Cooperative State Research, Education, and Extension Service (CSREES) Reports
Termination Year 2006
ACCESSION NO: 0152013 SUBFILE: CRIS
PROJ NO: NC06184 AGENCY: CSREES NC.
PROJ TYPE: HATCH PROJ STATUS: REVISED
START: 01 OCT 2001 TERM: 30 SEP 2006 FY: 2001
INVESTIGATOR: ANDERSON, K. E.
PERFORMING INSTITUTION:
POULTRY SCIENCE
NORTH CAROLINA STATE UNIV
RALEIGH, NORTH CAROLINA 27695
MAXIMIZATION OF LAYING HEN PERFORMANCE, ECONOMIC RETURN, AND EGG QUALITY
OBJECTIVES: Determine methods to optimize the productive potential, economic return, and egg quality of commercial strains of egg-type chickens by manipulation of pullet and layer nutrition, physical environment, husbandry practices, induced molt programs and shell egg processing factors. 1) Determine the relationship between body size and age at sexual maturity on subsequent layer performance and egg mass produced. 2) Determine the effects and interactions of strain, house type, density, husbandry practices, and layer nutrition on production and economics. 3) Determine the effects and interactions of induced molt programs on production, behavior, and economics. 4) Determine the effects of current egg processing technology on the physical quality and microbiological safety of shell eggs. 5) Examine alternative manure disposal systems.
APPROACH: Commercial strains of white and brown egg-type chickens and the eggs that they produce will be utilized. Pullets will be reared in environmental control housing with various combinations of nutritional or management programs which affect the productivity of the layers at the Poultry Research Unit of the Piedmont Research Station at Salisbury, NC. The factors could include: light programs, dietary regimen, body weight program, molt techniques, husbandry practices, environment control programs, and egg handling programs. Pullets and layers will be managed as close to commercial industry standards as possible except when deviations are needed to accomplish the research objectives. Pullet body weights, feed consumption and mortality as well as hen feed consumption, egg production, egg quality and livability data will be collected on each flock. Productivity will be defined by various measurements of body weight, skeletal structure, feed consumption, immune competence, physiological health, and egg production criteria. The welfare status will be defined by the behavioral profile and hormonal response of the birds. Egg quality will be defined by the USDA egg quality standards, physical shell and membrane measurements, and through microbiological testing of the shell and contents. The potential of poultry manure as an organic fuel source will be evaluated from commercial sources and based upon governmental standards for emissions. The general experimental design will be a factorial arrangement which will allow for examination of multiple factors simultaneously.
NON-TECHNICAL SUMMARY: NA
PROGRESS: 2000/10 TO 2001/09
The Pullet Rearing report for the 34th North Carolina Layer performance and management test indicates that there is divergence in growth parameters of the different commercial white egg layer strains available. The first cycle report is under preparation and a significant comparison of molt programs has been completed. The second full trial using the avian cancer model has been initiated. The current study has been establish to evaluate a new therapeutic application of a cancer treatment drug. This study will evaluate the drugs cancer inhibitory effect.
IMPACT: 2000/10 TO 2001/09
The NCLP&MT reports are utilized nationwide in the evaluation of pullet rearing nutrition and programs on growth and development of commercial layer strains. These programs are being used as guidelines by the food service industry to satisfy the welfare standards they are imposing on their egg suppliers.
PUBLICATIONS: 2000/10 TO 2001/09
1. Jones, D.R., K.E. Anderson, and G.S. Davis, 2001. The effects of genetic selection on production parameters of Single Comb White Leghorn hens. Poultry Sci. 80:1139-1143.
2. Bell, D. D., P. H. Patterson, K. W. Koelkebeck, K. E. Anderson, M. J. Darre, J. B. Carey, D. R. Kuney, and G. Zeideler, 2001. Egg marketing in national supermarkets: Egg quality-Part 1. Poultry Sci. 80:383-389.
3. Patterson, P. H., K. W. Koelkebeck, D. D. Bell, J. B. Carey, K.E. Anderson, and M.J. Darre, 2001. Egg Marketing in National Supermarkets: Specialty eggs - Part 2. Poultry Sci. 80:390-395.
4. Koelkebeck, K. W., D. D. Bell, J. B. Carey, K. E. Anderson, and M. J. Darre, 2001. Egg Marketing in National Supermarkets: Products, Packaging, and Prices - Part 3. Poultry Sci. 80:396-400.
5. Davis, G. S., K. E. Anderson, and A. S. Carrol, 2000. The Effects of Long Term Caging and Molt of Single Comb White Leghorn Hens on Herterophil to Lymphocyte Ratios, Corticosterone and Thyroid Hormones. Poultry Sci. 79:514-518.
6. McAvoy, K.C., P. A. Curtis, K. M. Keener, K. E. Anderson, and D. E. Conner, 2001. Comparison of quality and functionality of traditionally and cryogenically cooled shell eggs. Poultry Sci. Suppl. 80:55. (abstract)
7. Hughes, L. A., K. E. Anderson, and P. A. Curtis, 2001. The effect of cryogenic cooling with carbon dioxide on the USDA grade and microbial load of shell eggs in the commercial setting. Poultry Sci. Suppl. 80:54. (abstract)
8. Anderson, K. E., B. W. Sheldon, and K. Richardson, 2001. Effect of Termin-8 anti-microbial preservative on the growth of commercial white and brown egg type laying strains. Poultry Sci. Suppl. 80:88. (abstract)
9. K.E. Anderson, G.S. Davis, and S. Hudson. 2001. Effect of dietary chitosan on production characteristics and egg proportions and quality from commercial white egg laying strains. Poultry Sci. Suppl. 80:88. (abstract)
10. Fosnaught, M. H., and K. E. Anderson, 2001. Interaction of increased Ca and P regimens on commercial strains of layers housed at various densities. Poultry Sci. Suppl. 80:89. (abstract)
11. Anderson, K. E., 2001. Report on Pullet Rearing Period: 34th North Carolina Layer Performance and Management Test. Vol. 34, No. 2, February 2001.
PROJ CONTACT:
Name: Anderson, K. E.
Phone: 919-515-5527
Fax: 919-515-7070
Email: ken_anderson@ncsu.edu
ACCESSION NO: 0404010 SUBFILE: CRIS
PROJ NO: 6612-32000-026-00D AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW
START: 31 JAN 2001 TERM: 30 JAN 2006 FY: 2001
INVESTIGATOR: GAST R K; MITCHELL B W; SWAYNE D E
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
ATHENS, GEORGIA 30613
DETECTION AND CONTROL OF SALMONELLA ENTERITIDIS IN POULTRY
OBJECTIVES: Develop improved methods for detecting Salmonella enteritidis (SE) infections in laying flocks and SE contamination in eggs. Characterize how, where, when, and in what numbers egg contamination by SE occurs. Reduce airborne dust and SE in poultry hatching cabinets and breeder houses using an electrostatic space charge system (ESCS), and determine the mechanism by which ESCS kills airborne and surface SE.
APPROACH: Chickens will be infected with a range of doses of various SE strains by oral inoculation, horizontal contact with infected birds or contaminated environments, or exposure to contaminated air sources. Eggs and egg fractions will be experimentally contaminated with SE. Samples will be subjected to bacteriological and serological testing to evaluate the efficacy of SE detection technologies and to characterize SE deposition in eggs. Optimum designs for use of the ESCS to reduce dust and pathogens including SE in poultry hatchers will be determined in cooperation with commercial poultry integrators and breeders. The mechanism by which the ESCS inactivates airborne and surface SE, and biofilms, and the associated LD50 dose, will be determined. Effectiveness of the ESCS for reducing egg contamination and airborne transmission of SE will be determined in a small breeder house. Previously was 6612-42000-035-00D (05/01).
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving it? The objective of this project is to develop improved methods for preventing, detecting, and controlling infections of chickens with Salmonella enteritidis (SE) and the associated production of SE contaminated eggs. Among the specific goals of the research are determining the processes and mechanisms by which SE infects chickens, spreads vertically and horizontally, and is deposited in eggs; characterizing the nature of SE contamination of eggs; developing more sensitive and specific diagnostic tests for identifying SE infections of chickens and for detecting SE contamination of eggs; and developing electrostatic space charging technology to diminish airborne dust and pathogens - including SE throughout - poultry hatching and housing facilities. 2. How serious is the problem? Why does it matter? The association between human illness caused by SE and the consumption of contaminated poultry products is an important international public health and economic problem. In recent years, SE has been among the Salmonella serotypes most often reported to cause human illness. Eggs have been the most frequently implicated sources of human SE infections in the United States. As food-borne transmission of SE threatens both the safety of consumers and the ability of poultry producers to market their products, the formulation and implementation of effective control strategies for reducing the incidence of SE infections in chickens has been identified as an urgent priority by both government and industry. 3. How does it relate to the National Program(s) and National Component(s)? National Program 108, Food Safety (100%) This research contributes to the Microbial Pathogens component of the Food Safety National Program by providing scientists (veterinary, food, and agricultural), regulatory agency officials, and the poultry industry with urgently needed tools to understand, prevent, detect, and control SE infections of chickens. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2001. Replicated trials conducted in a small treatment chamber showed that an electrostatic space charge system (ESCS) reduced biofilms developed on stainless steel from poultry carcass rinses by 97.3% in 2 hours and 99.8% in 3 hours. These results are comparable to those obtained by existing disinfection chemicals and indicate that the ESCS could potentially be used as a non-chemical treatment to reduce pathogens in biofilms which develop in poultry egg or meat processing areas. B. Other significant accomplishments. Because bacteria grow much more rapidly in egg yolk than in albumen, the location at which SE cells are deposited is critical for determining how quickly refrigeration must lower internal egg temperatures. After experimentally infecting groups of laying hens with SE strains of different phage types, we found SE more often in yolk than in albumen. Most contaminated eggs contained fewer than 1 SE cell per ml. These results suggest that refrigeration standards must address the possibility that SE may sometimes be deposited in yolk. When contaminants were experimentally introduced into egg contents, we observed rapid multiplication in yolk and extended persistence in albumen for a diverse assortment of SE strains of several phage types. This suggests that egg refrigeration standards may not need to address much potential variability in growth and survival characteristics between SE strains, but preventing disease transmission by eggs requires rapid refrigeration in combination with other control measures. When we compared the levels of serum antibodies induced in experimentally infected hens to the frequency at which they laid eggs contaminated by SE, we found that the relationship between these two parameters was not very consistent. Accordingly, although antibody tests are useful tools for preliminary screening of laying flocks to detect SE infection, the magnitude of the antibody response by individuals hens does not predict the overall risk of egg contamination associated with the flock. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This project provided the first definitive experimental documentation that hens systemically infected with SE could produce internally contaminated eggs. Scientists found that SE infections can be highly persistent in both chicks and hens and elicit long-lasting antibody titers. The project developed and assessed the sensitivity and predictive value of methods for detecting specific antibodies in serum and egg yolks from infected hens. The project developed effective and practical bacteriological methods for consistently detecting very small numbers of SE contaminants in eggs. Efficient killed vaccines were developed and evaluated for reducing the susceptibility of laying hens to SE infection. Studies assessed the relationship between phage type of SE isolates and their virulence, infectivity, and invasiveness in chicks. One scientist determined that air movement can mediate the horizontal transmission of SE infection. An ESCS was developed under a CRADA to reduce airborne dust and microorganisms in hatching cabinets. The ESCS has been shown to have effectiveness comparable to a 95% media filter for removing dust in laboratory experiments in hatching cabinets and transmission cabinets and equal or better effectiveness for removing airborne bacteria and Salmonella. The ESCS reduced airborne SE in an isolation room with caged layers approximately 95%. The kill rate of the ESCS on airborne and surface SE at close range has been shown to be 98% or more. The ESCS has been patented and an exclusive license for poultry applications has been approved with BioIon, Inc. to manufacture and distribute the system. 6. What do you expect to accomplish, year by year, over the next 3 years? During FY2002, we will establish how the growth patterns of SE inside eggs affects the development and application of faster methods for detecting contamination. During this year we also will study the effectiveness of the ESCS for pathogen reduction in egg rooms and upsize pathogen reduction hatchery studies to include followup of chicks from ionizer treated hatching cabinets to full-sized production houses. We will also determine the effectiveness of the ESCS for reducing pathogens and airborne disease transmission in a small scale breeder house and investigate the mechanism by which ionization inactivates SE. Effectiveness of the ESCS for reduction of biofilms in large open areas will be determined. For FY2003, we will evaluate how the patterns of deposition and multiplication of SE isolates in egg contents affects the probable effectiveness of proposed standards for egg refrigeration. We will also design an ESCS for full sized breeder houses and test effectiveness for pathogen and dust reduction. In FY2004, we will seek to develop and apply improved models and methods for producing experimental SE contamination of eggs, detecting SE contaminants in eggs, and detecting specific antibodies in infected chickens. We will also study the effectiveness of self-cleaning ESCS devices in poultry production areas and test the effectiveness of electrostatically-enhanced high volume air samplers for airborne pathogens. 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 & durability of the technology product? Completed commercial hatcher tests of ESCS with two commercial poultry production companies. CRADA partner initiated patent applications for the ESCS for several foreign countries. Participated in the 2001 International Poultry and Egg Exhibition January 17-19, 2000 where the ESCS and related posters were displayed in a commercial exhibit for hatchery equipment. Recent developments in methods for culturing eggs and other poultry samples to detect SE contaminants have been incorporated into the testing protocols of a national flock testing program under development by FDA. 8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below) Report, "Electrostatic space charge system for reducing airborne pathogens and dust", FLC Awards Page, http://www.fda.gov/ohrms/dockets/dailys/00/oct00/102700/ts00002.pdf, May 2001. Report, "Researchers honored for transferring technologies." ARS News and Information, http://www.ars.usda.gov/is/pr/2001/010501.htm, May 2001. Presentation, "Reducing airborne dust and bacteria in the hatchery." Georgia International Poultry Course, Athens, Georgia 2001. Presentation, "Pathogen reduction in poultry housing." Nebraska Poultry Industries, Columbus, Nebraska, 2001. Presentation, "Epidemiology and ecology of Salmonella enteritidis in poultry: general issues and research needs." Public Meeting on Salmonella Enteritidis Research, Sponsored by U. S. Food and Drug Administration, Atlanta, Georgia, 2000. Presentation, "The evolving application of egg culturing for detecting Salmonella enteritidis infection in laying flocks." Salmonella Isolation and Identification Workshop. Sponsored by Georgia Poultry Laboratory and National Poultry Improvement Plan, Oakwood, Georgia, 2001.
PUBLICATIONS: 2000/10 TO 2001/09
1. Gast, R.K., Petter, J.G., Holt, P.S. Frequency and location of Salmonella enteritidis contamination in eggs associated with various routes of experimental infection of laying hens. Program of the annual meeting of the American Association of Avian Pathologists. 2001. p. 21. Abstract.
2. Gast, R.K., Holt, P.S. Deposition of phage type 4 and 13a Salmonella enteritidis strains in the yolk and albumen of eggs laid by experimentally infected hens. Avian Diseases. 2000. v. 44. p.706-710.
3. Gast, R.K., Holt, P.S. Multiplication in egg yolk and survival in egg albumen of Salmonella enterica serotype Enteritidis strains of phage types 4, 8, 13a, and 14b. Journal of Food Protection. 2001. v. 64. p.865-868.
4. Gast, R.K., Holt, P.S. The relationship between the magnitude of the specific antibody response to experimental Salmonella enteritidis infection in laying hens and their production of contaminated eggs. Avian Diseases. 2001. v. 45. p.425-431.
5. Gast, R.K., Nadir, M.S., Jolly, M.E., Holt, P.S., Stone, H.D. Serological detection of experimental Salmonella enteritidis infections in laying hens by fluorescence polarization and enzyme immunoassay. Poultry Science. 2001. v. 80. p.1044. (Abstract).
6. Mitchell, B.W. Emerging technology electrostatic space charge system for pathogen reduction. Institute of Food Technology Abstract. http://ift.confex.com/ift/2001/techprogram/ 2001.
7. Mitchell, B.W., Holt, P.S., Seo, K.H. Effectiveness of electrostatic space charge for reducing dust in a caged layer room. Journal of Applied Poultry Research. 2000. v. 9. p.292-296.
8. Ricke, S.C., Birkhold, S.G., Gast, R.K. Eggs and egg products. Downes, F. P., Ito, K., editors. American Public Health Association, Washington, DC. Compendium of methods for the microbiological examination of foods, 4th edition. 2001. p.473-481.
9. Seo, K.H., Mitchell, B.W., Holt, P.S., Gast, R.K. Bactericidal effects of negative air ions on airborne and surface Salmonella enteritidis from an artificially generated aerosol. Journal of Food Protection. 2001. v. 64. p.113-116.
ACCESSION NO: 0188875 SUBFILE: CRIS
PROJ NO: PEN03841 AGENCY: CSREES PEN
PROJ TYPE: HATCH PROJ STATUS: NEW
START: 01 JUL 2001 TERM: 30 JUN 2006
INVESTIGATOR: Mashaly, M. M.
PERFORMING INSTITUTION: POULTRY SCIENCE PENNSYLVANIA STATE UNIVERSITY 208 MUELLER LABORATORY UNIVERSITY PARK, PENNSYLVANIA 16802
OPTIMIZING MANAGEMENT PRACTICES TO ENHANCE PRODUCTION PERFORMANCE AND IMMUNITY IN ORDER TO IMPROVE POULTRY HEALTH UNDER STRESSFUL CONDITIONS
OBJECTIVES: 1) To investigate the role of photoperiod and melatonin in alleviating the negative economic impact of different stressors on broiler performance, immunity and health. 2) To study the influence of induced molting in laying hens on subsequent effects of different stressors on production performance, immunity, health and hormone profiles. 3) To determine the mechanisms involved in the initiation of immune responses, including the roles of cytokines and hormones, under different stressful conditions.
APPROACH: In studying the effect of photoperiod on broiler performance under stressful conditions, CobbxCobb broiler chicks will be used and exposed to either continuous light (23L:1D) or intermittent light (1L:3D). Half of the birds in each group will be exposed to 35C (heat stress) from 21 to 42 days of age, the other half will be exposed to 24C (control). Production parameters such as body weight, feed consumption, feed conversion, and mortality will be measured weekly. Immune parameters such as cytokines and antibody production will be measured at 3 and 6 weeks of age. In studying the effect of melatonin on broiler performance under stressful conditions, again CobbxCobb broiler chicks will be used. They will be kept under 23L:1D and receive in the feed either 0, 20, 40, or 60 ppm melatonin. The same heat stress treatment mentioned above will be applied as well as production and immune parameters. In order to study the influence of induced molting in laying hens on subsequent effects of different stressors on production performance, immunity, health and hormone profiles, 72-week-old laying hens will be induced molted and following period of egg laying, they will be exposed to different kinds of stresses. Percent hen-day egg production, percentage hen-housed egg production, egg shell quality (specific gravity method), albumin quality, albumin height, Haugh units, feed consumption, feed per dozen eggs, and percent mortality will be measured throughout the experiment. Corticosterone T3,T4, and prolactin will be measured in the plasma at different times during the experiment. Antibody production and cell mediated immunity will also be measured. In order to study the mechanisms involved in the initiation of immune responses under different stressful conditions, Cornell K-strain Single Comb White Leghorn immature male chickens will be used. The birds will be housed in batteries with food and water available ad libitum, and exposed to 16L:8D per day. Birds will be exposed to different environmental and management stressors. Different high temperatures will be used as the environmental stressor and different population densities will be used as the management stressor. Different T-independent and T-dependent antigens will be used to stimulate humoral immunity in these chickens. Blood and spleen samples will be collected at different time periods post-injection. Specific cytokines will be measured in all samples collected as appropriate. Antibodies against the different antigens will be measured in all plasma samples collected. Percentage of different lymphocyte subpopulations will be measured in all blood and spleen samples.
NON-TECHNICAL SUMMARY: Certain management practices can be optimized to reduce stress and improve poultry health. The purpose of this study is to determine and apply the optimum standards of certain management practices under stressful conditions in order to improve poultry health and production.
PROJ CONTACT:
Name: Mashaly, M. M.
Phone: 814-863-0533
Fax: 814-865-5691
Email: xn9@psu.edu
ACCESSION NO: 0188709 SUBFILE: CRIS
PROJ NO: VA-135638 AGENCY: CSREES VA.
PROJ TYPE: HATCH PROJ STATUS: NEW
START: 01 JUL 2001 TERM: 30 JUN 2006 FY: 2001
INVESTIGATOR: MCELROY, A. P.
PERFORMING INSTITUTION:
ANIMAL POULTRY SCIENCES
VIRGINIA POLYTECHNIC INSTITUTE
BLACKSBURG, VIRGINIA 24061
MORPHOLOGICAL CHARACTERIZATION AND DIETARY CAPSAICIN POTENTIATION OF INTESTINAL IMMUNITY IN CHICKENS.
OBJECTIVES: Describe the response of mast cells and eosinophils in the innate intestinal inflammatory immune response to coccidia or Salmonella infection in commercial broilers. Characterize the non-specific or specific effector functions of mast cells and eosinophils in protective intestinal immune responses to secondary exposure to homologous challenges with coccidia species or Salmonella. Determine the role of mast cells or eosinophils in immunopathology of coccidia. Evaluate the effect of dietary capsaicin administration on intestinal immune responses and resistance to coccidia or Salmonella.
