regional Research Center (S. Bhaduri), and APHIS (E. Bush), funded by the National Pork Board. Using real-time PCR assays, Y. enterocolitica was identified in tonsil
swabs and fecal samples on nearly half of the premises. Ultimately, four factors, with their accompanying odds ratio were identified: location in a non-central state, vaccination for E. coli, percent deaths due to scours, and presence of meat/bone meal in grower-finisher diet. Research was conducted to determine if dietary vitamin E (fed from the day of hatch to six weeks of age) protects turkeys against colonization with potential human bacterial foodborne pathogens and improves meat quality. Research was conducted through a specific cooperative agreement between ARS-NADC and Iowa State University (A. Mendonca), funded by the National Alliance for Food Safety. Dietary vitamin E (200 IU), increased serum levels of vitamin E, enhanced CD4+ and CD8+ immune cells, accelerated clearance of L. monocytogenes in live turkeys, prolonged shelf life by preserving meat color, and reduced off odors during irradiation. Taken together these studies suggest dietary interventions may both
contribute to on-farm pathogen reduction, a major objective of our inhouse project, and enhance meat quality. To determine environmental sources of E. coli O157:H7, five Texas feedlots in the Texas Panhandle were screened. Under a specific cooperative agreement between ARS and Texas AM University (G. Acuff), funded by the National Alliance for Food Safety, we detected E. coli O157:H7 in 5% of 800 feedlot samples (chutes, water and muck from lagoons, pen drainage, etc.). Positive samples were most often isolated from a single feedlot, which suggests that not all premises are equally at risk of contaminating beef. Recovery of a single strain, as determined by rep- PCR-DNA fingerprinting, from hides and from the feedlot environment on multiple visits suggests that its maintenance and transmission are enhanced by the cattle production environment. To compare genetic and virulence-related attributes of L. monocytogenes, we analyzed isolates from retail purchased foods. Research was
conducted under a specific cooperative agreement between NADC, North Carolina State University (S. Kathariou), and Cornell University (K. Boor), funded by the National Alliance for Food Safety. In this phase of the two-year study, nearly half of the strains of serotype 4b harbored genetic markers associated with major epidemic-associated lineage of L. monocytogenes. On the other hand, L. monocytogenes serogroup 1/2, known to be frequently found in foods, may also harbor unique genetic markers not encountered among clinical isolates of this serogroup which may confer competitive survival advantages in foods, but not human pathogenicity. Since cull sows are a major source of pork, we hypothesized that these older animals, when compared to younger market-weight hogs, may pose a greater public health risk for L. monocytogenes and Salmonella contamination. Research was conducted under a specific cooperative agreement between NADC, Iowa State University (J. McKean), and University of
Arkansas (M. Johnson), with the cooperation of an industry partner and funded by the National Alliance for Food Safety. For Listeria, there was no evidence of peri-marketing contamination, although the low isolation rates hampered this comparison. In contrast, the observed increase in Salmonella isolation rates and the serotype diversity in cull sows at slaughter compared to on-farm samplings were consistent with peri- marketing practices that increase contamination. C. Significant Activities that Support Special Target Populations: Collected data from three organic swine producers to estimate the prevalence of Salmonella in swine raised outdoors (non-confinement). 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Molecular-based detection methods are being used in field studies to detect Y. enterocolitica in hogs and to identify and speciate Campylobacter in turkeys. In addition, real-time PCR formats have been used to
monitor the levels of Salmonella in experimentally infected turkeys fed supplemental vitamin E, selenium, and conjugated linoleic acid. This fulfills the milestones for year 2002 of Objective 1 (Optimize Enzyme-Linked Immunosorbent Assay (ELISA) and PCR assays to detect Salmonella and other pathogens) of our project plan. Our studies indicate that the holding pen at the abattoir poses a significant risk factor for acute antemortem infection of hogs with Salmonella. This fulfills the milestones set for 2003 for Objective 3 (Correlate swine production and transportation practices with hog carcass contamination). A major program shift occurred in April 2003 when more emphasis was placed on elucidating the sources of Salmonella and Campylobacater contamination in turkeys. In response, we began intensive field studies during summer 2003 to determine the role of transportation and holding in turkeys immediately prior to slaughter. Achieving milestones projected for Objective 2 awaits
