Progress 10/01/08 to 09/30/13
Outputs
Target Audience: Animal and Poultry Science community Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest? Through publications in scientific journals and presentations at national conferences. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The interferon-regulated protein Mx has been identified in all species and has been reported to have anti-viral properties in most; however the antiviral activity of the chicken Mx protein is still unclear. Studies have indicated that a specific variant within exon 14 of this gene (amino acid S631N) modulates resistance to avian influenza virus infection. Sequencing of genomic DNA from 9 elite egg layer lines identified 29 SNP variants within the coding region and upstream 5’UTR and promoter region. Focusing on just the non-synonymous changes, these SNPs result in 12 haplotypes, yielding 12 different Mx proteins, with only 5 of these encoding the putative resistant N631 variant. Each elite line contained from 1 to 4 haplotypes, with many of these haplotypes being found in only one line. Intriguingly, there were significant changes in haplotype frequency in 6 of the 8 lines that were segregating, suggesting some unknown selective advantage for certain haplotypes. Previous studies in our laboratory using the direct-fed microbial (DFM) PrimaLac in broiler chicks have demonstrated an increase in thymus tissue respiration and a decrease in intestinal respiration following DFM supplementation, with no differences in whole-body respiration or growth. Additionally, we have previously reported an increased utilization of ATP in peripheral blood mononuclear cells isolated from DFM-fed animals, faster antigen specific antibody response, and differential expression of pro-inflammatory cytokines in various regions of the intestine following DFM supplementation. We have continued to develop methods and tools to better understand the biochemical signals responsible for the DFM affects on circulating PBMC energy utilization. Recent, preliminary studies suggest the production of some as yet unidentified soluble factor in circulation regulates the leukocyte activity. Studies are continuing to isolate and characterize this factor or factors.
Publications
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2013
Citation:
A. L. Ballou, R. Qiu, R. A. Ali, W. J. Croom, and M. D. Koci. Direct fed microbial supplementation affects host immune function and energy consumption. April 2013. Experimental Biology 2013. Boston, MA.
- Type:
Book Chapters
Status:
Other
Year Published:
2013
Citation:
S. Guix, N. Krishna, and M. D. Koci. Astrovirus Immunity . Astrovirus Research: Essential Ideas, Everyday Impacts, Future Directions. Ed S. Schultz-Cherry and E. Mendez. Springer Science. New York. 2013.
- Type:
Book Chapters
Status:
Other
Year Published:
2013
Citation:
M. Pantin-Jackwood, D. Todd, and M. D. Koci. Avian Astroviruses . Astrovirus Research: Essential Ideas, Everyday Impacts, Future Directions. Ed S. Schultz-Cherry and E. Mendez. Springer Science. New York. 2013.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2013
Citation:
J.E. Fulton, J. Arango, M. Koci, and C.M. Ashwell. Variation in the MX gene in commercial egg layer elite lines. September 2013. 8th European Symposium on Poultry Genetics. Venice, Italy.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2013
Citation:
W. J. Croom, C. W. Nash, M. Koci, B. W. McBride, B. Clemmons and J. T. Brake. Increasing growth in broilers by serotonin modulation: The effects of selective serotonin reuptake inhibitors. September 2013. Innovate 2013. Braselton, GA.
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Progress 10/01/12 to 09/30/13
Outputs
Target Audience: Animal scientists and allied industry Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Studies have been conducted to develop new methods for assessing the energetic cost of the immune response and understanding how changes in the microbiota affects immune function.
Publications
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Progress 10/01/11 to 09/30/12
Outputs
OUTPUTS: Our laboratory has worked for the past several years to understand the pathogenesis and host response the type-2 turkey astrovirus (TAstV-2). Previous studies in our laboratory have demonstrated TAstV-2 induces an acute diarrhea in turkey poults, and that this diarrhea is associated with changes in the apical expression of sodium/hydrogen exchanger-3 (NHE3). In more recent studies, and in collaboration with the Schultz-Cherry laboratory at St. Jude Children's Research Hospital, we have begun to investigate the specific elements of the TAstV-2 virus involved in the induction of diarrhea. In these studies we have determined that oral inoculation of turkey poults with recombinantly expressed TAstV-2 capsid protein is sufficient to induce diarrhea, changes in intestinal barrier function, and NHE3 activity. PARTICIPANTS: Stacey Schultz-Cherry, PhD St. Jude Children's Research Hospital TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
These findings are the first to our knowledge to demonstrate that a viral structural protein is all that is required to induce diarrhea. These findings provide a better understanding of how astroviruses induce disease in poultry, specifically turkeys, and are a major first step towards identifying cellular receptors which bind the capsid protein, and then activate cellular mechanisms which result in clinical signs. These findings are also very interesting in the fact that treatment with the recombinant capsid protein resulted in diarrhea, and induced physiological changes in both the ileum and jejunum (data not shown) however only ileum was observed to have a reduced barrier function. This suggests that while both regions of the intestine express similar receptors (both regions support virus replication) there are differences in the cellular physiology of the epithelial cells of the ileum and jejunum. A better understanding of these differences will likely be very important to our understanding of the pathological mechanisms and resistance to various enteric pathogens.