APPROACH: Naive or immunized commercial broiler chickens will be challenged with Salmonella or selected Eimeria species of coccidia. Intestinal tissue will be collected on selected days post-challenge for morphological evaluation and microbiologic culture or lesion scoring for Salmonella or Eimeria, respective to pathogen. Capsaicin will be included in commercial broiler diets for selected periods of time pre- or post-challenge. Intestinal tissues will be examined by light microscopy for alterations in structure or cellular influx and protection to infection by Salmonella or coccidia species will be evaluated by culture or lesion scoring.
NON-TECHNICAL SUMMARY: Although Salmonella and coccidia present problems to the commercial poultry industry from economic, food-borne illness, or bird productivity standpoints, relatively little is understood concerning the interactions between these pathogens and the host at the level of the intestinal immune response. The purpose of this research is to describe the effector functions of mast cells and eosinophils in innate intestinal immune responses to Salmonella and coccidia in broiler chickens.
PROGRESS: 2000/10 TO 2001/09
While live oocyst vaccination provides a viable alternative to anticoccidial usage and resulting drug-resistance in the commercial poultry industry, complete resistance to coccidia is difficult to achieve due to immunovariability between vaccine species and field strains. Immunovariability between coccidia species in vaccines and those found in poultry rearing facilities has emerged as a potential complication associated with vaccination. Additionally, there have been reports of coccidia-like impacts on performance with minimal appearance of coccidia lesions in the field environment. The purpose of the experiments conducted was to evaluate the host response to different field isolates of Eimeria acervulina (EA) by associating classical mucosal lesions with decreased body weight gain in infected broilers, to compare the host response of two commercial broiler breeds to the field isolates of EA, and to determine if other vectors (bacterial) may contribute to the a differential host response. Experiments were conducted comparing two isolates of EA, EA1 and EA2, obtained from commercial poultry rearing facilities. In three experiments, commercial broilers chicks were divided into control (non-challenged) and EA1 or EA2 challenged groups at 14 days of age. In all 3 experiments, EA1 resulted in significantly (P is less than 0.05) higher lesion scores than EA2, however weight gain of EA1 challenged birds was not significantly different from controls. EA2 challenged birds had significantly higher lesion scores than control birds in Expts 1 and 3, with no lesions characteristic of classical EA infection in Expt 2. EA2 resulted in significantly decreased weight gain as compared to EA1 or control in Expt 3. While EA1 resulted in classical EA lesions with no significant difference in weight gain, EA2 resulted in few classical lesions with significant depression of weight gain. Subjective observation of intestines from EA2 challenged birds was suggestive of a severe secretory intestinal response and weakened intestinal strength. In Expt 4, EA2 oocysts were cleaned with 5.25 percent sodium hypochlorite to evaluate the possibility of an external bacterial factor contributing to the observed detrimental affects in the presence of few lesions. Birds were challenged with bleached or non-bleached EA2. Although there was no significant difference in lesion scores between EA2 challenged groups, non-bleached EA2 resulted in significantly decreased weight gain as compared to bleached. These data are indicative of immunovariability between different isolates of the same coccidia species that result in differences in clinical lesions and performance. Furthermore, the results indicate differences in the host response that may contribute to the pathogenicity and suggest that standard coccidia lesion scoring procedures may not be applicable for evaluation of the severity of clinical infection with all Eimeria isolates.
IMPACT: 2000/10 TO 2001/09
These findings contribute to the understanding of the complex intestinal immune response to coccidia in commercial poultry and depict the diversity of the host response that may exist to different environmental isolates of the same coccidia species. Additionally, the findings suggest differential responsiveness exists in different breeds of commercial broilers. Continued description of the intestinal immune response to this pathogen will allow for more effective vaccination or control strategies.
PUBLICATIONS: 2000/10 TO 2001/09
Morris, B.C., H.D. Danforth, D.J. Caldwell, and A.P. McElroy, 2001. Differential Intestinal Response to Eimeria acervulina Challenge in Broiler Chickens. Poultry Sci. 80: Suppl. 1.
PROJ CONTACT:
Name: McElroy, A. P.
Phone: 540-231-8750
Fax: 540-231-3010
Email: amcelroy@vt.edu
ACCESSION NO: 0164533 SUBFILE: CRIS
PROJ NO: TEX08311 AGENCY: CSREES TEX
PROJ TYPE: HATCH PROJ STATUS: REVISED
START: 10 MAY 2001 TERM: 09 MAY 2006 FY: 2000
INVESTIGATOR: Ricke, S. C.
PERFORMING INSTITUTION:
POULTRY SCIENCE
TEXAS A&M UNIV
COLLEGE STATION, TEXAS 77843
FOODBORNE SALMONELLA SPP. REDUCTION IN POULTRY PRODUCTION
OBJECTIVES: A. Salmonella virulence expression in molted birds, objectives are 1) to determine if addition of dietary moderate zinc-low calcium will prevent colonization of Salmonella Enteritidis in the crop of chickens undergoing molting and what characteristics in the crop microenvironment are associated with this type of induced molt, and 2) to understand the potential role of that crop for Salmonella Enteritidis virulence expression while birds are undergoing molting and if key characteristics in the chicken crop microenvironment can be linked with limiting S. Enteritidis colonization and virulence expression. B. The role of methanogens in establishment of anaerobic microflora, the objectives are 1) to determine when methanogens become a part of the cecal microflora, what characteristics in the cecal microenvironment are associated with establishment of methanogens and enumerate the methanogen populations, and 2) to determine how stable the cecal methanogen population is by looking at whether feed deprivation alters methanogen activity while birds are undergoing fasting and if key characteristics in the chicken cecal microenvironment can be linked with decreased methanogen populations. The hypothesis is that the final stages of cecal obligate anaerobic microfloral development will coincide with the emergence of methanogens and their presence will signify completion of establishment of the dynamic complex anaerobic population characteristic of mature birds. In addition, we hypothesize that decreases in numbers of methanogens signifies the beginning of instability in the cecal anaerobic microflora.
APPROACH: A. Salmonella virulence expression in molted birds: the idea here is to compare the indigenous microbial response in crops of birds that have been molted either by feed deprivation or using the moderate Zn diet approach. The hypothesis is that feed intake will be retained in the birds molted via moderate Zn addition and the crop microflora will continue to be actively fermenting to the point of being inhibitory to Salmonella Enteritidis colonization and invasion. Based on our earlier work we hypothesize that the key indicators of an active fermentation in the crop are low pH and a high concentration of lactate. Therefore, in addition to enumerating Salmonella Enteritidis recovered after infection in the different treatment groups we plan to thoroughly examine the microenvironment of the crops from the birds in these different treatment groups. This will be done by quantitating fermentation products and enumeration of the crop Lactobacilli. Whether these key characteristics in the chicken crop microenvironment can be linked with limiting S. Enteritidis in vivo invasion will be determined by examining virulence activity of Salmonella Enteritidis (hilA-lacZY transcriptional operon fusion) with a gene fusion strain for in vitro assays of crop contents. B. The role of methanogens in establishment of anaerobic microflora: The overall experimental concept in this proposal is to determine when methanogens appear in the ceca as birds age on different diets and how stable the methanogen population is during drastic dietary changes. To accomplish this, methanogens will be enumerated from three different scenarios: 1) newly hatched chicks on different diets; 2) newly hatched chicks given a cecal transfer from adult birds and 3) feed deprived molted laying hens. These three scenarios represent the commercial settings where the cecal microflora are considered important in resisting colonization by foodborne pathogens, namely, young chicks with or without a probiotic and laying hens undergoing molting. In the proposed work we plan to thoroughly examine the microenvironment of the ceca in the three groups of birds. This will done by quantitating fermentation products (VFA, lactate, pH, and Eh) and in addition to quantifying methanogens, enumerate microbial components of the ceca including lactobacilli, bifidobacteria, Escherichia coli, total aerobic and anaerobic bacteria. The rationale for these choices is that aerobic bacteria and E. coli represent facultative populations that are generally early colonizers of the chick ceca, followed by lactobacilli and bifidobacteria and finally the strict anaerobes. These subgroups of organisms in addition to the fermentation products should give us a representative profile of the microbial population changes occurring in the ceca prior to and during the establishment of methanogens. Whether these characteristics in the chicken cecal microenvironment can be linked with establishment of methanogens will be the outcome of this component of the proposal.
NON-TECHNICAL SUMMARY: Specific research plans reflect an integrated approach for controlling Salmonella in the early stages of production of poultry. This project will provide A)ideas for management alternatives to reduce molting as a risk for S. Enteritidis contamination, and B) understanding of the role of methanogens in the development of the cecal microflora, the natural microflora being inhibitory to Salmonella.
PROGRESS: 2000/01 TO 2000/12
The incidence of foodborne disease is increasing yearly, despite the growing body of information regarding the most common foodborne pathogens. During its life cycle Salmonella spp. can encounter various environmental stress conditions which may have dramatic effects on their survival and virulence. One of the potential stress conditions that can be frequently encountered by foodborne pathogens is exposure to starvation stress. We have found that virulence expression of S. enteritidis and S. typhimurium to various stress conditions associated with food systems or host environments could be greatly increased by exposure to SCFA, and further enhanced by feed deprivation and the resulting microenvironment in the crop of molted laying hens. We continue to investigate the molecular response of Salmonella to gastrointestinal environmental conditions.
IMPACT: 2000/01 TO 2000/12
This research is expected to contribute to the understanding of virulence mechanisms of S. typhimurium and may provide a scientific basis for developing more optimal strategies to control the foodborne pathogen.
PUBLICATIONS: 2000/01 TO 2000/12
1. Durant, J.A., V.K. Lowry, D.J. Nisbet, L.H. Stanker, D.E. Corrier, and S.C. Ricke. 2000. Late logarithmic Salmonella typhimurium HEp-2 cell-association and invasion response to short chain volatile fatty acid addition. J. Food Safety 20: 1-11.
2. Durant, J.A., D.E. Corrier, and S.C. Ricke. 2000. Short-chain volatile fatty acids modulate the expression of the hilA and invF genes of Salmonella Typhimurium. J. Food Prot. 63: 573-578.
3. Durant, J.A., D.J. Nisbet, and S.C. Ricke. 2000. Response of selected poultry cecal probiotic bacteria and a primary poultry Salmonella typhimurium isolate grown with or without glucose in liquid batch culture. J. Environ. Sci. Health, B35: 503-516.
4. Durant, J.A., D.E. Corrier, L.H. Stanker, and S.C. Ricke. 2000. Expression of the hilA Salmonella typhimurium gene in a poultry S. enteritidis isolate in response to lactate and nutrients. J. Appl. Microbiol. 89: 63-69.
5. Durant, J.A., D.E. Corrier, L.H. Stanker, and S.C. Ricke. 2000. Salmonella enteritidis hilA gene fusion response after incubation in a spent media from either S. enteritidis or a probiotic Lactobacillus strain. J. Environ. Sci. Health, B35: 599-610.
6. Durant, J.A., V.K. Lowry, D.J. Nisbet, L.H. Stanker, D.E. Corrier, and S.C. Ricke. 2000. Short-chain fatty acids alter HEp-2 cell association and invasion by stationary growth phase Salmonella typhimurium. J. Food Sci.65: 1206-1209.
7. Kwon, Y.M., C.L. Woodward, S.D. Pillai, J. Pena, D.E. Corrier, J.A. Byrd, S.C. Ricke. 2000. Litter and aerosol sampling of chicken houses for rapid detection of Salmonella typhimurium using gene amplification. J. Industrial Microbiol. Biotech. 24: 379-382.
8. Kwon, Y.M., C.L. Woodward, D.E. Corrier, J.A. Byrd, S.D. Pillai, and S.C. Ricke. 2000. Recovery of a marker strain of Salmonella typhimurium in litter and aerosols from isolation rooms containing infected chickens. J. Environ. Sci. Health, B35: 517-525.
9. Kwon, Y.M. and S.C. Ricke. 2000. Efficient amplification of multiple transposon-flanking sequences. J. Microbiol. Methods 41: 195-199. 10. Kwon, Y.M., S.Y. Park, S.G. Birkhold, and S.C. Ricke. 2000. Induction of resistance of Salmonella typhimurium to environmental stresses by exposure to short-chain fatty acids. J. Food Sci. 65: 1037-1040 .
PROJ CONTACT:
Name: Ricke, S. C.
Phone: 979/862-1528
Fax: 979/845-1921
Email: sricke@poultry.tamu.edu
Termination Year 2003
ACCESSION NO: 0186918 SUBFILE: CRIS
PROJ NO: MD-VTMD-9130 AGENCY: CSREES MD.
PROJ TYPE: OTHER GRANTS PROJ STATUS: NEW
CONTRACT/GRANT/AGREEMENT NO: 00-51110-9739 PROPOSAL NO: 2000-05442
START: 15 SEP 2000 TERM: 14 SEP 2003 FY: 2001 GRANT YR: 2000
INVESTIGATOR: HECKERT, R. A.; LILLEHOJ, H.; BABU, U.; SONG, W.; RAYBOURNE,
R.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
UNIV OF MARYLAND
COLLEGE PARK, MARYLAND 20742
SALMONELLA IMMUNOBIOLOGY AND INTERVENTION STRATEGIES
OBJECTIVES: Our goals for this project are: Goal #1: Evaluate the host local immune responses to Salmonella infection and investigate the effects of various immunomodulating factors. Goal#2: Determine the effects of various vaccination and intervention strategies on the immune response and colonization, persistence and shedding of Salmonella in laying chickens. Goal #3: Examine the impact of forced molting on the immune status of commercial laying chickens.
APPROACH: The immune system is still the most powerful and effective system the host has in preventing and eliminating infections by microorganisms. Although SE infection may not result in serious illness in chickens, a carrier state may be established, suggesting that the immune system of the chicken is ineffective in completely preventing SE infection, invasion and shedding. We propose to systematically examine the immune system of the chicken before, during and after SE infection to gain a greater understanding of how the microorganism and the immune system interact. We will also examine mechanisms of altering this immune response in order to enhance the resistance to colonization, enhance the elimination of the microorganism and prevent shedding. Studies will not only take place under controlled laboratory conditions, but will also investigate the impact of the immune system on Salmonella infections of commercial laying chickens on farms. In recent years, SE has been the most commonly isolated Salmonella serotype in humans in the U.S.6 Between 1985 and 1991, eggs were the primary vehicle of SE infection in 82% of human food borne outbreaks.7 Control of SE in man will depend upon effective means of reducing SE in eggs. In order to effectively do this we must first understand the pathogenesis of Salmonella in poultry and the impact the immune system has on this infection. This could lead to strategies to modulate the immune system to better prevent and control Salmonella infections, thereby leading to effective pathogen reduction or elimination. We have established a consortium of researchers with expertise relevant to addressing the above research goals. This group will carry out a comprehensive evaluation of the role of the immune system in SE-elimination from various age groups of birds. In this proposal, we will investigate the potential of several intervention strategies for controlling SE infection in hens and determine their possible mechanisms of action. Additionally, the field component of this proposal is a unique feature, which has been identified as a key factor in the egg safety action plan (a federal initiative designed to reduce the threat of SE in shell eggs). Our proposed work includes examining the impact of forced molting on the immune system and SE in commercial laying hens. We have performed preliminary studies on various aspects of Salmonella-host interaction and have established the methodology necessary to carry out the investigations outlined in this proposal. This research should not only provide basic information on the pathogen-host cell interaction, but also give vital information on how the immune system of the chicken can be used to control or eliminate the spread of SE from animals to man.
NON-TECHNICAL SUMMARY: Salmonella enteritidis (SE) carried by chickens and shed into and onto shell eggs has become a major source of human intestinal infections. Despite a large amount of research, there are still no effective measures for preventing SE colonization, and we still lack an understanding of why Salmonella infected hens remain persistently infected. This study will investigate the immunobiology of Salmonella infections in laying chickens and develop novel immunomodulating strategies for the intervention of Salmonella colonization, persistence, shedding and egg contamination.
PROGRESS: 2001/01 TO 2001/12
In the last 20 years there has been an increase in human food-poisoning outbreaks attributable to Salmonella enteritidis (SE) in the United States. Epidemiological studies of this increase have indicated that grade A shell eggs are an important source of SE. Despite the tremendous efforts made by the poultry industry, no effective measures for elimination of SE colonization have been developed. The humoral immune responses after infection with SE have been extensively studied for diagnostic purposes. However, the fundamental mechanism of mucosal resistance to infection and clearance of SE from the gut has received scant attention. Protection from infection by SE through humoral mechanisms alone is unlikely, due to the organism being a facultative intracellular bacterium. There is enough evidence in various animal models that cell mediated immunity plays a major role in controlling Salmonella infection. In chickens, thus far, there is lack of detailed knowledge on the immune mechanisms involved in defense against Salmonella infection. Our studies thus far have supported earlier conclusions that natural infection produces weak and transient immune responses that are ineffective at preventing or clearing a Salmonella infection. We have found that immunization with a killed vaccine provides a better immune response than using a modified live vaccine. This immunity is better with respect to intestinal IgA (which is the main barrier to Salmonella colonization of the intestinal tract) and better in producing cellular immunity (that which is responsible for eliminating the infection). This is somewhat surprising, as traditional thinking indicated that killed vaccines were not effective at eliciting good cellular immunity. We are currently preparing several publications regarding these findings. A further goal in this research was to evaluate various methods of increasing the chick's natural resistance to infection. To this end we have been developing and evaluating various compounds that could be administered in ovo, to increase the innate immunity of the chick at hatch. One of the first substances we evaluated was a genetic adjuvant. This is a short stretch of nucleic acids containing many C and G bases in a particular motif. These sequences are bacterial in origin and have been shown in the mammalian system to up regulate the immune system. We have synthesized several of these sequences in various motifs and have shown that some of them are also powerful stimulants in activating avian cells. We have shown the CpG oligonucleotides to increase macrophage intracellular nitric oxide, increase expression of cell surface antigens and increase expression of IL-6 in vitro. Preliminary studies have also shown it to have similar effects when delivered in ovo. We are in the process of preparing a publication reporting this novel and important finding. As further goals in this project we are continuing to explore vaccination and other methods of increasing the immunity of the hatched chick, such that it is better able to resist Salmonella infection at hatch.
IMPACT: 2001/01 TO 2001/12
This research should not only provide basic information on the pathogen-host cell interaction, but also give vital information on how the immune system of the chicken can be used to control or eliminate the spread of Salmonella from animals to man.
PUBLICATIONS: 2001/01 TO 2001/12
No publications reported this period
PROJ CONTACT:
Name: Heckert, R. A.
Phone: 301-935-6083
Fax: 301-935-6079
Email: rh175@umail.umd.edu
ACCESSION NO: 0403095 SUBFILE: CRIS
PROJ NO: 3602-32000-003-02S AGENCY: ARS 3602
PROJ TYPE: USDA COOPERATIVE AGREEMENT PROJ STATUS: NEW
CONTRACT/GRANT/AGREEMENT NO: 58-3602-9-146
START: 15 SEP 1999 TERM: 14 SEP 2003 FY: 2001 GRANT YR: 1999
INVESTIGATOR: LAY JR D C; PAJOR E
PERFORMING INSTITUTION: ANIMAL SCIENCE PURDUE UNIVERSITY WEST LAFAYETTE, INDIANA 47907
ANIMAL WELFARE IN FOOD PRODUCING ANIMALS
OBJECTIVES: The objective of this cooperative agreement research project is to determine the effect of specific management practices on farm animal welfare and productivity in poultry, swine and dairy cattle.
APPROACH: The impact of specific management practices (induced-molting in poultry, housing environments in swine, and introductions to the milking parlor in dairy) on animal welfare will be scientifically quantified. Effects will be determined using ethological, physiological, immunological and neurological methodologies. Behavioral measures will be collected using live observation and advanced video/computer technology. Computer software will be used to collect and manipulate data in preparation for statistical analyses. Laboratory techniques will consist of immunological assays, high performance liquid chromatography, radioimmunoassays, immunohistochemistry and neuron mapping.
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving it? 2. How serious is the problem? Why does it matter? 3. How does it relate to the National Program(s) and National Component(s)? 4. What were the most significant accomplishments this past year? D. Progress Report This report documents research conducted under a specific cooperative agreement between ARS and Purdue University. Additional details of research can be found in the report for the parent CRIS 3602-33000-002- 00D Ethology of Food Producing Animals. A second postdoctoral associate has been hired under this agreement, bringing expertise in poultry behavior from Europe that is not available in the U.S. The use of gestation stalls is banned in Europe and questioned in the U.S. for animal welfare reasons. A project with Dr. Ed Pajor, Purdue University, comparing confinement in gestation stalls or access to group areas during gestation is under analysis. Gilt cortisol, immune function, and acuta phase response data is completed and will be presented at the 2002 National Animal Science Meeting. Gilt behavioral data and piglet immune function and behavioral data are being analyzed. This work will become part of a scientific data set that will be used to establish swine welfare standards by commercial users. With recent dairy expansions, the time that cattle are in holding areas waiting to enter the parlor has increased. A study in collaboration with Dr. Mike Schutz and Dr. Ed Pajor, Purdue University, investigated short term physiological and behavioral changes caused by the increased holding time (40 minutes or 2 hours). Data are being analyzed. This work will be useful to producers and extension agents during dairy expansions for advise on appropriate holding times prior to milking. A poultry behavior project in collaboration with Dr. Ed Pajor, Purdue University, that will investigate imprinting affects on chick behavior is underway. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. 6. What do you expect to accomplish, year by year, over the next 3 years? 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 & durability of the technology product? 8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below)
PUBLICATIONS: 2000/10 TO 2001/09
No publications reported this period.