the hiring of a new scientist in October 2003. These major accomplishments are consistent with the National Program Action Plan and milestones in the project plan. 6. What do you expect to accomplish, year by year, over the next 3 years? FY 2004: Optimize rapid and sensitive molecular-based techniques to detect, quantitate, and characterize bacterial foodborne pathogens in hogs and turkeys and their environment. FY 2005: Delineate the dynamics of foodborne pathogens in livestock and their environment and develop intervention strategies to reduce transmission. FY 2006: Identify on- farm risk factors by correlating management practices (feeding, live haul, holding, biosecurity, housing, etc.) with the presence of foodborne agents in hogs and turkeys. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the
adoption and durability of the technology products? Findings about the risk of abattoir holding pens are being transferred to abattoir management, veterinary practitioners, producers, and other researchers. In summer 2003, at the request of the USDA-ARS-National Program Staff and the turkey industry, we initiated studies to determine if live-haul and holding similarly impact the prevalence of Salmonella and Campylobacter in turkeys. 8. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: This does not replace your peer-reviewed publications listed below). Summarized Salmonella research and discussed future turkey research with Jennie O, Inc., the world's largest turkey producer and processor in Willmar, MN, August 2002. Summarized Salmonella and Campylobacter research and discussed future turkey research with president and CEO of West Liberty Foods, a turkey processor in West Liberty, IA, September
22, 2002. Present a seminar and six hours of lecture on Salmonella epidemiology to faculty and field veterinarians at the University of Parma, Parma, Italy, 2002. Highlighted swine Salmonella research at the National Center for Food Safety and Toxicology, Michigan State University, East Lansing, MI, 2002. Invited to Pharmacia Inc., to present swine Salmonella research and explore collaboration, Kalamazoo, MI, 2002. Invited by National Pork Board to draft a review paper, with Ray Gamble of the National Academy of Sciences, on the food safety effects of extensive swine management (e.g., outdoor housing), 2002-2003. Conferred with Center for Disease Control's Enteric Disease Branch as a member of the National Pork Board's Salmonella Working Group, Atlanta, GA, 2002. Consulted with International Ingredient Corporation on a Salmonella problem in feed, January 2003-present. Presented a seminar to faculty, students, and staff, highlighting our government-academic-private industry cooperative
projects at the Autonomous University of Nuevo Leon, Monterrey, Mexico, November 2002. Summarized food safety efforts at NADC at the ARS-Mexico Food Safety Workshop, Hermosillo, Sonora, Mexico, February 2003. Summarized major food safety issues in the U.S. at the First International Food Safety Workshop, Guadalajara, Mexico, May 2003. Presented a seminar detailing PCR-based methods for the detection and identification of human foodborne pathogens at the First PCR Workshop, Guadalupe, Mexico, October 2002. Interviewed by the ARS magazine staff on the role of vitamin E as a means of reducing foodborne pathogens in turkeys. Interview will appear in the November 2003 edition. Interviewed by the Food Safety Consortium to summarize role of cull sows as potential carriers of L. monocytogenes. Interview will appear in the 2003 winter quarter newsletter. Highlighted current vitamin E research to turkey producers at the annual meeting of the National Turkey Federation, San Antonio, Texas,
January 31, 2003.
Impacts
(N/A)
Publications
- HURD, H.S., MCKEAN, J.D., GRIFFITH, R.W., ROSTAGNO, M.H. LAIRAGE IS A HAZARD FOR SALMONELLA INFECTION IN MARKET SWINE. PROCEEDINGS OF ALLEN D LEMAN SWINE CONFERENCE. 2002. P. 223-225.
- ROSTAGNO, M.H., HURD, H.S., GAILEY, J.K., MCKEAN, J.D., LEITE, R.C. COMPARATIVE EVALUATION OF SALMONELLA DETECTION ASSAYS ON SWINE FECAL SAMPLES. CONGRESS OF THE INTERNATIONAL PIG VETERINARY SOCIETY. 2002. P. 311.
- ROSTAGNO, M.H., HURD, H.S., MCKEAN, J.D., ZIEMER, C.J., GAILEY, J.K., LEITE, R.C. PRESLAUGHTER HOLDING ENVIRONMENT IN PORK PLANTS IS HIGHLY CONTAMINATED WITH SALMONELLA ENTERICA. APPLIED AND ENVIRONMENTAL MICROBIOLOGY. 2003. V. 69. P. 4489-4494.
- HURD, H.S., MCKEAN, J., GRIFFITH, R., ROSTAGNO, M.H. MEASURING SALMONELLA PREVALENCE IN FINISH SWINE: EVALUATION OF THREE METHODS. PROCEEDINGS CONGRESS OF THE INTERNATIONAL PIG VETERINARY SOCIETY. 2002. P. 313.
- ROSTAGNO, M.H., HURD, H.S., MCKEAN, J.D., ZIEMER, C.J., GAILEY, J.K., LEITE, R.C. SALMONELLA INFECTION IN MARKET SWINE DURING PRE-SLAUGHTER HOLDING. CONGRESS OF THE PIG VETERINARY SOCIETY. 2002. P. 319.