Publications
- R. R. Meyerhoff, P. K. Nighot, R. A. Ali, A. T. Blikslager, and M. D. Koci. Characterization of turkey inducible nitric oxide synthase and identification of its expression in the intestinal epithelium following astrovirus infection. Comparative Immunology, Microbiology, and Infectious Diseases. 35:63-9, 2012.
- R. R. Meyerhoff, R. A. Ali, K. Liu, G. Q. Huang, M. D. Koci. Comprehensive analysis of commercially available mouse antichicken monoclonal antibodies for cross-reactivity with peripheral blood leukocytes from commercial turkeys. Poultry Science. 91:383-92, 2012.
- Regulation of the immune response through energy. What role does the microbiota play 2012 Eastern Nutrition Conference. Kitchener, Ontario, Canada. May 2012.
- Direct fed microbial supplementation in poultry results in changes in both mucosal and systemic immune activity. 2012 Eastern Nutrition Conference. Kitchener, Ontario, Canada. May 2012.
- S. Guix, N. Krishna, and M. D. Koci. Astrovirus Immunity. Astroviruses. Ed S. Schultz-Cherry and E. Mendez. Springer Science. (In Press) 2012.
- P. Nighot, R. Ali, L. A. Moser, P. Freiden, S. Schultz-Cherry, R. DuBois, A. Blikslager, and M. D. Koci. Turkey astrovirus capsid protein induces electrophysiological changes in the intestinal epithelium. August 2012. The 2012 American Association of Avian Pathologist/American Veterinary Medical Association Meeting. San Diego, CA.
- A. G. Seal, T. Helmy, R. Ali, E. Akers, M. Mendoza, E. Stafford, H. Hassan, and M. D. Koci. Development of a Salmonella specific ELISA to test the efficacy of a novel Salmonella vaccine. August 2012. 11th Annual NC State University Summer Undergraduate Research Symposium. Raleigh, NC.
- Y. T. Mak, M. Mendoza, T. Helmy, R. Ali, M. D. Koci, and H. Hassan. Efficacy of Salmonella detection in poultry: Evaluating different sampling methods to minimize number of animal use. August 2012. 11th Annual NC State University Summer Undergraduate Research Symposium. Raleigh, NC.
- J. Croom, C. Nash, M. D. Koci, J. Wock, A. Ballou, D. Carver, and J. Brake. Fluoxetine increases growth and feed efficiency in broilers. July 2012. Annual Meeting of the Poultry Science Association. Athens, GA.
- M. Pantin-Jackwood, D. Todd, and M. D. Koci. Avian Astroviruses. Astroviruses. Ed S. Schultz-Cherry and E. Mendez. Springer Science. (In Press) 2012.