ACCESSION NO: 0191950 SUBFILE: CRIS
PROJ NO: CALV-AH-198 AGENCY: CSREES CALV
PROJ TYPE: ANIMAL HEALTH PROJ STATUS: PENDING NEW
START: 01 NOV 2001 TERM: 31 OCT 2003
INVESTIGATOR: MCCOWAN, B.; JEFFREY, J.; CARDONA, C.
PERFORMING INSTITUTION:
POPULATION HEALTH & REPRODUCTION
UNIV OF CALIFORNIA (VET-MED)
DAVIS, CALIFORNIA 95616
THE EFFECTS OF INDUCED MOLTING AND CHICKEN WELL-BEING
OBJECTIVES: The objective of this research is (1) to examine differences in chicken behavior and physiology under conditions of no fast-induced molting, natural molting and fast-induced molting under control (no-molt) and experimental (molt) conditions using individual chickens as their own controls to assess the amount of pain, suffering and/or stress in chickens under various types of molting and (2) to develop protocols for inducing molt that reduce pain, suffering or stress in chickens if fast-induced molting is found to
comprise chicken well-being.
APPROACH: Eight groups of three chickens housed in standard cages at the VMTRC will be the subjects of this study. Four experimental groups will be subjected to fast-induced molting. Two experimental groups will be subjected to no fast-induced molting in which a low caloric diet will be given to the chickens. Two control groups will be unmanaged and allowed to undergo natural molting. During all experiments, feed and water consumption, egg production and physical appearance will be monitored on a per cage basis. Data will be collected daily to monitor au natural inanition and to follow the progress of the molt. Stage 1. Normative data on chicken behavior, both social and vocal interactions, will be collected on and analyzed from video and audio tape to develop a comprehensive ethogram for use in the subsequent stages. Dominance or pecking order will be determined for each group to account for its effects on differential rates of behaviors by each individual. These normative data are important because little is known on the types or rates of behaviors and vocalizations of chickens that are indicative of psychological or physiological pain, suffering or stress. Examples of possible types and changes in rates of behaviors that might indicate increased stress include aggressive pecking, cage (non-nutritive) pecking, stereotyped pacing, head flicking and alarm vocalizations. Stage 2. Chickens will be subjected to a housing and lighting program that typically would be used by the poultry industry during normal production. During this stage, behavioral and vocal data will be collected for one hour twice on three days per week on both the types and rates of aggressive and alarm behaviors and vocalizations. Stage 3. Chickens will be subjected to the different treatments and behavioral and vocal data will be collected as described above. Experimental groups will be molted by withdrawing feed, or by a low caloric diet feeding, under the housing and lighting conditions typical of an induced molting program used by the poultry industry. The control groups will be similarly subjected to the housing and lighting conditions but feed will not be withdrawn. For all groups, data will also be collected on body weight, feather status and body temperature during Stages II and III in order to monitor molting status. Blood samples will be drawn from subjects for subsequent hormonal analyses to measures cortisol levels, which is used as physiological indicator of chronic stress. Serum prolactin (PRL) levels will also be measured as a physiologic indicator of reproductive tract involution, which is an indicative of future reproductive performance and egg quality. Behavioral data will be analyzed from the videotapes using a focal animal and 10-second interval sampling regime. This regime will permit us to analyze both the types and rates of aggressive, alarm and other stress-related behaviors under control and experimental conditions with each chicken acting as her own control. Behavioral and physiological data will be statistically analyzed using mixed effects linear regression in S-Plus statistical software.
NON-TECHNICAL SUMMARY: There is deepening concern that the induced molting of egg-laying chickens is cruel. This management practice is widespread in the commercial industry of California and other states because it provides significant benefits to both the producer and the consumer, as well to the health of the chicken flock. The purpose of this study is to evaluate changes in the behavior and physiology of chickens under non-molt conditions and three types of molt conditions to assess the well-being of chickens subjected to induced molting.
PROJECT CONTACT:
Name: McCowan, B.
Phone: 559-688-1731
Fax: 559-686-4231
ACCESSION NO: 0181123 SUBFILE: CRIS
PROJ NO: NEB-13-146 AGENCY: CSREES NEB
PROJ TYPE: HATCH PROJ STATUS: NEW
START: 04 DEC 1998 TERM: 30 NOV 2003 FY: 2000
INVESTIGATOR: SCHEIDELER, S. E.
PERFORMING INSTITUTION:
ANIMAL SCIENCE
UNIVERSITY OF NEBRASKA
LINCOLN, NEBRASKA 68583
FACTORS AFFECTING CALCIUM UTILIZATION IN THE AVIAN AND EGG SHELL QUALITY
OBJECTIVES: Goal: To develop nutritional regimens that will improve eggshell quality in commercial egg flocks during the second cycle of egg production. 1. Test potential beneficial effects of new feed additives such as phytase enzymes or 25 OH cholecalciferol on eggshell quality specifically during late first cycle and during the second cycle of lay. 2. Measure potential effects of natural estrogen like compounds from lignans in flaxseed on calcium utilization during late lay in laying hens. 3. Investigate the effects of Zinc-source and level on carbonic anhydrase activity and subsequent effects on eggshell quality into late first cycle and second cycle hens.
APPROACH: Laying flocks will be maintained in the Poultry Research facilities at the University of Nebraska for the conduct of the planned research. Applied nutrition trials will be conducted with various age birds fed diets differing in nutrient content.
NON-TECHNICAL SUMMARY: This project address dietary modifications in laying hens to improve egg shell quality. Dietary modifications include adaptation of new technologies such as phytase enzyme, 25 dihydroxy Vitamin D, and trace mineral proteinates potential benefits to eggshell quality. The purpose of the project is to improve egg shell quality since 6-8gg loss occurs between the time of lay through processing and packaging to the consumer.
PROGRESS: 2000/10 TO 2001/09
A good deal of time was spent on the development of a novel monocalcium phosphorus feed ingredient utilizing eggshells as the calcium source for this product. This process was refined and a patent is pending for the process and product at UNL. Bioavailability studies in broiler chicks and laying hens were conducted indicating an availability of greater than 100% compared to dicalcium phosphorus. Further studies have also been conducted with hatchery waste to produce the reaction to produce monocalcium phosphorus. Studies were also conducted this year testing non-restrictive feeding programs for molting laying hens. Low salt, high fiber diets were fed to laying hens to induce molt and compared to typical feed molt feed restriction programs. Hens successfully went through their molt on the low sodium diets and then re-entered a 2nd cycle of egg production without total feed restriction.
IMPACT: 2000/10 TO 2001/09
The creation of a novel monocalcium phosphorus product from egg processing egg shell waste or hatchery waste has potentially tremendous positive benefits to the environment. The volume of this waste grows annually and disposal is difficult as fields become calcium saturated. Adding value to this waste to make it into a feed ingredient is economically and environmentally sound. The molting research helps address animal welfare concerns of consumers as the egg industry adopts the UEP Bird Welfare guidelines of non feed restrictive molts.
PUBLICATIONS: 2000/10 TO 2001/09
1. Scheideler, S.E., and P.R. Ferket. 2001. Phytase in broiler rations - Effects on carcass yields and incidence of tibial dyschondroplasia. J. Appl. Poultry Res. 9:469-475.
2. Puthpongsiriporn, U., S.E. Scheideler, J.L. Sell and M.M. Beck. 2001. Effects of vitamin E and C supplementation on performance and immune and antioxidant status of laying hens during heat stress. Poultry Science 80:1190-1200.
3. Jalal, M., and S.E. Scheideler. 2001. Effect of phytase supplementation on egg production parameters and amino acid digestibilities. Poultry Science 80:1463-1471.
4. Novak, C. and S.E. Scheideler. 2001. Long-term effects of feeding flaxseed based diets. 1. Egg production parameters, components and eggshell quality in 2 strains of laying hens.
5. Scheideler, S.E., N. Ward and M.Jalal, 2001. Effects of Roche Ronozyme CT on Hyline W-98 laying hen performance when fed low phosphorus diets. Poultry Science 80:478 (abstract).
6. Puthpongsiriporn, U. and S.E. Scheideler, 2001. Effects of ratios of dietary linoleic to linolenic acid on hen performance, mitogenic response and antibody production of White Leghorn hens against Newcastle disease vaccine. Poultry Science 80:169 (Abstract)
7. Scheideler, S.E.,M.A. Jalal and E. Pierson. 2001. Strain response of laying hens to varying dietary energy with and without enzyme. International Poultry Scientific Formum p. 20 (Abstract).
8. Ash, J.A., and S.E. Scheideler, 2001. Bioavailability assessment of eggshell derived monocalcium phosphate. International Poultry Scientific Forum p. 20 (Abstract).
PROJ CONTACT:
Name: Scheideler, S. E.
Phone: 402-472-6451
Fax: 402-472-6362
E-mail: Sscheideler1@unlinfo.unl.edu
Termination Year 2002
ACCESSION NO: 0177174 SUBFILE: CRIS
PROJ NO: IOWV-109-05-52 AGENCY: CSVM IOWV
PROJ TYPE: STATE PROJ STATUS: TERMINATED
START: 01 JUL 1997 TERM: 30 JUN 2002 FY: 1999
INVESTIGATOR: KRAMER, T.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
IOWA STATE UNIVERSITY
S. AND 16TH ELWOOD
AMES, IOWA 50011
LIVE VACCINE AGAINST EGG-TRANSMISSION OF SALMONELLA ENTERITIDIS
OBJECTIVES: To prevent spread of Salmonella between laying hens. To prevent food poisoning from Salmonella contaminated eggs.
APPROACH: Develop and test a live, heterophil attenuated Salmonella enteritidis vaccine. To determine safety of vaccine for mice as a model for human safety.
PROGRESS: 1997/07 TO 2002/06
Food poisoning caused by poultry meat and eggs contaminated with Salmonella enteritidis (SE) are the leading causes of food poisoning in the U.S. and in the developed world. We postulated that fecal shedding and egg transmission of SE can be reduced by vaccination with a live, attenuated SE vaccine. The goal of this project was to develop a safe and effective live Salmonella enteritidis vaccine as a safeguard against food poisoning of poultry origin. The objectives of the project were to: 1) improve the heterophil and adapted SE vaccine (HASE); 2) to test the improved HASE vaccine by long-term monitoring of chicken feces, egg shells and egg contents; 3) to assess cloacal vs. vertical (transovarian) SE contamination of eggs; and 4) to determine the virulence/avirulence of the HASE in mammalian animal models. Objectives 1-3 were fully met in 1997. Preliminary data were gathered on the mammalian safety of the HASE vaccine. Specific objectives for 1998-1999 research were: 1) Continue work on demonstration of human safety of HASE vaccine TK 605 using a mouse model; 2) Investigate cellular and molecular basis for mechanism of action of HASE TK 605 vaccine by probing into vaccine-heterophil interactions, by comparing phagocytic indices of SE and HASE at multiple adaptive steps by fluorescein activated cell sorting (FACS); 3) Expand the applicability of heterophil adaptation to other gram-negative bacteria of importance to the poultry industry. Scientific advances over the life of the project: 1) The vaccine adaptation steps were increased to 11. Fecal shedding of vaccine and of challenge were very satisfactorily reduced. The vaccine strain was given at a dose of 10(8)CFU for three consecutive days by gavage. It was shed by 2 hens on the 3rd day after vaccination, and by 1 hen on the 11th day after vaccination; 3) Fecal shedding of challenge ceased on the 9th day after challenge in the vaccinated group, and persisted for the 40-day duration after challenge in the challenge control group. 2) None of 525 eggs cultured after vaccination were infected. One egg isolation was made from 422 eggs in the vaccinated group after challenge. Of the 12 egg isolations in the challenge control group, 8 were egg shell (cloacal) isolates, and 4 were egg content isolates (transovarian). These limited data suggest the cloacal infection of eggs is the major risk factor. 3) A safe and effective live SE vaccine was developed in 1997. Many vaccines are effective in preventing disease, but few are also effective in preventing colonization and infection. The HASE TK 605 vaccine has met this stringent criterion. 4) It was shown that increasing the number of granulocyte (heterophil) passages from 6X to 11X has greatly improved the safety and effectiveness of the granulocyte adapted SE vaccine.
IMPACT: 1997/07 TO 2002/06
Salmonella enteritidis of poultry origin is the leading cause of foodborne illness in the U.S. and in the developed world. The HASE vaccine, developed earlier in this laboratory holds the promise of reduction (and perhaps elimination) of this source of foodborne disease. It is therefore important to know how this vaccine works.
PUBLICATIONS: 1997/07 TO 2002/06
1. Kramer, T., Reinke, C.R., and James, M. 1998. Reduction of fecal shedding and egg contamination of Salmonella enteritis by increasing the number of heterophil adaptations. Avian Dis 42:585:588.
2. Kramer, T.T. 1998. Effects of heterophil adaptation on Salmonella enteritidis fecal shedding and egg contamination. Avian Dis. 42:6-13.
PROJ CONTACT:
Name: Kramer, T. T.
Phone: 515-294-3090
Fax: 515-294-1401
Email: ttkramer@iastate.edu
ACCESSION NO: 0187249 SUBFILE: CRIS
PROJ NO: TEX08815 AGENCY: CSREES TEX
PROJ TYPE: NRI COMPETITIVE GRANT PROJ STATUS: NEW
CONTRACT/GRANT/AGREEMENT NO: 2001-35201-09946 PROPOSAL NO: 2000-02614
START: 15 NOV 2000 TERM: 30 NOV 2002 GRANT YR: 2001
INVESTIGATOR: RICKE, S. C.
PERFORMING INSTITUTION: POULTRY SCIENCE TEXAS A&M UNIV COLLEGE STATION, TEXAS 77843
MINIMIZING SALMONELLA enteriditis INVASION IN HENS DURING INDUCED MOLTING
OBJECTIVES: Determine if addition of dietary moderate zinc-low calcium will prevent colonization of Salmonella Enteritidis in the crop of chickens undergoing molting and what characteristics in the crop microenvironment are associated with this type of induced molt. Understand the potential role of the crop for Salmonella Enteritidis virulence expression while birds are undergoing molting and if key characteristics in the chicken crop microenvironment can be linked with limiting S. Enteritidis colonization and virulence expression.
APPROACH: The idea here is to compare the indigenous microbial response in crops of birds that have been molted either by feed deprivation or using a moderate Zn diet approach. The hypothesis is that feed intake will be retained in the birds molted via moderate Zn addition and the crop microflora will continue to be actively fermenting to the point of being inhibitory to Salmonella Enteritidis colonization and invasion. In addition to enumerating Salmonella Enteritidis recovered after infection in the different treatment groups we plan to quantitate fermentation products and enumerate crop lactobacilli. We will also examine virulence activity of Salmonella Enteritidis (hilA-lacZY transcriptional operon fusion) with a gene fusion strain for in vitro assays of crop contents.
NON-TECHNICAL SUMMARY: During the past 10-15 years, the number of cases of gastroenteritis due to Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) infections has greatly increased in the U.S. and Europe and by 1995, S. Enteritidis comprised 25% of all foodborne Salmonella isolates. Between 1985 and 1991, over 80% of S. Enteritidis infections in the U.S. were associated with table eggs and this may be linked to the specific stressful management practice of inducing a molt to stimulate multiple egg-laying cycles in hens. Feed withdrawal is the primary method used in the layer industry to induce molting. However, feed withdrawal dramatically enhances S. Enteritidis recovery from crops, increases invasion of organs in chickens and increases horizontal transfer in flocks. The poultry industry needs alternative molting procedures that do not require feed withdrawal but allow managers to keep the economic advantages of recycling laying hens by molting without causing a S. Enteritidis contamination problem. In this proposal we plan to determine whether molt induction diets will minimize S. Enteritidis and if key characteristics in the chicken crop microenvironment can be linked with limiting S. Enteritidis colonization and pathogenesis.
PROJ CONTACT:
Name: Ricke, S. C.
Phone: 979-862-1528
Fax: 979-845-1921
Email: sricke@poultry.tamu.edu
Termination Year 2001
ACCESSION NO: 0400561 SUBFILE: CRIS
PROJ NO: 6612-32000-017-00D AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: TERMINATED
START: 16 APR 1996 TERM: 30 JAN 2001 FY: 2001
INVESTIGATOR: GAST R K; PETTER J G; MITCHELL B W; SWAYNE D E
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
ATHENS, GEORGIA 30613
PATHOGENESIS, DETECTION, AND CONTROL OF SALMONELLA ENTERITIDIS AND OTHER SALMONELLAE IN CHICKENS
OBJECTIVES: Determine how Salmonella enteritidis (SE) spreads within and between poultry flocks and is deposited in eggs. Determine how phenotypic and genetic diversity influences invasion of host organs and egg contamination by SE. Identify environmental factors that influence the emergence of virulent forms of SE. Develop sensitive and specific diagnostic tests for SE in chickens and eggs. Develop effective SE vaccines. Develop methods to reduce surface and airborne dispersal of SE by dust reduction.
APPROACH: Chickens will be infected with SE by oral inoculation or exposure to environmental sources. Environmental factors and management practices that influence persistence and vertical or horizontal transmission of SE will be determined. Rapid and sensitive bacteriological and serological methods for detecting SE infection or contamination will be applied. Vaccine preparations will be tested for their efficacy. The pathogenic effects of SE phage types and strains will be compared. Chromosomal heterogeneity in virulence factor regulatory regions will be studied as it relates to egg contamination. The identity of protein and carbohydrate receptors that determine phage type will be established. Stages of the SE life cycle will be defined by coupling continuous cell culturing to analysis of cell surface protein and carbohydrate variability. Dust and bacterial counts will be measured in poultry production areas and techniques including ionization for reducing airborne dust and disease transmission will be evaluated. Athens, GA-SEPRL-main lab & bldg 3.11/4/99.
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving it? This project focuses on explaining the pathogenesis of Salmonella enteritidis (SE) infections in chickens and developing improved methods for prevention, detection, and control. Among the principal goals of the research are determining the processes and mechanisms by which SE infects chickens, spreads vertically and horizontally, and is deposited in eggs; assessing the influence of strain variations on the ability of SE to invade host organs and contaminate eggs; evaluating environmental factors that result in the emergence of virulent forms of SE from avirulent populations; developing more sensitive and specific diagnostic tests for identifying SE infections of chickens and for detecting SE contamination of eggs; developing and evaluating effective killed and live vaccines for controlling SE infections in chickens and associated egg contamination; and developing electrostatic space charging technology to diminish the airborne spread of SE throughout poultry hatching and housing facilities. 2. How serious is the problem? Why does it matter? The association between human illness caused by SE and the consumption of contaminated poultry products is an important international public health and economic problem. In recent years, SE has been among the Salmonella serotypes most often reported to cause human illness. Eggs have been the most frequently implicated sources of human SE infections in the United States. As food-borne transmission of SE threatens both the safety of consumers and the ability of poultry producers to market their products, the formulation and implementation of effective control strategies for reducing the incidence of SE infections in chickens has been identified as an urgent priority by both government and industry. 3. How does it relate to the National Program(s) and National Component(s)? National Program 108, Food Safety (100%) This research contributes to the Microbial Pathogens component of the Food Safety National Program by providing scientists (veterinary, food, and agricultural), regulatory agency officials, and the poultry industry with urgently needed tools to understand, detect, and control SE infections of chickens. 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY2000 year. No activity for this fiscal year. This project has been terminated and replaced by CRIS number 6612-32000-026. All activity is reported under that CRIS number. B. Other Significant Accomplishment(s), if any. No activity for this fiscal year. This project has been terminated and replaced by CRIS number 6612-32000-026. All activity is reported under that CRIS number. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This project provided the first definitive experimental documentation that hens systemically infected with SE could produce internally contaminated eggs. Scientists found that SE infections can be highly persistent in both chicks and hens and elicit long-lasting antibody titers. The project developed and assessed the sensitivity and predictive value of methods for detecting specific antibodies in serum and egg yolks from infected hens. The project developed effective and practical bacteriological methods for consistently detecting very small numbers of SE contaminants in eggs. Efficient killed vaccines were developed and evaluated for reducing the susceptibility of laying hens to SE infection. Studies assessed the relationship between phage type of SE isolates and their virulence, infectivity, and invasiveness in chicks. One scientist determined that air movement can mediate the horizontal transmission of SE infection. An ESCS was developed under a CRADA to reduce airborne dust and microorganisms in hatching cabinets. The ESCS has been shown to have effectiveness comparable to a 95% media filter for removing dust in laboratory experiments in hatching cabinets and transmission cabinets and equal or better effectiveness for removing airborne bacteria and Salmonella. The ESCS reduced airborne SE in an isolation room with caged layers approximately 95%. The kill rate of the ESCS on airborne and surface SE at close range has been shown to be 98% or more. The ESCS has been patented and an exclusive license for poultry applications has been approved with BioIon, Inc. to manufacture and distribute the system. 6. What do you expect to accomplish, year by year, over the next 3 years? No activity for this fiscal year. This project has been terminated and replaced by CRIS number 6612-32000-026. All activity is reported under that CRIS number. 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 & durability of the technology product? No activity for this fiscal year. This project has been terminated and replaced by CRIS number 6612-32000-026. All activity is reported under that CRIS number. 8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below) No activity for this fiscal year. This project has been terminated and replaced by CRIS number 6612-32000-026. All activity is reported under that CRIS number.
