Progress 05/15/02 to 12/15/05
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? This project is aligned with National Program 108, Food Safety. The presence of Salmonella is ubiquitous in livestock and livestock environments. This constant threat to food safety must be addressed. When swine are shipped to slaughter a high percentage of them start to shed Salmonella; this reaction is thought to be associated with the stress of transportation and mixing inherent in the process. As few as 4% may be positive for Salmonella on the farm but that percentage can increase up to 40% at the slaughter house. Our lab is investigating the role of stress hormones and their interactions with Salmonella in the living pig. Thus far our research shows that Salmonella can monitor their host's physiology and respond to the stress hormones it releases. This ability may allow
Salmonella to take advantage of the low immune function typically found in animals under stress. 2. List by year the currently approved milestones (indicators of research progress) The milestones are as follows: 2003, Establish Salmonella strain, (Implementation & Analysis); 2004, Transport Model Intranasal Infection (Planning, Implementation, and Analysis); 2005, Transport Model with commingling (Planning, Implementation, and Analysis; 2006, Finalize all manuscripts from conducted research 5. Describe the major accomplishments to date and their predicted or actual impact. This project aligns with NP 108 Component 1.1, Pathogens, Toxins and Chemical Contaminants Preharvest; and it addresses Problem Statement 1.1. 3: Ecology, Host Pathogen and Chemical Contaminants Relationships and 1.1. 4: Intervention Strategies. Successful completion of the project will allow us to develop effective interventions that can be easily instituted by the livestock producer. We found that swine that are
constant shedders of Salmonella have lower levels of cortisol, a stress hormone, than those swine that do not constantly shed Salmonella. This is a surprising finding because it goes directly against the accepted belief that high stress levels allow bacteria to invade their host. The impact of this finding is that management strategies can be developed to temporarily elevate cortisol if it proves to be effective in preventing Salmonella transmission during transport. In addition, by identifying characteristics of individuals that pose a health risk we can develop management tools such as separation during transport to keep these individuals from passing Salmonella to healthy animals. This accomplishment impacts Priority Objective 1.3.1.1. and 1.3.1.2. Our research has also found that Salmonella can monitor and respond to elevated concentrations of norepinephrine, a stress hormone, and more easily infect swine. This is the first time that the ability of bacteria to monitor their
host has been shown to occur in swine. This finding provides a mechanism by which Salmonella can become more virulent and it allows researchers a focus to develop strategies by which producers can lower the stress of mixing and transportation and thus reduce norepinephrine concentrations which will reduce Salmonella's ability to infect swine. This accomplishment impacts Priority Objective 1.3.1.1. and 1.3.1.2. Our lab has recently developed a tool, using biophotonics and Salmonella engineered to express luciferase (this enables the bacteria to emit light) , to enable visual monitoring of Salmonella in the living piglet. To date it is still unclear how Salmonella travel in the living pig and where it resides when it can not be detected in many of the pig's organs. Using biophotonics we will be able to see the answer many of the basic biological questions of Salmonella infection in addition to being able to follow the progress of infection in the same living pig. To date pigs must
be killed. The impact of this research is that it has provided competitive funding from the USDA, National Research Initiative (NRI) program and upon successful completion of this three year grant we will provide a method to monitor bacteria in the living pig in 'real time'. This diagnostic approach will be the next generation of tools to track Salmonella. This accomplishment impacts Priority Objective 1.3.1.1 and 1. 3.1.2. 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? Our findings have been presented at scientific meetings and are likely the topic of research in other laboratories. Our development of biophotonics to track Salmonella will require an additional 1 year to impact animal agriculture. We have expanded this work to optimize the
technology by developing techniques to more easily allow light to pass through the skin of the pig. 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). Visualizing Salmonella. 2005. MicrobeWorld Radio Program. http://www. flpradio.com/microbeworld/2005-JUL-1-15.htm Who knows what lurks? Technology tracks Salmonella in live pigs. ARS Research Magazine. Special Food Safety Edition.