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Progress 10/01/10 to 09/30/11
Outputs
OUTPUTS: 1.Identify and characterize genes and their relationships to disease resistance in poultry with an emphasis on the major histocompatibility complex as well as other genes encoding alloantigens, communication molecules and their receptors and other candidate systems. The NCSU laboratory will investigate innate immune gene expression following viral infection. Studies will examine in vitro infections of primary cells from distinct genetic lineages and differences in the innate gene response kinetics and profiles following infection to different viral families. Additional studies will examine the localized expression of innate antiviral genes of the poult intestine following enteric viral infection. 3.Develop, evaluate and characterize methodologies, reagents and genotypes to assess immune function and disease resistance to enhance production efficiency through genetic selection in poultry. Turkey-specific assays and reagents which include recombinant protein, antibodies, and molecular reagents for real-time RT-PCR assays to assess cytokine expression and immune function will be developed by NC. Nucleotide analysis of chicken cytokine genes available in the GeneBank database will be used to design degenerate primers to amplify homologous genes from turkey mRNA. Resulting RT-PCR product will be confirmed by sequence analysis. Complete gene sequence will be used to design specific diagnostic tests PARTICIPANTS: Collaborators: Chris Ashwell, Jim Croom, Hosni Hassan, Anthony Blikslager TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Identification and characterization of turkey iNOS and its expression in the intestinal epithelium following infection with turkey astrovirus type-2 (TAstV-2). These results help to establish the utility of the anti-tkiNOS Ab in studies to understand the role of iNOS in the response to infection in turkeys. In addition, these results are the first, to our knowledge, to specifically demonstrate the expression of iNOS in the intestinal epithelium following infection with astrovirus. Characterization of turkey CD8α gene. These data further characterize the turkey CD8α gene, and provide evidence for the potential level of allelic variations. Recombinant expression method for characterizing cross-species activity of anti-leukocyte monoclonal Abs. The development of this approach will allow for the systematic analysis and confirmation of cross-reactivity of other mAbs, and potentially other avian species. Additionally, this approach will be useful in assessing how allelic variation affects the ability of a specific mAbs to bind a given antigen. Recombinant cell line for the detection of avian CXCL8 (IL-8) bioactivity. CXCL8 is a very important chemokine whose expression is upregulated in the very early stages of most innate immune responses. The development of this assay will allow for the detection of changes in CXCL8 bioactivity in chicken and turkey (potentially other avian species) cells, tissues, or serum allowing for a more detailed understanding of its role in initiating and directing the overall immune response.
Publications
- R. Meyerhoff, R. A. Ali, K. Liu, G-Q. Huang, and M. D. Koci. Comprehensive Analysis of Commercially Available Mouse Anti-Chicken Monoclonal Antibodies for Cross-Reactivity with Peripheral Blood Leukocytes from Commercial Turkeys. Poultry Science. 2012 91(2):383-392.
- R. Qiu, J. Croom, R. A. Ali, C. Ashwell, H. M. Hassan, C. C. Chiang, and M. D. Koci. Direct Fed Microbial Supplementation Repartitions Host Energy to the Immune System. Journal of Animal Science. In Press. 2012.
- R. R. Meyerhoff, P. K. Nighot, R. A. Ali, A. T. Blikslager, and M. D. Koci. Characterization of Turkey Inducible Nitric Oxide Synthase and Identification of its Expression in the Intestinal Epithelium Following Astrovirus Infection. Comparative Immunology, Microbiology, and Infectious Diseases. 2012 35(1):63-9.
- N. M. Leandro, R. Ali, M. Koci, V. Moraes, R. Malheiros, M. J. Wineland, and E. O. Oviedo-Rondon. Effects of Broiler Breeder Genetics, Diet Type, and Feeding Program on Maternal Antibody Transfer and Development of Lymphoid Tissues. Journal of Applied Poultry Research. 20:474-484. 2011.
- N. M. Leandro, R. Ali, M. Koci, V. Moraes, P. E. Eusebio-Balcazar, J. Jornigan, R. D. Malheiros, M. J. Wineland, J. Brake, and E. O. Oviedo- Rondon. Maternal Antibody Transfer to Broiler Progeny Varies Among Strains and Is Affected by Grain Source and Cage-Density. Poultry Science. 90:2730-2739. 2011.
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Progress 10/01/09 to 09/30/10
Outputs
OUTPUTS: Development of methods to assess how changes in intestinal flora affect the energy requirements of the chicken immune system Development of turkey specific anti-iNOS antibodies as well as the establishment of methods using these reagents to detect the expression of iNOS in fixed turkey tissues. Development of turkey specific anti-CXCL8 antibodies and expression systems to produce turkey CXCL8 recombinantly in bacteria cells. PARTICIPANTS: Chris Ashwell, NC State Anthony Blikslager, NC State Jim Croom, NC State Jesse Grimes, NC State TARGET AUDIENCES: Scientific community North Carolina Poultry Industry PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The reagents and methods developed above will be critical to our ability to assay how the host responds to disease challenge and how these changes affect animal growth and performance.