PUBLICATIONS: 2000/10 TO 2001/09
No activity for this fiscal year. This project has been terminated and replaced by CRIS number 6612-32000-026. All activity is reported under that CRIS number.
ACCESSION NO: 0400984 SUBFILE: CRIS
PROJ NO: 6612-42000-022-00D AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: TERMINATED
START: 01 DEC 1996 TERM: 30 JAN 2001 FY: 2001
INVESTIGATOR: HOLT P S; GAST R K; PETTER J G; SWAYNE D E; MITCHELL B W
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
ATHENS, GEORGIA 30613
EPIDEMIOLOGY AND ECOLOGY OF SALMONELLA ENTERITIDIS IN COMMERCIAL POULTRY FLOCKS
OBJECTIVES: Identify the sources of introduction of Salmonella enteritidis (SE) and other salmonellae into commercial poultry flocks and the reservoirs where they persist in the poultry housing environment. Determine how SE spreads within and between commercial poultry flocks. Identify unique characteristics of Salmonella isolates, strains, serotypes, and phage types and apply this information to determine the epidemiological relationships between isolates from different sources.
APPROACH: Samples taken from diverse locations in commercial poultry housing facilities, and from the birds themselves, will be tested for S.enteritidis and other enteroinvasive salmonellae. Different sampling and testing methods will be selected or developed and then evaluated for their effectiveness in detecting SE. Samples will be obtained from flocks of different ages and housed using different management systems. Experiment- ally infected chickens will be used to establish or verify fundamental aspects of the host-pathogen relationship between SE and poultry. Statistical modeling tools will be applied to establish the significance of and relationships between individual sources and reservoirs of salmonellae in commercial poultry flocks. Molecular analytical methods will be applied to characterize and differentiate Salmonella isolates.
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving it? Salmonella enteritidis (SE)is the causative agent of pandemic salmonellosis in people, which is most often acquired by consumption of contaminated eggs. The reservoir involved in transmission of SE to people is well identified, namely the egg-laying hen. It can be difficult to identify infected flocks, because they do not often become ill and because the organism can be difficult to detect in the hen-house environment even when the organs of birds are positive. It is also accepted that mice are important vectors for transmission of SE to hens. Thus, the basic biology of SE appears to have unique parameters as compared to what happens when other types of Salmonella are in the on-farm environment. This research program addresses how the biology and genetics of SE differs from that other Salmonella. 2. How serious is the problem? Why does it matter? Increased illness in people from SE has existed for at least 20 years and it is currently the most prevalent serotype causing salmonellosis in the world and the second most prevalent in the U.S. The concern is that the incidence of illness in the U.S. from SE has the potential to increase to levels seen in Western Europe and other countries. 3. How does it relate to the National Program(s) and National Component(s)? This projects is in the Microbial Pathogens component of National Program 108, Food Safety (100%). The research evaluates how the biology and genetics of SE can be used to halt its, growth and survival in chickens and in the hen house, because these links in the infection route to humans precede egg contamination. 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2001 year: SE had been thought to be a single clone with minimal difference between isolates from different geographic regions. This project studied surface carbohydrate variation of SE, especially from mice in chickens houses, and showed that SE has a significantly more variable cell surface than did Salmonella typhimurium, which is another important cause of human illness not associated with egg contamination. These findings supported the concept that the house mouse plays a unique role in the pandemic associated with SE. B. Other Significant Accomplishment(s), if any: The genetic basis for why SE is infectious was poorly understood. A series of infection studies were conducted that revealed that loss of a molecule called flagella enhances the ability of SE to be infectious when given by mouth. This information may indicate that vaccines directed against the flagella may prevent SE infections. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This research identified that it is possible to use the structure of the lipopolysaccharide molecule of SE and other Salmonella serotypes to conduct epidemiological investigations of how strain heterogeneity potentially contributes to emerging patterns of salmonellosis in people and animals. The outcome of this research is improved epidemiological methods for monitoring emergence of SE and other potential pathogens on-farm by increasing knowledge of how bacteria can change to cause sustained illness in people through the food supply 6. What do you expect to accomplish, year by year, over the next 3 years? This project was terminated. 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 & durability of the technology product? The science and technology of how to assess strain heterogeneity of SE has been transmitted to epidemiologists, infectious disease experts, and public health officials through publications broadly available in peer-reviewed journals. It is being made available to farmers and industry through vaccine studies funded by CRADAS that will deliver improved products to market. One aspect of the science has been made available to industry by patenting. The technology is durable, because the general knowledge about how to grow and analyze SE can be applied to other bacteria. 8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below) No activity during FY2001
PUBLICATIONS: 2000/10 TO 2001/09
Parker,C.T., Liebana,E., Henzler,D.J., Guard-Petter,J. Lipopolysaccharide O-chain microheterogeneity of Salmonella serotypes Enteritidis and Typhimurium. Environmental Microbiology. 2001. V.5.p.332-342.
ACCESSION NO: 0404088 SUBFILE: CRIS
PROJ NO: 6612-42000-036-00D AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW
START: 31 JAN 2001 TERM: 30 JUN 2001
INVESTIGATOR: HOLT P S; VACANT; SWAYNE D E
PERFORMING INSTITUTION: AGRICULTURAL RESEARCH SERVICE ATHENS, GEORGIA 30613
EVALUATION OF STRATEGIES FOR INDUCED MOLTING ON SALMONELLA ENTERITIDIS PROBLEMS IN LAYING FLOCKS
OBJECTIVES: Define the physiological, neuroendocrinological and psychological alterations that occur in the hen during feed withdrawal to induce molt vs. alternative molt procedures. Evaluate whether experimental observation of molt effects on Salmonella enteritidis infection also occur under commercial conditions. Evaluate whether alternative molt procedures can reduce or eliminate the Salmonella enteritidis problem normally observed during an experimental molt.
APPROACH: Examine Salmonella enteritidis situation in commercial operations at various times pre, during and post molt. Develop molting procedures alternative to feed withdrawal such as low energy feeds or hormone therapy which do not cause exacerbated Salmonella enteritidis infection. Examine the effects of molting on hormone levels in brain, serum, and gut and the impact of molting on intestinal integrity. Examine effect of molting on reproductive tract function and morphology.
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving it? Induced molting is an important management tool used by 70-80% of the layer industry to achieve a second egg lay from aging hens (an estimated 168-192 million hens are molted annually). Previous experimental studies showed that molting via long term feed removal dramatically increased problems with Salmonella enterica serotype Enteritidis (SE). Because of the studies, molting has come under increased scrutiny by federal regulatory agencies and animal welfare groups as a potential food safety issue. Information is still too limited to determine the legitimacy of such claims and the current project will provide additional information by determining whether molting in a commercial setting similarly increases SE problems, examining alternative molting procedures with respect to their effects on SE infection, studying intervention strategies such as vaccination to ameliorate the problem, investigating in detail the degree of stress in birds subjected to molt and whether alternative molt procedures are less stressful, and studying the physiological effects of molting to determine potential causes of the exacerbated infection observed in the hens during molt. 2. How serious is the problem? Why does it matter? Eggs produced by molted hens may be a food safety risk to the consuming public. It is estimated that fully a third of the profits derived from a flock during its life span comes from molted hens. The producer could lose this very important management tool unless studies are conducted to assess the problem, develop intervention strategies to reduce the problem, and generate new methods to recycle flocks. 3. How does it relate to the National Program(s) and National Component(s)? This project is in the National Programs 108 on Food Safety (75%) and 103 on Animal Health (25%). In Food Safety, developing alternatives to molting of laying hens that will prolong their productive egg laying and not be conducive to SE infection supports the Microbial Pathogens Component. In addition, the project's objectives and goals are aligned with the President's Council on Food Safety 1999 Action Plan document that directs further research to provide data on molting to federal action agencies that will support regulation of industry practices based on logical, science- based information. In Animal Health, the stress component of the studies addresses Disease Control Strategies and Host/Pathogen Interactions components. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2001? This was a temporary bridging CRIS, please see progress reported under project 6612-32000-031-00D. NPS changed the project number. B. Other significant accomplishment(s),if any. Please see accomplishments listed under project 6612-32000-031-00D. C. Significant accomplishments/activities that support special target populations. Please see accomplishments/activities noted under project 6612-32000-031-00D. D. Progress Report. This was a temporary 3 month bridging CRIS for the final CRIS number 6612-32000-031-00D. All progress is reported under 6612-32000-031-00D. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This was a 3 month temporary bridging CRIS. Previous accomplishments of the CRIS dealing with molting include - 1) molting depressed cell-mediated immunity in hens; 2) molting increased the severity of SE infections in hens; 3) molting made the hens 1,000-10,000-fold more susceptible to an SE infection which resulted in the rapid transmission SE to uninfected, but exposed, molted hens. As a result of these studies, the USDA/FSIS Salmonella enteritidis Risk Assessment Team, in their 1998 Risk Assessment document, placed molting as a central flock variable for increasing the risk of eggs produced which contain SE. 4) developed alternative molting procedure using wheat middlings which dramatically decreased SE levels in hens. As a result of these studies, researchers have moved to field studies to examine the economic utility of using wheat middlings in a commercial situation. 6. What do you expect to accomplish, year by year, over the next 3 years? This project was a 3 month temporary bridging project and is terminated. 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 & durability of the technology product? This project number was changed by NPS to 6612-32000-031-00D. Project now terminated. 8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below) See project 6612-32000-031-00D.
PUBLICATIONS: 2000/10 TO 2001/09
No publications reported this period.
ACCESSION NO: 0178904 SUBFILE: CRIS
PROJ NO: IOWV-405-25-09 AGENCY: CSVM IOWV
PROJ TYPE: STATE PROJ STATUS: TERMINATED
START: 09 MAR 1998 TERM: 31 MAR 2001 FY: 2000
INVESTIGATOR: KRAMER, T. T.; HARRIS, R. S.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
IOWA STATE UNIVERSITY
S. AND 16TH ELWOOD
AMES, IOWA 50011
A LIVE SALMONELLA ENTERITIDIS (SE) VACCINE AGAINST EGG TRANSMITTED SALMONELLOSIS
OBJECTIVES: Salmonella enteritidis (SE) is the most common source of food poisoning worldwide. The primary source of SE contaminated food is the chicken egg. The objective of this research is to validate the safety for humans, and to explore the marketability of a live, heterophil attenuated Salmonella enteritidis (HASE) vaccine for the prevention of egg-transmitted salmonellosis.
APPROACH: Selection of live vaccine candidates by heterophil adaptation.
PROGRESS: 1998/03 TO 2001/03
Fecal shedding and egg transmission of a heterophil attenuated, vaccine candidate strain of Salmonella enteritidis (SE) was evaluated in laying hens. Fecal shedding and egg transmission of vaccine and challenge SE were used to assess, safety and efficacy of the heterophil attenuated SE. Fecal shedding was very significantly reduced in vaccinated hens by comparison to challenge controls (p < 0.001). None of 1,019 eggs cultured from vaccinated and challenged hens were infected with SE. Twelve of 479 (2.5%) of control eggs were infected
IMPACT: 1998/03 TO 2001/03
Further evidence was obtained for the suitability of heterophil adapted Salmonella enteritidis as a live attenuated vaccine against salmonellosis of poultry.
PUBLICATIONS: 1998/03 TO 2001/03
No publications reported this period
PROJ CONTACT:
Name: Kramer, T. T.
Phone: 515-294-3090
Fax: 515-294-1401
Email: ttkramer@iastate.edu
ACCESSION NO: 0178167 SUBFILE: CRIS
PROJ NO: IOWV-416-23-73 AGENCY: CSREES IOWV
PROJ TYPE: NRI COMPETITIVE GRANT PROJ STATUS: TERMINATED
CONTRACT/GRANT/AGREEMENT NO: 97-35201-4608
START: 01 SEP 1997 TERM: 31 AUG 2001 FY: 2000 GRANT YR: 1997
INVESTIGATOR: MINION, F. C.; TRAMPEL, D. W.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
IOWA STATE UNIVERSITY
S. AND 16TH ELWOOD
AMES, IOWA 50011
SALMONELLA ENTERITIDIS HETEROPHIL RESISTANCE
OBJECTIVES: 9702755. Obtain a better understanding of Salmonella enteritidis-chicken interactions at the molecular level. This will be accomplished using unique mutants that are resistant to chicken heterophil killing and by completing the following specific aims: 1)We will complete the assessment of the SE mutants in chicken in order to assess their pattern of colonization of infected tissues and affinity for egg laying tissues; 2) Genes involved in immune cell resistance will be identified by screening complemented strains in cell cultures; 3) Mutations will be constructed in these genes in the wild type strain in order to confirm their role in immune cell resistance; 4) Mutants will be assessed for virulence and egg transmissibility in chickens. For unknown reasons, immune cell resistance is directly linked to lowered virulence in SE. These studies will shed light on the possible mechanisms involved and add to our understanding of salmonella pathogenesis.
APPROACH: First develop mutant strains expressing green fluorescent protein to enhance our ability to follow S. enteritidis in host tissues and eggs. These strains will then be used to infect chickens and bacteriological and histopathological studies undertaken. Cosmid libraries of wild type S. enteritidis will be constructed and used to complement the mutant strains. Complementing clones with wild type phenotype will then be analyzed by subcloning and transposon mutagenesis, and the genes involves in heterophil resistance (htr) sequenced and analyzed. Knockout mutations in putative htr genes will be constructed in wild type strains, and the resulting strains analyzed for sensitivity in heterophil cultures.
PROGRESS: 1997/09 TO 2001/08
The overall goal of this proposal was to determine the genetic basis for heterophile resistance of a Salmonella enteritidis (SE) mutant (TK605) and to characterize this mutant in terms of its persistence and vaccine potential. The latter aim was accomplished in the first two years of the grant. A third aim was to develop green fluorescent protein (GFP) producing strains of SE strain TK474 and follow their transmission through the chicken to learn more about the cell types and tissue locations for SE during persistence. After integrating the GFPuv structural gene downstream of the lac promoter in the chromosome, colonies were unexpectedly fluorescently unstable in their fluorescence after in vitro passage. We then obtained another derivative of GFP on a R6 vector used in Salmonella typhimurium studies, which was stable in SE after prolonged passage in vitro. Studies of this construct in SE in mature hens showed that the numbers of fluorescent salmonella in tissues was low, making it difficult to follow the organism in specific tissues, and we did not pursue this avenue further. To determine the genetic basis for heterophile adaption, we sought to complement the mutations in SE strain TK605 using cosmid based approaches. The differences in survival between the mutant and wild type strains in chicken heterophile cultures, however, were not great enough to allow direct selection for a complemented mutant strain. Instead, we focused the last 18 months on invasion studies and signature tagged mutagenesis (STM) to identify genes needed for SE cell invasion with the hypothesis that invasion is critical for heterophile survival, tissue dissemination and persistence in the reproductive tract. STM invasion negative mutants were identified using Vero cell cultures. Assays using MTT or XTT to quantify the number of Salmonella present after lysis of the Vero cells had high background and variability. Invasion was then assessed by a growth/no-growth assay on brilliant green agar plates containing kanamycin, which led to the identification of 46 invasion-negative mutants. These mutants were then tested in a day-old chick model for virulence. The livers of all birds were sampled for Salmonella mutants using brilliant green agar plates with kanamycin. Virulence varied among the invasion-negative mutants with some low pathogenicity and some retaining high virulence, rapidly killing all chicks in a group. Further, STM experiments were also performed, and the mutants displayed variability in their ability to persist in chickens in comparison to wild type salmonella. Identification of the STM mini-transposon insertion site by DNA sequencing is currently in progress. Direct cloning of the transposon and flanking sequences, and direct sequencing of chromosomal DNA had little success. The most successful method for obtaining junction site sequences was using inverse PCR. The insertion site for eleven of the forty-six mutants has been identified. At the conclusion of this stage, we will have identified different loci in SE involved in Vero cell invasion and correlated that phenotype with persistence in mature chickens and virulence in a day-old chick model.
IMPACT: 1997/09 TO 2001/08
Salmonella enteritidis remains an important world-wide food safety pathogen. Identification of genes involved in tissue invasion and the correlation of specific genes with virulence and persistence in chickens will provide important information needed for a better understanding of the pathogenic mechanisms of this organism. These genes might also provide therapeutic targets for development of better intervention strategies both at the pre-harvest and post-harvest stages.
PUBLICATIONS: 1997/09 TO 2001/08
1. Kramer, T. T. 1998. Effects of heterophil adaption on Salmonella enteritidis fecal shedding and egg contamination. Avian Dis. 42:6-13.
2. Kramer, T.T., C. R. Reinke, and M. James. 1998. Reduction of fecal shedding and egg contamination of Salmonella enteritidis by increasing the number of heterophil adaptions. Avian Dis. 42:585-588.
3. Kramer, T.T. and J. Vote. 2000. Granulocyte selected live Salmonella enteritidis vaccine is species specific. Vaccine 18:2239-2243.
PROJ CONTACT:
Name: Minion, F. C.
Phone: 515-294-6347
Fax: 515-294-1401
Email: fcminion@iastate.edu
ACCESSION NO: 0400221 SUBFILE: CRIS
PROJ NO: 6612-32000-019-00D AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW
START: 09 APR 1996 TERM: 08 APR 2001 FY: 2000
INVESTIGATOR: TUMPEY T; MITCHELL B W; HOLT P S; SWAYNE D E
PERFORMING INSTITUTION: AGRICULTURAL RESEARCH SERVICE ATHENS, GEORGIA 30613
STIMULATION OF MUCOSAL IMMUNITY IN CHICKENS TO PROTECT AGAINST ENTERIC AND RESPIRATORY PATHOGENS
OBJECTIVES: Examine the development of local humoral immune response at mucosal surfaces in chickens and compare this response with systemic immunity. Develop vaccines for mucosal immunity against intestinal and respiratory pathogens in poultry and diagnostic tests that will predict effectiveness. Determine the mechanisms for generation of airborne pathogens. Develop controls to improve poultry health and enhance mucosal vaccine effectiveness by reducing airborne pathgens and dust.
APPROACH: Birds will be orally infected with salmonella enteritidis (SE) and serum and intestinal anti-SE antibody levels will be ascertained over time. The birds will be re-infected to determine the development of serum and intestinal immunological memory. Immune recognition of different components of SE in serum and the intestinal tract will be compared. The protective role of serum and mucosal antibodies will be ascertained by passive administration of antibodies to naive birds and following the progression of the infection. The development of immunity in the intestinal tract will be delineated by immunoassay of intestinal contents and elispot analysis of purified lamina propria lymphocytes. Dust and bacterial counts will be measured in hatching cabinets and other poultry production areas. Dust reduction techniques studied will include lowering air velocity and using an electrostatic space charge with a grounded collection system. Experiments will be conducted to characterize airborne transmission of SE and to explore treatments for reducing it.
