NC228: Avian Respiratory Diseases: Pathogenesis, Surveillance, Diagnosis and Control

(Multistate Research Project)

Status: Inactive/Terminating

Project Menu

SAES-422 Reports

Annual/Termination Reports:

[01/21/2002] [03/06/2003] [12/10/2003] [02/28/2005]

Date of Annual Report: 01/21/2002

Report Information

Annual Meeting Dates: 11/09/2001 - 11/10/2001
Period the Report Covers: 10/01/2000 - 10/01/2001

Participants

AL AES V. S. Panangala

CT AES Report submitted by M. I. Khan

DE AES J. E. Dohms and J. Gelb, Jr.

IA AES D. L. Reynolds

IL AES D. N. Tripathy

IN AES C. C. Wu

MD AES Report submitted by V. N. Vakharia and R. A. Heckert

MN AES V. Kapur (report included work by D. N. Foster, S. M. Goyal,
D. A. Halvorson, K. V. Nagaraja, M. K. Njenga, S. Noll, and J. M. Sharma)

NC AES Report submitted by D. H. Ley

OH AES Y. M. Saif (report included work by D. J. Jackwood)

Administrative Advisor J. Klausner (U. Minnesota)

CSREES Representative W. Wagner (USDA)

Brief Summary of Minutes

The NC228 business meeting was held Nov. 9, 2001, in the Atrium C Room

of the Millennium Hotel, St. Louis, Missouri. Dr. Jack Gelb, Chair of

NC228 opened the meeting at 2 PM. Dr. Gelb will continue to serve as

Chair and Dr. Wu as Secretary for 2001-2002. For completion of the final

composite annual report, Dr. Gelb requested that each Station to submit the

report by objectives via e-mail. In keeping with the regional research mission,

cooperative activities among project members were to be highlighted. Dr. Klausner stated that the NE138 regional project had officially been merged

into the NC228 project. A subcommittee consisting of Drs. Kapur, Reynolds and Gelb will create a NC228 website that will post the membership list, project proposal, annual station and composite reports. Drs. Saif and Dohms agreed to identify and invite new members to the NC228 project. Drs. Saif and Kapur will finish a position paper on genotypes/serotypes and submit to the journal, Avian Diseases. Dr. Saif proposed that the NC228 co-sponsor a symposium on poultry respiratory diseases with the North Central Avian Diseases Committee for the 2003 meeting in Ohio. Dr. Wagner indicated that conference grants are available from the National Research Initiative. Dr. Wagner summarized federal legislative activities as follows; funding for FY 2002 has passed; the NRI received $120 million for FY ‘02; the Initiative for Future Agriculture and Food Systems was cancelled for FY ‘02; Formula, Hatch and Extension funding is the same as FY‘01; Animal Health and Disease funding is similar to last year (~$5 million); indirect cost allowable charges to 35% are under consideration. The business meeting concluded at 4 PM. The remainder of the meeting was devoted to presentation and discussion of the station reports. The meeting journed Nov. 10 at 3:30 PM.

Minutes Attachment:

Attachment

Accomplishments

Objective 1. Determine the pathogenesis and interactions of specific agents. <o:p></o:p> Alabama determined that M. gallisepticum sequences both upstream and downstream of the GAA repeats in the promoter region of the pMGA gene are important for regulation and that the number of repeats affects expression by altering the spacing between the flanking sequences. They have hypothesized that a hemagglutinin-activator protein (HAP) binds to the GAA repeat region and stimulates pMGA gene transcription if and only if 12 copies of the GAA repeat are present. <o:p></o:p> Delaware. Vaccination of 1-day-old SPF chicks with Marek‘s disease virus (MDV) vaccines HVT + SB1 is highly effective against very virulent plus MDV (584A) challenge and did not suppress immunity to infectious bronchitis virus (IBV) vaccination at 1 and 14 days of age. Dr. Robin Morgan contributed to this work. <o:p></o:p> Delaware. Continued work on the molecular pathogenesis of Mycoplasma gallisepticum, resulting in further characterization of the cytadhesin operon consisting of mgc1, mgc2, mgc3, and mgc4. MGc3 was surface-exposed as determined by protease digestion and partitions into the insoluble triton shell suggesting that it is also associated with the cytoskeleton, a structure involved in cell division, gliding motility, and bleb organization. A tn916 transposon library containing approximately 700 MG mutants was prepared by optimizing the plasmid (pAM120) DNA and the percent PEG concentrations. A summary of all the insertions will soon be available at http://udgenome.ags.udel.edu/mgall/mutants/html. <o:p></o:p> Illinois. A-type inclusion body protein was found not to be essential for replication of fowlpox virus in cultured avian cells. Replication of photolyase-deficient fowlpox virus in the chicken, was impaired when compared to that of the parent virus, but still was immunogenic and protective and thus may be a vaccine candidate. Insertional inactivation of a hemagglutinin (HA)-like gene homologue in fowlpox virus had no effect on the growth kinetics of the recombinant and parent virus. Previously unrecognized condorpox virus was determined to be a pox virus based on EM yet was biologically, antigenically, and genetically distinct from fowlpox virus and thus may represent a new species of the avipoxvirus genus. <o:p></o:p> Iowa. The Colorado strain of avian pneumovirus (APV) was found to be a mild/low virulent pathogen for egg-type laying chickens based on clinical respiratory signs and egg production. <o:p></o:p> Minnesota. Transmission of APV was demonstrated via vertical an airborne roues. <o:p></o:p> Minnesota completed the genomic sequence of Pasteurella multocida, the first major veterinary bacterial pathogen sequenced in its entirety. The filamentous hemagglutinin protein homolog, which is expressed in virulent but not avirulent isolates and whose inactivation results in more than a million-fold reduction in ability to cause mortality, was identified. <o:p></o:p> Ohio. Serotype 2 infectious bursal disease virus (IBDV) that was pathogenic for chicken embryos was found to be infectious, but not pathogenic in chickens or turkeys. <o:p></o:p> Ohio. Uncomplicated APV infection did not persist in birds for a long period and was usually limited to the respiratory tract. APV was shown to have limited transmissibility. <o:p></o:p> Objective 2. Surveillance, occurrence and consequences of agents and host variation on disease susceptibility <o:p></o:p> Minnesota. Wild passerine birds were found to be sero- and virus-positive for APV. Infectious virus persisted on premises inhabited by APV inoculated birds for a period of up to 6 weeks after initial exposure. APV infection increased T and B cell infiltration into the gland of Harder, upregulated Type I and II interferon synthesis, and reduced splenic T-cell responsiveness. Disease in young poults was more severe and associated with reduced T-cell function compared to older birds. Microarray studies showed that lymphoid tissues undergo altered gene expression in response to APV infection. Transcripts representing proteins involved in the activation of T cells, T- and B-cell signal transduction and efficient antigen processing and presentation were found to be upregulated. <o:p></o:p> North Carolina. The random amplification of polymorphic DNA (RAPD) fingerprint database consisting of all Mycoplasma gallisepticum (MG) reference strains and more than 200 poultry isolates, plus ~100 songbirds isolates has been completed. The application of a computer-assisted analysis system using Gene Profiler (Scanalytics, Inc., version 4.0) is ongoing. Since January 2001, MG was isolated from only one broiler breeder and two turkey farms in the state. Each isolate was a new and unique RAPD type, which suggested that they were introduced to the farms from separate external sources (evidence suggests backyard flocks), and there was no evidence of transmission to or from other commercial poultry. <o:p></o:p> Ohio. Selection for increased body weight in turkeys was associated with increased mortality following vaccination with live LaSota Newcastle disease virus (NDV) vaccine. Objective 3. Develop new and improved methods for the diagnosis, prevention, and control of avian respiratory diseases. Connecticut, having developed IBV DNA vaccines for chicks, is now extending the application to in ovo administration. Future work will focus on in ovo DNA and viral vector (fowlpox) vaccination and protection studies using vaccines containing the IBV surface glycoprotein S and S1 genes. Delaware. Following live IBV vaccine priming, inactivated oil emulsion IBV vaccination containing the PA/Wolgemuth/98 strain provided better protection than a heterologous inactivated vaccine containing Massachusetts + Arkansas strains based upon challenge with the virulent nephropathogenic strain, PA/Wolgemuth/98. Dr. Conrad Pope contributed to this study. Illinois. Monoclonal antibodies against fowlpox virus (FPV) were used to differentiate between vaccine and field strains of fowlpox virus and may be used for diagnosis. Illinois and Delaware. A recombinant (r) infectious laryngotracheitis virus (ILTV) lacking the ability to produce glycoprotein C (gC) was generated to circumvent possible reversion to a more virulent form and also to "genetically mark" a vaccine virus. The vaccine potential of the rILTV is being evaluated by Dr. Calvin Keeler, Indiana. Three weekly DNA vaccinations of chickens using plasmids containing various fragments of large segment genome (VP2) of serotype 1 IBDV protected chickens against clinical signs and mortality following virulent challenge. Iowa. Anti NDV specific immunoglobulin administered subsequent to virulent NDV challenge protected chickens against clinical ND if the immunoglobulin was given prior to the onset of clinical signs. Maryland. Using the cRNA-based reverse genetics system developed for IBDV, residues involved in virulence and pathogenesis were identified that could lead to the development of a marked, attenuated, multi-spectrum vaccine against IBDV. Maryland. Dermal DNA vaccination of chickens using the HN gene of NDV and the VP2 of IBDV induced IgG, IgM, IgA antibody responses to the viruses in serum and tears. Plasmid DNA persisted in kidney, bone marrow and muscle. DNA vaccination in ovo using combinations of plasmid DNA, neutral lipid and DMSO resulted in gene expression in liver and muscle. Production of insert-specific mRNA and protein in a variety of tissues was evident, resulting an immune response that can provide partial protection from viral challenge with IBDV. Minnesota. Turkey turbinate and kidney cells were successfully life-span-extended to produce higher APV titers for the purposes of vaccine production. APV sequencing lead to the development of RT-PCR and Taqman diagnostic tests. ELISA assays for APV were developed using recombinant viral M and N proteins. APV isolates from the upper Midwest were sequenced and the N, P, M, F, and M2 genes were analyzed. Nucleotide substitution rates across provided evidence that only a single clone of APV exists and is widely disseminated. Formalin and UV-inactivated APV vaccines were determined to be safe but ineffective whilst a live-attenuated passage 63 vaccine was both safe and effective. In ovo vaccination did not adversely affect hatchability or livability of the hatched poults and induced earlier and longer lasting protection compared to post-hatch vaccination. Minnesota. A temperature-sensitive (Ts) mutant of the respiratory pathogen Ornithobacterium rhinotracheale (ORT) was shown to elicit an immune response in turkeys vaccinated via the drinking water and resulted in reduced clinical signs, gross lesions, and re-isolation rates following experimental challenge with pathogenic ORT. Ohio. Single and multiple nucleotide polymorphisms of IBDV were detected using real-time RT-PCR. Analysis revealed the presence of multiple genetic sub-populations of virus within a vaccine. Studies will be conducted to determine if plaque purification creates single and stable genetic viral populations. Real-time RT-PCR will be used to examine vaccines and laboratory samples for polymorphisms encoding major neutralizing epitiopes. A genetic marker for wild-type potentially pathogenic IBDV strains was identified making it possible to differentiate between strains that are the potential cause of the disease and those viruses that are attenuated. Monoclonal antibodies to the very virulent IBDV will be developed.