- LARSEN, S.T., MCKEAN, J.D., HURD, H.S., ROSTAGNO, M.H., GRIFFITH, R.W., WESLEY, I.V. IMPACT OF COMMERCIAL PREHARVEST TRANSPORTATION AND HOLDING ON THE PREVALENCE OF SALMONELLA ENTERICA IN CULL SOWS. JOURNAL OF FOOD PROTECTION. 2003. V. 66. P. 1134-1138.
- FRANK, G.H., BRIGGS, R.E., DUFF, G.C., HURD, H.S. EFFECTS OF INTRANASAL EXPOSURE TO LEUKOTOXIN-DEFICIENT MANNHEIMIA HAEMOLYTICA AT THE TIME OF ARRIVAL AT THE FEEDYARD ON THE SUBSEQUENT ISOLATION OF M. HAEMOLYTICA FROM NASAL SECRETIONS OF CALVES. AMERICAN JOURNAL OF VETERINARY RESEARCH. 2003. v. 64(5). p. 580-585.
- LARSEN, S.T., MCKEAN, J.D., HURD, H.S., ROSTAGNO, M.H., GRIFFITH, R.W., WESLEY, I.V. IMPACT OF COMMERCIAL PRE-HARVEST TRANSPORTATION AND HOLDING ON THE PREVALENCE OF SALMONELLA ENTERICA IN CULL SOWS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2002. ABSTRACT P. 50P.
- LARSEN, S.T., MCKEAN, J.D., HURD, H.S., GRIFFITH, R.W., WESLEY, I.V. THE EFFECTS OF ABATTOIR HOLDING PENS ON THE PREVALENCE OF SALMONELLA ENTERICA IN CULL SOWS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2002. P. 82.
- HURD, H.S., MCKEAN, J.D., GAILEY, J.K., GRIFFITH, R.W., O'CONNOR, A.C. THE DIRECT EFFECT OF HOLDING PENS ON SALMONELLA ISOLATION RATES FROM MARKET SWINE. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2002. ABSTRACT P. 81.
- LEVIS, I.M., HURD, H.S., MCKEAN, J.D., GAILEY, J.K., LARSEN, S.T., HARBAUGH, E., GRIFFITH, R.W. USE OF FRUCTOOLIGOSACCHARIDE TO PREVENT RAPID SALMONELLA INFECTION IN MARKET WEIGHT SWINE. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2002. ABSTRACT P. 54P.
- O'CONNOR, A.M., MCKEAN, J.D., GAILEY, J.K., HURD, H.S. SAMPLING STRATEGIES FOR QUANTIFYING SALMONELLA SPP. IN SWINE LAIRAGE PENS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2002. ABSTRACT P. 78.
- WESLEY, I.V., LARSON, D., HARMON, K., LUCHANSKY, J.B., SCHWARTZ RAMOS, A. A CASE REPORT OF SPORADIC OVINE LISTERIAL MENIGOENCEPHALITIS IN IOWA WITH AN OVERVIEW OF LIVESTOCK AND HUMAN CASES. JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION. 2002. V. 14. P. 314-321.
- DU, M., AHN, D.U., MENDONCA, A.F., WESLEY, I.V. QUALITY CHARACTERISTICS OF IRRADIATED READY-TO-EAT BREAST ROLLS FROM TURKEYS FED CONJUGATED LINOLEIC ACID. POULTRY SCIENCE. 2002. V. 81. P. 1378-1384.
- KANUGANTI, S.R., WESLEY, I.V., REDDY, P.G., MCKEAN, J., HURD, H.S. DETECTION OF LISTERIA MONOCYTOGENES IN PIGS AND PORK. JOURNAL OF FOOD PROTECTION. 2002. V. 65. P. 1470-1474.
- ZHU, M., WESLEY, I.V., MENDONCA, A., AHN, D.U. THE ROLE OF DIETARY VITAMIN E IN EXPERIMENTAL LISTERIA MONOCYTOGENES INFECTIONS IN TURKEYS. FOOD SAFETY CONSORTIUM PROCEEDINGS. 2002. P. 178-185.
- WESLEY, I.V., LARSEN, S., HURD, H.S., MCKEAN, J., FRANA, T., COX, M., JOHNSON, M.G., NANNAPANENI, R., HOGNESS, A.L., RIVERA, F. THE PREVALENCE OF L. MONOCYTOGENES IN CULL SOWS. FOOD SAFETY CONSORTIUM PROCEEDINGS. 2002. P. 193-197.