PROGRESS: 1999/01 TO 1999/09
1. What major problem or issue is being resolved and how are you resolving it? Pathogens such as Salmonella enteritidis (SE), avian influenza virus, and Newcastle disease virus initiate their infections at mucosal surfaces such as the intestinal or respiratory tract. Procedures which protect these sites from infection, either by inducing active immunity or by reducing the challenge dose reaching the animal would significantly reduce the morbidity, mortality, and egg contamination resulting from infection. Vaccination regimens against these infections generally involve parenteral injections which induce a good systemic immune response. While this regimen does evoke a certain degree of protection at mucosal surfaces, it is not complete - a certain percentage of the birds remain infected and continue to shed organism into the environment and, in the case of SE, produce eggs contaminated with the organism. This scenario is observed regularly in the field and, as a result, vaccination against avian influenza virus is not performed in chickens in this country and only moderate vaccination against SE occurs. Recent serious outbreaks of highly pathogenic avian influenza in poultry in Mexico and in Hong Kong underscore the need for improved vaccination regimens to combat this problem organism. The continuing problem of SE infections in humans traced to contaminated eggs and the limited use by industry of currently available vaccines indicates the need for newer and better immunization methods to prevent infections. If immunity were to be elicited via mucosal vaccination at the site where infection was initiated, there would be a greater chance of stopping the organism before it has a chance to become firmly established within the tissue. Our laboratory is taking a multiple-prong attack at this problem by examining development of immunity at mucosal surfaces following infection and comparing this response with that in serum. We are also developing vaccination regimens which activate protective immunity at these surfaces. A third area of research is the use of procedures to reduce the levels of the pathogen in the air environment and therefore reduce the challenge dose reaching the bird. 2. How serious is the problem? Why does it matter? With highly pathogenic avian influenza virus, the situation is grave and the primary remedy is massive flock slaughter and quarantine of poultry products leaving the area, both of which put a severe financial burden on the producer, the region, and the industry as a whole. For SE, the situation is very serious. This organism is the leading Salmonella serovar causing human food borne infections in the U.S. and, while the number of outbreaks of food borne infections by this organism is decreasing, the number of sporadic cases continues to increase. The significance of this is two-fold: 1) a large number of individuals are becoming ill, and some die, due to consumption of eggs contaminated with this organism; 2) this has dramatic repercussions on the egg industry due to reduced egg consumption and the involvement of regulatory agencies in diverting eggs to pasteurization. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? National Program 103, Animal Health (75%) National Program 108, Animal Health (25%) Because of the disastrous nature of poultry infections by avian influenza virus and Newcastle disease virus, regimens which prevent the initiation and dissemination of the organisms within flocks fit well within the National Program animal health mandate. Similarly, food safety is a major component of the National Program agenda and intervention strategies which help prevent the dissemination of a human disease organism within flocks and block the entry of the organism into the human food chain are important areas of investigation. 4. What were the most significant accomplishments this past year? Studies were conducted with an electrostatic space charge system in a caged layer room to determine dust reduction capabilities and potential of the system for reducing airborne Salmonella enteritidis. Duplicated experiments in a caged layer room with artificially and naturally generated dust indicated that electrostatic space charge system treatment reduced dust concentration by 52-91%. Reductions in dust level of 50% reduce airborne bacterial counts by 100 fold or more. A prototype electrostatic space charge system that included an automatic rinse system was developed and installed in several commercial chicken hatcheries. The system reduced dust by 82% and Salmonella positive cabinets by as much as 83%. Hatchability with the system improved an average of 2.7%. The electrostatic space charge system will decrease transmission of disease-causing organisms and improve food safety. One-2 day-old chicks were unable to mount a significant antibody response in either the serum or intestinal tract. In adult chickens, a strong systemic and intestinal response to SE occurred within a week post challenge and the chickens cleared the infection within several weeks. These results demonstrated that the age when birds are exposed to infection is a very important variable with regards to their ability to respond to challenge and also possibly to vaccination. An ELISPOT assay was developed to detect specific antibody producing cells in the spleen and cecal tonsil in birds either infected or vaccinated against SE. This assay will assess the role of different intestinal tissues in the elicitation of an intestinal immune response. Determined that certain compounds added to Salmonella enteritidis killed vaccine emulsions resulted in an enhanced serum and mucosal antibody response to the SE immunogen. This will improve vaccines for use in SE control. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This a new CRIS, in its third year, so a large proportion of the information was presented in Question 4. However, we found that chicks infected with SE at 1-2 days post hatch cannot clear the infection and remain persistently infected for over 20 weeks. Serum and mucosal responses in these birds were severely depressed, with many birds not having any detectable SE-specific antibodies. When these individuals were vaccinated or re-infected, they were only minimally able to respond immunologically to the new stimulus, indicating that very little immunological memory occurred. These studies point to the hatchery as a very important area to prevent SE infection since challenge at this age can have serious repercussions on the health of the chicks. In the previous CRIS we examined the effect of stress on immunity in poultry and there were numerous accomplishments. We found that induced molting, a procedure used by 70% of the layer industry to achieve a second egg lay from aging flocks (affecting approximately 140-166 million hens annually) depressed the immunity in the birds and substantially increased the severity of, susceptibility to, and transmission of SE infections. Airborne transmission of SE, not normally a route of spread of SE, was also observed in molted hens indicating that the procedure could substantially affect the spread of the organism through a flock. As a result of this work, molting was identified by USDA/APHIS as a major risk factor for the production of eggs contaminated with SE. 6. What do you expect to accomplish, year by year, over the next 3 years? Develop ELISPOT assays to further delineate the immune response in various locations in the intestinal tract to identify the most important areas for targeting vaccines. Determine whether the cecal tonsil is an important organ for the development of an intestinal immune response. Evaluate more compounds with respect to their adjuvanticity in the intestinal tract and examine whether these vaccines can be adapted for use in ovo vaccination protocols. 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 & durability of the technology product? A patent application is pending for the electrostatic space charge system technology and three license applications have been received for manufacture of the system. Working under a CRADA with a manufacturer, we developed a panel system, using monoclonal specific for SE flagella developed in our laboratory, for detecting SE in poultry samples within 15 minutes. A Small Business Innovative Research grant was awarded by USDA to further evaluate the capabilities of the panel in depth. 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below). HOLT, P.S. Induced molting and possible Salmonella enteritidis problems in layer flocks. Foodonics Incorporated Egg Quality Assurance Producer's Meeting, Blackshear, GA, 1999. HOLT, P.S. Induced molting and possible Salmonella enteritidis problems in layer flocks. Southeastern Egg Producers Conference, Tallahassee, FL, 1999. HOLT, P.S. Risk factors for a Salmonella enteritidis infection. SE Research Forum, Atlanta, GA, 1999 sponsored by the United Egg Producers.
PUBLICATIONS: 1999/01 TO 1999/09
1. HOLT, P.S., MITCHELL, B.W., SEO, K-H.,and GAST, R.K. 1999. Use of negative air ionization for reducing airborne levels of Salmonella enterica serovar Enteritidis ... Journal of Applied Poultry Research, in press.
2. DODSON, S.V., MAURER, J.J.. HOLT, P.S., and LEE, M.D. 1999. Temporal changes in the population genetics of Salmonella pullorum. Avian Diseases 43, in press.
3. HOLT, P.S., GAST, R.K. ,... 1999. Hyporesponsiveness of the systemic and mucosal humoral immune systems in chickens infected with Salmonella at one day of age. Poultry Science, in press.
4. CHAUBAL, L.H. and HOLT, P.S. 1999. Characterization of motility and identification of flagella proteins in the avian pathogen Salmonella pullorum. American Journal of Veterinary Research, in press.
5. MITCHELL, B.W. 1999. Electrostatic space charge system for dust and pathogen removal in commercial hatching cabinets. Poultry Science 78(S1):143.
6. MITCHELL, B.W. 1999. Performance of an electrostatic dust reduction system in a commercial hatchery. Dust Control in Animal Production Facilities International Symposium, CIGR EurAgEng, Jutland, Denmark.
7. HOLT, P.S., MITCHELL,B.W. , SEO, K.-H., and GAST, R.K. 1999. Use of negative air ionization for reducing airborne levels of Salmonella Enteritidis in a room. Proc. Western Poultry Disease Conference, page 31.
8. HOLT, P.S., GAST,R.K., STONE, H.D.. 1999. Infection of chicks with Salmonella enteritidis at one day of age results in persistent infection and hyporesponsiveness humoral immune system. Immunology Letters 69:65.
9. HOLT, P.S. 1999. Possible application of the alpha-1 acid glycoprotein assay as an indicator of stress during an induced molt. Poultry Science 78(S1):29.
Termination Year 2000
ACCESSION NO: 0178835 SUBFILE: CRIS
PROJ NO: ILLU-35-0226 AGENCY: SAES ILLU
PROJ TYPE: STATE PROJ STATUS: TERMINATED
START: 01 AUG 1998 TERM: 31 JUL 2000 FY: 2000
INVESTIGATOR: BAHR, J. M.; MALOY, S. R.; EDWARDS, R. A.
PERFORMING INSTITUTION:
ANIMAL SCIENCES
UNIVERSITY OF ILLINOIS
URBANA, ILLINOIS 61801
DEVELOPMENT OF SALMONELLA VACCINES FOR CHICKENS
OBJECTIVES: C-FAR 1999 Internal. Salmonella enteritidis is a pathogenic bacteria responsible for most cases of human food poisoning acquired from eating chickens or eggs. Because this bacterium does not usually cause symptoms in chickens, it is difficult to detect contaminated flocks. The long term aim of this project is to design a vaccine against S. enteritidis that can be easily administered to young chicks to protect chickens and their eggs from infection.
APPROACH: Virulence factors that have formed the basis of other vaccines include fimbriae, long proteinaceous molecules on the bacterial surface. The SEF fimbriae are unique to S. enteritidis virulence and seem to play a pivotal role in the ability of this particular bacteria to infect chickens and eggs. Therefore, we will use a molecular genetic approach to characterize the role of SEF fimbriae in virulence and to test the role of SEF fimbriae as a protective immunogen.
PROGRESS: 1998/08 TO 2000/07
C-FAR 1999 Internal. The objective was to design an effective, inexpensive vaccine against Salmonella enterica sv. enteriditis that can be administered to young chicks to protect chickens and their eggs from infection. Preliminary studies suggested that a type of fimbriae restricted to enteriditis may provide a useful vaccine target. Fimbriae are long, proteinaceous structures that extend from bacterial cells and mediate attachment to specific surfaces. Fimbriae have been shown to be critical virulence factors in enteric bacteria and play multiple roles in the course of infection. Salmonella infections are initiated by adherence to intestinal epithelium using multiple fimbriae. Fimbriae involved in this common step of Salmonella infections are found in all Salmonella serovars. Enteritidis also produce a unique fimbriae encoded by the sef genes (Salmonella Enteritidis fimbriae). SEF are not found in S. Typhimurium or more distantly related Salmonella serovars, but all Enteritidis isolates express SEF fimbriae. Results implicated SEF as a unique virulence factor for Enteritidis. Immunization of mice with purified SEF protects mice from Enteritidis infection, indicating that SEF are expressed in the host and elicit a strong immune response. Properties of these fimbriae suggest that this may be a novel vaccine target. We showed that mutations in sef reduce virulence of Enteritidis in mice by 10 4-fold and 10 5-fold. In contrast, mutations in sef increase virulence in chickens, suggesting that these fimbriae may play an important role in establishment of chronic, asymptomatic infections in chickens. We have characterized Enteritidis infections in chickens to develop a model system for subsequent characterization of disease. Our research showed that day old Leghorn chicks are susceptible to Enteritidis infections and a dose of 10 4-fold bacteria is lethal to young chicks. At lower doses, Enteritidis colonizes the ceca, cloaca, liver, spleen and reproductive tissue of chicks. Older chickens effectively clear the infection. Competition experiments between the sefA mutant and wild-type Enteritidis confirm that loss of SEF results in increased virulence in chickens. A simple explanation of these results may be that SEF acts as an effective immunogen, stimulating an protective cell-mediated immune response. This would enhance the clearance of the wild-type strain while allowing a mutant lacking SEF to persist in higher numbers. This conclusion is supported by previous studies indicating that SEF induces a strong T-lymphocyte response. Why would SEF promote virulence in mice and decrease virulence in chickens? In BALB/c mice, SEF promotes uptake of Enteritidis into macrophages which may limit the direct exposure of this potential antigen to the host immune system. These results imply that SEF may play different roles in the pathogenesis of Enteritidis depending on the host. To test the effectiveness of SEF as a vaccine, we developed methods to overproduce and purify SEF protein and we plan to use this protein to immunize chickens. The immunized chickens will be subsequently tested for protection against Enteritidis infections.
IMPACT: 1998/08 TO 2000/07
Salmonella enteriditis infects a wide variety of animals, including chickens and humans. enteriditis typically initiates an infection through the fecal oral route where it colonizes the intestinal wall and causes diarrhea. In adult chickens these infections are usually asymptomatic, but seemingly healthy chickens can carry and shed enteriditis for long periods of time after the initial infection. enteriditis can also colonize the developing egg in an otherwise asymptomatic chicken, making enteriditis infections very insidious. Recently, the occurrence of enteriditis infections in humans has increased dramatically, largely due to the consumption of infected chicken eggs. Despite the impact of enteriditis infections on agriculture and human health, the virulence determinants (i.e., the mechanisms responsible for causing disease) that mediate the unique aspects of enteriditis infections in chickens are poorly understood. Understanding the unique virulence mechanisms of enteriditis may lead to new, better ways of preventing infections in chickens and thereby alleviating this major source of food poisoning. Although enteriditis is the principle causes of Salmonella food poisoning, most research on Salmonella pathogenesis has focused on Salmonella Typhimurium. Work in many labs is beginning to elucidate the Typhimurium virulence determinants. Our work identified a virulence factor that is unique to Salmonella Enteritidis which has potential for use as a cheap, effective vaccine for chickens.
PUBLICATIONS: 1998/08 TO 2000/07
1. Edwards, R., Schifferli, D. and Maloy, S. 2000. A novel role for Salmonella enteritidis fimbriae in resistance to macrophage killing. Proc. Natl. Acad. Sci. USA 97: 1258-1262.
2. Edwards, R. and Maloy, S. 2001. Inside or outside: Detecting the cellular location of Salmonella during infection. Biotechniques 30: 304-311.
3. Townsend, S. 2001. Salmonella enterica serotype Typhi possesses a unique repertoire of fimbrial genes. Infect. and Immun. 69 (In Press).
4. Edwards, R., Matlock, B., Heffernan, B. and Maloy, S. 2001. Regulation of SEF fimbriae expression in Salmonella enteritidis (Submitted).
5. Helm, R.A., Sanderson, S. and Maloy, S. 2001. A rapid approach to determine the rrn arrangement in Salmonella serovars (Submitted).
6. Matlock, B., Heffernan, B., Edwards, R., Bahr, J. and Maloy, S. 2001. The role of SEF14 fimbriae in the pathogenesis of Salmonella enteritidis in day-old chicks (In Preparation).
PROJ CONTACT:
Name: Bahr, J. M.
Phone: 217-333-2900
Fax: 217-333-8286
Email: j-bahr@uiuc.edu
ACCESSION NO: 0149377 SUBFILE: CRIS
PROJ NO: 6406-32000-005-00D AGENCY: ARS 6406
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW
START: 21 MAR 1995 TERM: 20 MAR 2000 FY: 2000
INVESTIGATOR: BRANTON S L; BEARSON S M; LOTT B D; SIMMONS J D
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
MISSISSIPPI STATE, MISSISSIPPI 39762
DIAGNOSIS AND CONTROL OF MYCOPLASMOSIS IN POULTRY
OBJECTIVES: Improve both the diagnosis and identification of Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG) using an antigen-based ELISA and selected monoclonal antibodies. Strain specificity will be pursued for MG. Determine effects of common poultry mycoplasmal commensals on serology and egg quality. Determine effects of commercially available live MG vaccines on layer chickens. Isolate the receptor gene(s) from F strain Mycoplasma gallisepticum and insert them into Mycoplasma gallinarum.
APPROACH: Commercial hens maintained in fiberglass isolation units will provide both production and egg quality data. They will further serve as both the sera and culture source for use in selecting from currently existing monoclonal antibodies which will be used to produce the antigen-based ELISA. BALB/c mice will be purchased and used to produce other needed monoclonal antibodies which will be screened and selected against both direct hen mycoplasmal isolates and against multiple-passaged mycoplasmal cultures. Selection should result in a bank of monoclonals having specificity towards mycoplasmal-common, species-specific as well as strain-specific epitopes. The antigen-based ELISA will be utilized to determine the contribution of common avian mycoplasmal commensals on MG and MS serology. Using molecular approaches, a non-pathogenic, life-long protective immunity to respiratory disease caused by Mycoplasma gallisepticum will be developed.
PROGRESS: 1999/01 TO 1999/09
1. What major problem or issue is being resolved and how are you resolving it? The objectives of this research are: a) to improve the diagnosis and identification of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), b) to identify virulence factors that enable F strain MG to displace S6 strain MG, c) to characterize the effects of the three currently available live MG vaccines on egg production, egg and eggshell quality parameters, d) to characterize the antibody responses of infected poultry to mycoplasma antigens by using monoclonal antibodies and molecular techniques, and e) to evaluate the impact of Mycoplasma infection in poultry. 2. How serious is the problem? Why does it matter? Fully 80% of commercial egg laying chickens are believed infected with Mycoplasma gallisepticum (MG) which results in decreased egg production, increased pharmaceutical costs, and mortality. MG infection of broiler breeders is sporadic and results in either condemnation and slaughter of infected hens and/or infection of progeny which in turn, results in increased mortality, feed costs and slaughter house condemnation. "Backyard" flocks exist as primary reservoirs of infection. Numerous other species of Mycoplasma have been found in commercial poultry. Fully 20% of the approximately 125 known mycoplasmas can be found in the class Aves and of these about one-half (12) are found in poultry. Of these 12, only four have been investigated to any depth. At least five of the 12 mycoplasmas found in poultry can be isolated in commercial layer chickens. We do not know the effect(s) of these other mycoplasmal species on poultry and, coupled with the fact that mycoplasmal infections in poultry are not cleared with antibiotics (once infected, a chicken remains infected for the duration of its life) mycoplasmal infection presents a dread scenario for the poultry industry. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? This research is assigned to National Program 103 Animal Diseases. It is appropriate for the sections on "pathogen detection," "epidemiology of disease," and "disease prevention/control through vaccine and novel strategies." 4. What were the most significant accomplishments this past year? Although the temperature sensitive (ts-11) live vaccine strain of MG has been available for use in commercial layers for the past nine years, there has been NO information available to the producer concerning the vaccines effects on egg production, egg weight distribution, egg weight, blood/meat spot incidence, pimpling incidence, Haugh unit determination and eggshell strength despite the fact that the vaccine was developed to reduce egg production losses associated with MG infection. The research included measuring, recording and analysis of the aforementioned parameters together with hen mortality was conducted at the Mid South Area Poultry Research Unit and involved collaborators from Mississippi State University's College of Veterinary Medicine as well as from the Mid South Area Office of the Director. We determined that the ts-11 MG vaccine does not detrimentally affect any of the parameters measured when the hens are vaccinated at ten weeks of age. The outcome of this research is that now egg producers can make an informed decision regarding the use of this vaccine in their flocks. Most commercial layer chickens are infected with MG and most of these chickens remain "on the farm" for approximately 100 weeks of age; this becomes important when attempting to eradicate MG from multi-age layer operations, particularly in view of the fact that "the influence of age on resistance to disease has not been adequately examined." Chickens were maintained in biological isolation units at the Mid South Area Poultry Research Unit and chickens were infected at either 10 or 66 weeks of age whereupon recovery of the organism was attempted involving collaboration from Mississippi State University's College of Veterinary Medicine. Organism recovery data indicated that older infected hens are more efficient at suppressing the replication of the organism than are hens infected at a younger age. The ramification of this research impacts epidemiological investigations in that it suggests that older infected hens do not shed (implying that transmission to other hens is decreased) the organism as readily as younger infected hens. Most commercial layers are infected with MG and remain "on the farm" through 100 weeks of age during which time they are molted at about 65 weeks of age. Choanal cleft swabs were taken before and after molting during which time the birds were maintained in biological isolation units at the Mid South Area's Poultry Research Unit and involved collaboration from Mississippi State University's College of Veterinary Medicine. Molting was shown to result in a decreased ability to isolate MG from known infected chickens. This result has epidemiological ramifications in that it suggests that confirmatory MG isolation tests may be impaired when conducted on hens either in molt or immediately after molt. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. Major accomplishments over the life of the project include: a) the finding that the chicken remains infected over its life and that neither isolation nor disinfection of its environment will rid the chicken of the organism, impact - unless you can depopulate an infected farm, you cannot get rid of the organism from the flock, b) that the cellular profile of the chicken's blood reflects an acute infection with mycoplasma; however, the cellular blood profile of the chronically infected chicken is similar to control hens, despite the fact that the organism can still be isolated from the chronically infected hen, impact - the chicken's immune system is apparently "tricked" by the organism into believing there is no need to continue mounting an immune response, c) that the addition of the commonly used therapeutic agent ammonium chloride in layer chickens hinders the bacteriological recovery of Mycoplasma gallisepticum from chickens, impact - a diagnostician needs to know that a flock is being treated with ammonium chloride when he is attempting to isolate the organism and therefore take a greater number of swabs in order to assure the organism's isolation when the flock is actually infected, d) that vaccination of chickens at 10 weeks of age with the F strain of Mycoplasma gallisepticum does not reduce egg production over a 44-week laying cycle, impact - since 1979, flock managers have used the F strain of Mycoplasma gallisepticum with the understanding that the vaccine reduces egg production by approximately 7 eggs/hen over a 45-week laying cycle. 6. What do you expect to accomplish, year by year, over the next 3 years? In 2000, we expect to demonstrate the effects of an available live MG vaccine (6/85) on egg production, egg size distribution, and egg and eggshell quality parameters of layer chickens. Further, we expect to sequence the genetic code and location of a putative gene associated with the ability of the MG organism to adhere to a host cell (cytadhesin gene) thereby inciting infection of the host. In 2001, we expect to investigate the effect of one of the most prevalent Mycoplasma species (Mycoplasma gallinarum) found in commercial layer chickens on egg production, egg size distribution, and egg and eggshell quality parameters. Also, we expect to show the impact of MG infection on the lipoprotein content of eggs and further, to evaluate the virulence of the cytadhesin gene sequenced in 2000. In 2002, we expect to demonstrate the impact of Mycoplasma gallinaceum, the second most prevalent Mycoplasma species found in commercial layer chickens, on egg production, egg size distribution, and egg and eggshell quality parameters. We also expect, contingent upon a determination of non-virulence in 2001, to insert the cytadhesin gene sequenced in 2000 into Mycoplasma gallinarum to result in the initiation of the development of a novel vaccine against MG. This novel vaccine will be capable of preventing colonization by wild or field strains of MG and will also result in enhanced interstate movement and export of commercial poultry by not resulting in positive seroconversion for both MG and MS tests. Finally, this vaccine should be more acceptable to the broiler and turkey industries as it will be protective against MG and MS yet cause no pathology. 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 & durability of the technology product? Information pertaining to the effect of ts-11 MG vaccine on egg production, egg size distribution, Haugh unit measurement, egg shell breaking strength, blood meat spot incidence, pimpling incidence, and average egg weight has been made available to egg producers, veterinarians, and diagnosticians. Similarly, information regarding both molting and age of the chicken at the time of MG vaccination has been made available as it pertains to these factors potential for hindrance to isolation of the MG organism. The foregoing information is relevant immediately as approximately 80% of the commercial layer chickens are either infected or vaccinated with MG. This technology has been delivered to commercial layer managers via telephone call, reviewed journal articles, and conference talks. The value will continue as long as this vaccine is used in commercial layers. The only constraints to adoption of the technology rest with the individual states and whether they permit use of the vaccine within their borders. 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below). Becker, Hank. "Timing of vaccination could increase egg output." Poultry Times, March 22, 1999, p. 14. Becker, Hank. "Vaccinating hens at the right time saves eggs." Agricultural Research, March 1999, p. 17. Becker, Hank. "Vacc boost for egg production." ANIMAL PHARM World Animal Health and Nutrition News, July 2, 1999, p. 19.