Publications

Abdel-Alim, G.A. and Y. M. Saif: Immunogenicity and antigenicity of very virulent strains of infectious bursal disease viruses. Avian Dis. 45:92-101, 2001. <o:p></o:p> Alkhalaf, A.N., R.N. Dearth, and Y.M. Saif.  Pathogenicity, infectivity, and tissue distribution of avian pneumovirus in turkey poults. Proc. 52nd North Central Avian Dis. Conf., p. 38, 2001. <o:p></o:p> Bautista DA, Elankumaran S, Heckert RA, Oshop GL, Moura LC, Wilson JD. Interaction of Salmonella typhimurium and infectious bursal disease virus (IBDV) in broiler chickens. Proc. 138th Am. Vet. Med. Assn. Mtg, Boston, MA, July 14-18, 2001. <o:p></o:p> Brandt, M., Yao, K., Liu, M., Heckert, R.A., and Vakharia,V.N. Molecular determinants of virulence, cell tropism and pathogenic phenotype of infectious bursal disease.  J. Virol.  74. In press. <o:p></o:p> Chang, H. C., Lin, T. L., and C. C. Wu. DNA-mediated vaccination against infectious bursal disease in chickens. Vaccine 20: 328-335, 2002. <o:p></o:p> Chiang, S., A. M. Dar, S. M. Goyal, M. A. Sheikh, J. C. Pedersen, B. Panigrahy, D. Senne, D. A. Halvorson, K. V. Nagaraja, and V. Kapur. A modified enzyme-linked immunosorbent assay for the detection of avian pneumovirus antibodies J Vet Diagn Invest. 12:381-4. 2000. <o:p></o:p> Dar, A. M., K. Tune, S. Munir, B. Panigrahy, S. M. Goyal, and V. Kapur. PCR-based detection of an emerging avian pneumovirus in US turkey flocks J Vet Diagn Invest. 13:201-5. 2001. <o:p></o:p> Dar, A. M., S. Munir, S. M. Goyal, M. S. Abrahamsen, and V. Kapur. Sequence analysis of the nucleocapsid and phosphoprotein genes of avian pneumoviruses circulating in the US Virus Res. 79:15-25. 2001. <o:p></o:p> Davison, S., A. F. Ziegler, J. Gelb, Jr., P. A. Dunn, and R. J. Eckroade. Infectious bronchitis in Pennsylvania. Proc. AAAP Symposium. Respiratory Diseases of Poultry. AVMA/AAAP Ann. Mtg. Boston, Massachusetts, July 14-18, 2001. <o:p></o:p> Elankumaran, S, Heckert, RA, Moura, L. Persistence and tissue distribution of a variant strain of infectious bursal disease virus in commercial broiler chickens. Proc. 2nd International symposium on infectious bursal diseases and chicken infectious anemia. Rauischholzhausen, Germany, June 2001. <o:p></o:p> Gelb, J., Jr.,  C. L. Keeler, B. S. Ladman, and B. F. Kingham. S1 sequence analysis; a tool for understanding IBV outbreaks.  Proc. AAAP Symposium. Respiratory Diseases of Poultry. AVMA/AAAP Ann. Mtg. Boston, Massachusetts, July 14-18, 2001. <o:p></o:p> Goyal, S. M., S. J. Chiang, A. M. Dar, K. V. Nagaraja, D. P. Shaw, D. A. Halvorson, and V. Kapur. Isolation of avian pneumovirus from an outbreak of respiratory illness in Minnesota turkeys J Vet Diagn Invest. 12:166-8. 2000. <o:p></o:p> Gulati, B. R., D. P. Patnayak, A. M. Sheikh, P. E. Poss, and S. M. Goyal. Protective efficacy of high-passage avian pneumovirus (APV/MN/turkey/1- a/97) in turkeys Avian Dis. 45:593-7. 2001. <o:p></o:p> Gulati, B. R., K. T. Cameron, B. S. Seal, S. M. Goyal, D. A. Halvorson, and M. K. Njenga. Development of a highly sensitive and specific enzyme-linked immunosorbent assay based on recombinant matrix protein for detection of avian pneumovirus antibodies J Clin Microbiol. 38:4010-4. 2000. <o:p></o:p> Gulati, B. R., S. Munir, D. P. Patnayak, S. M. Goyal, and V. Kapur. Detection of antibodies to US. isolates of avian pneumovirus by a recombinant nucleocapsid protein-based sandwich enzyme-linked immunosorbent assay J Clin Microbiol. 39:2967-70. 2001. <o:p></o:p> Hartup B. K., Kollias GV, Ley DH. Mycoplasmal conjunctivitis in songbirds from New York.  J. Wildlife Dis., 36:257-264.  2000. <o:p></o:p> Hartup BK, Bickal JM, Dhondt AA, Ley DH,  Kollias GV. Dynamics of conjunctivitis and Mycoplasma gallisepticum infections in house finches.  The Auk,118:327-333, 2001. <o:p></o:p> Jackwood, D. J.  Diagnosis of infectious bursal disease viruses using the RT/PCR-RFLP assay: Advantages, limitations and comparison to other molecular assays. Proc. of the Partnerships in Poultry Symposium.  Fort Dodge Animal Health. Paris, France. July 9-11. 2001. <o:p></o:p> Jackwood, D. J.  Genotypic and phenotypic diversity among wild-type IBDV strains. Proc. of the Partnerships in Poultry Symposium.  Fort Dodge Animal Health. Paris, France. July 9-11. 2001. <o:p></o:p> Jackwood, D. J.  Molecular diagnosis of infectious bursal disease viruses: Practical applications and significance of the results. Proc. of the Symposium on Molecular identification and epidemiology of avian pathogens. Amer. Assn. of Avian Pathol.. 137th Am. Vet. Med. Assn.. Mtg., Salt Lake City, Utah.  2000. <o:p></o:p> Jackwood, D. J.  Molecular diagnosis of infectious bursal disease viruses. Proc. of the Intervet Workshop on Immune Suppression and Emerging Diseases in Broilers.  Baltimore, Maryland. 2000. <o:p></o:p> Jackwood, D. J.  New molecular techniques for the diagnosis and control of infectious bursal disease virus.  Proc. of the 51st North Central Avian Dis. Conf., Columbus, Ohio. 2000. <o:p></o:p> Jackwood, D. J.  Standardization and quality control of diagnostic reverse transcription polymerase chain reaction (RT-PCR) assays: Detection and identification of single nucleotide polymorphisms in poultry pathogens using new fluorescence hybridization based technology.  Proc.of the X International Symposium of Vet. Laboratory Diagnosticians and OIE Seminar on Biotechnology. Salsomaggiore-Parma, Italy, July 4-7. 2001. <o:p></o:p> Jackwood, D. J. and S. Sommer.  Detection of single and multiple nucleotide polymorphisms in infectious bursal disease viruses using real-time RT/PCR.  Abstr. 138th AVMA Mtg. 2001. <o:p></o:p> Jackwood, D. J., E. H. Byerley and S. E. Sommer.  Molecular marker for attenuation in infectious bursal disease viruses.  Avian Dis.  45:701-705. 2001. <o:p></o:p> Jackwood, D. J., S. E. Sommer and H. V. Knoblich.  Amino acid comparison of infectious bursal disease viruses placed in the same or different molecular groups using RT/PCR-RFLP.  Avian Dis. 45:330-339. 2001. <o:p></o:p> Jirjis, F. E., S. L. Noll, D. A. Halvorson, K. V. Nagaraja, and D. P. Shaw. Immunohistochemical detection of avian pneumovirus in formalin-fixed tissues J Vet Diagn Invest. 13:13-6. 2001. <o:p></o:p> Jirjis, F. E., S. L. Noll, D. A. Halvorson, K. V. Nagaraja, E. L. Townsend, A. M. Sheikh, and D. P. Shaw. Avian pneumovirus infection in Minnesota turkeys: experimental reproduction of the disease Avian Dis. 44:222-6. 2000. <o:p></o:p> Khan, M. I. Avian Pathogenic Mycoplasmas. PCR detection of Microbial Pathogens. Methods in Molecular Biology. eds. J. Frey and K. Sachse. Humana Press Inc. Totowa, NJ. In press. <o:p></o:p> Khan, M. I. Avian respiratory tract infections and their control strategies. World Poultry Science Assn. in collaboration with Pakistan Poultry Assn., Islamabad, Pakistan, July 7, 2001. <o:p></o:p> Khan, M. I. Recombinant fowlpox virus containing the S1 gene of Massachusetts 41 strain of infectious bronchitis virus, Institute of Poultry Science, Shandong Academy of Agricultural Science, Jinan City, Shandong, Peoples Republic of China, March 26, 2001. <o:p></o:p> Khan, M., Wang, X., Schnitzlein, W. and Tripathy, D.N. A recombinant fowlpox virus containing IBV-S1 gene and its potential for a vaccine. Abst. AAAP/AVMA Scientific Program, Boston, MA. p. 39. 2001. <o:p></o:p> Khan. M. I. Are your turkeys healthy? 37th Annual New England Turkey Growers Conf., Sturbridge, Massachusetts. March 8, 2000. p. 9. <o:p></o:p> Kim, H., S. You, B. W. Kong, L. K. Foster, J. Farris, and D. N. Foster, Necrotic cell death by hydrogen peroxide in immortal DF-1 chicken embryo fibroblast cells expressing deregulated MnSOD and catalase Biochim Biophys Acta. 1540:137-46. 2001. <o:p></o:p> Kim, H., S. You, I. J. Kim, J. Farris, L. K. Foster, and D. N. Foster. Increased mitochondrial-encoded gene transcription in immortal DF-1 cells Exp Cell Res. 265:339-47. 2001. <o:p></o:p> Kim, H., S. You, I. J. Kim, L. K. Foster, J. Farris, S. Ambady, F. A. Ponce de Leon, and D. N. Foster. Alterations in p53 and E2F-1 function common to immortalized chicken embryo fibroblasts Oncogene. 20:2671-82. 2001. <o:p></o:p> Kim, H., S. You, J. Farris, L. K. Foster, Y. J. Choi, and D. N. Foster. Gonad-specific expression of two novel chicken complementary DNA isoforms Biol Reprod. 64:1473-80. 2001. <o:p></o:p> Kim, H., S. You, L. K. Foster, J. Farris, and D. N. Foster. The rapid destabilization of p53 mRNA in immortal chicken embryo fibroblast cells Oncogene. 20:5118-23. 2001. <o:p></o:p> Kim, H., S. You, L. K. Foster, J. Farris, Y. J. Choi, and D. N. Foster. Differential expression of chicken dimerization cofactor of hepatocyte nuclear factor-1 (DcoH) and its novel counterpart, DcoHalpha Biochem J. 354:645-53. 2001. <o:p></o:p> Kim, I. J., and J. M. Sharma. IBDV-induced bursal T lymphocytes inhibit mitogenic response of normal splenocytes Vet Immunol Immunopathol. 74:47-57. 2000. <o:p></o:p> Kim, I. J., S. K. You, H. Kim, H. Y. Yeh, and J. M. Sharma. Characteristics of bursal T lymphocytes induced by infectious bursal disease virus J Virol. 74:8884-92. 2000. <o:p></o:p> Kim, T-J. and Tripathy, D.N. Reticuloendotheliosis virus integration in the fowlpox virus  genome: not a recent event.  Avian Dis. 45: 663-669. 2001.   <o:p></o:p> Kim, T-J., Pessier, A.P. and Tripathy, D.N. Condor poxvirus with biological differences from fowlpox virus. Poster Presentation at the AAAP/AVMA Scientific Program, Boston, MA. p. 54. 2001. <o:p></o:p> Kinde, H., H. L. Shivaprasad, B. M. Daft, D. H. Read, A. Ardans, R. Breitmeyer, G. Rajashekara, K. V. Nagaraja, and I. A. Gardner. Pathologic and bacteriologic findings in 27-week-old commercial laying hens experimentally infected with Salmonella enteritidis, phage type 4 Avian Dis. 44:239-48. 2000. <o:p></o:p> Knoblich, H. V., S. E. Sommer and D. J. Jackwood.  Antibody titers to infectious bursal disease virus in broiler chicks following vaccination at one day-of-age with infectious bursal disease virus and Mareks disease virus.  Avian Dis. 44:874-884.  2000. <o:p></o:p> Kwon, H. M., D. K. Kim, T. W. Hahn, J. H. Han and D. J. Jackwood.  Sequence of precursor polyprotein gene (segment A) of infectious bursal disease viruses isolated in Korea.  Avian Dis. 44:691-696.  2000. <o:p></o:p> Ley DH, Martinez A, Vaillancourt J-P, Smith C.  DNA fingerprints of Mycoplasma gallisepticum isolates from the 1999-2000 North Carolina experience. Proc. 35th Natl. Mtg. on Poultry Health and Processing. p. 31-33, 2000. <o:p></o:p> Ley DH, Vaillancourt J-P, Martinez A.  Molecular and field epidemiological investigations of Mycoplasma gallisepticum outbreaks in commercial poultry. Proc. XXI Worlds Poultry Congress, Abstracts and Proc. CD. 2000. <o:p></o:p> Ley DH, Vaillancourt J-P, Martinez A. Mycoplasma gallisepticum in North Carolina: 1999-2000. Convention Notes from the 138th Ann. Convention of the Amer. Vet. Med. Assn., CD-ROM produced by Veterinary Software Publishing, Inc., OFallon, IL., July 14-18, 2001. <o:p></o:p> Ley DH, Vaillancourt J-P, Martinez A. Mycoplasma gallisepticum in North Carolina: 1999-2000 (and beyond). Proc. Respiratory Diseases of Poultry, Amer. Assn. of Avian Pathol. Symposium 2001, Amer. Vet. Med. Assn Ann. Convention, Boston, MA (4 pp.). July 15, 2001. <o:p></o:p> Ley DH, Vaillancourt J-P, Martinez A. Mycoplasma gallisepticum in North Carolina: 1999-2000. Amer. Assn. Avian Pathol./Amer. Vet. Med. Assn. Scientific Program, Boston, MA; (abstract, p. 10). July 2001. <o:p></o:p> Ley DH. Identification of Avian Mycoplasma Strains by Random Amplification of Polymorphic DNA (RAPD). Zootechnica No. 6, pp. 46-48. June 2001. <o:p></o:p> Li, Z., K. E. Nestor, Y. M. Saif, J. W. Anderson, and R. A. Patterson. Effect of selection for increased body weight in turkeys on lymphoid organ weights, phagocytosis, and antibody responses to fowl cholera and Newcastle Disease-inactivated vaccines. Poultry Sci. 80:689-694, 2001. <o:p></o:p> Liu, L., K. Dybvig, V. S. Panangala. Sequences 5‘ of the GAA repeats and the spacing between GAA repeats and the transcription start site are important for pMGA gene expression in Mycoplasma gallisepticum. 101" General Mtg. Amer. Soc. Micro., (abstract # G23). May-June, Orlando, Florida. 2001. <o:p></o:p> Liu, M, Brandt, M., and Vakharia,V.N.  (2001).  Phylogenetic analysis of infectious bursal disease virus strains of different pathotypes.  20th Ann. Mtg. of Amer. Soc. for Virol. July 21-25, Madison, WI. 2001. <o:p></o:p> Liu, M., and Vakharia,V.N.  (2001).  Expression of infectious bursal disease virus structural and Newcastle disease virus HN protein genes in a baculovirus insect/cell system.  73rd Northeastern Conf. on Avian Dis., College Park, MD, June 2001. <o:p></o:p> Liu, Y., and Vakharia,V.N.  (2001).  Molecular determinants of virulence in variant infectious bursal disease.  73rd Northeastern Conf. on Avian Dis., College Park, MD, June 2001. <o:p></o:p> Lopes, V., G. Rajashekara, A. Back, D. P. Shaw, D. A. Halvorson, and K. V. Nagaraja. Outer membrane proteins for serologic detection of Ornithobacterium rhinotracheale infection in turkeys Avian Dis. 44:957-62. 