Impacts
(N/A)
Publications
- Rostagno, M.H., Hurd, H.S., Mckean, J.D. 2006. Prevalence of Salmonella enterica and S. enterica serotype typhimurium in swine at slaughter. Research Workers in Animal Diseases Conference Proceedings. p. 439-440.
- Rostagno, M.H., Wesley, I.V., Trampel, D.W., Hurd, H.S. 2006. Prevalence of Salmonella enterica in market-age turkeys. Research Workers in Animal Diseases Conference Proceedings. Paper No. 84.
- Rostagno, M.H., Hurd, H.S., Mckean, J.D. 2006. Resting pigs on transport trailers: potential intervention to reduce salmonella prevalence at slaughter? Research Workers in Animal Diseases Conference Proceedings. Paper No. 85.
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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? This project addresses the goals of the National Program 108, Food Safety (Animal and Plant Products). The presence of Salmonella is ubiquitous in livestock and livestock environments. This constant threat to food safety must be addressed. When swine are shipped to slaughter a high percentage of them start to shed Salmonella; this reaction is thought to be associated with the stress of transportation and mixing inherent in the process. As few as 4% may be positive for Salmonella on the farm but that percentage can increase up to 40% at the slaughter house. Our lab is investigating the role of stress hormones and their interactions with Salmonella in the living pig. Thus far our research shows that Salmonella can monitor their host's physiology and respond to the stress hormones it releases. This
ability may allow Salmonella to take advantage of the low immune function typically found in animals under stress. 2. List the milestones (indicators of progress) from your Project Plan. The milestones are as follows: 2003, Establish Salmonella strain, (Implementation & Analysis); 2004, Transport Model Intranasal Infection (Planning, Implementation, and Analysis); 2005, Transport Model with commingling (Planning, Implementation, and Analysis); 2006, Finalize all manuscripts from conducted research 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. Transport Model with commingling (Planning, Implementation, and Analysis) Milestone Substantially Met 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? Finalize all manuscripts
from conducted research. This project will terminate in FY 2006. Thus we expect to complete manuscript submissions during this time. We have initiated a new project proposal to continue our work on pre-harvest food safety. 4a What was the single most significant accomplishment this past year? The Stress Response Alters Salmonella Biology: In collaboration with researchers at NADC, we discovered that Salmonella can can grow in an acid environment after they have been exposed to the hormone norepinephrine. This is the first evidence to suggest that transportation stress in swine can actually enhance the virulence of salmonella infection, and thus increase the opportunity of post-harvest contamination. A complete understanding of this phenomenon will allow the development of strategies to decrease the amount of Salmonella found on the carcasses at slaughter. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. We found that
swine that are constant shedders of Salmonella have lower cortisol, a stress hormone, than those swine that do not constantly shed Salmonella. This is a surprising finding because it goes directly against the accepted belief that high stress levels allow bacteria to invade their host. The impact of this finding is that management strategies can be developed to temporarily elevate cortisol if it proves to be effective in preventing Salmonella transmission during transport. In addition, by identifying characteristics of individuals that pose a health risk we can develop management tools such as separation during transport to keep these individuals from passing Salmonella to healthy animals. This accomplishment impacts Priority Objective 1.3.1.1. and 1.3. 1.2. Our research has also found that Salmonella can monitor and respond to elevated concentrations of norepinephrine, a stress hormone, and more easily infect swine. This is the very first time that the ability of bacteria to
monitor their host has been shown to occur in swine. The impact of this finding is that it provides a mechanism by which Salmonella can become more virulent and it allows researchers a focus to develop strategies by which producers can lower the stress of mixing and transportation and thus reduce norepinephrine concentrations which will reduce Salmonella's ability to infect swine. This accomplishment impacts Priority Objective 1.3.1.1. and 1.3.1.2. Our lab has recently developed a tool, using biophotonics and Salmonella engineered to express luciferase (this enables the bacteria to emit light) , to enable visual monitoring of Salmonella in the living piglet. To date it is still unclear how Salmonella travel in the living pig and where it resides when it can not be detected in many of the pig's organs. Using biophotonics we will be able to see the answer many of the basic biological questions of Salmonella infection in addition to being able to follow the progress of infection in
the same living pig. To date pigs must be killed. The impact of this research is that it has provided $300, 000 in competitive funding from the USDA, NRI program and upon successful completion of this three year grant we will provide a method to monitor bacteria in the living pig in 'real time'. This diagnostic approach will be the next generation of tools to track Salmonella. This accomplishment impacts Priority Objective 1.3.1.1 and 1.3.1.2. 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? To date it has been too early for our findings to have been transferred to the industry. However, our findings have been presented at scientific meetings and are likely the topic of research in other laboratories. Our development of biophotonics to track
Salmonella will require an additional 2 years to impact animal agriculture.