- WESLEY, I.V., BORUCKI, M.K., LARSON, D., SCHROEDER-TUCKER, L. LISTERIA AND LISTERIOSIS IN LIVESTOCK. ISAACSON, R.E., TORRENCE, M.E. EDITORS. IOWA STATE UNIVERSITY PRESS, AMES, IA. CURRENT TOPICS IN FOOD SAFETY AND ANIMAL AGRICULTURE. 2003. P. 223-241.
- WESLEY, I.V., PINKERTON, A.L., RIVERA, F., LARSEN, S.T., HURD, H.S., MCKEAN, J., GRIFFITH, R., FRANA, T., NANNAPANENI, R., WAGNER, D. THE PREVALENCE OF L. MONOCYTOGENES IN CULL SOWS. CONFERENCE OF RESEARCH WORKERS IN ANIMAL DISEASES. 2002. ABSTRACT P. 69.
- NAM, K.C., MIN, B.R., YAN, H., LEE, E.J., MENDONCA, A., WESLEY, I.V., AHN, D.U. EFFECT OF DIETARY VITAMIN E AND IRRADIATION ON LIPID OXIDATION, COLOR, AND VOLATILES OF FRESH AND PREVIOUSLY FROZEN TURKEY BREAST PATTIES. MEAT SCIENCE. 2003. V. 65. P. 513-521.
- ZHU, M., WESLEY, I.V., NANNAPANENI, R., COX, M., MENDONCA, A., JOHNSON, M. G., AHN, D.U. THE ROLE OF DIETARY VITAMIN E IN EXPERIMENTAL LISTERIA MONOCYTOGENES INFECTIONS IN TURKEYS. UNITED STATES JAPAN NATURAL RESOURCES ANIMAL AND AVIAN HEALTH PANEL. 2002. ABSTRACT P. 37.
- BOYAPALLE, S., WESLEY, I.V., MENDONCA, A.F. EVALUATION OF REVERSE TRANSCRIPTASE 5' NUCLEASE PCR ASSAY FOR THE DETECTION OF VIABLE HEAT- INJURED AND RESUSCITATED LISTERIA MONOCYTOGENES IN GROUND PORK. UNITED STATES JAPAN NATURAL RESOURCES ANIMAL AND AVIAN HEALTH PANEL. 2002. ABSTRACT P. 24.
- WESLEY, I.V., LARSEN, S., HURD, H.S., MCKEAN, J.D., FRANA, T., COX, M., JOHNSON, M.G., NANNAPANENI, R., HOGNESS, A.L., RIVERA, F. THE PREVALENCE OF L. MONOCYTOGENES IN CULL SOWS. UNITED STATES JAPAN NATURAL RESOURCES ANIMAL AND AVIAN HEALTH PANEL. 2002. ABSTRACT P. 31.
- HARRIGAN, K.A., HARKINS, K.R., WESLEY, I.V. EVALUATING PNA (PEPTIDE NUCLEIC ACID) RRNA SPECIFIC PROBES FOR USE IN THE DETECTION OF LISTERIA SPP. AND SALMONELLA SPP. IN POULTRY. AMERICAN SOCIETY FOR MICROBIOLOGY. 2003. ABSTRACT P. 502.
- HAMIR, A.N., SONN, R.J., FRANKLIN, S.L., WESLEY, I.V. SMALL INTESTINAL INTUSSUSCEPTION ASSOCIATED WITH CAMPYLOBACTER JEJUNI INFECTION IN A RACCOON (PROCYON LOTOR). 2003. DISEASES AT THE INTERFACE BETWEEN DOMESTIC LIVESTOCK AND WILDLIFE SPECIES. ABSTRACT P. 37.