2000. <o:p></o:p> Martinez A, Vaillancourt J-P, Ley DH. The epidemiology of Mycoplasma gallisepticum in North Carolina: an update. Convention Notes from the 138th Ann. Convention of the Amer. Vet. Med. Assn. CD-ROM produced by Veterinary Software Publishing, Inc., OFallon, IL., July 14-18, 2001. <o:p></o:p> Martinez A, Vaillancourt J-P, Ley DH. The epidemiology of Mycoplasma gallisepticum in North Carolina: an update. Amer. Assn. Avian Pathol./Amer. Vet. Med. Assn. Scientific Program, Boston, MA; (abstract, p. 25). July 2001. <o:p></o:p> May, B. J., Q. Zhang, L. L. Li, M. L. Paustian, T. S. Whittam, and V. Kapur. Complete genomic sequence of Pasteurella multocida,Pm70 Proc Natl Acad Sci U S A. 98:3460-5. 2001. <o:p></o:p> Meir, R., D. J. Jackwood and Y. Weisman.  Molecular typing of infectious bursal disease virus of Israeli field and vaccine strains by the reverse transcription/polymerase chain reaction/restriction fragment length polymorphism assay.  Avian Dis. 45: 223-228. 2001. <o:p></o:p> Mikaelian I, Ley DH, Claveau R, Lemieux M.  Mycoplasma gallisepticum from evening grosbeaks (Coccothraustes vespertinus) and pine grosbeaks (Pinicola enucleator) with conjunctivitis in Quebec, Canada.  Proc. 49th Ann. Wildlife Dis. Assn. Conf., 91, Abstr. 2000. <o:p></o:p> Moura L, Elankumaran S, Oshop GL, Bautista DA, Wilson JD, Heckert RA. Determination of the most effective route for in ovo delivery of DNA vaccines. Proc. 138th Am. Vet. Med. Assn. Mtg., Boston, MA, July 14-18, 2001. <o:p></o:p> Oshop GL, Elankumaran S, Vakharia VN, Wilson JD, Moura LC, Bautista DA, Heckert RA. In ovo nucleic acid immunization of the chicken against infectious bursal disease and Newcastle disease, Proc. 138th Am. Vet. Med. Assn. Mtg, Boston, MA, July 14-18, 2001. <o:p></o:p> Oshop GL, Heckert RA, Elankumaran S. DNA and mRNA persistence and tissue distribution following topical delivery of a DNA vaccine in chickens. 73rd Northeastern Conf. on Avian Dis., College Park, MD, June 2001. <o:p></o:p> Pang, Yao-shan., M. I. Khan, H. Wang, Z. Xie and T. Girshick. Multiplex PCR and its application in experimentally infected SPF chickens with respiratory pathogens. 50th  Western Poultry Dis. Conf.,.March 23-26, Davis, California p. 141. 2001. Paustian, M. L., B. J. May, and V. Kapur. Pasteurella multocida gene expression in response to iron limitation Infect Immun. 69:4109-15. 2001. <o:p></o:p> Pedersen J. C., D. A. Senne, B. Panigrahy and D. L. Reynolds. Detection of avian pneumovirus in tissues and swab specimens from infected turkeys. Avian Dis. 45:581-92. 2001. <o:p></o:p> Pitts, G. R., S. You, D. N. Foster, and M. E. El-Halawani. Evidence for multiple prolactin receptor transcripts in the turkey Poult Sci. 79:355-62. 2000. <o:p></o:p> Qureshi, M. A., Y. M. Saif, R. A. Ali, F. W. Edens, C. L. Heggen-Peay, and G. B. Havenstein. Comparison of PEMS-associated and classical astroviruses-mediated effects on performance and immune functions of turkey poults.  Poultry Sci., Vol. 80, supplement 1, abstract 538, 2001. <o:p></o:p> Rajashekara, G., E. Haverly, D. A. Halvorson, K. E. Ferris, D. C. Lauer, and K. V. Nagaraja. Multidrug-resistant Salmonella Typhimurium DT104 in poultry J Food Prot. 63:155-61. 2000. <o:p></o:p> Rajashekara, G., S. Munir, M. F. Alexeyev, D. A. Halvorson, C. L. Wells, and K. V. Nagaraja. Pathogenic role of SEF14, SEF17, and SEF21 fimbriae in Salmonella enterica serovar enteritidis infection of chickens Appl Environ Microbiol. 66:1759-63. 2000. <o:p></o:p> Rautenschlein, S., and J. M. Sharma. Immunopathogenesis of haemorrhagic enteritis virus (HEV) in turkeys Dev Comp Immunol. 24:237-46. 2000. <o:p></o:p> Rautenschlein, S., M. Suresh, and J. M. Sharma. Pathogenic avian adenovirus type II induces apoptosis in turkey spleen cells Arch Virol. 145:1671-83. 2000. <o:p></o:p> Rautenschlein, S., R. L. Miller, and J. M. Sharma. The inhibitory effect of the imidazoquinolinamine S-28828 on the pathogenesis of a type II adenovirus in turkeys Antiviral Res. 46:195-205. 2000. <o:p></o:p> Reiness, C. G., M. J. Seppa, D. M. Dion, S. Sweeney, D. N. Foster, and R. Nishi. Chick ciliary neurotrophic factor is secreted via a nonclassical pathway Mol Cell Neurosci. 17:931-44. 2001. <o:p></o:p> Reynolds, D. and S. Akinc.  Passive immunization protects birds following challenge with virulent NDV. Oral presentation. AVMA / AAAP annual meeting.  Boston, MA. July 14-18, 2001. <o:p></o:p> Reynolds, D., J. Oesper and S. Akinc.  The effects of avian pneumovirus on laying chickens.  Oral presentation. Proc. North Central Avian Dis. Conf.. Grand Rapids, Michigan. p. 42. September 30 - October 2, 2001. <o:p></o:p> S. Elankumaran, R.A. Heckert, and L. Moura. Persistence and tissue distribution of a variant strain of infectious bursal disease virus in commercial broiler chickens. Avian Dis. In press. Saif, Y. M. Experiences with infectious bursal disease.  Proc. 45th Ann. Mtg. Korean Society of Vet. Sci., pages 6-7, Korea, October 12, 2001. <o:p></o:p> Sanei B, Ley DH, Barnes HJ, Vaillancourt J-P. Pathogenicity of Mycoplasma gallisepticum field isolates for chickens and turkeys. Convention Notes from the 138th Ann Convention of the Amer. Vet. Med. Assn.,. CD-ROM produced by Veterinary Software Publishing, Inc., OFallon, IL. July 14-18, 2001. <o:p></o:p> Sanei B, Ley DH, Barnes HJ, Vaillancourt J-P. Pathogenicity of Mycoplasma gallisepticum field isolates for chickens and turkeys. Amer. Assn of Avian Pathol./Amer.Vet. Med. Assn. Scientific Program, Boston, MA; (abstract, p. 25) July 2001. <o:p></o:p> Schnitzlein, W. M., Singh, P., and Tripathy, D. N. Genetic variability of reticulo-endotheliosis provirus in the genome of fowlpox virus. p. 17. AAAP/AVMA Scientific Program, Boston, MA, p. 53. 2001. <o:p></o:p> Sharma, J. M., I. J. Kim, S. Rautenschlein, and H. Y. Yeh. Infectious bursal disease virus of chickens: pathogenesis and immunosuppression Dev Comp Immunol. 24:223-35. 2000. <o:p></o:p> Shin, H. J., B. McComb, A. Back, D. P. Shaw, D. A. Halvorson, and K. V. Nagaraja. Susceptibility of broiler chicks to infection by avian pneumovirus of turkey origin Avian Dis. 44:797-802. 2000. <o:p></o:p> Shin, H. J., G. Rajashekara, F. F. Jirjis, D. P. Shaw, S. M. Goyal, D. A. Halvorson, and K. V. Nagaraja. Specific detection of avian pneumovirus (APV) US isolates by RT-PCR Arch Virol. 145:1239-46. 2000. <o:p></o:p> Shin, H. J., G. Rajashekara, F. F. Jirjis, D. P. Shaw, S. M. Goyal, D. A. Halvorson, and K. V. Nagaraja. Specific detection of avian pneumovirus (APV) US isolates by RT-PCR Arch Virol. 145:1239-46. 2000. <o:p></o:p> Shin, H. J., M. K. Njenga, B. McComb, D. A. Halvorson, and K. V. Nagaraja. Avian pneumovirus (APV) RNA from wild and sentinel birds in the United States has genetic homology with RNA from APV isolates from domestic turkeys J Clin Microbiol. 38:4282-4. 2000. <o:p></o:p> Shin, H. J., M. K. Njenga, B. McComb, D. A. Halvorson, and K. V. Nagaraja. Avian pneumovirus (APV) RNA from wild and sentinel birds in the United States has genetic homology with RNA from APV isolates from domestic turkeys J Clin Microbiol. 38:4282-4. 2000. <o:p></o:p> Shin, H. J., M. K. Njenga, D. A. Halvorson, D. P. Shaw, and K. V. Nagaraja. Susceptibility of ducks to avian pneumovirus of turkey origin Am J Vet Res. 62:991-4. 2001. <o:p></o:p> Singh, P. Schnitzlein, W. and Tripathy, D.N. A caveat in FPV vaccines and their derived recombinants, a need for better vaccines.   Abst. AAAP/AVMA Scientific Program, Boston, MA. p. 41. 2001. <o:p></o:p> Singh, P., Schnitzlein, W. M., and Tripathy, D. N. Analysis of integrated reticuloendotheliosis virus in the genome of fowlpox virus.  Amer. Soc. for Virol., Madison, WI. 2001. <o:p></o:p> Singh, P., Schnitzlein, W.M. and Tripathy, D.N. The role of integrated reticuloendotheliosis virus in the reemergence of fowlpox..  Abst. 4th Ann. Conference on New and Re-emerging Infectious Diseases. p. 12-13. 2001. <o:p></o:p> Smiley, J. R., and D. J. Jackwood.  Genetic stability of the VP2 hypervariable region of four infectious bursal disease virus isolates after serial passage in specific-pathogen-free chicken embryos.  Avian Dis. 45:1-8, 2001. <o:p></o:p> Sprenger, S. J., D. A. Halvorson, D. P. Shaw, and K. V. Nagaraja. Ornithobacterium rhinotracheale infection in turkeys: immunoprophylaxis studies Avian Dis. 44:549-55. 2000. <o:p></o:p> Sprenger, S. J., D. A. Halvorson, K. V. Nagaraja, R. Spasojevic, R. S. Dutton, and D. P. Shaw. Ornithobacterium rhinotracheale infection in commercial laying-type chickens Avian Dis. 44:725-9. 2000. <o:p></o:p> Srinivasan, V., Schnitzlein, W. and Tripathy, D.N. Homologous fowlpox virus derived promoters for the development of recombinant vaccines.  Abst. AAAP/AVMA Scientific Program, Boston, MA. p. 39. 2001. <o:p></o:p> Srinivasan, V., Schnitzlein, W. M., and Tripathy, D.  N. Fowlpox virus encodes for a novel DNA repair enzyme that restores infectivity of UV-light damaged virus. J. Virol., 75; 1681-88. 2001. <o:p></o:p> Srinivasan, V., Schnitzlein, W.M. and Tripathy, D.N. An unique poxvirus bidirectional promoter for the development of polyvalent recombinant vaccines.  Abst. 4th Ann. Conf. on New and Re-emerging Infectious Diseases. p.11-12. 2001. <o:p></o:p> St Hill, C. A., and J. M. Sharma. Viral pathogenesis in chicken embryos and tumor induction in chickens after in ovo exposure to serotype 1 Marek‘s disease virus Avian Dis. 44:842-52. 2000. <o:p></o:p> Townsend, E., D. A. Halvorson, K. V. Nagaraja, and D. P. Shaw. Susceptibility of an avian pneumovirus isolated from Minnesota turkeys to physical and chemical agents Avian Dis. 44:336-42. 2000. <o:p></o:p> Tripathy, D.N. , Schnitzlein, W.M. Singh, P. and Srinivasan, V. Fowlpox, a re-emerging disease of chickens:  need for a new generation of vaccines.  Proc. 50th Western Poultry Dis. Conf. Davis, California, pp. 50-52. March 24-26, 2001. <o:p></o:p> Tripathy, D.N., Kim, T-J., Shivprasad, H.L. and Woolcock, P.R. Genetic and antigenic characterization of a poxvirus from ostrich. Abst. AAAP/AVMA Scientific Program, Boston, MA. p. 41. 2001. <o:p></o:p> Vaillancourt JP, Ley D, Martinez A. Les Mycoplasmes Aviaires: Les Caracteristiques; Les Enjeux; Les Moyens de Lutte et Les Diagnostics (Avian Mycoplasma: Characteristics; Diagnoses; Control; and Issues). Proc. UCAAB Poultry Pathology Session, Chateau-Thierry, France, 8 pp.. 2000. <o:p></o:p> Vaillancourt JP, Martinez A, Carver D, Ley DH.  The epidemiology of Mycoplasma gallisepticum in North Carolina: what we know, what we dont know, and why.  72nd Northeastern Conf. on Avian Dis., p. 2-3, Abstr. 2000. <o:p></o:p> Vaillancourt J-P, Martinez A, Ley D, Carver D, Quinn J, Wages D, Smith C.  Actualizacion Sobre Mycoplasma.  Curso Enfermedades emergentes: Criterios generales sobre problemas respiratorios de las aves. Proc.ANECA, p. 22-28, 2000. <o:p></o:p> Vaillancourt J-P, Martinez A, Ley D. Disease Prevention and Treatment: MG/MS. Production and Health Seminar. Proc. US Poultry & Egg Assn.  Alabama, 6 pp. 2000. <o:p></o:p> Vaillancourt J-P, Martinez A, Smith C, Ley DH.  The epidemiology of Mycoplasma gallisepticum in North Carolina. Proc. 35th Natl. Mtg. on Poultry Health and Processing, p. 34-36. 2000. <o:p></o:p> Wellehan, J. F., M. Calsamiglia, D. H. Ley, M. S. Zens, A. Amonsin, and V. Kapur. Mycoplasmosis in captive crows and robins from Minnesota J. Wildl. Dis. 37:547-55. 2001. <o:p></o:p> Wellehan, J. F., M. S. Zens, M. Calsamiglia, P. J. Fusco, A. Amonsin, and V. Kapur. Diagnosis and treatment of conjunctivitis in house finches associated with mycoplasmosis in Minnesota J. Wildl Dis. 37:245-51. 2001. <o:p></o:p> Wooming B, Huyan S, Ley DH. Observations on the use of the random amplified polymorphic DNA analysis test on Mycoplasma gallisepticum cases in Colorado. Amer. Assn. of Avian Pathol./Amer. Vet. Med. Assn. Scientific Program, Boston, MA; (abstract, p. 25) July 2001. <o:p></o:p> Wu, C. C., Chang, H. C., and T. L Lin.. Protection of chickens against IBD by DNA-mediated vaccination. Proc. 2nd International Symposium on Infectious Bursal Disease and Chicken Infectious Anemia. Rauischolzhausen, Germany, June, 2001. <o:p></o:p> Wu, C. C., Chang, H. C., and T. L. Lin. DNA vaccination with plasmids containing various fragments of large segment genome of infectious bursal disease virus. Proc. 138th Ann. Mtg. of the Amer. Vet. Med. Assn. Boston, Massachusetts, Avian Medicine Section. July, 2001. <o:p></o:p> Yao, K., and Vakharia, V.N. Induction of apoptosis in vitro by the 17-kDa nonstructural protein of infectious bursal disease virus: possible role in viral pathogenesis.  Virology 285, 50-58. <o:p></o:p> Yu, L., Liu, W., Schnitzlein, W.M., Tripathy, D.N. and Kwang, J. Study of protection by recombinant fowlpox virus expressing C-terminal nucleocapsid protein of infectious bronchitis virus against challenge.  Avian  Dis.  45:340-348. 2001. <o:p></o:p> Zhang, Y., and J. M. Sharma. Early posthatch protection against Marek‘s disease in chickens vaccinated in ovo with a CVI988 serotype 1 vaccine Avian Dis. 45:639-45. 2001.