PUBLICATIONS: 1999/01 TO 1999/09
1. BRANTON, S.L., LOTT, B.D., MAY, J.D., MASLIN, W.R., PHARR, G.T., BROWN, J.E. and BOYKIN, D.L. 1999. The effects of F strain ... ..... II. Egg size distribution. Avian Diseases 43:326-330.
2. BRANTON, S.L., MAY, J.D., LOTT, B.D. and PHARR, G.T. 1999. Effects of age at inoculation and induced molt on .... Mycoplasma gallisepticum from layer chickens. Avian Diseases 43:516-520.
3. BRANTON, S.L., LOTT, B.D., MAY, J.D. and MASLIN, W.R. 1999. The effects of ts-11 strain of Mycoplasma gallisepticum in commercial layer hens. Poultry Science (Suppl. 1):17.
4. BRANTON, S.L., SIMMONS, J.D., PHARR, G.T. and BROWN, J.E. 1999. Mycoplasma isolates in layer chickens. Poultry Science (Suppl. 1):107.
ACCESSION NO: 0177392 SUBFILE: CRIS
PROJ NO: MOR-9702552 AGENCY: CSREES MO.R
PROJ TYPE: NRI COMPETITIVE GRANT PROJ STATUS: TERMINATED
CONTRACT/GRANT/AGREEMENT NO: 97-35201-4936
START: 01 OCT 1997 TERM: 30 SEP 2000 FY: 2000 GRANT YR: 1997
INVESTIGATOR: CURTIS III, R.; WILMES-RIESENBERG, M.
PERFORMING INSTITUTION:
BIOLOGY
WASHINGTON UNIV
#1 NORTH BROOKINGS DRIVE, CAMPUS BOX 1137
ST LOUIS, MISSOURI 63130
ADHESINS FOR COLONIZATION OF CHICKENS & THEIR USE IN PREVENTION OF SALMONELLOSIS
OBJECTIVES: An understanding of the mechanism of Salmonella adherence to chicken cells could be particularly valuable when developing strategies to eliminate or reduce Salmonella colonization of poultry, and consequently, the shedding of Salmonella in feces, its transmission to eggs, and the cross-contamination of chicken carcasses which occurs during processing. The specific aims of this proposal are: to identify the gene(s) encoding the iron-induced adhesin, to evaluate the role of the iron-induced adhesin in the adherence of the Salmonella to avian cells in culture, the colonization of the chicken intestine, and the attachment of Salmonella to the surface of chicken carcasses and to determine if the iron-induced adhesin is made by other serotypes of S. enterica which colonize chickens.
APPROACH: The goals of this proposal are to identify the iron-induced adhesin and to evaluate its role in the adherence of the Salmonella to avian cells in culture, the colonization of chicken intestine, and in the attachment of Salmonella to the surface of chicken carcasses. This information may ultimately be used to design an avirulent S. typhimurium vaccine strain which will express the gene(s) encoding the Iia constitutely, and to evaluate its potential to induce an immune response that would lessen the ability of S. typhimurium and other serotypes to colonize the intestinal tract of chickens. The experiments proposed involve using a variety of genetic and molecular biology techniques. Mutants will be evaluated using tissue culture and animal models.
PROGRESS: 1999/10 TO 2000/09
Our long-term objective is to reduce or eliminate Salmonella colonization of poultry resulting in a reduction in the shedding of Salmonella in feces, transmission to eggs and cross contamination which occurs during processing. Toward this end, we initially constructed a S. typhimurium strain possessing knockout mutations in the genes encoding type 1 fimbriae (Fim), long polar fimbriale (Lpf), plasmid encoded fimbriale (Pef) and thin aggregative fimbriae (Agf). A derivative of this strain was also generated but lacking the ability to synthesize flagella, which can also serve as an adhesin. These strains were constructed to facilitate better identification of mutants defective in the expression of an iron-induced adhesin using TnphoA mutagenesis. Such a mutation, iia-8::TnphoA was identified in a S. typhimurium strain with a phoP allele to block synthesis of acid phosphatase which would then enable screening for strains that could or could not express alkaline phosphatase encoded by the phoA gene in the presence rather than in the absence of high iron concentrations. This Iia-defective mutant adhered to tissue culture cells with only 5 to 10 percent of the efficiency of wild-type cells under high iron conditions and with equal but low efficiency under low iron conditions. Using genomic subtractive hybridization and selective capture of transcribed sequences (SCOTS), we have identified another unique fimbrial operon, termed stf, in the S. typhimurium genome flanking the ferrochrome uptake operon. This stf fimbrial operon is similar to sequences encoding a mannose- resistant fimbrial type in Proteus mirabilis. Although the stf fimbrial operon is absent from the human pathogen S. typhi, it is present in the genomes of broad host range serovars that are specifically adapted to avian hosts. A lacZ reporter fusion was constructed, and studies demonstrated that the stf fimbrial operon was not regulated by iron concentration or by other environmental stimuli or stresses that affect expression of other fimbrial operons. A mutant with constitutive expression of Stf fimbriae has been constructed as well as a knockout mutant unable to synthesize Stf fimbriae. These mutants, as well as those with combination of these mutations and those affecting synthesis of other adhesins, including Iia fimbriae, will be evaluated for ability to colonize day-of-hatch chicks and to cause disease. Future studies will explore the potential effectiveness of the immune responses against these fimbrial adhesins in blocking the ability of Salmonella of diverse serotypes to colonize the avian intestinal track and thus to reduce the likelihood that the Salmonella will be transmitted through the food chain to humans.
IMPACT: 1999/10 TO 2000/09
We have identified six fimbrial adhesins, two newly identified by our efforts, that permit Salmonella typhimurium strains to effectively colonize the intestinal tract of chickens and thus become invasive. Flagella can also facilitate adherence and contribute to colonization of the avian intestinal tract. Using mutant strains with various constellations of mutations, colonization of the intestinal tract and virulence can be reduced significantly, if not eliminated. The results obtained permit us to postulate that effective immune responses against the 7 adhesins would likely abolish the ability of Salmonella strains to colonize the intestinal tract and be invasive and thus reduce the likelihood for Salmonella transmission through the food chain to humans.
PUBLICATIONS: 1999/10 TO 2000/09
1. Morrow, B.J., J.E. Graham, and R. Curtiss III. 1999. Genomic subtractive hybridization and selective capture of transcribed sequences identify a novel Salmonella typhimurium fimbrial operon and putative transcriptional regulator that are absent from the Salmonella typhi genome. Infect. Immun. 67:5106-5116.
2. Abstracts of poster presentations have been submitted for the 2001 general meeting of the American Society for Microbiology. Full-length papers describing our results are in preparation.
ACCESSION NO: 0402941 SUBFILE: CRIS
PROJ NO: 6612-32000-017-05T AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW
START: 26 APR 1999 TERM: 31 MAR 2000 FY: 2000
INVESTIGATOR: HOLT P S
PERFORMING INSTITUTION: AGRICULTURAL RESEARCH SERVICE, ATHENS, GEORGIA 30613
COMPARISON OF THE IMPACT OF ALTERNATIVE METHODS TO INDUCE INFECTION IN WHITE LEGHORN HENS
OBJECTIVES: Examine the impact of different molting procedures on a Salmonella enteritidis infection in chickens
APPROACH: Hens will be molted via feed removal, skip feeding, or low nutrition diets and then infected with s. enteritidis. The course of the infection in these hens will be compared with normal-fed hens. Trust Agreement with US Poultry an Egg Association
PROGRESS: 1999/01 TO 1999/09
1. What major problem or issue is being resolved and how are you resolving it? Induced molting is a prevalent procedure used by the layer industry to achieve a second egg lay from aging flocks. It is estimated that 70% of the flocks nationwide are molted. There are a variety of methods to induce a molt but taking the birds off feed until they drop 25-30% body weight is the preferred method. Early studies on molting showed that the effects of molting on a flock were primarily positive but we showed that the procedure depressed the immune system of the birds and dramatically exacerbated a Salmonella enteritidis (SE) infection. This increased the chance for the production of eggs contaminated with SE and therefore presented a food safety problem. Procedures need to be developed which will allow the producers access to this important economic tool while, at the same time, not putting the consuming public at risk for an SE infection. One procedure which we examined was the use of alternative molt procedures in the place of chronic feed removal to reduce or eliminate the exacerbated SE infection. We submitted a grant which was funded by the U.S. Poultry and Egg Association to examine the efficacy of using these alternative molt procedures to decrease or eliminate the SE problem. 2. How serious is the problem? Why does it matter? A 70% prevalence of molting means the procedure affects between 144-168 million hens annually - a tremendous number of birds. It is estimated that approximately one third of the egg industry's profits is derived from molted birds which means that this is a very valuable industry tool. Successful completion of these studies could mean that the egg industry could use this important procedure without putting the consuming public at risk. The impact would therefore be dramatic. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? National Program 103, Animal Health (25%) National Program 108, Food Safety (75%) Food safety is a major component of the National Program agenda and intervention strategies which help prevent the dissemination of a human disease organism within flocks and block the entry of the organism into the human food chain are important areas of investigation. 4. What were the most significant accomplishments this past year? We wanted to see if feeding the hens a nonnutritive filler such as soybean hulls, which would provide the hens with bulk materials to keep the intestinal tract filled but yet provide no nutrition and therefore put the hens into a egg-laying pause, would result in a less severe intestinal infection compared with hens totally without feed. We found that the use of soybean hulls had a minimal effect on the SE infection at day 3 post challenge, compared with the fasted birds but there was a significant, 1000-fold SE decrease in the soybean hull group at day 10 post challenge, indicating that use of a nonnutritive diet can affect the SE situation during a molt. In a second group of experiments, we examined whether a shortened feed removal period, 6 days, followed by 3 alternating periods of 3 days on feed/1 day off, would result in lower intestinal SE levels compared with total feed withdrawal. As was seen in the soybean hull experiment, no significant effects were observed at day 3 post challenge but by day 10, there was a significantly lower number of SE in the intestinal tract of the birds receiving the skip feed procedure compared with the birds off feed for the entire time. These studies show that there are molting strategies which will allow producers to recycle their birds and not increase the risk of contaminated egg production. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This CRADA has been going 0.5 years and question 4 is the data to date. 6. What do you expect to accomplish, year by year, over the next 3 years? This grant terminates on March 31, 2000. 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 & durability of the technology product? None 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work (NOTE: this does not replace your peer reviewed publications which are listed below).
PUBLICATIONS: 1999/01 TO 1999/09
No publications reported this period.
ACCESSION NO: 0149083 SUBFILE: CRIS
PROJ NO: 6202-42000-008-00D AGENCY: ARS 6202
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW
START: 01 MAY 1995 TERM: 30 APR 2000 FY: 2000
INVESTIGATOR: KOGUT M H; VACANT; VACANT
PERFORMING INSTITUTION: AGRICULTURAL RESEARCH SERVICE COLLEGE STATION, TEXAS 77845
CYTOKINE-MEDIATED MODULATION OF THE INNATE IMMUNE RESPONSE TO PREVENT SALMONELLOSIS IN POULTRY
OBJECTIVES: 1) Produce cytokines or immune lymphokines in poultry by stimulation of spleen cells to proliferate in vitro; 2) identify functional activity of cytokines; 3) elucidate the mechanism of action of cytokines in poultry to aid in identification; 4) encapsule and/or bind cytokines to cell or other support matrix for delivery to neonatal chicks; and 5) purify cytokine(s) and cloning of gene for possible mass production.
APPROACH: Research will be focused on the identification and purification of cytokines which potentiate the innate immune responses of poultry that effectively prevent organ invasion of Salmonella enteritidis. Experiments will be conducted to separate and determine any effector cytokines present in our T cell supernatants and any host-derived cytokines induced endogenously in response to the injected material. Studies will be conducted to identify the specific components of a protective response and to elucidate the mechanisms of protection induced by a prophylactic administration of cytokines in neonatal chicks. Emphasis will be placed on the development of a cost-effective delivery system that can be incorporated into the poultry industry.
PROGRESS: 1999/01 TO 1999/09
1. What major problem or issue is being resolved and how are you resolving it? Despite the endeavors of researchers and public health agencies, the incidence of human salmonellosis has continued to increase over the past 20 years. Salmonellosis is now one of the most common food-borne causes of gastroenteritis. The number of reported cases of human salmonellosis exceeds 40,000 per year. The CDC estimates the true annual incidence of human salmonellosis in the US may be as high as 2 to 4 million cases/year. Animal food products are the principal source of human infections. Traditional management of infectious diseases in poultry has for the large part depended on the use of either broad-spectrum antibiotics and/or vaccines. Inappropriate use of antibiotics and concerns of antibiotic residues in meat causing microbial resistance in humans disease, has caused a public outcry demanding the removal of many antibiotics from animal feed and ultimately limiting future introduction of antibiotics for animal use. Vaccinations have limitations, at least 7-10 days are required for the stimulation of the acquired immune response for protection, and thus early protection for chicks from diseases is attempted through maternal antibodies. We propose an alternative immunological method for the control of infectious diseases, cytokine immunomodulation. Using avian cytokines as natural enhancers of the functional activity of the avian immune system results in direct protection against infectious diseases and provides an adjuvant-like promotion to any vaccine administered. Additionally, cytokines would be given during the first few days of life when the animals' immune system is functionally immature. Cytokines induce a very rapid protective response (within hours) which would last through this susceptible period of life. 2. How serious is the problem? Why does it matter? More than 8 billion broiler chickens are produced and commercially processed in the US each year. Research has indicated that 35% or more of all processed poultry meat products purchased by consumers may be contaminated by salmonellae. Because those Salmonella serotypes most often associated with human illness do not cause serious disease or grossly identifiable lesions in poultry, it has not been possible to eliminate contaminated poultry carcasses by inspection procedures during processing. Induced molting of laying hens is a common practice used by the US layer industry for the continual productivity of aging hens. However, induced molting has been linked to the decreased resistance and increased severity of Salmonella enteritidis (SE) in these hens. In fact, table eggs have been shown to be major sources of SE infections in humans with the risk of eggs becoming positive for SE contamination increasing threefold in flocks molted within the previous 10 weeks. Since 60% of the hens undergo a forced molt, approximately 200 million hens have an increased risk to SE infection. Salmonellosis in swine has been estimated to cost swine producers over $100 million dollars/year in losses. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? Food safety belongs to a cross cutting program area. A major research direction in the Food Safety program (NP-108) is to decrease pathogenic bacteria. Targeted microbes include Salmonella spp. There is a need to develop integrated strategies for the prevention of Salmonella colonization and organ invasion in poultry and swine. One such strategy is the modulation of the host immune system. Immunological research on the control of intestinal and tissue colonization of poultry by invasive Salmonella enteritidis (SE) has largely focused on the development of live, attenuated vaccines. Regardless of the efficacy of a vaccine, at least 7-10 days are required for the stimulation of the acquired immune response for protection. Unfortunately for the poultry industry, neonatal poultry are most susceptible to Salmonella infections during the first 4 days post-hatch, after which they become increasingly more resistant to infection. During this first 4 days post-hatch we hypothesized immunopotentiation of the innate defense mechanism(s) would prevent Salmonella organ infectivity. 4. What were the most significant accomplishments this past year? This year tests were undertaken to determine the effects of induced molt on basal functional activities of heterophils from aging hens. The results indicated that feed withdrawal to induce molt alters the number and function of peripheral blood heterophils. This decreased efficiency of heterophil functional activity appears to play a role in the increased susceptibility of molting hens to Salmonella enteritidis infections. Preliminary investigations conducted during the last year demonstrated that the oral administration of Salmonella enteritidis-immune lymphokines (SILK) to hens 24 h before feed withdrawal for molting will prevent SE infections in the hens. These results suggest that SILK could become a commercial product for hens during induced molt that can convey protection against Salmonella enteritidis infections. During the last year, we conducted experiments that clearly showed that SILK delivered to day-old turkey poults orally, intranasally, and subcutaneously is effective in protecting the birds from SE infections. This protection is mediated by the activation of peripheral blood heterophils. These results are significant because we have now demonstrated in both chickens and turkeys multiple cost-effective industry friendly delivery systems for SILK. We have also conducted experiments to evaluate the effect of SILK on the incidence of horizontal transmission of S. arizonae in turkey poults and S. gallinarum in broiler chickens. Our results demonstrated that the immunoprophylactic administration of SILK to young turkey poults and broiler chicks significantly reduces the horizontal transmission of Salmonella in poultry. These results further suggest the possibility of using a non-vaccine immunologically-based preventive strategy against Salmonella in poultry. We also evaluated the effects of SILK on the phagocytic and bactericidal activities of heterophils from chicks during the first 1-7 days of life. We demonstrated that following the administration of SILK, the functional activities of heterophils from 1-7 day-old birds is comparable to that of an immunologically mature (2-3 weeks of age) bird. Mechanistically, these functionally mature heterophils are responsible for the protective inflammatory response, which protects the birds from salmonellae infections. These data demonstrate that SILK augments the innate host defenses in chicks during the greatest period of susceptibility to Salmonella infections. We have also demonstrated that recombinant chicken interferon-gamma (rChIFN-gamma) is capable of enhancing the functional activities of heterophils from day-of-hatch chicks. These results demonstrate that the heterophils from neonatal chicks possess the receptor for IFN-gamma; thus, this cytokine could possibly be used either as an immunopotentiator for day-old chicks or as an adjuvant for vaccines. We conducted studies to identify the specific protein in SILK responsible for the protective effect in chickens. SILK was fractionated by ammonium sulfate precipitation and the protective activity was recovered in the 40- 60% ammonium sulfate saturation fraction. Monoclonal antibodies against SILK were then linked to a solid matrix to form an immunoaffinity column over which the 40-60% fraction was run. The recovered activity was then subjected to size exclusion, ion exchange, and hydrophobic interaction chromatography. Isolation of the SILK protein will enable us to clone the gene for this protein that will be a useful tool for the development of economical and effective immunologically-based treatments for the reduction of Salmonella in poultry products. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. Our laboratory has been evaluating the practicality of potentiating the endogenous innate host defenses of poultry using immune and inflammatory cytokines. Cytokines, chemical messengers secreted by various immune and non-immune cells, are some of the most effective mediators of natural host defenses. Specifically, we have found that the prophylactic administration of cytokines derived from T cells (SILK) isolated from Salmonella enteritidis-immune chickens have a favorable effect in controlling or eliminating salmonellosis in neonatal poultry and hens undergoing induced molting. This resistance is associated with a dramatic peripheral blood leukocytosis after the injection of the SILK followed by a marked infiltration of bactericidal inflammatory heterophils in the lamina propria of the ceca. Heterophils are highly phagocytic, polymorphonucleated white blood cells which are important mediators of innate resistance in poultry; especially in young birds that have not yet developed an acquired immune response. Under normal conditions, invasion of the intestinal mucosa by Salmonella spp. initiates the recruitment of heterophils to the lamina propria which control bacterial numbers in the bird until the development of acquired immunity. Administration of SILK into neonatal chicks induces a significant increase in the production and release of heterophils into the peripheral blood and also augments the effector functions of these phagocytes. Thus, salmonellae infections are not only controlled but eliminated from chicks following the administration of SILK. Further studies indicate that the delivery of SILK either in ovo at 18- days of embryogenesis or orally and as an aerosol at hatch can be used by the poultry industry to confer protection to chickens against a localized enteric SE organ invasion by potentiating the systemic heterophilic innate response. 6. What do you expect to accomplish, year by year, over the next 3 years? In FY-00, we hope to identify and purify the effector cytokine(s) in SE- ILK and clone the gene for mass production. We also hope to identify and clone genes for many avian cytokines for use in the immunopotentiation of host immune responses in neonatal poultry. Genes for interferon-gamma and the putative homologue of human IL- 8, 9E3/CEF4 have been cloned in our laboratory and they will be evaluated for their protective abilities in our Salmonella models. This CRIS will terminate in April FY-00. 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 & durability of the technology product? SILK has been sent to scientists all over the world for use in research in immunology enhancement of the avian and porcine immune systems, as well as for evaluation in the prevention and control of various viral, bacterial, and protozoal diseases of food animals. The main constraint for the transfer of this technology is the purification and identification of the effector molecules in the SILK. Once we can fully identify the components of our SILK soup the pharmaceutical industry will license this material. In addition, we have developed a CRADA with Cobb-Vantress Poultry Breeders to evaluate the effect of SILK in commercial birds. 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below). Heterophils to the Rescue! Agricultural Research p. 9. July, 1997.