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Progress 10/01/01 to 09/30/02
Outputs
1. What major problem or issue is being resolved and how are you resolving it? Food animals are the primary source of bacterial human foodborne pathogens, such as Campylobacter, Salmonella, Listeria, and Yersinia. Following Food Safety and Inspection Service (FSIS) -Hazard Analysis and Critical Control Points (HACCP) regulation and consumer concerns, livestock producers are attempting to reduce the on-farm prevalence of these pathogens. We are designing molecular-based strategies to detect, quantitate, and characterize bacterial foodborne pathogens in livestock. We are using these assays to study the epidemiology of human bacterial foodborne pathogens on-farm, during transport, and at the abattoir. This study will examine animal-to- animal transmission, environment-to-animal transmission, as well as environmental sources and reservoir hosts of bacterial foodborne agents. Rapid methods combined with the ecological perspective will identify management practices to
reduce the prevalence of foodborne pathogens in hogs and turkeys. 2. How serious is the problem? Why does it matter? In the U.S., approximately 5 million cases of bacterial foodborne illness occur annually at a cost of approximately $3.2 billion. In the U.S., over 300,000 swine per day are slaughtered. Approximately 3% to 50% of those animals harbor Salmonella when they reach the kill floor. Hogs are also a major animal reservoir for Yersinia enterocolitica, a bacterial agent, which is transmitted to humans by pork. The U.S. turkey industry is valued at $7.8 billion with per capita consumption 18 pounds annually. Of the 458,000 pounds of whole or cut-up turkey parts exported, nearly 10% is destined for Russia. Thus, the recent month-long Russian embargo against U.S. poultry attributed to Salmonella contamination, impacted the industry. Turkeys, which represent the second commodity addressed in this project, are a natural reservoir for Salmonella and C. jejuni. Reducing the
prevalence of these bacteria in hogs and turkeys may lower human foodborne illnesses and deaths. 3. How does it relate to the national Program(s) and National Program Component(s) to which it has been assigned? National Program 108, Food Safety (animal products) (100%). Sampling isolation, identification, characterization, and quantification of foodborne pathogens in livestock and their environment are major research priorities for ARS pre-harvest food safety efforts (Action Plan Priority Objective 1.1.1). In-house efforts are augmented by specific cooperative agreements (funded through the National Alliance for Food Safety). The in-house research and extramural project (National Pork Board) will use statistically-based methods to identify pre-harvest food safety risk factors in livestock by correlating on-farm management practices with the prevalence of foodborne pathogens. Management practices include feeding, transport, holding, biosecurity, housing, etc., as summarized in
Action Plan Priority Objectives 1.3.1.1 and 1.5.1.1. Stress, commingling, and environmental contamination will also be considered (Action Plan Priority Objective 1.2.1.2). In-house efforts are complemented by external support (USDA-CSREES-NRI-CGP, USDA Program 406, and National Alliance for Food Safety funding). This research is central to achieving effective intervention strategies to reduce on-farm transmission (Action Plan Priority Objective 1.4.1). 4. What was your most significant accomplishment this past year? A. Single Most Significant Accomplishment during FY 2002: Research has progressed to identify sources of Salmonella infection in market pigs during transport and holding prior to slaughter. In collaboration with J. McKean and R. Griffith, Iowa State University, and various Midwest producers and abattoirs, we tested market swine, culled sows, and holding pen environments for Salmonella and Listeria. We have demonstrated that holding pens pose a significant risk for
Salmonella contamination of pork. This work could significantly impact the management of swine and other livestock during transport and holding. B. Other Significant Accomplishment(s), if any: We have developed a statistically validated method for spatial sampling of the abattoir holding pens, funded by a four-year USDA-CSREES-NRI-CGP award ($783,400 "Perimarketing Control Strategies to Reduce Salmonella Contamination of Pork Carcasses"). We completed the first trial to determine if pigs killed immediately after unloading from the transport trailers harbor less Salmonella than those held for 2 hours on solid or slatted concrete floors. To evaluate the impact of immediate kill on meat quality, samples collected from market swine in one commercial packing plant (n = 450 hogs) were cultured for Salmonella and data on meat quality (pH, color, drip loss) were obtained. If meat quality is maintained in the absence of the 2 h rest, then the holding pen and associated risks of pork
contamination may be eliminated. To determine the on-farm prevalence of Yersinia enterocolitica in hogs, which are the major animal reservoir for this foodborne pathogen, we sampled pen fecal (n = 2,664) and tonsil swab samples (n = 1,180) collected on farms (n = 127) located in the 17 pork-producing states during the Animal and Plant Health Inspection Service (APHIS)-National Animal Health Monitoring System (NAHMS) Swine 2000 survey. This is a collaboration with ARS-Eastern Regional Research Center (S. Bhaduri) and APHIS (E. Bush), funded by the National Pork Board. Using real-time polymerase chain reaction (PCR) assays, Y. enterocolitica was found in up to 10.1% of tonsil swabs and up to 13% of fecal samples with an overall on-farm prevalence of 48% (58 of 120 premises). We are now correlating prevalence data (individual animal and farm) with management practices in order to design on-farm pathogen reduction strategies. Research was conducted to determine if dietary vitamin E