Impact Statements

See <a href: "http://www.wisc.edu/ncra/nc228annualreport2000.htm"> http://www.wisc.edu/ncra/nc228annualreport2000.htm</a>.

Date of Annual Report: 03/06/2003

Report Information

Annual Meeting Dates: 11/09/2002 - 11/10/2002
Period the Report Covers: 10/01/2001 - 10/01/2002

Participants

AL AES Victor S. Panangala
CT AES Mazhar I. Khan
DE AES John E. Dohms and Jack Gelb, Jr.
GA, USDA, SEPRL David L. Suarez
IA AES Donald L. Reynolds
IL AES Deoki N. Tripathy
IN AES Ching Ching Wu
MD AES Report submitted by Vikram Vakharia
MN AES Report submitted by Vivek Kapur
NC AES David H. Ley
OH AES Y. Mo Saif
Administrative Advisor Jeffrey Klausner (University of Minnesota)
CSREES Representative William Wagner (USDA)

Brief Summary of Minutes

Summary of minutes of annual meeting: The NC228 business meeting was convened by Dr. Gelb, Chair of NC228 at 8:30 am on Saturday, Nov. 9, 2002, in the Jefferson A Room, Millennium Hotel, St. Louis, MO. He welcomed the Station Representatives, Dr. Klausner, Dr. Wagner, and new member, Dr. Suarez of Southeast Poultry Research Laboratory (SEPRL), USDA, Athens, GA.

Dr. Wagner summarized federal legislative activities as follows; although the farm bill has passed, the funding appropriation was still pending. For NRI, an additional $20-25 million was proposed for agricultural bio-security issues, bringing the total NRI budget to $130-164 million. Allocations for improving air and seaport security and diagnostic lab capabilities to deal with homeland security were explained. Dr. Wagner announced effective Jan. 2003, Dr. Robert Smith would coordinate animal and plant CSREES educational programs. Dr. Wagner also announced his retirement from CSREES at the end of Dec. 2002. The Committee recognized him for his contributions and service to NC228.

The Committee agreed to begin working on a renewal NC228 project for 2004. Dr. Klausner presented a timeline for new project submission as well as distributed information on the project approval process. A writing subcommittee (Drs. Dohms, Saif, and Wu) agreed to coordinate the renewal project resubmission activities in 2003. All members will be working with the subcommittee to finalize the preparation of the proposal. The first step was to provide project objectives to members in January 2003.

Dr. Klausner reported that the current project received an overall favorable mid-project review in a memo dated March 22, 2002 from the Multistate Research Committee (MRC). However, the MRC did encourage a greater commitment to interdependence among the participants. At Dr. Klausners request, the Committee updated project participant information. The Committee also agreed to invite new project participants to strengthen our project renewal request. A list of potential participants was made and Drs. Dohms and Saif agreed to contact the individuals.

Dr. Gelb was re-elected as Chair for 2003-4 and Dr. Khan was elected as Secretary for the same period. The Committee thanked Dr. Wu for serving as Secretary the past two years.

The ongoing development of a NC228 website was discussed. Suggestions were made to add a visitors counter and to add links for the current project and the renewal project, which would be accessible to the writing subcommittee.

For the 2003 NC228 meeting, the Committee agreed to meet in Chicago (Congress Hotel) from Friday Nov. 7 from 2-5 pm and from 8 am-5 pm Saturday Nov. 8. The 2003 CRWAD meeting will be held November 911.

The business meeting concluded at 11 am. The remainder of the meeting was devoted to presentation and discussion of the station reports. The meeting adjourned Nov. 10 at noon.

Accomplishments

Accomplishments and Impacts: Objective 1. Determine the pathogenesis and interactions of specific agents. Alabama studied the regulation of the pMGA responsible for hemagglutinin adhesin protein in Mycoplasma gallisepticum (MG) by deleting or inserting nucleotides at the 5 or 3 positions of the GAA repeats. The findings suggested the existence of a hemagglutinin activator binding protein that regulates transcription in the pMGA promoter region. Proteins that regulate transcription in Mycoplasma species have not been reported. Delaware (Drs. Keeler and Dohms) and Minnesota (Dr. Kapur) are sequencing the Mycoplasma synoviae (MS) genome. MS sequencing will be conducted at U. Minnesota and data will be made available on the Internet (http://udgenome.ags.udel.edu/). Delaware. Compared to the virulent MG S6 strain, attenuated strains demonstrated varied abilities of to enter chicken embryo fibroblasts suggesting that attenuation may limit intracellular invasion. Illinois. A genetically modified light-sensitive (photolyase deficient) fowlpox virus was found to replicate less efficiently than the parental FPV and, upon exposure to UV light, its infectivity was drastically reduced compared to the parent. The light-sensitive virus has a potential as an eco-friendly vaccine against fowlpox. Minnesota. In response to avian pneumovirus (APV) infection in turkeys, B- and T-lymphocytes in the Harderian gland were highly activated and secreted high levels of cytokines, but that T cells were not an absolute requirement in turkey resistance against infection. Molecular analysis showed that APV infection resulted in up-regulation of interferon-associated, pro-inflammatory leukocyte chemo-attractant, adhesion molecule, and complement-associated genes. Avian E. coli genome sequencing is nearly completed and will help identify important virulence factors. North Carolina. Studies with S. J. Geary (U. Connecticut) reaffirmed that high frequency phenotypic variation occurs in MG shed following homologous challenge of chickens vaccinated with whole cell killed bacterins. This underscores the need for live attenuated vaccines which will ultimately present to the host (vaccinated birds) the full array of diverse phenotypes required to develop complete protective immunity without the risk of developing persistent shedders. B. Sanei and D. Ley compared the pathogenicity of a RAPD type B field strain of MG from North Carolina with the S6 reference strain in chickens and turkeys. Mean gross air sac lesion scores were highest in type B-infected chickens. Tracheas of type B-inoculated chickens demonstrated severe lesions 2 wk PI. Objective 2. Surveillance, occurrence and consequences of agents and host variation on disease susceptibility Delaware. Although antigenic differences of infectious bronchitis virus (IBV) were observed by VN, S1 sequencing and challenge of immunity findings suggest that Georgia 98 and Alabama isolates AL/2975/00 and AL/3701/00 obtained from the 1999-2000 outbreak were related to the DE/072/92 strain. Illinois. A poxvirus isolate from ostriches was found to be similar to fowlpox virus based on antigenic (Western blotting), genetic (RFLP) and biologic (infection of susceptible chickens) studies. Hawaiian goose poxvirus (HGPV) and an avipoxvirus from finches on the Galapagos Island were identified as new species of the avipoxvirus genus. In addition, poxvirus infection in two Palila (Loxioides bailleui), an endangered Hawaiian forest bird, was confirmed using either chicken embryos or avian cell cultures for virus isolation. USDA SEPRL. Turkeys were found to be highly susceptible to wild waterfowl-origin AIV, requiring 5 to 10,000 times less virus to become infected than chickens. AIVs from the US and around the world were characterized by sequence analysis and animal studies for the purposes of assessing the risk of influenza outbreaks and to study mechanisms enabling low pathogenic AIV to become highly pathogenic. Researchers at NVSL, TX, CA, and NJ Dept. of Agriculture, the Korean National Vet. Res. and Quarantine Service, and the Chile Dept. of Agriculture collaborated on this work. Minnesota. Wild birds harbor APV suggesting that they may be important reservoirs. A partially attenuated APV strain (APV/MN/1A-97) shown to be efficacious as vaccine in turkeys was approved by the USDA. North Carolina and Georgia and collaborators from other laboratories. MG isolates suspected of being of vaccine origin recovered from unvaccinated chicken or turkey flocks were shared and characterized. A publication resulted from this effort. In a related study. scientists from North Carolina and Georgia and collaborators K. Whithear and G. Browning (U. Melbourne, Australia) found the molecular properties of MG isolates from an unvaccinated broiler breeder and a layer were essentially identical to a vaccine strain. Since the1999-2000 MG epornitic in commercial poultry in North Carolina, MG has been isolated from only one broiler breeder and two turkey farms, both in January 2001. Each of these isolates was a new and unique RAPD type, suggesting separate external sources of infection that has been linked to backyard flocks. Objective 3. Develop new and improved methods for the diagnosis, prevention, and control of avian respiratory diseases. Alabama and North Carolina. Fifty-five isolates from house finches infected with MG were compared to 11 strains from poultry. The finch isolates were found to be more similar to each other than to the poultry isolates. Three RFLP groupings and 16 genotypes were found among the MG finch isolates. Connecticut. SPF chickens given a spike gene (Mass-type) DNA vaccine in ovo did not produce IBV serum antibodies but did show resistance to Mass 41 challenge at 6 weeks of age, but not at 2 or 4 weeks of age. The vaccine did not have an adverse effect on embryo development. Illinois. Recombinant fowlpox vaccines in which REV sequences have been deleted or only REV-envelope gene is incorporated provided complete protection against fowlpox virus. These vaccines will minimize the chances of future REV integration in FPV genome. Homologous fowlpox virus promoters can be modified by specific mutations in order to develop strong promoters. Specific PCR primer sets have been designed for rapid diagnosis of avianpox virus infection. Indiana. Chickens vaccinated with plasmid encoding infectious bursal disease virus (IBDV) genes VP243-VE alone or both plasmids (VP243-VE and VP243-STC) conferred protection against classical or variant IBDV challenge. Iowa. A composting study was initiated to evaluate the inactivation of viruses (Newcastle disease virus and avian encephalitis virus), odor emissions, and the effect of cover materials (silage, ground cornstalks, finished yard waste compost) on animal carcass degradation. Maryland. An attenuated, marked, multi-spectrum vaccine candidate, derived from full-length and chimeric cDNA clones of segments A and B of the D78 and GLS strains protected against classical and variant strains of IBDV. A transcutaneous method utilizing dimethylsulfoxide (DMSO) for delivery of DNA vaccines for IBDV and Newcastle disease virus (NDV) gave promising results. Neutral lipid encapsulation may be useful for delivering DNA to the avian embryo for immunization. Minnesota and Ohio. Avian pneumovirus was shown to differ antigenically from avian paramyxoviruses, PMV-1, PMV-2, PMV-3 and PMV-7 by ELISA, virus-neutralization, and hemagglutination-inhibition. A publication resulted from this effort. North Carolina and Georgia are evaluating various methods of MG strain identification. Ohio. Real-time RT-PCR was used to identify nucleotide sequence homology at the hydrophilic B region of VP2 for each virus encoding a neutralizing epitope of IBDV. The technique has the potential to rapidly provide information for selecting the most appropriate vaccine strain. USDA SEPRL. In collaboration with APHIS NVSL, a real time RT-PCR test was developed and validated for detecting and subtyping influenza virus. A real time RT-PCR test for avian influenza virus (AIV) diagnosis was developed, validated and adopted for clinical use by state veterinary diagnostic labs in VA, PA, and NC. The test was widely employed to rapidly identify positive flocks in the 2002 H7N2 AIV outbreak in VA that lead to control of the outbreak. Pasteurization temperature inactivated low path AI virus H7N2 LPAI virus in homogenized egg, liquid egg whites and salted egg yolks. This information re-assures our trading partners that pasteurization destroys AIV. Minnesota. A partially attenuated APV strain (APV/MN/1A-97), shown to be efficacious as a vaccine in turkeys, was approved by USDA. Work Planned for the Coming Year Avian influenza. USDA SEPRL will continue to investigate AIV. Avian paramyxoviruses (pneumovirus and Newcastle disease virus). Minnesota will continue to investigate APV. Delaware and Maryland will evaluate Newcastle disease virus as a vector for delivering IBV S antigen in chickens. Infectious bronchitis. Connecticut will continue to evaluate DNA and fowl pox viral vector S1 gene vaccines. Delaware will characterize genotype PA/1220/98 related isolates from geographically widespread regions in the USA and evaluate the replication interference potential of NDV and IBV vaccine strains. Indiana will send IBDV field isolates to Delaware for S1 genotyping. Infectious bursal disease. Indiana will study immune responses, immunomodulation, and delivery system regarding DNA vaccination against IBDV. Iowa will continue composting studies regards to biosecurity. Maryland will evaluate the efficacy of an attenuated recombinant IBDV vaccine in ovo, as well as an IBDV/Newcastle DNA vaccine in ovo Ohio will continue evaluating interactions of vaccine and mild strains of IBDV, develop monoclonals specific for very virulent IBDV, and perform epidemiologic studies using real-time RT-PCR to determine sequence diversity among wild-type IBDV strains. Mycoplasmosis. Delaware will complete the M. synoviae genome sequence project. North Carolina will continue the development and application of RAPD fingerprinting of pathogenic avian Mycoplasma spp., the development of new capabilities by application of a software system for DNA fragment analysis and databasing, assessing the pathogenic potential of emerging avian Mycoplasma species or isolates for commercial poultry, will continue research on the ongoing MG outbreak in house finches and will continue investigations related to avian mycoplasma vaccines.