PUBLICATIONS: 1999/01 TO 1999/09
1. KOGUT M.H., HOLTZAPPLE C., LOWRY V.K., GENOVESE, K. and STANKER, L.H. 1998. Comparison of the functional and turkey heterophils in response to stimulation by inflammatory agonists. Am. J. Vet. Res. 59:1404-1408.
2. GENOVESE, K.J., LOWRY, V.K., STANKER, L.H. and KOGUT, M.H. 1998. Administration of Salmonella enteritidis-immune lymphokine blood heterophilia, and heterophil activation. Avian Pathol. 27:597-604.
3. LOWRY, V.K., NISBET, D.J., STANKER, L.H. and KOGUT, M.H. 1999. Efficacy of Salmonella enteritidis-immune turkeys and S. gallinarum in chickens. Int. J. Food Microbiol. 48:139-148.
4. GENOVESE, L.L., LOWRY, V.K., GENOVESE, K.J. and KOGUT, M.H. 1999. Enhancement of phagocytosis and bacterial administration of Salmonella enteritidis-immune lymphokines. Vet. Microbiol. 65:133-143.
ACCESSION NO: 0153343 SUBFILE: CRIS
PROJ NO: IND073045V AGENCY: CSREES IND
PROJ TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START: 07 MAY 1996 TERM: 30 SEP 2000 FY: 2000
INVESTIGATOR: SAEED, A. M.; ASEM, E. K.; CSONCO, L.
PERFORMING INSTITUTION:
VETERINARY PATHOBIOLOGY
PURDUE UNIVERSITY
WEST LAFAYETTE, INDIANA 47907
VIRULENCE FACTORS OF SALMONELLA ENTERITIDIS AND INFECTIVITY FOR THE AVIAN REPRODUCTIVE SYSTEM
OBJECTIVES: To characterize the virulence factors in Salmonella enteritidis isolates from poultry, eggs and human cases of food poisoning to study the relatedness among Salmonella isolates from different sources and to establish pathotypes for the organism that may be associated with disease outbreaks in commercial poultry flocks, which result in the production of Salmonella infected eggs.
APPROACH: To use the characteristics of Salmonella pathotypes in experimental infection of egg-laying hens and study the course of the disease microbiologically, pathologically and serologically to identify useful markers for the diagnosis of the disease and characterize Salmonella infectivity to the avian reproductive system.
PROGRESS: 1996/05 TO 2000/09
Salmonella Enteritidis is among the most virulent Salmonella Serotypes in humans and animals. In the US. More than 25 percent of all cases of salmonellosis in humans are associated with eating raw or undercooked eggs. In previous reports we demonstrated that egg contamination may take place transovarially. Our recent investigation targeted the virulence factors of Salmonella Enteritidis that enable the organism to contaminate the preovulatory follicles of the laying hen that leads to the contamination of laid eggs. We produced knockout mutants of three phage types of Salmonella Enteritidis such that their flagellar expression was suppressed. We studied the role of flagella in the in vitro attachment and invasion of the avian ovary cells maintained in primary tissue culture. We found that flagellar suppression was associated with the failure of the organism to attach or invade the avian ovary cells compared to the parent or wild flagellated strains. We also found that incubating the inoculated cells at 42C, which is closer to the normal temperature of the avian species, enhanced flagellar expression. These results may suggest the preference of hens by the salmonella organisms as a major reservoir.
PUBLICATIONS: 1996/05 TO 2000/09
Popiolarczyk. M., Asem E.,., Koons. C., Csanco. L., Kazocos. E., Thacker. L. Saeed. A.M.. The role of flagella in the virulence of Salmonella Enteritidis. pp 425-439. in Proceedings of the 103rd Annual Meeting of the United States Animal Health Association. San Diego, CA, October 7-14. 1999
Termination Year 1999
ACCESSION NO: 0401397 SUBFILE: CRIS
PROJ NO: 6612-32000-019-01T AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: TERMINATED
START: 01 DEC 1997 TERM: 30 NOV 1999
INVESTIGATOR: HOLT P S; GAST, R. K.
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
ATHENS, GEORGIA 30613
IMPLEMENTATION OF INTERVENTION MEASURES TO REDUCE SALMONELLA ENTERITIDIS INFECTIONS IN CHICKENS
OBJECTIVES: Examine the utility of using different licensed products as intervention measures to reduce the severity of Salmonella enteritidis infections in birds exposed during periods of high susceptibility.
APPROACH: Subject hens to induced molting procedure and infect with Salmonella enteritidis. Use antibiotics in the presence or absence of probiotics to eliminate infection. Subject hens to induced molting procedure and infect select hens with Salmonella enteritidis. Treat part of the unchallenged hens with probiotics while the others remain untreated to examine the effects of the probiotics on horizontal transmission of the Salmonella enteritidis. Infect one day old chicks with Salmonella enteritidis in the presence or absence of probiotics to examine the effects of treatment on long term persistent infections and immunodepression. CRADA with Bayer Corporation.
PROGRESS: 1999/01 TO 1999/11
1. What major problem or issue is being resolved and how are you resolving it? Induced molting is a prevalent procedure used by the layer industry to achieve a second egg lay from aging flocks. It is estimated that 70% of the flocks nationwide are molted. There are a variety of methods to induce a molt but taking the birds off feed until they drop 25-30% body weight is the preferred method. Early studies on molting showed that the effects of molting on a flock were primarily positive but we showed that the procedure depressed the immune system of the birds and dramatically exacerbated a Salmonella enteritidis (SE) infection. This increased the chance for the production of eggs contaminated with SE and therefore presented a food safety problem. Procedures need to be developed which will allow the producers access to this important economic tool while, at the same time, not putting the consuming public at risk for an SE infection. One procedure which we examined was the use of antibiotics, in combination with reconstituting the intestinal bacterial flora with competitive exclusion cultures, to eliminate the SE problem. We established a CRADA with Bayer Corporation, marketers of the fluoroquinolone antibiotic enrofloxacin and a competitive exclusion culture , to examine the efficacy of treating with these products to decrease or eliminate the SE problem 2. How serious is the problem? Why does it matter? A 70% prevalence of molting means the procedure affects between 144-168 million hens annually - a tremendous number of birds. It is estimated that approximately one third of the egg industry's profits is derived from molted birds which means that this is a very valuable industry tool. Successful completion of these studies could mean that the egg industry could use this important procedure without putting the consuming public at risk. The impact would therefore be dramatic. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? National Program 103, Animal Health (25%) National Program 108, Food Safety (75%) Food safety is a major component of the National Program agenda and intervention strategies which help prevent the dissemination of a human disease organism within flocks and block the entry of the organism into the human food chain are important areas of investigation. 4. What were the most significant accomplishments this past year? We wanted to see if treatment of molted infected birds with antibiotics and competitive exclusion cultures would decrease the SE infection after the molt. Birds were molted, infected with SE 4 days into the feed removal, and then put back on feed after 14 days. They were put on a regimen of antibiotics for 10 days followed by two days of competitive exclusion administration. We showed that antibiotic treatment after the birds went back on feed reduced the percentage of infected birds from 33% and 25% to 4% and 0% in experiments one and two, respectively. The addition of the competitive exclusion cultures were critical in the protection. These studies showed that there are intervention strategies which will allow producers to molt their birds and not increase the risk of contaminated egg production. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This CRADA has been going 1.5 years and question 4 is the data to date. 6. What do you expect to accomplish, year by year, over the next 3 years? This CRADA will be terminated at the end of the calendar year. 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 & durability of the technology product? N/A 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below). HOLT, P.S. Possible intervention strategies for Salmonella enteritidis problems in layer hens. Poultry Diagnostic and Research Center, University of Georgia, January 1999.
PUBLICATIONS: 1999/01 TO 1999/11
SEO, K.-H., HOLT, P.S., ... 1999. Combined effect of antibiotic and competitive exclusion treatment on Salmonella enteritidis fecal shedding in molted laying hens. Proceedings of the 86th IAMFES meeting, pp 55-56.
ACCESSION NO: 0401397 SUBFILE: CRIS
PROJ NO: 6612-32000-019-01T AGENCY: ARS 6612
PROJ TYPE: USDA INHOUSE PROJ STATUS: TERMINATED
START: 01 DEC 1997 TERM: 30 NOV 1999
INVESTIGATOR: HOLT P S; GAST, R. K.
PERFORMING INSTITUTION: AGRICULTURAL RESEARCH SERVICE ATHENS, GEORGIA 30613
IMPLEMENTATION OF INTERVENTION MEASURES TO REDUCE SALMONELLA ENTERITIDIS INFECTIONS IN CHICKENS
OBJECTIVES: Examine the utility of using different licensed products as intervention measures to reduce the severity of Salmonella enteritidis infections in birds exposed during periods of high susceptibility.
APPROACH: Subject hens to induced molting procedure and infect with Salmonella enteritidis. Use antibiotics in the presence or absence of probiotics to eliminate infection. Subject hens to induced molting procedure and infect select hens with Salmonella enteritidis. Treat part of the unchallenged hens with probiotics while the others remain untreated to examine the effects of the probiotics on horizontal transmission of the Salmonella enteritidis. Infect one day old chicks with Salmonella enteritidis in the presence or absence of probiotics to examine the effects of treatment on long term persistent infections and immunodepression. CRADA with Bayer Corporation.
PROGRESS: 1999/01 TO 1999/11
1. What major problem or issue is being resolved and how are you resolving it? Induced molting is a prevalent procedure used by the layer industry to achieve a second egg lay from aging flocks. It is estimated that 70% of the flocks nationwide are molted. There are a variety of methods to induce a molt but taking the birds off feed until they drop 25-30% body weight is the preferred method. Early studies on molting showed that the effects of molting on a flock were primarily positive but we showed that the procedure depressed the immune system of the birds and dramatically exacerbated a Salmonella enteritidis (SE) infection. This increased the chance for the production of eggs contaminated with SE and therefore presented a food safety problem. Procedures need to be developed which will allow the producers access to this important economic tool while, at the same time, not putting the consuming public at risk for an SE infection. One procedure which we examined was the use of antibiotics, in combination with reconstituting the intestinal bacterial flora with competitive exclusion cultures, to eliminate the SE problem. We established a CRADA with Bayer Corporation, marketers of the fluoroquinolone antibiotic enrofloxacin and a competitive exclusion culture , to examine the efficacy of treating with these products to decrease or eliminate the SE problem 2. How serious is the problem? Why does it matter? A 70% prevalence of molting means the procedure affects between 144-168 million hens annually - a tremendous number of birds. It is estimated that approximately one third of the egg industry's profits is derived from molted birds which means that this is a very valuable industry tool. Successful completion of these studies could mean that the egg industry could use this important procedure without putting the consuming public at risk. The impact would therefore be dramatic. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? National Program 103, Animal Health (25%) National Program 108, Food Safety (75%) Food safety is a major component of the National Program agenda and intervention strategies which help prevent the dissemination of a human disease organism within flocks and block the entry of the organism into the human food chain are important areas of investigation. 4. What were the most significant accomplishments this past year? We wanted to see if treatment of molted infected birds with antibiotics and competitive exclusion cultures would decrease the SE infection after the molt. Birds were molted, infected with SE 4 days into the feed removal, and then put back on feed after 14 days. They were put on a regimen of antibiotics for 10 days followed by two days of competitive exclusion administration. We showed that antibiotic treatment after the birds went back on feed reduced the percentage of infected birds from 33% and 25% to 4% and 0% in experiments one and two, respectively. The addition of the competitive exclusion cultures were critical in the protection. These studies showed that there are intervention strategies which will allow producers to molt their birds and not increase the risk of contaminated egg production. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. This CRADA has been going 1.5 years and question 4 is the data to date. 6. What do you expect to accomplish, year by year, over the next 3 years? This CRADA will be terminated at the end of the calendar year. 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 & durability of the technology product? N/A 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below). HOLT, P.S. Possible intervention strategies for Salmonella enteritidis problems in layer hens. Poultry Diagnostic and Research Center, University of Georgia, January 1999.
PUBLICATIONS: 1999/01 TO 1999/11
SEO, K.-H., HOLT, P.S., ... 1999. Combined effect of antibiotic and competitive exclusion treatment on Salmonella enteritidis fecal shedding in molted laying hens. Proceedings of the 86th IAMFES meeting, pp 55-56.
ACCESSION NO: 0167506 SUBFILE: CRIS
PROJ NO: IOWV-400-25-27 AGENCY: CSVM IOWV
PROJ TYPE: STATE PROJ STATUS: TERMINATED
START: 01 JUL 1994 TERM: 30 JUN 1999 FY: 1995
INVESTIGATOR: REYNOLDS, D. L.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
IOWA STATE UNIVERSITY
S. & 16TH ELWOOD
AMES, IOWA 50011
STUDIES ON THE EFFICACY OF A SWINE SALMONELLA VACCINE FOR PREVENTION OF SALMONELLA COLONIZATION IN
OBJECTIVES: The objective of this study is to evaluate the feasibility of using a swine salmonella vaccine for use in poultry.
APPROACH: The experimental design and approach for evaluating safety and efficacy of the swine vaccine for use in poultry is very direct and straight-forward and is as follows: All birds used will be specific pathogen free (SPF) leghorn type. These birds will be hatched and reared at our facility designed for maintaining SPF status. The SC-54 modified live vaccine will be supplied by a commercial laboratory and used at 10/8 CFUs/chick administered intranasally. A nalidixic acid resistant strain of Salmonella enteritidis will be used at a challenge dosage of 10/8 CFUs per bird. Four groups of birds will be used as follows: 1) unvaccinated, unchallenged; 2) unvaccinated, challenged; 3) vaccinated, unchallenged; 4) vaccinated, challenged. Day-old birds will be intranasally vaccinated with SC-54 and will be challenged at 2 weeks of age. Birds will be monitored daily for the presence of clinical signs of disease. At 8 to 10 days following inoculation birds will be euthanized, data and samples will be collected for evaluation. Parameters of evaluation include 1) number of salmonella (CFUs) per gram of cecal content, 2) clinical signs and/or lesions of disease, 3) body weights, 4) serologic antibody to pullorum antigen (plate test), 5) organ cultures of liver and spleen will be done.
PROGRESS: 1995/01 TO 1995/12
A commercially licensed live Salmonella swine vaccine was evaluated for efficacy in protecting chickens from Salmonella enteritidis infections. It was found that there was no efficacy in protecting chickens from SE infections.
PUBLICATIONS: 1995/01 TO 1995/12
No publications reported this period.
PROJ CONTACT:
Name: Reynolds, D. L.
Phone: 515-294-0914
Fax: 515-294-1401
Email: dlr@iastate.edu
ACCESSION NO: 0149221 SUBFILE: CRIS
PROJ NO: 3602-32000-002-00D AGENCY: ARS 3602
PROJ TYPE: USDA INHOUSE PROJ STATUS: NEW MULTISTATE PROJ NO: W-173
START: 10 NOV 1994 TERM: 09 NOV 1999 FY: 1999
INVESTIGATOR: SHEA MOORE M M; VACANT; VACANT; PRUIETT S D
PERFORMING INSTITUTION: AGRICULTURAL RESEARCH SERVICE WEST LAFAYETTE, INDIANA 47907
ETHOLOGY OF FOOD PRODUCING ANIMALS
OBJECTIVES: Identify internal states of animal cognition and learning. Establish objective indicators of well-being (behavioral, immunological, neuroendocrinological and molecular). Determine how well-being in food producing animals relates to behavior, health and production efficiencies.
APPROACH: Develop physiological measures as indicators of stress and well-being including immune and neuroendocrine responses as well as alterations in gene products in food producing animals. Measure behavior and learning in livestock to identify animal interactions with the production environment and identify conditions that both cause and alleviate stress. Combine physiological and behavioral measures to define cause and effect relationships between well-being and production efficiency in food animals.
PROGRESS: 1999/01 TO 1999/09
1. What major problem or issue is being resolved and how are you resolving it? Determination of objective indicators of animal cognition and well-being in food producing animals. We have developed a team consisting of expertise in ethology, immunology and neuroscience combined with highly skilled technicians to ensure a multidisciplinary approach to answering complex whole-animal questions. 2. How serious is the problem? Why does it matter? Animal welfare is an international issue and consumers of food products in this country are beginning to demand to know how the animals are raised and cared for in production systems. If information on the well- being of food-producing animals is not provided by scientifically assessing the animal's welfare, then emotional, subjective opinions will be used to provide this information to the consumer. This will in turn, be harmful to the American Producer with no scientific evidence to support or contradict the opinions. Ultimately, the market for U.S. livestock products will be affected both locally and internationally. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? This project is in direct concurrence with National Program 105. All of the research conducted in this project directly addresses the "Animal Stress and Well-Being, 105" National Program. 4. What were the most significant accomplishments this past year? As this project progresses, advances in determining indicators of well- being are being made in the species studied. Behavioral and immunological data continue to show advantages to all tail-banding first calf heifers over young calves. Calves show higher levels of agitative behaviors and elevations in acute phase proteins (liver proteins released in response to tissue trauma) if docked in close proximity to banding date. Additionally, a new study has shown improvements in immunological & productivity measures when first-calf heifers are exposed to a pre-milking experience prior to calving. The implications of this study suggest that gradual habituation of the animal to a management procedure, such as milking, improves the well-being of the animal. Further progress was also made with the different genetic lines of pigs selected for lean gain. The most significant accomplishment in this area of research was to combine the genetic lines with a transportation stressor. Results indicated that pigs selected for lean growth show significantly higher levels of aggressive activity when mixed during transportation. These data compliment the previous data describing greater handling problems in these pigs selected for lean growth. Salivary stress hormones, although significantly elevated due to transportation stress, were not higher in pigs selected for lean gain over other genotypes. Furthermore, genotype did not affect the meat quality scores after transportation. Finally, significant progress was made in the area of cognition work this year. It was shown that piglets are capable of perceiving and remembering olfactory and gustatory cues presented in their prenatal environment, affecting postnatal behavior. By adding volatile compounds to diets during gestation, it may be possible to increase feed consumption and reduce stress at weaning. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. The major accomplishment over the life of this project is the building of an interdisciplinary team to address complex questions about animal welfare across three species, dairy, swine and poultry. To combine neuroscience, immunology, and ethology in comprehensive approaches to answering animal welfare questions is very difficult. We have made significant progress in this area. Furthermore, we have built teams with scientists at Purdue University to incorporate genetics, nutrition and production to strengthen our approach to welfare questions. This is an essential component to the success of this project. 6. What do you expect to accomplish, year by year, over the next 3 years? Year 1: Addition of 2 new scientists to expand our interdisciplinary team. One neuroscientist with expertise in gross morphology/structure/function, and one new post-doctoral scientist to focus in the area of neuroethology (relationship between the brain and behavior)in dairy, swine and poultry. These two people will establish a quantifiable link between management systems and animal welfare by incorporating the role of the brain into new and ongoing projects in our unit. Year 2: Both the first and second year will also determine both chronic and/or acute 'pain', associated with common industry practices (beak trimming, induced molting, tail docking)in conjunction with the expression/lack of expression of behavior in these animals. There is a politically sensitive concern between forced molting and food safety (Salmonella enteritidis) currently under discussion in the U.S. Year 2 and 3 will try to connect this information with animal welfare (neuroscience, stress physiology, immunology and behavior). Year 3: We should have a full-team of scientists hired into our unit at this point. This will include: 2 ethologist (one of whom will address food safety concerns in connection with behavior), 1 immunologist, 1 neuroscientist (1 neuroethologist, postdoc), 1 animal scientist, and possibly a gut microbiologist. This unit should be up to full-speed by year 3 with a significant scientific contribution to the area of animal welfare science. 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 & durability of the technology product? Data from three research projects on tail docking in dairy cattle were summarized in a letter published in Hoard's Dairyman (May 25, 1999, Vol. 144 #10, p. 390). Currently being disseminated to the dairy industry, are results from a study using pre-milking in dairy heifers to decrease the stress of first milking in first-calf heifers. This information has been shared with and presented by Purdue Dairy Extension Specialists to producers in Indiana. One of our ARS scientists will present this information in an invited seminar at the University College Dublin, Animal Science and Production Department in September, 1999. A primer for the porcine IL-1 receptor antagonist was developed and will be listed as available for other scientists as earlier as October, 1999. This work will permit us to look at the microenvironment of the pig for early cell signals in response to stressors (environmental, physiological, social). The receptor antagonist modulates the early response of cytokines to stress. 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed
below).
PUBLICATIONS: 1999/01 TO 1999/09
1. BUSSE, C. and SHEA-MOORE, M. 1999. Behavioral and physiological responses to transportation stress on three genetic lines of pigs. J. Anim. Sci. 77 (Suppl. 1):147.
2. EICHER-PRUIETT, S.D. and MORROW-TESCH, J.L. 1999. Tail-docking alters behavior but not acute phase proteins of young dairy calves. J. Dairy Sci. 82:842.
3. EICHER-PRUIETT, S.D. and SCHUTZ, M.M. 1999. Prepartum milking of Holstein heifers: II Effect on acute phase proteins and immune activation. J. Dairy Sci. 82 (Suppl. 1):60.
4. EICHER-PRUIETT, S.D. 1999. Data on Docking. p. 390 IN: Hoard's Dairyman Vol.144 N o. 10. Eds.:Knox and Larson.
5. SCHUTZ, M.M. and EICHER, S.D. 1999. Prepartum milking of Holstein heifers: I Effects on production, parturition, edema, and SCC. J. Dairy Sci. 82 (Suppl. 1):60.