(fed from the day of hatch to six weeks of age) protects turkeys against colonization with potential human bacterial foodborne pathogens and improves meat quality. Research was conducted through a specific cooperative agreement between ARS (National Animal Disease Center (NADC)) and Iowa State University (A. Mendonca) funded by the National Alliance for Food Safety. Dietary vitamin E (200 IU) did not alter the colonization of turkeys with L. monocytogenes, but increased serum levels of vitamin E, enhanced CD4+ and CD8+ immune cells, prolonged shelf life by preserving meat color, and reduced off-odors during irradiation. Taken together these studies suggest dietary interventions may contribute to on-farm pathogen reduction and enhance meat quality. To determine environmental sources of E. coli O157:H7, July 2001 through March 2002, we screened five Texas feedlots in the Texas Panhandle using a sensitive immunomagnetic separation technique. Under a specific cooperative agreement
between ARS (NADC and Bushland, Texas) and Texas A&M University (G. Acuff) funded by the National Alliance for Food Safety, we detected E. coli O157:H7 in 5% of 800 feedlot samples (chutes, water and muck from lagoons, pen drainage, etc.). Prevalence of E. coli O157:H7 was stable July to September (5.1%-6.6%), declined in October through February (1.2%-1.7%), but peaked in November (13.5%). Positive samples ( 30%) have most often been isolated from a single feedlot, which suggests that not all premises are equally at risk of contaminating beef. To compare genetic and virulence-related attributes of L. monocytogenes, we analyzed isolates from foods implicated in clinical cases (n = 15) and ready-to-eat products not linked to human illness (n = 67). Research was conducted under a specific cooperative agreement between ARS (NADC), North Carolina State University (S. Kathariou), and Cornell University (K. Boor), funded by the National Alliance for Food Safety. Genetic profiles and
surface antigens of the listeriosis-related food isolates were typical of clinical isolates. In contrast, only three strains from control ready-to-eat foods ( 5%) exhibited some genetic markers typical of epidemic-associated strains, which suggests that not all L. monocytogenes food isolates pose the same threat to human health. Since cull sows are a major source of pork, we hypothesized that these older animals, when compared to younger market-weight hogs, may pose a greater public health risk for L. monocytogenes. Research was conducted under a specific cooperative agreement between ARS (NADC), Iowa State University (J. McKean), and University of Arkansas (M. Johnson), with the cooperation of an industry partner (n = 340 hogs), and funded by the National Alliance for Food Safety. Overall, L. monocytogenes was detected in 0.14% of the total samples (n = 2,858); isolates were of either serotype 1 (hog gut isolates) or 4b (hog carcass swabs), which is the serotype associated with
human foodborne outbreaks, were highly virulent (> 90% killing), and similar in antimicrobial resistance profiles. Since the L. monocytogenes prevalence in the cull sow is lower than that of a market-weight hog, the cull sow may not present any additional risk of introducing L. monocytogenes into the pork chain. C. Significant Activities that Support Special Target Populations: none. 5. Describe your major accomplishments over the life of the project, including their predicted or actual impact? Real-time molecular-based detection methods are being used in field studies. This fulfills the milestones for year 2002 of Objective 1 (Optimize Enzyme Linked Immunoassay (ELISA) and PCR assays to detect Salmonella and other pathogens) of our Project Plan. Our studies indicate that the holding pen at the abattoir is a significant risk factor for acute antemortem infection of hogs with Salmonella. This fulfills the milestones set for 2003 for Objective 3 (Correlate swine production and
transportation practices with hog carcass contamination). Achieving milestones projected for Objective 2 await the hiring of a new SY. 6. What do you expect to accomplish, year by year, over the next 3 years? FY 2003: Optimize rapid and sensitive molecular-based techniques to detect, quantitative, and characterize bacterial foodborne pathogens in hogs and turkeys and their environment. FY 2004: Delineate the dynamics of foodborne pathogens in livestock and their environment and develop intervention strategies to reduce transmission. FY 2005: Identify on- farm risk factors by correlating management practices (feeding, transport, holding, biosecurity, housing, etc.) with the presence of foodborne agents in hogs and turkeys. 7. What technologies have been transferred and to whom? When is the technology likely to become available to the end user (industry, farmer other scientist)? What are the constraints, if known, to the adoption durability of the technology? Findings about the
risk of abattoir holding pens are just beginning to be transferred to abattoir management, veterinary practitioners, producers, and other researchers. Reportedly, Denmark has already modified their national Salmonella control program based on our findings. 8. List your most important publications and presentations, and articles written about your work (NOTE: this does not replace your review publications which are listed below) Interviewed by the Iowa Farmer Today for the March 2002 issue. Invited by major turkey processing company and the Iowa Turkey Federation to discuss and initiate projects to reduce Salmonella in pre- harvest birds, March 2002, May 2002, August 2002. The role rapid infection with Salmonella in lairage has become a research priority for funding by the National Pork Board, June 2002. Presented research results to quality assurance managers of pork sausage and market hog slaughter plants. Presented research results at Texas A&M University to faculty and ARS
collaborators, March 2002. Met with Chinese delegation of renderers to discuss Salmonella in pork, July 2002. Met with Practical Farmers of Iowa regarding Salmonella in sustainable agricultural production and potential research projects. Presented a seminar highlighting rapid methods for the detection of bacterial foodborne pathogens to faculty, students, and staff, University of Nuevo Leon, Monterrey, Mexico, March 1, 2002. Invited to present a seminar detailing rapid methods for the detection and identification of Campylobacter at the International Meeting of the Association of Analytical Chemists, Kansas City, MO, September 13, 2001. Interviewed by the Food Safety Consortium to summarize role of vitamin E and the immune response of turkeys. Interview will appear in the August 2002 newsletter. Met with representatives from The Willmar Poultry Group to discuss food safety research in turkeys, April 2002. Presented research summary of Food Safety Research to Midwest Turkey Producers
in Des Moines, IA, on December 5, 2001.