Publications

Alkahalaf, AN, Halvorson DA, Saif YM. Comparison of enzyme-linked immunosorbent assays and virus neutralization test for detection of antibodies to avian pneumovirus. Avian Dis.:46:700-703. 2002. Alkhalaf, A.N., D.A. Halvorson, and Y.M. Saif: Comparison of enzyme-linked immunosorbent assays and virus neutralization test for detection of antibodies to avian pneumovirus. Avian Dis. 46:700-703, 2002. Alkhalaf, A.N., L.A. Ward, R.N. Dearth, and Y.M. Saif: Pathogenicity, transmissibility, and tissue distribution of avian pneumovirus in turkey poults. Avian Dis. 46:650-659, 2002. Amonsin, A, Wellehan JF, Li LL, Laber J, Kapur V. DNA fingerprinting of Pasteurella multocida recovered from avian sources. J Clin Microbiol. 40:3025-3031. 2002. Berinstein, A., B. Seal and D.L. Suarez. Heteroduplex mobility assay (HMA): its use for detection of new avian influenza virus (AIVs) variants. Avian Dis. 46:393-400. 2002. Chang, H. C.; Lin, T. L.; Wu, C. C. DNA vaccination with plasmids containing various fragments of large segment genome of infectious bursal disease virus. Vaccine, 2002. In press. Chang, H. C.; Lin, T. L.; Wu, C. C. DNA-mediated vaccination against infectious bursal disease in chickens. Vaccine 20:328-335, 2002. Chary, P, Rautenschlein S, Njenga MK, Sharma JM. Pathogenic and immunosuppressive effects of avian pneumovirus in turkeys. Avian Dis. 46:153-161. 2002. Chary, P, Rautenschlein S, Sharma JM. Reduced efficacy of hemorrhagic enteritis virus vaccine in turkeys exposed to avian pneumovirus. Avian Dis. 46:353-359. 2002. Dar AM, Munir S, Goyal SM, Abrahamsen MS, Kapur V. Sequence analysis of the nucleocapsid and phosphoprotein genes of avian pneumoviruses circulating in the US. Virus Res. 79:15-25. 2001. Dar AM, Tune K, Munir S, Panigrahy B, Goyal SM, Kapur V. PCR-based detection of an emerging avian pneumovirus in US turkey flocks. J Vet Diagn Invest.13:201-205. 2001. Dar, AM, Munir S, Goyal SM, Kapur V. 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Role of intrabursal T cells in infectious bursal disease virus (IBDV) infection: T cells promote viral clearance but delay follicular recovery. Arch Virol. 147:285-304. PMID:11890524. 2002. Rautenschlein, S., H.-Y. Yeh, and J. M. Sharma. The role of T cells in protection by an inactivated infectious bursal disease virus vaccine. Vet. Immunolo. Immunopathol. 89:159. 2002. Saif, Y.M. and K.E. Nestor: Increased mortality in turkeys selected for increased body weight following vaccination with a live Newcastle disease virus vaccine. Avian Dis. 46:505-508, 2002. Saif, Y.M. Infectious bursal disease vaccines and vaccinations. Proc. 45th Annual Meeting, AAAP, Poultry Vaccines and Vaccination Practices, Nashville, TN, July 14, 2002. Saif, Y.M. Recent research findings on infectious bursal disease. Proc. XII Intl. Congress of the World Veterinary Poultry Association, pages 23-29, Cairo, Egypt, January 28-February 2, 2002. Sanei B, Barnes H J, Vaillancourt J-P, Ley DH. Pathogenicity of Mycoplasma gallisepticum field isolates for turkeys. 4th International Symposium on Turkey Diseases, Berlin Germany; May 15-18, 2002. Sato N, Matsuda K, Sakuma C, Foster DN, Oppenheim RW, Yaginuma H. Regulated gene expression in the chicken embryo by using replication-competent retroviral vectors. J Virol. 76:1980-1985. 2002. Schultz-Cherry S., Dybdahl-Sissoko N., Neumann G., Kawaoka Y., Hinshaw V.S. Influenza virus ns1 protein induces apoptosis in cultured cells. J Virol ;75:7875-81. 2001. Sharma JM, Zhang Y, Jensen D, Rautenschlein S, Yeh HY. Field trial in commercial broilers with a multivalent in ovo vaccine comprising a mixture of live viral vaccines against Marek‘s disease, infectious bursal disease, Newcastle disease, and fowl pox. Avian Dis. 46:613-622. 2002. Shin HJ, Cameron KT, Jacobs JA, Turpin EA, Halvorson DA, Goyal SM, Nagaraja, KV, Kumar MC, Lauer DC, Seal BS, Njenga MK. Molecular epidemiology of subgroup C avian pneumoviruses isolated in the United States and comparison with subgroup A and B viruses. J Clin Microbiol. 40:1687-1693. 2002. Shin HJ, Jirjis FF, Kumar MC, Njenga MK, Shaw DP, Noll SL, Nagaraja KV, Halvorson DA. Neonatal avian pneumovirus infection in commercial turkeys. Avian Dis. 46:239-244. 2002. Shin HJ, Nagaraja KV, McComb B, Halvorson DA, Jirjis FF, Shaw DP, Seal BS, Shin HJ, Njenga MK, Halvorson DA, Shaw DP, Nagaraja KV. Susceptibility of ducks to avian pneumovirus of turkey origin. Am J Vet Res. 62:991-994. 2001. Shivprasad, H.L., Kim, T-J., Woolcock, P.R. and Tripathy, D.N. Genetic and Antigenic Characterization of a Poxvirus Isolate from Ostriches. Avian Dis. 46: 429-436. 2002. Singh, P. and Tripathy, D.N. Evaluation of recombinant vaccines for protection against fowlpox and reticuloendotheliosis. Poster presentation, XIV International Poxvirus and Iridovirus Workshop, Lake Placid, NY. p 157. 2002. Singh, P., Kim, T-J and Tripathy, D.N. Identification and Characterization of Fowlpox Virus Strains Utilizing Monoclonal Antibodies. J. Vet. Diag.Ivest. In press. 2002. Singh, P., Schnitzlein W.M. and Tripathy, D.N. (2002), Evaluation of a recombinant vaccine for protection against fowlpox in chickens. Poster Presentation (Best Poster)American Association of Avian Pathologists, American Veterinary Medical Association, Nashville, TN, July 14-17, 2002 Spalding, B. D. Identification of multiple genetic infectious bursal disease virus populations (quasispecies) in commercial vaccines following plaque purification. M.S. Thesis, The Ohio State University. 2002. Spalding, B. D. and D. J. Jackwood. Identification of multiple genetic infectious bursal disease virus populations (quasispecies) in commercial vaccines following plaque purification. Abstr. 139th AVMA Mtg. 2002. Srinivasan, V., Schnitzlein, W.M. and Tripathy, D.N. (2002). 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Impact Statements

Please see "Accomplishments" section of the report.

Date of Annual Report: 12/10/2003

Report Information

Annual Meeting Dates: 11/07/2003 - 11/08/2003
Period the Report Covers: 10/01/2002 - 11/01/2003

Participants

CT AES M. Khan; DE AES J. E. Dohms and J. Gelb, Jr.; IA AES D. L. Reynolds;
IL AES D. N. Tripathy; IN AES C. C. Wu; MD AES V. N. Vakharia; MN AES M. K. Njenga; NC AES D. H. Ley; OH AES Y. M. Saif; USDA SEPRL D. Suarez

Brief Summary of Minutes

The NC 228 technical committee meeting was held on Friday November 7, 2003 at Grant Park Room in the Congress Hotel, Chicago, IL. Dr. Jack Gelb, Chair of NC228 opened the meeting at 2 PM. Welcomes and introductions followed.

Dr. Khan, the elected NC-228 Secretary this year, was unable to attend the meeting due to another commitment. Dr. Wu kindly agreed to record the minutes.

Dr. Klausner (Administrative Advisor to NC-228) reviewed the time line for the renewal project submission (December 1, 2003) and approval process.

Dr. Peter Johnson (USDA, CSREES) updated the Committee on the following:

1. Personnel updates.

2. A new issue-based CSREES web site will be launched in March 2004.

3. Competitive Programs. Discussed the FY2003 appropriations and the scope of the NRI with special reference to the Integrated Projects programs. Discussed the FY2004 NRI program dates, award sizes and the new Coordinated Agricultural Projects (CAP) program. Avian Influenza was identified for poultry in FY2003; the proposals submitted for that disease were not funded, but one larger consortia proposal approach received a favorable review. An AI proposal will again be eligible for submission in FY2004. Dr. Gelb agreed to e-mail the poultry disease (AIV) scientific community in order to initiate a planning meeting in January 2004. A detailed CSREES budget summary for FY2001-2004 was also discussed."

4. Future Poultry Disease/Issue Priorities. CSREES has begun soliciting input from federal, state and local partners to advise the CAP program. For example, animal agricultural coalition membership, were asked to identify 3-5 issues/diseases from coalition membership for the animal component of the Animal Biosecurity Program. Dr. Johnson requested that NC-228, as a multi-state committee, provide consensus feedback by identifying and prioritizing poultry diseases for future consideration. The committee discussed many poultry diseases and prioritized them. The following diseases were prioritized. 1. Newcastle disease 2. Avian coronaviral diseases 3. Infectious bursal disease. Dr. Gelb agreed to inform Dr. Johnson of the NC-228 recommendations.

Elections of officers were not held this year. For 2004, Dr. Gelb will continue as Chair and Dr. Khan as Secretary.

The committee spent considerable time and effort developing the renewal project. Additional work on the final revision was assigned. The revisions of various sections were to be returned to Dr. Gelb for compilation of the final draft of the proposal. Dr. Gelb agreed to e-mail the participants the final draft for one last review prior to the submission deadline.

All participants in the renewal project were asked to contact their Experimental Station Directors to update the Participant List (Appendix E) of the renewal project.

The 2002-2003 Annual Station Reports were discussed from 8 AM-5 PM, Saturday, November 8. Dr. Gelb requested the Station Reports be e-mailed to him for preparing the final composite report for 2002-2003. The meeting was adjourned at 5 PM.

Accomplishments

Objective 1. Determine the pathogenesis and interactions of specific agents. AVIAN FOWLPOX Immunization of chickens with two avian poxviruses (Hawaiian goose pox and Palila pox) from Hawaiian endangered forest birds did not provide protection against fowlpox virus indicating that these viruses are biologically different from fowlpox virus (Illinois). AVIAN PNEUMOVIRUS (APV) Epidemiologic studies showed the incidence of APV in Minnesota between 1999 and 2002 still high (> 36%), and that the virus was spreading, albeit slowly, to neighboring states (Minnesota). INFECTIOUS BRONCHITIS VIRUS (IBV) Quantitative RT-PCR was found to be a suitable procedure for measuring IBV and NDV following coinfection with vaccine strains. IBV replication interfered with the growth of a commercial B1 NDV and highly attenuated CA B1 NDV strains in embryonated eggs. In chickens, Ca strain NDV replication was greatly diminished by IBV. NDV did not affect IBV growth. QRT-PCR can be used to determine the interference potential of NDV and IBV strains in combination vaccines (Delaware). INFECTIOUS BURSAL DISEASE VIRUS (IBDV) Suppression of bursal B cell proliferation after inoculation with DNA constructs expressing the IBDV VP243 polyprotein gene from either classical STC or variant E strain identified IBDV VP243 polyprotein as a mediator of virus-induced immune suppression (Indiana). Amino acid residues at positions 253 and 284 in VP2 protein are important for viral entry and virulence, and VP1 protein is involved in the efficiency of viral replication and virulence of IBDV in vivo (Maryland). Mild and intermediate vaccine strains of IBDV interfered with the in vivo replication of a virulent IBDV (Ohio). Knowledge of host-virus interactions is important for designing control strategies. MYCOPLASMOSIS Additional experiments in chick embryo fibroblast (CEF) cells confirmed that attenuated strains of MG penetrated as or more efficiently than pathogenic strains, thus indicating that intracellular invasion is not necessarily linked to virulence (Delaware). Preliminary findings indicated that in a health survey of Mycoplasma gallisepticum-free wild house finches, other biotic cofactors, such as West Nile virus and feather mites, may influence host susceptibility to M. gallisepticum infection and sustain outbreaks during periods of low disease transmission. Under laboratory conditions, a high survival rate and recovery of individually caged house finches following experimental infection with Mycoplasma gallisepticum was associated with the use of controlled environmental conditions, acclimatization, a high plane of nutrition, and low stocking (housing) density (North Carolina). NEWCASTLE DISEASE VIRUS (NDV). Using the embryonated chicken egg as a model system to evaluate virulence properties of NDV, embryos from eggs inoculated with different NDV isolates were compared for virus distribution in the tissues and membranes. Low-virulence strains were detected exclusively in the cells of the chorioallantoic membrane, whereas more virulent NDV isolates and mutated strains that had been demonstrated to have acquired virulence for chickens were widely disseminated in chicken embryo tissues (USDA, ARS, SEPRL). PASTEURELLA MULTOCIDA Differential gene expression profiles for Pasteurella multocida under natural infection and in various experimentally-induced stresses continue to be examined (Minnesota). Objective 2. Surveillance, occurrence and consequences of agents and host variation on disease susceptibility. AVIAN PNEUMOVIRUS APV infection was diagnosed for the first time in Ohio in a turkey breeder flock and was associated with a severe drop in egg production. The diagnosis of APV will necessitate initiation of surveillance and control measures (Ohio). APV was identified from 12 APV antibody-positive wild birds and it was determined through sequence analysis of the glycoprotein, matrix and fusion genes that the wild bird viruses are closely related to viruses found in domestic poultry (USDA, ARS, SEPRL). MYCOPLASMOSIS Using the putative cytadhesin protein pvpA gene of M. gallisepticum, three different RFLP groups and 16 genotypes were evident from 55 house finch isolates evaluated suggesting a greater degree of polymorphism than previously recognized by random amplification of polymorphic DNA (RAPD) studies. Isolates from five MG outbreaks in commercial poultry flocks during 2001, the tail end of the epidemic that began in 1999, were unique RAPD types and could be associated with a backyard flock or live bird market. These sporadic cases did not result in widespread epidemic transmission that occurred previously. It appears that backyard flocks and/or live bird markets were important reservoirs of MG infections (North Carolina). Objective 3. Develop new and improved methods for the diagnosis, prevention, and control of avian respiratory diseases. INFECTIOUS BRONCHITIS VIRUS Chickens vaccinated at hatching have T and B-lymphocytes reactant to the IBV-S protein that should elicit some level of cellular and humoral response to IBV infection. An IBV DNA S vaccine was expressed in all lymphoid organs (Connecticut). AVIAN FOWLPOX Chickens immunized with genetically modified fowlpox virus (i) lacking any sequences of reticuloendotheliosis virus (REV) in its genome or (ii) containing only REV envelope gene in its genome provided protection against two field strains of fowlpox virus (Illinois). AVIAN PNEUMOVIRUS An APV vaccine licensed by the USDA and several other vaccine viruses are being evaluated. In addition, the genome sequencing of APV has just been completed, paving way for generation of reverse genetic system for vaccine development and further characterization of the virus. Host gene expression patterns following APV infection and immunosuppressive effects were assessed. (Minnesota). INFECTIOUS BURSAL DISEASE VIRUS Two plasmids encoding chicken IFN- or large segment protein of IBDV given at separate sites did not enhance protection of chickens against IBD by DNA vaccination, but two plasmids given at the same site showed an adverse effect on protection of chickens against IBD by DNA vaccination (Indiana). Amino acid mutations were identified in a neutralizing epitope of an IBDV quasispecies and molecular epidemiology of genetic mutations in a region of the VP2 gene encoding a neutralizing epitope of IBDV was initiated. Determining how quasispecies contribute to the antigenicity and pathogenicity of IBDV will aid in the development of better vaccines and control measures. (Ohio). MYCOPLASMOSIS Pure M. gallisepticum cultures were isolated from fast-growing mixed contaminating mycoplasma cultures using an inexpensive gentomycin invasion assay taking only about two-weeks compared to months needed using colony picking procedures. The technique will increase the number of MG isolates to be recovered and therefore improve diagnostic identification efforts during future outbreaks. Sequencing of the Mycoplasma synoviae genome with Minnesota is continuing. The information will be used to develop better diagnostic, prevention (vaccine) and control measures (Delaware). Polymerase chain reaction test sensitivity was increased by sampling the choanal cleft compared to conjunctiva for detecting MG infections in house finches with no clinical signs or lesions. However, either site could be used for sampling in finches with lesions (North Carolina). NEWCASTLE DISEASE VIRUS. The delivery of DNA to the avian embryo for immunization has been optimized. Using the reverse genetics technology, a recombinant NDV vector carrying the VP2 gene of IBDV protected chickens against both viruses (Maryland). Composting resulted in the inactivation of NDV and avian encephalomyelitis virus. The methodology used has contained the viruses within the compost pile. These preliminary findings indicate that composting is proving to be a safe and environmentally friendly and feasible way to dispose of large amounts of contaminated animal carcasses that may occur in such catastrophic disease events as avian influenza and Newcastle disease (Iowa). A real-time reverse transcription-polymerase chain reaction test (RRT-PCR) was developed for exotic Newcastle disease and validated through the cooperation of SEPRL, APHIS National Veterinary Services Laboratories, and the California Veterinary Diagnostic Laboratory System. The test was adopted for use by the National Animal Health Laboratory Network (NAHLN) (USDA, ARS, SEPRL). ORNITHOBACTERIUM RHINOTRACHEALE A temperature sensitive mutant vaccine against Ornithobacterium rhinotracheale was developed (Minnesota). Work Planned for the Coming Year. INFECTIOUS BRONCHITIS VIRUS In ovo IBV DNA, viral vector vaccination trials and protection studies will continue (Connecticut). Evaluate the replication interference potential of NDV and IBV vaccine strains. Characterize genotype PA/1220/98 and related isolates from geographically widespread regions in the USA. Evaluate NDV as a vector for delivering IBV S antigen (Delaware). INFECTIOUS BURSAL DISEASE VIRUS Indiana will study the long-term effect of VP243 polyprotein on immune response of chickens, and other chicken cytokine genes on immunomodulation and protection of chickens against IBD by DNA vaccination. NEWCASTLE DISEASE VIRUS Iowa will continue the planned work with regards to biosecurity and composting. Maryland will introduce other avian genes in NDV to evaluate the efficacy of an attenuated recombinant NDV vaccine, and also evaluate IBDV vaccine in ovo, using broiler chickens. MYCOPLASMAS Complete the M. synoviae genome sequence project (Delaware). INFECTIOUS BURSAL DISEASE VIRUS Studies will continue on IBDV interference. Surveillance for APV will be intensified. Production of monoclonal antibodies to IBDV will continue (Ohio). Molecular epidemiology studies on IBDV will continue using real-time RT-PCR.