6. SCOTT, K.A. and SHEA-MOORE, M. 1999. Influence of sow gestation diet on piglet behavior. J. Anim. Sci. 77 (Suppl. 1):145.
7. SHEA-MOORE, M. 1999. Social behavior of Pekin ducks raised in a production environment. 33rd Intl. Congress of the Intl. Soc. for Appl. Ethology. P. 183.
ACCESSION NO: 0173189 SUBFILE: CRIS
PROJ NO: GEO00812 AGENCY: CSREES GEO
PROJ TYPE: HATCH PROJ STATUS: TERMINATED
START: 01 NOV 1996 TERM: 31 OCT 1999 FY: 1999
INVESTIGATOR: WEBSTER, A. B.
PERFORMING INSTITUTION: POULTRY SCIENCE UNIVERSITY OF GEORGIA ATHENS, GEORGIA 30602
WELFARE ASPECTS OF INDUCED MOLTING PROGRAMS
OBJECTIVES: To assess the welfare impact on hens of extended feed withdrawal typical of commercial induced molting programs for commercial layers.
APPROACH: To obtain quantitative measures of behavior, plasma corticosterone and heterophil:lymphocyte ratios during an induced molt and second cycle of egg production. To assess the capacity of hens to react to an additional stressor during a fast by recording behavior and plasma corticosterone of fasting hens in open field tests. To determine if hens experience increasing levels of hunger during a fast, and to determine if hens have sufficient cognitive capacity to suffer during a fast by recording the behavior of trained, fasting hens in a standardized test in which a food reward which might be expected is withheld.
PROGRESS: 1996/11 TO 1999/10
The first phase of this project has been wound up. Behavioral changes of hens during the molt fast reflected conservation of bodily reserves rather than debilitation. A small increase in aggression occurred on the first day of feed withdrawal. Increased attentiveness and stimulus generalization in foraging behavioral patterns was evident on the second day of feed withdrawal. Afterward, hens appeared to adjust expectations in regard to feeding opportunity although behavior associated with foraging and feeding did not disappear entirely. Hens which experienced feed withdrawal had significantly better survival during the second production cycle than did control hens. Preparations are being made to develop the next phase, which will involve investigation of the variation of motivational strengths in relation to hunger. This research will shed light on changes in a hen's cognitive status during feed withdrawal and determine if the potential for a hen to suffer during feed deprivation reaches a steady state or increases continuously with time off feed.
PUBLICATIONS: 1996/11 TO 1999/10
Webster, A.B., 2000. Behavioral changes of laying hens during feed withdrawal. Poultry Science: (in press)
Termination Year 1998
ACCESSION NO: 0149691 SUBFILE: CRIS
PROJ NO: 6612-32000-017-03S AGENCY: ARS 6612
PROJ TYPE: USDA COOPERATIVE AGREEMENT PROJ STATUS: TERMINATED
CONTRACT/GRANT/AGREEMENT NO: 58-6612-6-012
START: 08 JAN 1996 TERM: 30 SEP 1998 GRANT YR: 1997
INVESTIGATOR: PETTER, J. G.; MATIN, A.
PERFORMING INSTITUTION:
MICROBIOLOGY & IMMUNOLOGY
STANFORD UNIV
PALO ALTO, CALIFORNIA 94305
PRODUCTION OF SALMONELLA ENTERITIDIS SUBUNIT VACCINES
OBJECTIVES: To produce stabilized populations of S. enteritidis that hyperexpress virulence factors using continuous culture technology.
APPROACH: Specific nutrients will be titrated into cell populations at defined dilution rates. Once stabilized, samples of cells will be assayed for the production of virulence factors, which include but are not limited to lipopolysaccharide, flagella, invasomes, and pili. Those conditions that are found to produce copious amounts of virulence factors will be used to obtain enough cells for the production of a pilot killed subunit vaccine.
PROGRESS: 1998/01 TO 1998/09
1. What major problem or issue is being resolved and how are you resolving it? Vaccines made from killed cells have been made by industry for years as aids to prevent colonization and organ invasion by Salmonella enteritidis and other salmonellae. However, these products lack the level of efficacy (greater than 90% protection) that is the usual hallmark of a highly successful vaccine. We believe that partial protection by vaccines can in some cases perpetuate problems with infection by this organism rather than solve them. Our research indicates current products do not provide a high degree of protection against S. enteritidis, possibly because new information about subpopulation biology is not incorporated into current methods for preparing vaccines. The research of many investigators already indicates that Salmonella isolates vary in their ability to survive in the environment, to colonize the intestinal tract, to invade the mucosa, to gain access to internal organs via the lymphatic or circulatory systems, and to survive intracellularly. Indeed, mutations introduced into Salmonella produces defined strains that target one niche versus another, but do not complete a cycle of pathogenesis because an essential ability to undergo change has been deleted. At SEPRL, scientists have concentrated on understanding what characteristics enable Salmonella enteritidis to be so invasive that eggs become routinely contaminated with this pathogen. Contamination of eggs is the final outcome of infection in hens that affects the food safety, because eggs are the most important source of S. enteritidis for people. Research at SEPRL indicates that the ability of S.enteritidis to contaminate eggs can be tracked by following the structure of the predominant outer membrane molecule lipopolysaccharide (LPS), and indeed, some strains produce a structure much like that of S. typhi, the causative agent of typhoid fever. This phenomena is known as LPS O-chain phase variation. The importance of this phenomena to manufacturers of vaccines is that production of immunogenic cell-surface proteins change in coordination with changes in LPS O-chain structure. In addition, S. enteritidis appears to be able to produce S. typhi-like LPS while it grows to very high cell densities. Because these biological capabilities are so remarkable, scientists at SEPRL have developed special strains and methods to improve second generation bacterins intended to aid in the prevention of egg contamination specifically and organ invasion by any salmonellae in general. 2. How serious is the problem? Why does it matter? Salmonella enteritidis has emerged within the last fifteen years to become the leading cause of salmonellosis around the world, with overall incidence of human illness increased from 5 to 40 fold depending upon the country. The most frequently identified food source for this pathogen is the hen's egg, and wherever isolation of this organism from eggs has increased, so has human salmonellosis. While the United States does not yet have a problem of epidemic proportions, with risk of consuming a contaminated egg estimated at about 1 in 5,000 to 10,000 eggs consumed, the European problem is at least 10 times worse, with risk of eating a contaminated egg estimated at between 1 in 100 (Germany) to 1 in 700 eggs consumed (United Kingdom). Since incidence of infection with S. enteritidis in Central America, South America and Pacific Rim countries is also estimated to be much worse than within the USA, the potential for eggs produced in this country to become contaminated at epidemic levels is ever present. Infection of people by S. enteritidis is indeed a serious problem for USDA due to its association with poultry. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? This research allowed scientists to develop novel methods for increasing the efficacy of bacterins against Salmonella enteritidis. This project ended 9/30/98. 4. What were the most significant accomplishments this past year? A patent award was received on a special medium and associated intellectual property that describes enhanced recovery of proteins from Salmonella enteritidis that produces Typhi-like lipopolysaccharide. This patented medium in general supports high cell density growth and cellular differentiation of the salmonellae. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact? . The actual impact of this research is that it broadens our knowledge about how Salmonella enteritidis became a world wide problem. The predicted impact is that the information can be used to understand how sustained epidemics (greater than 10 years) due to bacterial pathogens come into existence. Specific accomplishments are as follows: 1) A strain of Salmonella enteritidis was developed that efficiently contaminates eggs and that greatly improved our ability to assess the efficacy of vaccines, both killed and modified live products. This new strain produces a plethora of cell-surface proteins and grows to cell densities two logs higher than the parent strain. Surprisingly, use of this strain in animals also indicated that parenteral adaptation is required to achieve efficient egg contamination. This finding confirmed previous results that indicated an orally invasive phenotype was required to be mixed with parenterally adapted phenotypes in order to achieve egg contamination following oral infection. 2) Collaboration with Dr. A.C. Matin of Stanford University helped expose yet another problem associated with production of bacterins. Dr. Matin grew a wildtype attenuated strain of Salmonella enteritidis in continuous culture, but found he could get good protein production only in diluted medium. Since economical production of bacterins requires growth of bacteria to high cell yields, this collaboration confirmed that SE-HCD, which can produce lots of cell-surface proteins at high-cell-densities in rich broth, was a crucial development. Dr. Matin has been asked for a final report on this collaborative research, which is still pending. 3) At least 3 manuscripts were published that described findings from these studies as they were pursued for their relevance to general objectives of CRIS 6612-32000-017. One of these papers describes the intellectual property associated with the patented medium. Two others describe the recovery from farms of invasive Enteritidis that grows to high cell density and the unique growth properties of a genetically modified strain of Salmonella enteritidis that grows to high-cell-density (SE-HCD). 4) Use of the patented growth medium revealed that serotyping can be used to do more than just name salmonellae according to the LPS O-chain and flagellar antigens they produce. It appears that the patented medium produces unexpected changes in serotype that can be used to assess invasive potential and important aspects of epidemiology. 5) The patented medium also detects metabolites present in hen houses that can cause cells to undergo differentiation into potentially more invasive forms. New lines of research with relevance to major CRIS objectives are originating from this use. 6. What do you expect to accomplish, year by year, over the next 3 years? This proj. has ended. However, we are now in a new stage of applying these findings. We will be growing SE-HCD in continuous culture to begin defining the exact conditions for maximizing both cell-surface protein and cell yields. These two factors are crucial to producing pilot vaccines with enhanced efficacy that can be produced en masse. 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 & durability of the technology product? The patented medium is now available for non-exclusive licensing. The technology is available now. One major constraint to the adoption of the technology is a lack of understanding of the new information and how it addresses practical issues. Another major constraint is that this research has been dependent upon the efforts of one scientist who has no authority to scientific support staff to build upon this research. To educate USDA staff and other scientists about the impact this new information could have on issues of food safety, ARS News is planning an article on how bacterial cells communicate when they grow to high cell density, how high cell density growth changes gene regulation, and how novel research conducted at SEPRL provides the first clear evidence that these factors are important to improving food safety. 8. List your most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below) . Presentations: Approved Patent: "A complex growth supplement for maintenance of bacterial cell viability and induction of bacterial cell differentiation." Uses of the medium include improving bacterins and refining diagnostic serotyping to detect pandemic strains of S. enterica ser Enteritidis. Jean Guard-Petter. 1998. Variants of smooth Salmonella enterica serovar Enteritidis that grow to higher cell density than the wild type are more virulent. Appl. Environ. Microbiol. 64:2166-2172. American Soc. for Microbiology (ASM), S.E. Branch Meeting, Montgomery, Alabama. Symposium on emerging human disease: The specialized virulence of Salmonella enterica serovar Enteritidis. Oct. 1998.
PUBLICATIONS: 1998/01 TO 1998/09
HOGUE, A., WHITE, P., GUARD-PETTER, J., SCHLOSSER, W., GAST, R.K., et al.
1998. Invited Review Article: Epidemiology and control of egg-associated S.E. in the U.S. Rev. Sci. et Tech., Intnl Office of Epizootics 16:542-553.
ACCESSION NO: 0161480 SUBFILE: CRIS
PROJ NO: IND073058V AGENCY: CSREES IND
PROJ TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START: 01 OCT 1993 TERM: 30 SEP 1998 FY: 1998
INVESTIGATOR: PORTER, R. E.; BOWERSOCK, T. L.
PERFORMING INSTITUTION:
VET SCIENCE & ANIMAL DISEASE DIAGNOSTIC LAB
PURDUE UNIVERSITY
WEST LAFAYETTE, INDIANA 47907
ORAL VACCINATION OF CHICKENS WITH HYDROGELS TO PROMOTE RESISTANCE TO ENTERIC SALMONELLA ENTERITIDIS
OBJECTIVES: To stimulate systemic and mucosal immunity in chickens by oral vaccination with hydrogels; to determine whether a defined oral vaccination regimen with SE flagellin-hydrogels can promote resistance of the intestine to colonization and invasion by Salmonella enteritidis.
APPROACH: Hydrogels containing flagellin of Salmonella enteritidis will be orally administered to white leghorn chickens. The dose regimen will be varied to determine what regimen promotes the greatest humoral, mucosal, and cellular immune responses. Alimentary fluid and serum will be analyzed for IgA and IgG anti-SE activity by ELISA. Cell-mediated immunity will be measured by delayed-type hypersensitivity of the toeweb to intradermal injection of purified protein subunits of SE, such as flagellin or fimbriae. In an additional study, a defined oral vaccination regimen of hydrogels containing flagellin will be administered to chickens. Two weeks later the chickens will be infected with Salmonella enteritidis phage type 13. At one week after infection chickens will be euthanized and cecum, colon, liver, and spleen will be cultured for Salmonella enteritidis to determine the effect of the vaccine.
PROGRESS: 1993/10 TO 1998/09
Vaccination of poultry against various infectious disease etiologies using parenteral routes is cumbersome and costly because it involves handling the birds during the vaccination process. This can be associated with significant stress which may compromise the immune status of the birds. We have started the evaluation of the Alginate hydrogels as carrier particles for Salmonella enteritidis proteins that may offer protection to the hens against natural infection with these organisms. We evaluated several lectins from: Arachis hypogea, Conavalia ensiformis, Lens culinaris, Ricinus communus, Triticum esculentum (wheat germ lectin) and Triticum vulgaris. These lectins were conjugated with Fluoresin isothiocyanate before oral administration into the White Leghorn hens. Results of these experiment suggested that there are specific pattern of attachment and engulfemnet of these lectins by the different parts of the alimentary tract of the hens. These results suggested the heterogenous composition of the villus epithelium of the alimentary tract in chickens. Wheat germ lectin was found to be the best stimulant when used to coat the Hydrogel particles uptake by the intestinal and liver and spleen tissues of the hens. The mean diameter of less that 5 microns appeared to be optimal in the tissue uptake of these microspheres.
PUBLICATIONS: 1993/10 TO 1998/09
1. T. L. Bowersock, K. Park, R.E. Porter, Jr. 1997. Patent No. 08-394, 802 awarded 9/97. Alginate based vaccine composition
2. R. E. Porter, Jr, N.P. Macri, T.L. Bowersock. 1997. Uptake of aliginate microspheres by the intestinal tract of white leghorn chickens. Proceedings of the North Central Avian Disease Conference, Des Moines, IA., p.11-12.
Termination Year 1997
ACCESSION NO: 0169155 SUBFILE: CRIS
PROJ NO: PENV-9503113 AGENCY: CSREES PENV
PROJ TYPE: NRI COMPETITIVE GRANT PROJ STATUS: TERMINATED
CONTRACT/GRANT/AGREEMENT NO: 95-37201-2007
START: 15 SEP 1995 TERM: 30 SEP 1997 FY: 1998 GRANT YR: 1995
INVESTIGATOR: KELLER, L. H.
PERFORMING INSTITUTION:
PATHOBIOLOGY
NEW BOLTON CENTER
KENNETT SQUARE, PENNSYLVANIA 19348
IMMUNE RESPONSE TO SALMONELLA ENTERITIDIS COLONIZATION OF 2 MHC-CONGENIC LINES OF CHICKENS
OBJECTIVES: This laboratory has previously identified two MHC-congenic lines of White Leghorn chickens that respond differentially to an oral challenge with S. enteritidis. The project will examine the role of the immune response in this differential susceptibility in 2 day old and 5 day old chicks from these lines. The study will also determine the effect of the MHC locus differences on the shed of S. enteritidis into the eggs of adult laying hens from these two lines of chickens, and the involvement of the immune response in that shed. Therefore, the objectives of this project are to: (1) define immune cell populations that respond to S. enteritidis infection of neonatal chicks from the MHC-congenic lines, G-B1 and G-B2. (2) Compare S. enteritidis tissue colonization and shed into the eggs between the adult G-B1 and G-B2 laying hens. (3) Define immune cell populations that relate to S. enteritidis colonization of tissues and eggs from adult G-B1 and G-B2 laying hens.
APPROACH: Objective 1 will be carried out by isolation of intestinal intra-epithelial or lamina propria lymphocytes, and lymphoid cells from the spleen, cecal tonsils from chicks infected with S. enteritidis at either 2 or 5 days of age. Subtype specificities of the immune cell populations responding to S. enteritidis challenge will be compared with the specificities of immune cell populations in naive chickens by flow cytometric analysis after staining isolated lymphoid cells with avian immune-cell-specific monoclonal antibodies. Tissue samples will be taken 24, 72, 120 and 144 hours post-infection (pi). Objective 2 will be accomplished through bacterial culture for S. enteritidis isolation of tissues from the liver, spleen, intestine, ovary and oviduct, including eggs both pre-and post-oviposition, from infected and un-infected adult G-B1 and G-B2 laying hens. Objective 3 will utilize adult G-B1 and G-B2 laying hens, who will be inoculated with S. enteritidis at 24 weeks of age to compare immune cell populations that may relate to a differential shed of the pathogen into eggs. The same methods as described under objective 1 will be employed except the immune cell populations will also be identified from the oviduct and the cloaca of these hens.
Termination Year 1996
ACCESSION NO: 0174233 SUBFILE: CRIS
PROJ NO: PENV-5-24358 AGENCY: CSVM PENV
PROJ TYPE: STATE PROJ STATUS: NEW
START: 01 JAN 1996 TERM: 31 DEC 1996 FY: 1998
INVESTIGATOR: KELLER, L.
PERFORMING INSTITUTION:
SCHOOL OF VETERINARY MEDICINE
UNIV OF PENNSYLVANIA
PHILADELPHIA, PENNSYLVANIA 19104
SHED OF S. ENTERITIDIS INTO THE EGGS OF SUSCEPTIBLE AND RESISTANCE CHICKENS
OBJECTIVES: To compare the shed of S. enteritidis into the eggs of adult hens from two MHC-congenic lines of chickens with differential susceptibility to the pathogen and to monitor antibody titers and examine phagocytic cell functions that may relate to the differential response to S. enteritidis of the two MHC-congenic lines of chickens.
APPROACH: One-hundred twenty chickens from lines G-B1 and G-B2 will be used. At 3 days of age the chicks will be orally inoculated with organisms of the invasive strain of 575 S. enteritidis. Males will be culled within 10 weeks. As the matured hens begin to lay, their eggs will be collected daily. The eggs will be tested by bacteriology for the presence of S. enteritidis contamination for up to 12 months. Serum blood samples will be taken from the chickens every week for 1 month post-inoculation and every 2 weeks thereafter to test for anti-Salmonella antibody titers. Samples of egg yolks will be taken from each freshly laid egg as it is cracked for culturing. Yolk samples will be pooled in groups of 10 for ELISA testing.
ACCESSION NO: 0165256 SUBFILE: CRIS
PROJ NO: LAB03087 AGENCY: SAES LA.B
PROJ TYPE: STATE PROJ STATUS: TERMINATED
START: 01 JUL 1994 TERM: 31 DEC 1996 FY: 1997
INVESTIGATOR: LOE, L. C.; INGRAM, D. R.
PERFORMING INSTITUTION:
CALHOUN RESEARCH STATION
LOUISIANA STATE UNIVERSITY
BATON ROUGE, LOUISIANA 70893
EVALUATION OF MOLTING PROGRAMS FOR LAYING HENS
OBJECTIVES: Evaluate the effectiveness of molting programs on subsequent egg production and egg shell quality in laying hens.
APPROACH: Six molting treatments will be examined: 1) control (no treatment); 2)10-day fast; 3) 4-day fast; 4) feeding 150,000 ppm zinc for 10 days; 5) feeding 150,000 ppm zinc for 4 days; 6) fasting hens until they achieve a 30% body weight loss. 240, 72-week old hens will be subjected to 7 days of 24 hour photostimulation prior to the start of the study. Egg production, egg specific gravity, egg weight, feed consumption, and livability will be measured during the treatment period and for 9, 28-day periods.
PROGRESS: 1994/01 TO 1996/12
Feed withdrawal (FW) and .15% dietary zinc (ZN) were evaluated for inducing molting laying hens. Six treatments (trt) included: 1) control- no FW or ZN (CON); 2) 4-day FW (4FW); 3) 10-day FW (10FW); 4) 4-day ZN (4ZN); 5) 10-day ZN (10ZN); and 6) feed withdrawal until a 30% loss in body weight was achieved 14-18 days (30BW). Each trt was applied to ten, individually cages hens, 74 wks old. The experiment had 4 replicates. Seven days prior to trt application, photoperiod was increased from 18L:6D to 24L:OD. Photoperiod was reduced to 17L:7D at trt initiation and was maintained at this level for the duration of the study. During the molt period, ZN trt proved inhibitory to feed consumption. No differences between FW and ZN trt of the same length were noted for days to pause, number of shell-less eggs or time to 50% production. Increasing length of molt trt increased hen weight loss and days to 50% production. Cumulative feed consumption per hen (36 wk post trt) was affected in a like manner. Hen weight at the end of the study was the same. No differences in cumulative production indices were found (egg production, egg weight, egg specific gravity, and feed efficiency) although initial trt effects were noted.
PUBLICATIONS: 1994/01 TO 1996/12
Loe, Linda C. and LeRon Robbins. 1996. Evaluation of dietary zinc or fasting in molting laying hens. Louisiana Academy of Sciences 70th Annual Meeting. (Poster presentation).
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October 23, 2002