Impacts
(N/A)
Publications
- O'Connor, A., Gailey, J., Hurd, H.S. Development of a spatially valid sampling technique for the enumeration of Salmonella in the swine abattoir holding pen. Annual Meeting of International Association of Food Protection. 2002. Abstract p. 127.
- Wesley, I.V. Campylobacter: Genomic methods for identification. Association of Analytical Chemists International Meeting and Exposition. 2001. Abstract p. 115.
- Ziemer, C.J. Evaluation of culture methods used for investigation of Salmonella enterica serovar ecology in feces. International Symposium on Microbial Ecology. Abstract p. 319.
- Ziemer, C.J., Dang, E. Presence and expression of tetracycline resistance genes in Salmonella. General Meeting of the American Society for Microbiology. 2002. Abstract p. 515.
- Boyapalle, S., Wesley, I.V., Hurd, H. S., Reddy P.G. Comparison of culture, multiplex, and 5' nuclease polymerase chain reaction assays for the rapid detection of Yersinia enterocolitica in swine and pork products. Journal of Food Protection. 2001. v. 64(9). p. 1352-1361.
- Dickins, M.A., Franklin, S., Stefanova, R., Schutze, G.E., Eisenach, K.D., Wesley, I., Cave, M.D. Diversity of Campylobacter isolates from retail poultry carcasses and from humans as demonstrated by pulsed-field gel electrophoresis. Journal of Food Protection. 2002. v. 65(6). p. 957-962.
- Hurd, H.S., McKean, J.D., Griffith, R.W., Wesley, I.V., Rostagno, M.H. Salmonella enterica infections in market swine with and without transport and holding. Applied and Environmental Microbiology. 2002. v. 68(5). p. 2376-2381.
- Hurd, H.S., McKean, J.D., Griffith, R.W., Wesley, I.V., Rostagno, M.H., Gailey, J.K., Karriker, L.A. Lairage is a hazard for Salmonella in the pork chain. Proceedings of Pork Quality Summit. 2002. p. 463-469.
- Jourdan, A., Marichal, M.D., Byl, L., Wesley, I.V. Detection of Yersinia species in pig tonsils by a 5' nuclease fluorogenic (TaqMan) PCR assay specific for the 16S rRNA gene. Proceedings of Annual Meeting of the Food Safety Consortium. 2001. p. ISU27-ISU33.
- Jourdan, A., Marichal, M.D., Byl, L., Wesley, I.V. Detection of Yersinia species in pig tonsils by a 5' nuclease fluorogenic (TaqMan) polymerase chain reaction assay specific for the 16S rRNA gene. CD-ROM. Swine Reports 1998-2001. Ames, IA: Iowa State University Extension Service. AD-646:27-33. 2002.
- Hurd, H.S., Gailey, J.K., McKean, J.D., Rostagno, M.H. Experimental rapid infection in market swine following exposure to a Salmonella contaminated environment. Berliner Muchener Tierarztliche Wochenschrift. 2001. v. 114(9/10). p. 382-384.
- McKean, J.D., Hurd, H.S., Larsen, S., Rostagno, M., Griffith, R.W., Wesley, I.V. Impact of commercial pre-harvest processes on the prevalence of Salmonella enterica in cull sows. Berliner Muchener Tierarztliche Wochenschrift. 2001. v. 114(9/10). p. 353-355.
- Wesley, I.V., Harmon, K.M., Dickson, J.S., Ramos, A. Application of a multiplex polymerase chain reaction assay for the simultaneous confirmation of Listeria monocytogenes and other Listeria species in turkey sample surveillance. Journal of Food Protection. 2002. v. 65(5). p. 780-785.
- Wesley, I.V. Arcobacter and Helicobacter. Labbe, R.G., Garcia, J.S., editors. John Wiley & Sons, Inc., New York, NY. Guide to Foodborne Pathogens. 2001. p. 23-34.