Publications

Abdel-Alim, G.A. and Y.M. Saif: Pathogenicity of embryo-adapted serotype 2 OH strain of infectious bursal disease virus in chickens and turkeys. Avian Dis. 46:1001-1006, 2002. Alkahalaf AN, Halvorson DA, Saif YM. Comparison of enzyme-linked immunosorbent assays and virus neutralization test for detection of antibodies to avian pneumovirus. Avian Dis 46:700-703. 2002. Alkhalaf, A.N. and Y.M. Saif. Lack of antigenic relationships between avian pneumovirus and four avian paramyxoviruses. Avian Dis. 47:175-179, 2003. Alvarez R, Lwamba H., Kapczynski DR, Njenga MK, Seal B. Nucleotide and predicted amino acid sequence-based analysis of the avian metapneumovirus type C cell attachment glycoprotein gene: Phylogenetic analysis and molecular epidemiology of U.S. Pneumoviruses. J Clin Microbiol. 41:1730-1735. 2003. Bennett RS, McComb B, Shin HJ, Njenga MK, Nagaraja KV, Halvorson DA. Detection of avian pneumovirus in wild Canada (Branta canadensis) and blue-winged teal (Anas discors) geese. Avian Dis. 46:1025-1029. 2002. Boyce JD, Wilkie I, Harper M, Paustian ML, Kapur V, Adler B. Genomic scale analysis of Pasteurella multocida gene expression during growth within the natural chicken host. Infect Immun. 70:6871-6879, 2002. Campion, L.. Assessment of interference between live infectious bronchitis virus and Newcastle disease virus vaccines using quantitative polymerase chain reaction. Proc. Seventy-fifth Ann. Mtg. Northeastern Conference on Avian Diseases. University of Maine, Orono, Maine. June 11-13, 2003. Caterina, K., M. I. Khan, T. Grishick and S. Frasca. Development of multiplex PCR for avian enteric pathogens.52nd Western Poultry Disease Conference, Sacramento, California. March 8-11, 2003. Chang, HC, Lin, TL, and Wu, CC. DNA vaccination with plasmids containing various fragments of large segment genome of infectious bursal disease virus. Vaccine, 21: 507-513. 2003. Chary, P., S. Rautenschlein, M. K. Njenga, and J. M. Sharma. Pathogenic and immunosuppressive effects of avian pneumovirus in turkeys. Avian Dis. 46:153-161. 2002. Chen, H., Matsuoka, Y., Swayne, D., Chen, Q., Cox, N.J., Murphy, B.R., Subbarao, K. Generation and characterization of a cold-adapted influenza A H9N2 reassortant as a live virus vaccine candidate. Vaccine. 21:1974-1979. 2003. Cook W, Charlton K, Ley D. Testing recommendations for wild turkeys. 52nd Annual Wildlife Disease Association Conference, Program and Abstracts (pp. 84-86), Saskatoon, Saskatchewan, August 11-14, 2003. Dar, A.M., Munir, S., Goyal, S.M., and Kapur, V. 2003. Sequence analysis of the matrix (M2) protein gene of avian pneumovirus recovered from turkey flocks in the United States. J. Clin. Microbiol. 41:2748-2751. 2003. Dar, A.M., Munir, S., Goyal, S.M., and Kapur, V. A single subtype of avian pneumovirus circulates among Minnesota turkey flocks. J. Vet. Diag. Invest. 14:371-376. 2002. Fabis, J. J., Z. Xie and M. I. Khan. Delineation of IBV-Recombinant plasmid DNA in embryonating and hatched chicks. 140th annual American Veterinary Medical Association Convention and Meeting, & XIII Congress of the World Veterinary Poultry Association. Denver, Colorado. July 19-23, 2003. Gelb, J., Jr. and B. S. Ladman, Infectious bronchitis update-Current thinking on controlling IB. Is it working or is it the lull before the next storm? Proc. North Atlantic Poultry Health and Management Conference. Portsmouth, New Hampshire. March 26-27, 2003. Gelb, J., Jr. Future challenges and opportunities in poultry disease epidemiological methodology. Proc. Seventy-fifth Ann. Mtg. Northeastern Conference on Avian Diseases. University of Maine, Orono, Maine. June 11-13, 2003. Gelb, Jr., J., L. Campion and B. S. Ladman. Replication interference associated with infectious bronchitis virus and Newcastle disease virus co-infections. Proc. AVMA/AAAP Ann. Mtg. AVMA Convention Notes (Poultry section) compact disc. Denver, Colorado. July 19-23, 2003. Glanville, T. D., T. L. Richard, J. D. Harmon, D. L. Reynolds, S. S. Sadaka and S. Akinc. Environmental Impact & Biosecurity of Composting for Emergency Disposal of Livestock Mortalities. Paper No. 032262. Presented at the American Society of Agricultural Engineers 2003 Annual International Meeting. Las Vegas, NV July 27-30, 2003. Goyal, S.M., Lauer, D., Friendshuh, K. and Halvorson, D.A. Seroprevalence of avian pneumovirus in Minnesota turkeys. Avian Dis. 47:244-250, 2003. Hartup BK, Stott-Messick B, Guzy M, Greiner EC, Ley DH. Health Survey of Mycoplasma gallicepticum-free house finches (Carpodacus mexicanus) from Wisconsin. Journal of Wildlife Diseases. 2003 (in press). Huang, Z., Y. Elankumaran, A. Panda, Y. S. Abdul, and S. K. Samal. Recombinant Newcastle disease virus expressing the VP2 protein of infectious bursal disease virus (IBDV) provides protection from IBDV challenge. Twenty-second Annual Meeting of American Society for Virology, July 12-16, Davis, CA. 2003. Jackwood D. J., and S. E. Sommer. Identification of infectious bursal disease virus quasispecies in commercial vaccines and field isolates of this double-stranded RNA virus. Virology. 304:105-113. 2002. Jackwood, D. J. and S. E. Sommer. Characterization and sequence comparison of infectious bursal disease virus quasispecies. XIII Congress of the World Vet. Poultry Assoc. 2003. Jackwood, D. J. and S. E. Sommer. Molecular epidemiology of infectious bursal disease viruses: Use of real-time RT/PCR to detect new variants. XIII Congress of the World Vet. Poultry Assoc. 2003. Jackwood, D. J., and S. E. Sommer. Virulent vaccine strains of infectious bursal disease virus not distinguishable from wild-type viruses with the use of a molecular marker. Avian Dis. 46:1030-1032. 2002. Jackwood, D. J., B. D. Spalding, and S. E. Sommer. Real-time reverse transcriptase-polymerase chain reaction detection and analysis of nucleotide sequences coding for a neutralizing epitope on infectious bursal disease virus. Avian Dis. 47:738-744. 2003. Jackwood, D. J., Q. Zhang and Y. M. Saif. Effect of infectious bursal disease on Campylobacter shedding in chickens. XIII Congress of the World Vet. Poultry Assoc. 2003. Jacobs AJ, Njenga MK, Seal BS, Kavanaugh, D. Subtype B metapneumovirus resembles subtype A more closely than C or human metapneumovirus with respect to the phosphoprotein, and second matrix and small hydrophobic proteins. Virus Res. 92:171-178. 2003. Jacobs, J.A., Njenga, M.K. Alvarez, R., Mawditt, K., Britton, P., Cavanagh, D., Seal, B.S. Subtype B avian metapneumovirus resembles subtype A more closely than subtype C or human metapneumovirus with respect to the phosphoprotein, second matrix and small hydrophobic proteins. Virus Res. 92:171-178.2003. Jirjis FF, Noll SL, Halvorson DA, Nagaraja KV, Shaw DP. Pathogenesis of avian pneumovirus infection in turkeys. Vet Pathol.300-310. 2002. Jirjis, F.F., Noll, S.L., Halvorson, D.A., Nagaraja, K.V., Townsend, E.L., Goyal, S.M., and Shaw, D.P. Rapid detection of avian pneumovirus in tissue culture by microindirect immunofluorescence test. J. Vet. Diag. Invest. 14:172-175. 2002. Kapczynski, D.R., Hilt, D., Shapiro, D., Sellers, H.S., Jackwood, M.W. Protection of chickens from infectious bronchitis by in ovo and intramuscular vaccination with a DNA vaccine expressing the S1 glycoprotein. Avian Dis. 47:272-285. 2003. Kapczynski, D.R., Sellers, H.S., Rowland, G., Jackwood, M.W. Detection of in ovo inoculated infectious bronchitis virus by in situ hybridization in epithelial cells of the lung and bursa in chickens. Avian Dis. 46: 679-685. 2002. Kapczynski, D.R., Sellers, H.S., Simmons, V., Schultz-Cherry, S. Sequence analysis of the S3 gene from a turkey reovirus. Virus Genes. 25: 95-100. 2002. Khan, M. I. Molecular biology of bacteria. Symposium on "Molecular biology made easy". American College of Poultry Veterinarian. Sacramento California. March 8, 2003. Khan, M. I. Avian Pathogenic Mycoplasmas. PCR detection of Microbial Pathogens. Methods in Molecular Biology. eds. J. Frey and K. Sachse. Humana Press Inc. Totowa, NJ. Inc. Totowa, NJ. pp.203-229, 2003. Kim, T-J, Schnitzlein, W.M., McAloose, D., Pessier, A.P. and Tripathy, D.N. Characterization of an avianpox virus isolated from an Andean condor (Vultur gryphus). Veterinary Microbiology 96: 237-246. 2003. Kollias GV, Sydenstricker KV, Kollias HW, Ley DH, Hosseini PR, Connolly V, Dhondt AA. Experimental infection of individually caged house finches with Mycoplasma gallisepticum. Journal of Wildlife Diseases. 2003 (in press). Kommers, G.D., King, D.J., Seal, B.S., Brown, C.C. Pathogenesis of chicken-passaged Newcastle disease viruses isolated from chickens and wild, and exotic birds. Avian Dis. 47:319-329. 2003. Kommers, G.D., King, D.J., Seal, B.S., Brown, C.C. Virulence of six heterogenous-origin Newcastle disease virus isolates before and after sequential passage in domestic chickens. Avian Pathology. 32:81-93. 2003. Ladman, B. S., C. R. Pope, A. F. Ziegler, T. Swieczkowski, and J. M. Callahan, and J. Gelb, Jr. Protection of chickens following live and inactivated virus vaccination against challenge with nephropathogenic infectious bronchitis virus PA/Wolgemuth/98. Avian Dis. 46:938-944. 2002. LaFleur, D., H. Kim, and D.N. Foster. Expression of the chicken homologue of the mouse double minute 2 gene. Biochem. Biophys. Acta 1574:277-282. 2002. Ley DH, Martinez A, Vaillancourt J-P. Mycoplasma gallisepticum outbreaks in North Carolina, 1999-2001: Lessons learned from the tail end of the epidemic curve. XIII Congress of the World Veterinary Poultry Association, Co-sponsored by the American Association of Avian Pathologists and the American Veterinary Medical Association, Program and Abstracts (pp. 203), and 2003 AVMA Convention Notes CD, Denver, CO; July 19-23, 2003. Ley DH, Swarthout E, Sydenstricker K, Kollias GV, Dhondt AA. Mycoplasma gallisepticum conjunctivitis in house finches (Carpodacus mexicanus). Correlations among clinical signs and detection by polymerase chain reaction from conjunctival and choanal swabs. 52nd Annual Wildlife Disease Association Conference, Program and Abstracts (pp. 32-33), Saskatoon, Saskatchewan, August 11-14, 2003. Ley DH. Mycoplasma gallisepticum infection. In: Diseases of Poultry, 11th ed., BW Calnek, HJ Barnes, CW Beard, LR McDougald, YM Saif, eds. Ames, Iowa: Iowa State University Press, pp. 722-744. 2003. Lin, TL, Wu, CC, Chang, HC, and Hsieh, MK. DNA vaccination against classical and variant infectious bursal disease virus. Proc. of the DNA Vaccines 2002 Conference. p. 51. 2002 Lopes V, Back A, Halvorson DA, Nagaraja KV. Minimization of pathologic changes in Ornithobacterium rhinotracheale infection in turkeys by temperature-sensitive mutant strain. Avian Dis. 46:177-185. 2002. Lopes VC, Back A, Shin HJ, Halvorson DA, Nagaraja KV. Development, characterization, and preliminary evaluation of a temperature-sensitive mutant of Ornithobacterium rhinotracheale for potential use as a live vaccine in turkeys. Avian Dis. 46:162-168. 2002. Lopes VC, Velayudhan B, Halvorson DA, Nagaraja KV. Survival of Ornithobacterium rhinotracheale in sterilized poultry litter. Avian Dis 46:1011-1014. 2002. Lwamba HC, Bennett RS, Lauer DC, Halvorson DA, Njenga MK. Characterization of avian metapneumoviruses isolated in the USA. Animal Health Res. Rev. 3:107-117. 2002. Lwamba HC, Halvorson DA, Nagaraja KV, Turpin EA, Swayne D, Seal BS, Njenga MK. Antigenic cross-reactivity among avian pneumoviruses of subgroups A, B, and C at the matrix but not nucleocapsid proteins. Avian Dis 46:725-729. 