Publications
- P. K. Nighot, A. Moeser, R. A. Ali, A. T. Blikslager, and M. D. Koci. Astrovirus infection induces sodium malabsorption and redistributes sodium hydrogen exchanged expression. Virology. In press. 2010
- J. L. Grimes, M. D. Koci, C. R. Stark, D. P. Smith, P. K. Nighot, and T. Middleton. Biological Effect of Naturally Occurring Mycotoxins Fed to Poults Reared to 21 Days of Age. International Journal of Poultry Science. 9: 871-874, 2010.
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Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: Book Chapters. M. D. Koci and S. Schultz-Cherry. "Avastrovirus, Astroviridae". The Springer Index of Viruses. 2nd Edition. Ed C. Tidona and G. Darai. Springer Science. (In Press) 2009. Refereed Journals. J. Croom, M. Chichlowski, M. Froetschel, B. W. McBride, R. Qui and M. D. Koci. The effects of direct-fed microbial, Primalac, or salinomycin supplementation on intestinal lactate Isomers and cecal volatile fatty acid concentrations in broilers. International Journal of Poultry Science. 8: 128-132, 2009. Abstracts R. R. Meyerhoff, R. A. Ali, and M. D. Koci. Characterization of turkey iNOS and development of assays to study its role in poult enteritis. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. R. A. Ali, and M. D. Koci. Development of a CXCL8 (IL-8) bioassay to determine its role in turkey poult viral enteritis. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. R. Qiu, J. Croom, G. Wu and M. D. Koci. Effects of the DFM Primalac on crop structure and intestinal development in broilers. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. P. K. Nighot, A. Moeser, R. A. Ali, A. T. Blikslager, and M. D. Koci. Astrovirus infection induces sodium malabsorption and redistributes sodium hydrogen exchanger expression. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. M. D. Koci, R. A. Ali, M. D. Quiles, E. Strain, C. M. Ashwell. Evidence of Mx independent genetic resistance to viral infection. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. R. Qiu, J. Croom, R. A. Ali, and M. D. Koci. The Effects of the DFM Primalac on energy repartitioning to the immune system of broilers. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. W. L. Richards, R. A. Ali, C. M. Ashwell, and M. D. Koci. Determination of the effect genetic polymorphisms have on subcellular localization of the chicken Mx protein. July 2009. Annual Meeting of the Poultry Science Association. Raleigh, NC. R. R. Meyerhoff, R. A. Ali, and M. D. Koci. Characterization of turkey iNOS and development of assays to study its role in poult enteritis. April 2009. The 18th Annual NC State University Undergraduate Research Symposium. Raleigh, NC. W. L. Richards, R. A. Ali, C. M. Ashwell, and M. D. Koci. Determination of the effect genetic polymorphisms have on subcellular localization of the chicken Mx protein. April 2009. The 18th Annual NC State University Undergraduate Research Symposium. Raleigh, NC. PARTICIPANTS: Chris M. Ashwell Dept of Poultry Science NC State Jim Croom Dept of Poultry Science NC State Anthony Blikslager Clinical Sciecnes NC State TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
OBJECTIVE 1. The majority of work involving the genetic basis of disease resistance, vaccine response, and antibody production has focused on the MHC, yet these are very complex processes involving many different pathways and responses to antigens. By utilizing this population we will be able to identify genes outside of the MHC that are involved in the response to antigen in the chicken with the goal of more clearly developing the mechanisms regulating antibody production and perhaps provide alternative markers for selection and breeding of stocks with desired traits. OBJECTIVE 2. The mechanism by which DFM supplementation augments host immunity is poorly understood. The results of these experiments suggest that DFM-mediated changes in energy allocation within the animal may play a key role in its immunomodulation activity and increase the ability of birds to respond to specific antigens. OBJECTIVE 3. The development of turkey specific iNOS reagents allows for the detection of iNOS protein in turkey samples. This will allow us to begin to assess the role of iNOS in the host response to infection.
Publications
- J. Croom, M. Chichlowski, M. Froetschel, B. W. McBride, R. Qui and M. D. Koci. The effects of direct-fed microbial, Primalac, or salinomycin supplementation on intestinal lactate Isomers and cecal volatile fatty acid concentrations in broilers. International Journal of Poultry Science. 8: 128-132, 2009.
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Progress 10/01/08 to 12/01/08
Outputs
OUTPUTS: This is a new multistate project which began 10/01/08. No outputs to report at this time. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
This is a new multistate project which began 10/01/08. No outputs to report at this time.
Publications
- No publications reported this period
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