- Byl, L., Wesley, I.V. Analysis of National Animal Health Monitoring System tonsil samples for the presence of ail-bearing Yersinia enterocolitica by using a 5' nuclease fluorogenic PCR (TaqMan) assay. CD-ROM. Swine Reports 1998-2001. Ames, IA: Iowa State University Extension Service. 2002.
- Byl, L., Wesley, I.V. Analysis of NAHMS tonsil samples for the presence of ail-bearing Yersinia enterocolitica using a 5' nuclease fluorogenic PCR (TaqMan) assay. Proceedings of Annual Meeting Food Safety Consortium. 2001. p. ISU34-ISU37.
- Hurd, H.S., McKean, J.D., Griffith, R.D., Wesley, I.V., Rostagno, M.H. Estimation of the on-farm Salmonella enterica prevalence in market swine. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 521-523.
- Hurd, H.S., McKean, J.D., Griffith, R.W., Rostagno, M.H., Wesley, I.V. Salmonella epidemiology - transmission in transport and holding. Proceedings of the Iowa State University Swine Conference. 2001. p. 123- 127.
- Hurd, H.S., McKean, J.D., Wesley, I.V., Karriker, L.A. The effect of lairage on Salmonella isolation from market swine. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 289-291.
- Hurd, H.S., Gailey, J.K., McKean, J.D., Rostagno, M.H. Experimental rapid infection in market swine following exposure to a Salmonella contaminated environment. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 462- 464.
- McKean, J.D., Hurd, H.S., Larsen, S., Rostagno, M., Griffith, R.W., Wesley, I.V. Impact of commercial pre-harvest processes on the prevalence of Salmonella enterica in cull sows. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 295-297.
- McKean, J.D., Hurd, H.S., Rostagno, M.H., Griffith, R.W., Wesley, I.V. Transport and holding at the abattoir: A critical control point for Salmonella in market swine? Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 292-294.
- Rostagno, M., Hurd, H.S., Gailey, J.K., McKean, J., Leite, R. Comparative evaluation of Salmonella detection assays in swine feces. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 531-533.
- Rostagno, M., Hurd, H.S., McKean, J., Ziemer, C.J., Gailey, J., Leite, R. Abattoir holding pens as a source of Salmonella for swine. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 298-300.
- Rostagno, M.H., Hurd, H.S., McKean, J.D., Ziemer, C.J., Gailey, J.K., Leite, R.C. Salmonella infections in swine during pre-slaughter holding. Proceedings of the Brazilian Meeting of Swine Veterinarians. 2001. p. 119- 120.
- Rostagno, M.H., Hurd, H.S., Gailey, J.K., McKean, J.D., Leite, R.C. Acute experimental infection by Salmonella Typhimurium in swine. Proceedings of the Brazilian Meeting of Swine Veterinarians. 2001. p. 125-126.
- Wesley, I.V., Jourdan, A.D., Hurd, H.S., Pinkerton, A.L. 5' fluorogenic PCR assay for the iap gene of Listeria species. Proceedings Annual Meeting of the Food Safety Consortium. 2001. p. ISU38-ISU43.
- Wesley, I.V., Jourdan, A.D., Hurd, H.S., Pinkerton, A.L. 5' fluorogenic PCR assay for the iap gene of Listeria species. CD-ROM. Swine Reports 1998- 2001. Ames, IA: Iowa State University Extension Service. 2002.
- Wesley, I.V., Ransom, G., Franklin, S.K. Identification of Campylobacter species by PCR-based methods. Proceedings of Annual Meeting of the Food Safety Consortium. 2001. p. ISU44-ISU48.
- Zhu, M., Wesley, I.V., Wang, C., Ahn, D.U. Dietary Vitamin E improved the immune responses of turkeys inoculated with Listeria monocytogenes. Proceedings of Annual Meeting of the Food Safety Consortium. 2001. p. ISU49-ISU57.
- Ziemer, C.J. Evaluation of culture methods used for investigation of Salmonella enterica serovar ecology in feces. Proceedings of International Symposium on Epidemiology and Control of Salmonella and Other Foodborne Pathogens in Pork. 2001. p. 537-539.
- Cotta, M.A., Ziemer, C.J., Whitehead T.R. Differentiation between microbial populations of swine feces and stored manure by group specific amplified rDNA restriction analysis. International Symposium on Microbial Ecology. 2001. Abstract p. 349.
- Hurd, H.S., McKean, J.D., Gailey, J.K., Rostagno, M.H., Griffith, R.W. Salmonella infections in market swine before and after transport and holding at the abattoir. Annual Meeting of the Conference of Research Workers in Animal Disease. 2001. Abstract No. 99.
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