2002. Malik, Y.S., Olsen, K., Kumar, K., and Goyal, S.M. In vitro antibiotic resistance profiles of Ornithobacterium rhinotracheale strains isolated from Minnesota turkeys during 1996-2002. Avian Dis. 47:588-593. 2003. Mauro, L.J. and D.N. Foster. Regulators of Telomerase Activity. Am. J. Respitory Cell Molec. Biol. 26:521-524. 2002. Moura, L., and V. N. Vakharia. Development and evaluation of an in ovo DNA vaccine against Newcastle disease virus. 8th Congress of the World Veterinary Poultry Assn. and the American Association of Avian Pathologist, July 19-23, 2003, Denver, CO. 2003. Munir S, Kapur V. Regulation of host cell transcriptional physiology by the avian pneumovirus provides key insights into host-pathogen interactions. J Virol. 77:4899-4910. 2003. Njenga MK, Lwamba HCM, Seal BS. Metapneumoviruses in birds and humans. Virus Res: 91:163-169. 2003. Oshop, G. L., S Elankumaran, V. N. Vakharia, and R.A. Heckert. In ovo delivery of DNA to the avian embryo. Vaccine 21, 1275-1281. 2003. Patnayak, D.P., Munir, K. and Goyal, S.M. The role of vaccine inoculation routes on protective immunity against avian pneumovirus. J. Appl. Res. Vet. Med. 1:122-126. 2003. Patnayak, D.P., Sheikh, A.M., Gulati, B.R. and Goyal, S.M. Experimental and field evaluation of a live vaccine against avian pneumovirus. Avian Pathol. 31:377-382. 2002. Paustian ML, May BJ, Cao D, Boley D, Kapur V. Transcriptional response of Pasteurella multocida to defined iron sources. J Bacteriol.184:6714-6720. 2002. Perkins, L.E.L., Swayne, D.E. Susceptibility of laughing gulls (Larus atricilla) to H5N1 and H5N3 highly pathogenic avian influenza viruses. Avian Dis. 46: 877-885. 2002. Perkins, L.E.L., Swayne, D.E., Varied pathogenicity of a Hong Kong-origin H5N1 avian influenza virus in four passerine species and budgerigars. Veterinary Pathology. 40:14-24. 2003. Peters, M, Wu, CC, and Lin, TL. Immune suppression induced by infectious bursal disease virus polyprotein independently of virus infection. Proc. of the 54th North Central Avian Disease Conference. p. 26. 2003. Pillai, SR, Mays HL, Jr., Ley DH, Luttrell P, Panangala VS, Farmer KL, Roberts SR. Molecular variability of house finch Mycoplasma gallisepticum isolates as revealed by sequencing and restriction fragment length polymorphism analysis of the pvpA gene. Avian Dis. 47:640-648. 2003. Rautenschlein, S., A. M. Sheikh, D. P. Patnayak, R. L. Miller, J. M. Sharma, and S. M. Goyal. Effect of an immunomodulator on the efficacy of an attenuated vaccine against avian pneumovirus in turkeys. Avian Dis. 46:555-561. 2002. Reynolds, D. L., S. Akinc and T. Glanville. Biosecurity of Large-Scale Composting. Poster presentation. AAAP / AVMA annual meeting. Denver Colorado. July 19 23, 2003. Reynolds, D. L., S. Akinc and T. Glanville. Biosecurity of Large-Scale Composting. Poster presentation. Conference of Research Workers in Animal Diseases. Chicago, IL. November 9 -11, 2003. Saif, Y. M. Molecular Methodologies: Challenges and Opportunities. Proc. 75th Northeastern Conference on Avian Diseases (NECAD), Orono, Maine, June 12, 2003. Sato, N., K . Mastuda, C. Sakuma, D.N. Foster, R.W. Oppenheim, H.Yaginuma. Regulated gene expression in the chicken embryo by using replication-competent retroviral vectors. J. Virol 76:1980-1985. 2002. Shin HJ, Nagaraja KV, McComb B, Halvorson DA, Jirjis FF, Shaw DP, Seal BS, Njenga MK. Isolation of avian pneumovirus from mallard ducks that is genetically similar to viruses isolated from neighboring commercial turkeys. Virus Res 83:207-212. 2002. Shin, H.J., Cameron K.T., Jacobs, J.A., Turpin, E.A., Halvorson, D.A., Goyal, S.M., Nagaraja, K.V., Kumar, M.C., Lauer, D.C., Seal, B.S., and Njenga, M.K. Molecular epidemiology of subgroup C avian pneumoviruses isolated in the United States and comparison with subgroup A and B viruses. J. Clin. Microbiol. 40:1687-1693. 2002. Singh, P. and Tripathy, D.N. Fowlpox virus infection causes a lymphoproliferative response in chickens. Viral Immunology 16: 223-227. 2003. Singh, P. and Tripathy, D.N. Generation and evaluation of recombinant vaccines for protection against fowlpox and reticuloendotheliosis in chickens. Poster No. 49. Program and Abstracts of XIII Congress of the World Veterinary Poultry Association, Denver, CO. p. 136. 2003. Singh, P., Kim, T-J and Tripathy, D.N. Identification and characterization of fowlpox virus strains using monoclonal antibodies. J. Vet. Diag.Ivest.15:50-54. 2003. Singh, P., Schnitzlein, W.M. and Tripathy, D.N. Reticuloendotheliosis virus sequences within the genomes of field strains of fowlpox virus display variability. J. Virology 77:5855-5862. 2003. Srinivasan, V., Schnitzlein, W.M. and Tripathy, D.N. A consideration of previously uncharacterized fowlppox virus unidirectional and bi-directional promoters for inclusion in homologous recombinant vaccines. Avian Dis. 47: 286-295. 2003. Subbarao, K., Chen, H., Swayne, D., Mingay, L., Fodor, E., Brownlee, G., Xu, X., Lu, X., Katz, J., Cox, N., Matsuoka, Y. Evaluation of a genetically modified reassortant H5N1 influenza A virus vaccine candidates generated by plasmid-based reverse genetics. Virology. 305:192-200. 2003. Swayne, D.E. and Halvorson, D.A. Influenza pp. 135-160. In Saif, Y.M., Barnes, H.J., Fadly, A., Glisson, J.R., McDougald, L.R. and Swayne, D.E. (eds.) Diseases of Poultry, 11th ed., Iowa State University Press, Ames, IA. 1231 pp. 2003. Swayne, D.E., King, D.J.. Zoonosis update: avian influenza and Newcastle disease. J. American Veterinary Medical Assn. 222:1534-1540. 2003. Tripathy, D. N. and Reed, W. M. Pox. In: Diseases of Poultry. 11th ed. Eds..Saif, M. et al. Iowa State University Press , Ames, Iowa. 2003. Tripathy, D.N. Impact of vaccines and future of genetically modified vaccines for poultry. 3rd International Veterinary Vaccines and Diagnostics Conference, July 13-18, Guelph, Canada. 2003. Tripathy, D.N., Schnitzlein, W.M., Srinivasan, V. Kim, T.J. and Singh, P. Molecular Strategies towards improvement of vaccines against fowlpox. U.S. Animal. Health Assn. San Diego, CA. 2003. Vakharia, V.N. and M. Liu. Both VP1 and VP2 proteins contribute to the virulence of infectious bursal disease virus in vivo. 8th International Symposium on ds-RNA viruses. September 13-18, Lucca, Italy. 2003. Velayudhan BT, Lopes VC, Noll SL, Halvorson DA, Nagaraja KV. Avian pneumovirus and its survival in poultry litter. Avian Dis 47:764-768. 2003. Wang, X, Schnitzlein, W. M, Tripathy, D. N., Girshick T. and Khan, M.I. Construction and immunogenicity studies of recombinant fowlpox virus containing the S1 gene of Massachusetts 41 strain of IBV. Avian Dis. 46 831-838. 2002. Wu, C. C., M. K. Hsieh, and T. L. Lin. The effect of chicken IFN- on protection of chickens against infectious bursal disease by DNA vaccination. In: Proc. of Modern Vaccine Adjuvants and Vaccine Delivery Systems Conference. p. 83. 2003. Zanetti, F., Rodriquez, M., King, D.J., Capua, I., Carillo, E., Seal, B.S. Berinstein, A. Matrix protein gene sequence analysis of avian paramyxovirus 1 isolates obtained from pigeons. Virus Genes. 26: 199-206. 2003. Ziegler, A. F., B. S. Ladman, P. A. Dunn, A. Schneider, S. Davison, P. G. Miller, H. Lu, D. Weinstock, M. Salem, R. J. Eckroade, and J. Gelb, Jr. Nephropathogenic infectious bronchitis in Pennsylvania chickens 1997-2000. Avian Dis. 46:847-858. 2002.

Impact Statements

Date of Annual Report: 02/28/2005

Report Information

Annual Meeting Dates: 10/01/1999 - 09/30/2004
Period the Report Covers: 10/01/1999 - 09/01/2004

Participants

CT AES -M. Khan (Secretary);
DE AES -J. E. Dohms and J. Gelb, Jr. (Chair);
IA AES -D. L. Reynolds;
IL AES -D. N. Tripathy;
IN AES -C. C. Wu;
MN AES -M. K. Njenga;
OH AES -Y. M. Saif;
USDA ARS SEPRL -D. Suarez;

New member for the new/renewal project NC-1019:
B. Buckles (New York);

Administrative Advisor: Jeffrey Klausner (University of Minnesota);
USDA CSREES Representative: Peter Johnson

Brief Summary of Minutes

The NC-228 annual Technical Committee meeting was held on Friday, November 12, 2004 at Grant Park Room in the Congress Hotel, Chicago, IL. Jack Gelb, Chair of NC-228, opened the meeting at 2 PM. Welcomes and introductions followed.

New Project NC-1019 and the Annual and Termination Reports for Current Project NC-228
Dr. Klausner congratulated the Committee for submitting an excellent proposal for the renewal project NC-1019. He and Dr. Gelb also indicated that the NCRA Multistate Research Committee that had reviewed the project had made two recommendations: to develop more stakeholder interaction with representatives of the animal health industry and practitioners; and to make additional efforts to leverage financial support.

Dr. Gelb agreed to coordinate the preparation of the NC-228 2003-2004 annual report and will prepare a draft and circulate it to the participants for review. After making all changes, Dr. Gelb will submit the final version of the NC-228 2003-2004 annual report by January 13, 2005, the 60-day deadline from the date of the 2004 annual meeting.

Dr. Klausner indicated that this years 2003-2004 annual report, the last report for the NC-228 project, could serve as the termination report with the addition of the impact and accomplishments sections. The Committee agreed to the best way to prepare the NC-228 termination report was to take all the highlighted bullets from the annual reports and assemble them, as well as the publications and a summary of the impact statements into a draft for circulation to the participants for review. Dr. Gelb, the out-going chair of NC-228, agreed to assist the newly elected chair of the NC-1019 project with the preparation of the NC-228 termination report. The deadline for the NC-228 termination report is March 15, 2005.

Given that the new project, NC-1019 had just commenced on October 1, 2004, the first annual report will be prepared after next years 2005 annual meeting.

Update from Dr. Peter Johnson (USDA CREES)
Dr. Johnson (USDA, CREES) also congratulated the Committee for their the efforts in successfully developing a new proposal on respiratory diseases of poultry. He reiterated his strong support for multi-state regional projects. Dr. Johnson distributed a handout entitled, USDA Cooperative State Research, Education, and Extension Service (CREES) Report, Chicago IL November 2004. He reported on the following.
1. CREES personnel changes.
2. A new CSREES website (www.csrees.usda.gov).
3. Competitive Programs (www.csrees.usda.gov/fo/funding.cfm). Updated FY2005 competitive grant program with emphasis on the ones of interest to our regional project. Discussed the FY2004 NRI program dates and the funding within various programs. Noted the Postdoctoral Fellowship program and the restriction to US citizens and the need to make these awards. Noted the Animal Well-Being Assessment and Improvement and Veterinary Immunological Reagents programs. Noted the identification of Avian Coccidia, Mareks Disease, and Poult Enteritis Mortality Syndrome as diseases receiving high priority species-specific status in FY 2005. Some concern was expressed on the part of the Committee as to how these diseases were identified. Indicated that proposals on exotic ND and AI would be considered under non-species specific high priority areas.
4. The projected Presidents non-defense budget from 2004-2009 detailing anticipated reductions in USDAs as well as other agencys budgets.