Impacts
(N/A)
Publications
- Willard, S.T., Baily, R.H., Rybolt, M.L., Gandy, B.S., Ryan, P.L., Lay Jr., D.C. Real-Time monitoring of Salmonella in swine: Specificity and sensitivity of bacterial detection through the gastrointestinal tracts of juvenile and market weight pigs. Journal of Animal Science. Supplement. 2003.
- Lay Jr., D.C., Stabel, T.J., Toscano, M.J. Effect of mixing and transportation on behavior and cortisol response in relation to Salmonella infection in swine. Journal of Animal Science. 2003. v. 81(Suppl. 1): p. 323.
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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? The presence of Salmonella is ubiquitous in livestock and livestock environments. This constant threat to food safety must be addressed. When swine are shipped to slaughter a high percentage of them start to shed Salmonella; this reaction is thought to be associated with the stress of transportation and mixing inherent in the process. As few as 4% may be positive for Salmonella on the farm but that percentage can increase up to 40% at the slaughter house. Our lab is investigating the role of stress hormones and their interactions with Salmonella in the living pig. Thus far our research shows that Salmonella can monitor their host's physiology and respond to the stress hormones it releases. This ability may allow Salmonella to take advantage of the low immune function typically found in animals
under stress. 2. List the milestones (indicators of progress) from your Project Plan. The milestones are as follows: 2003, Establish Salmonella strain, (Implementation & Analysis); 2004, Transport Model Intranasal Infection (Planning, Implementation, and Analysis); 2005, Transport Model with commingling (Planning, Implementation, and Analysis; 2006, Finalize all manuscripts from conducted research 3. Milestones: A. List the milestones 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 or why not, and your plans to do so. Our milestone in 2004 was to develop a transportation model using intranasal infection of salmonella. During 2004 we conducted two experiments in which we infected swine with salmonella and transported them during Experiment 1, or did not transport them for Experiment 2. The reason for these comparative experiments was to
determine the role that stress of transportation can have on Salmonella infection. This year we have shown that Salmonella exposed to norepinephrine, a stress hormone, in vitro are more capable of colonizing the stomach of swine. This is significant because the stomach is a hostile environment in which Salmonella usually does not colonize, thus stress has the potential to alter the infection path of Salmonella. This is a major finding and one of the first in swine. A manuscript is in preparation on this research. During 2005 we wish to pursue the findings to determine how pigs transmit salmonella amongst themselves. However, currently we have 2 new vacancies in this area and will need to fill these prior to conducting this research. B. List the milestones that you expect to address over the next 3 years (FY2005, 2006, 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? In 2005, we hope to conduct research on the effects of co-mingling
during transport (Milesetone for 2005) but this may get moved to 2006, once new hires are established. During 2005, 2006, and 2007 we will be working to validate a novel method of tracking Salmonella using biophotonics. The development of this new method of tracking will allow us to more effectively identify how Salmonella is transmitted during transportation and mixing and it will provide a tool for other bacteriologist to answer questions that have been unattainable. The development of biophotonics as a tool in tracking Salmonella in the living pig will progress the research of Salmonellosis by 20 years. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004 (one per research project): In collaboration with researchers at NADC, we discovered that Salmonella can have an enhanced ability to colonize the gastrointestinal tract of swine after exposure to the stress hormone norepinephrine. This discovery is important
because it provides a mechanism to explain the fact that we find a low rate of animals testing positive at the farm and the high rate of positive swine at the slaughter plant. These data indicate that bacteria are able to monitor their host and take advantage of the host when it is most susceptible to infection. A complete understanding of this phenomenon will allow the development of strategies to decrease the amount of Salmonella found on the carcasses at slaughter. B. Other significant accomplishments(s), if any. None. 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. We found that swine that are constant shedders of Salmonella have lower cortisol, a stress hormone, than those swine that do not constantly shed Salmonella. This is a surprising finding because it goes directly against the accepted belief that high stress levels allow bacteria to
invade their host. The impact of this finding is that management strategies can be developed to temporarily elevate cortisol if it proves to be effective in preventing Salmonella transmission during transport. In addition, by identifying characteristics of individuals that pose a health risk we can develop management tools such as separation during transport to keep these individuals from passing Salmonella to healthy animals. This accomplishment impacts Priority Objective 1.3.1.1. and 1.3. 1.2. Our research has also found that Salmonella can monitor and respond to elevated concentrations of norepinephrine, a stress hormone, and more easily infect swine. This is the very first time that the ability of bacteria to monitor their host has been shown to occur in swine. The impact of this finding is that it provides a mechanism by which Salmonella can become more virulent and it allows researchers a focus to develop strategies by which producers can lower the stress of mixing and
transportation and thus reduce norepinephrine concentrations which will reduce Salmonella's ability to infect swine. This accomplishment impacts Priority Objective 1.3.1.1. and 1.3.1.2. Our lab has recently developed a tool, using biophotonics and Salmonella engineered to express luciferase (this enables the bacteria to emit light) , to enable visual monitoring of Salmonella in the living piglet. To date it is still unclear how Salmonella travel in the living pig and where it resides when it can not be detected in many of the pig's organs. Using biophotonics we will be able to see the answer many of the basic biological questions of Salmonella infection in addition to being able to follow the progress of infection in the same living pig. To date pigs must be killed. The impact of this research is that it has provided $300, 000 in competitive funding from the USDA, NRI program and upon successful completion of this three year grant we will provide a method to monitor bacteria in
the living pig in 'real time'. This diagnostic approach will be the next generation of tools to track Salmonella. This accomplishment impacts Priority Objective 1.3.1.1 and 1.3.1.2. 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? To date it has been to early for our findings to have been transferred to the industry. However, our findings have been presented at scientific meetings and are likely the topic of research in other laboratories. Our development of biophotonics to track Salmonella will require an additional 3 years to impact animal agriculture. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. 2004. Fire fly gene guides researchers on path to unlock
salmonella mystery. Indiana AgriNews.
Impacts
(N/A)
Publications
- Willard, S.T., Baily, R.H., Rybolt, M.L., Gandy, B.S., Ryan, P.L., Lay Jr., D.C. Real-Time monitoring of Salmonella in swine: Specificity and sensitivity of bacterial detection through the gastrointestinal tracts of juvenile and market weight pigs. Journal of Animal Science. Supplement. 2003.
- Lay Jr., D.C., Stabel, T.J., Toscano, M.J. Effect of mixing and transportation on behavior and cortisol response in relation to Salmonella infection in swine. Journal of Animal Science. 2003. v. 81(Suppl. 1): p. 323.
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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? The presence of Salmonella is ubiquitous in livestock and livestock environments. This constant threat to food safety must be addressed. Dr. Tom Stabel, USDA-ARS-NADC, Ames, IA is an expert in the biology of Salmonella infection in swine. By conducting collaborative research with Dr. Stabel, the USDA-ARS Livestock Behavior Research Unit will be able to tie together the biology of stress with the biology of pathogen infection to provide an understanding of the interaction between livestock stress and pathogen biology that will enable the threat of pre-harvest food safety to be minimized. An understanding of the interaction between livestock stress and pathogen biology will enable this threat to be minimized. 2. How serious is the problem? Why does it matter? Pathogens such as Salmonella are prevalent in livestock herds across the U.S. This reservoir of pathogens is a constant threat to the
health of the American public. This threat is compounded when animals are shipped to slaughter because the stress of handling and transportation increases the shedding of these organisms, thereby increasing the chance of contaminating meat products. 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%). The proposed project is a direct application of the National Program 108 Action Plan for food safety and animal health. Successful completion of the project will allow us "to develop effective interventions that can be easily instituted by the livestock producer." It will also produce information that is critical to understanding how stress effects pathogen biology and thus identification of strategies to decrease the load which livestock possess at the slaughter plant. 4. What were the most significant accomplishments this past year? A. Single Most
Significant Accomplishment during FY 2003. We conducted this research to determine if stress could reliably induce Salmonella shedding by swine. In Ames IA, at the NADC, in collaboration with Dr. Tom Stabel, we were able to experimentally infect swine with Salmonella in cause some individuals to shed Salmonella in response to mixing and transportation stress. Limited data indicate that those swine that consistently shed Salmonella also had low 'stress hormones' (cortisol). This is in direct contrast to what would be predicted and indicates that specific characteristics of some individual swine may pre- dispose them to be vulnerable to Salmonella infection. B. Other Significant Accomplishments if any during FY 2003. Laying hens molted by food withdrawal are more susceptible to pathogen infection, which become potential risks to human foodborne illness by laying Salmonella Enteritidis contaminated eggs, and has led to widespread public concern regarding to public health.We conducted
research to develop alternative methods for molting in the Livestock Behavior Research Unit. Our preliminary data showed that molting can be done without the need to withhold feed. The second experiment had been planned to replicate the first, and if it successful the new method can be adopted by the egg industry in developing new managerial practices to improve animal well-being in chickens and food safety in human. C. Significant Activities that support target populations: None. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. The proposed research is a continuation of project 3602-32000-004-00D, "Prevention of Pathogen Contamination in Food." In it, we used our stress model of transportation, and created the ability to cause swine to shed Salmonella. This will aid in our ability to identify characteristics of individuals that do shed compared to infected, but non-shedding herd mates. Understanding these critical
differences in individuals will allow interventions to be developed that will help ensure food safety. 6. What do you expect to accomplish, year by year, over the next 3 years? Year 1 and Year 2: We expect to be able to identify those major components of transportation and shedding that contribute to shedding of Salmonella. Year 3: We expect to have more clearly identified characteristics of animals that shed and have developed management strategies to help control and decrease the number of animals shedding Salmonella. We also expect to determine if specific characteristics of Salmonella are responsible for their ability to invade their host. 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? Within three years, we expect to develop techniques in
biophotonics that will enable other researchers, as well as ourselves to more fully understand the infective ability of Salmonella in swine.
Impacts
(N/A)
Publications
- Willard, S.T., Baily, R.H., Rybolt, M.L., Gandy, B.S., Ryan, P.L., Lay Jr., D.C. Real-Time monitoring of Salmonella in swine: Specificity and sensitivity of bacterial detection through the gastrointestinal tracts of juvenile and market weight pigs. Journal of Animal Science. Supplement. 2003.
- Lay Jr., D.C., Stabel, T.J., Toscano, M.J. Effect of mixing and transportation on behavior and cortisol response in relation to Salmonella infection in swine. Journal of Animal Science. 2003. v. 81(Suppl. 1): p. 323.
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