Election of officers for 2004-2005
A Nomination Subcommittee consisting of Drs. Wu, Saif and Gelb recommended Dr. David Suarez (USDA, ARS, SEPRL) for Chair and Dr. Ching-Ching Wu (Indiana) for Secretary for two-year terms (2005-2006). Both were unanimously elected and thanked by the Committee for taking on these important responsibilities.

Stakeholders participation in NC-1019
Dr. Gelb initiated a discussion about bringing in stakeholders from the poultry industry to seek their input for the research direction and support for our regional project. It was proposed that one or two persons could be invited from the industry. The group discussed several names of stakeholders and agreed that Dr. Bruce Stewart-Brown of Perdue Farms, Inc. should be invited to our next meeting. We will ask Dr. Stewart-Brown to report on the status of current poultry diseases in general and the specific nature and impact of respiratory diseases on production in layers, broilers and turkeys.

Potential new location for the NC-1019 annual meeting
The Committee discussed the possibility of moving the NC-1019 annual meeting to another location in conjunction with another meeting to encourage better attendance. Members polled at this years meeting indicated a preference in the following order: Southern Conference on Avian Diseases meeting and the US Poultry and Egg Assn. trade show in Atlanta, Georgia in January; US Animal Health Assn. meeting in various locations in October, Western Poultry Disease Conference in various locations in March, and lastly, the current Chicago location in conjunction with the Conference of Research Workers in Animal Diseases in November. Given that several members were not in attendance, Dr. Gelb agreed to poll all participants via e-mail so that this matter could be discussed further before any decision was made.

Discussion of the annual station reports
The Committee discussed research activities during the period from October 1, 2003 - September 30, 2004.

Work planned for the coming year

AVIAN INFLUENZA VIRUS
Avian influenza reservoirs will be identified in the New England states (CONNECTICUT). Develop improved diagnostic capabilities including real time PCR as well as other rapid on-farm tests for economically important respiratory diseases (CONNECTICUT). Develop and optimize of real time multiplex-PCR tests for avian influenza (CONNECTICUT).

Characterize AIV isolates from the 2004 Delmarva outbreak (DELAWARE).

Continue surveillance for AIV. Studies will be initiated on the molecular changes in influenza viruses associated with crossing the species barrier (OHIO)

AVIAN METAPNEUMOVIRUS
Develop a continuous cell line capable of growing higher titers of APV (MINNESOTA).

Continue surveillance for avian pneumoviruses (OHIO).

ESCHERICHIA COLI
Expand studies to evaluate the effect of various strains of avian pathogenic E. coli (APEC) on avian macrophage gene expression and to compare these responses to those observed when macrophage are exposed to viral pathogens (DELAWARE).

Continue sequencing the genome of an avian clone of E. coli (MINNESOTA).

INFECTIOUS BRONCHITIS VIRUS
IBV S gene specific recombinant DNA vaccine and its application in-ovo using interferon Type 1 as an adjuvant will continue. Develop and optimize of real time multiplex-PCR tests for infectious bronchitis for differentiation of serotypes (CONNECTICUT).

Characterize Delmarva isolates from 2004 (DELAWARE).

INFECTIOUS BURSAL DISEASE VIRUS
Examine the pathogenesis of IBDV using reverse genetically engineered strains for the purpose of understanding the molecular events and mechanisms by which the virus interacts with bursa of Fabricius. Study effect of chicken cytokine genes on immunomodulation and protection of chickens against IBD by DNA vaccination. Study the effect of boosting with transgenic algae expressing IBDV VP2 on DNA vaccination of chickens against IBD (INDIANA).

Continue studies on the genetic drift observed in IBDV and its relationship to antigenic variations in these viruses. Produce and characterize monoclonal antibodies to the vvIBDV (OHIO).

ORNITHOBACTERIUM RHINOTRACHEALE (ORT)
Investigate the role of ORT on peritonitis in laying chickens (MINNESOTA)

NEWCASTLE DISEASE VIRUS
Characterize Delmarva isolates from 2004 (DELAWARE).

MYCOPLASMAS
Additional work with the unusual M. synoviae strain will continue (DELAWARE).

The meeting was adjourned at 4:30 pm on November 13, 2004.

Minutes submitted by M. Khan.

Accomplishments

Objective 1. Determine the pathogenesis and interactions of specific agents. AVIAN FOWLPOX The hemagglutinin gene homologue in the genome of fowlpox virus is not essential for virus multiplication (ILLINOIS). AVIAIN INFLUENZA VIRUS (AIV) Low path H7N2 AIV isolate from Delmarva broiler chickens were more pathogenic for broilers and turkeys than for SPF white leghorn chickens and suggest that host species plays an important role in susceptibility to disease (DELAWARE). Pathogenesis of the highly pathogenic H5N1 avian influenza viruses from Asia was performed to identify the host ranges for these viruses, sequence analysis of viruses from the region, and vaccination studies to determine what vaccines will be effective if the virus was ever introduced into the U.S. (USDA ARS SEPRL). AVIAN METAPNEUMOVIRUS (APV) Studies suggested that secondary bacterial infection of APV-infected turkeys may aggravate disease (MINNESOTA). INFECTIOUS BURSAL DISEASE VIRUS (IBDV) A reverse genetically engineered IBDV is being generated for the study of pathogenesis of infectious bursal disease. Preliminary data revealed the virus grew in Vero cells and had positive immunofluorescence (INDIANA). Interference in the replication of a virulent strain of IBDV was shown to occur when a vaccine strain was present in the host (OHIO). IBDV induced immune suppression increased the colonization and shedding of the food-borne pathogen, Campylobacter jejuni in chickens (OHIO). ESCHERICHIA COLI Gene expression of avian macrophage activation was conducted using a 4906 element macrophage-specific microarray. Of the elements on AAM, 981 (20%) exhibited significant (>2-fold, p<0.01) changes in expression changes during phagocytosis of Escherichia coli and 243 (5%) exhibited significant expression for both phagocytosis and LPS stimulation, representing a set of core response elements. (DELAWARE). NEWCASTLE DISEASE VIRUS (NDV) The effect of virulent NDV strains varies among host species. Project scientists and collaborators from the University of Georgia infected chickens, turkeys, and pigeons with an isolate (California 2002) of exotic Newcastle disease (END) virus and monitored for clinical disease and gross and microscopic tissue lesions. All species were readily infected and shed virus that could be readily transmitted to other susceptible birds, but only the chickens and disease free turkeys experienced severe clinical disease with mortality and extensive tissue lesions (USDA ARS SEPRL). SARS CORONAVIRUS Five common poultry species could not be easily infected with the SARS virus and therefore likely played no role in either the origin or spread of the virus (USDA ARS SEPRL in collaboration with the Centers for Disease Control). Objective 2. Surveillance, occurrence and consequences of agents and host variation on disease susceptibility. AVIAN INFLUENZA VIRUS A H3N2 influenza A virus was isolated from turkey flock with a drop in egg production. Preliminary sequence analysis of the isolated virus confirmed that it was influenza A virus with 99% (235/236) identity with the matrix gene of a swine influenza A virus, A/Swine/Illinois/100084/01 (H1N2) (OHIO collaborating with USDA Southeast Poultry Research Laboratory, USDA National Veterinary Service Laboratory, and University of Maryland.). INFECTIOUS BRONCHITIS VIRUS Viruses other than low path H7N2 avian influenza were recovered from cases of respiratory disease in commercial broilers during surveillance for AIV in 2004 on the Delmarva Peninsula. IBV was the most frequently isolate virus from the respiratory tract (52%) followed by NDV (38%), and avian adenovirus (10%). The isolation of IBV, NDV and Adenovirus was more frequent from older broiler flocks (>28 days of age) than from younger (<28 days of age) flocks. Arkansas S1 genotype type IBVs were much more often isolated than were other genotypes (Massachusetts and Connecticut (DELAWARE). INFECTIOUS BURSAL DISEASE VIRUS The presence of a serotype 2 IBDV strain in penguins from a zoological park in the United Kingdom was confirmed using RT-PCR and nucleotide sequence analysis (OHIO). Studies on 38 field isolates of IBDV revealed at least one substitution mutation in every amino acid except one across the hydrophilic B epitope region of VP2 suggesting that antigenic diversity due to genetic drift may be responsible for continuing problems with IBDV infections in the U.S (OHIO). MYCOPLASMOSIS An atypical strain of M. synoviae was isolated from broiler flocks hatched from young breeder hens in Arkansas. The strain appears to be highly bird adapted. Growth in Frey broth is extremely slow and the strain does not grow on PPLO plates with the appropriate supplements. The M. synoviae strain should be compared with more typical M. synoviae strains (DELAWARE). NEWCASTLE DISEASE VIRUS Viruses other than low path H7N2 avian influenza were recovered from cases of respiratory disease in commercial broilers during surveillance for AIV in 2004 on the Delmarva Peninsula. IBV was the most frequently isolate virus from the respiratory tract (52%) followed by NDV (38%), and avian adenovirus (10%). The isolation of IBV, NDV and Adenovirus was more frequent from older broiler flocks (>28 days of age) than from younger (<28 days of age) flocks (DELAWARE). NDV isolates from the outbreak of virulent Newcastle disease (v-ND) in southern California, Nevada, Arizona and Texas during 2002-03 were compared to Mexican and Honduras 1996/2000 v-ND V isolates as well as reference avian paramyxovirus type-1 isolates. Phylogenetic analysis demonstrated that the CA 2002/03, AZ, TX and NV viruses were closely related to the Mexican and Honduras viruses. However, the TX/2003 NDV represented a separate introduction of v-NDV as this virus was more closely related to the Mexican 2000 isolates than the CA, AZ, and NV/2002-03 viruses (USDA ARS SEPRL). Avian paramyxovirus-1 (APMV-1), also referred to as NDV, variants of low virulence were isolated from chickens, ducks and other unidentified species found in live-bird markets of the northeastern U.S. These isolates were characterized as APMV-1 by the hemagglutination-inhibition (HI) assay utilizing NDV-specific polyclonal antisera. However, the isolates failed to react with a monoclonal antibody that has specificity for a wide variety of APMV-1 isolates. Genetically, the isolates were found by project scientists and collaborators at the USDA, APHIS, National Veterinary Services Laboratories, to be unique NDV isolates designated as a lineage 6 genotype. This lineage had not been previously reported in North America and further substantiates the heterogeneous genetic nature of these commercially important pathogens found worldwide (USDA ARS SEPRL). Objective 3. Develop new and improved methods for the diagnosis, prevention, and control of avian respiratory diseases. AVIAN INFLUENZA VIRUS Multiple outbreaks in the U.S. were evaluated using molecular epidemiology methods for H1, H5, H6, and H7 viruses, which helped establish the origin of viruses and helped establish the risk of these viruses to the poultry industry (USDA ARS SEPRL). An egg yolk antibody test was developed to perform AIV surveillance in layers (USDA ARS SEPRL). AVIAN METAPNEUMOVIRUS Sequencing of the APV genome was completed thus paving way for generation of reverse genetic system for vaccine development and further characterization of the virus. The genome of another recent APV isolate from wild geese was also sequenced and was shown to have a large attachment (G) protein, a characteristic that could be used as a natural marker (MINNESOTA and USDA ARS SEPRL). A real-time RT PCR test capable of detecting the major APV subtypes has been developed for use in surveillance against introduction of European subtypes into the U.S. turkey population (MINNESOTA). A recent APV isolate from wild geese was determined to be avirulent for turkeys, thus making it a good vaccine candidate. Field evaluation of the efficacy of this wild bird isolate as vaccine in turkeys is ongoing (MINNESOTA). A virosome vaccine for APV was developed and evaluated. Turkeys vaccinated with APV virosomes had decreased viral load in the lungs and clinical signs of disease following a virulent challenge infection (USDA ARS SEPRL). INFECTIOUS BURSAL DISEASE VIRUS Plasmids encoding the chicken IL-2 gene and the large segment gene of IBDV given two times at separate sites enhanced protection of chickens against IBD. Immunostimulating modulators such as chicken IL-2 may be used to enhance immunity and protection of chickens against IBD by DNA vaccination (INDIANA). NEWCASTLE DISEASE VIRUS Six composting studies either have been, or are in the process of being, conducted to evaluate biosecurity of large-scale composting of animal carcasses. These trials were initiated at various times of the year to explore seasonal effects. The duration of the trials were well over one year, lasting from 15-18 months. The results of these trials provide evidence that composting resulted in the inactivation of NDV and avian encephalomyelitis virus. The composting methods used also contained the viruses within the compost pile (IOWA).

Publications

Abdel-Alim, G.A., M.H.H. Awaad, and Y.M. Saif: Characterization of Egyptian field strains of infectious bursal disease virus. Avian Dis. 47:1452-1457, 2003. Al-Natour, M.Q., L.A. Ward, Y.M. Saif, B. Stewart-Brown and L.D. Keck. Effect of different levels of maternally derived antibodies on protection against infectious bursal disease virus. Avian Dis. 48:177-182, 2003. Alvarez R, Njenga MK, Scott M, Seal BS. Development of a nucleoprotein-based enzyme-linked immunosorbent assay utilizing a synthetic peptide antigen for detection of avian metapneumovirus antibodies in turkey sera. Clin Diag Lab Immunol. 11: 245-249, 2004. Alvarez, R., Lwamba, H.M., Kapczynski, D.R., Njenga, M.K., Seal, B.S. Nucleotide and predicted amino acid sequence-based analysis of the avian metapneumovirus type C cell attachment glycoprotein (G) gene: Phylogenetic analysis and molecular epidemiology of U.S. pneumoviruses. 2003. Journal of Clinical Microbiology. v. 41. p. 1730-1735. Alvarez, R., Njenga, M.K., Scott, M., Seal, B.S.: Development of a nucleoprotein-based enzyme-linked immunosorbent assay utilizing a synthetic peptide antigen for detection of avian metapneumovirus antibodies in turkey sera. Clin. Diagn. Lab. Immunol. 2004. v. 11. p. 245-249. Beck J.R., Swayne D.E., Davison S., Casavant S., Gutierrez C. Validation of egg yolk antibody testing as a method to determine influenza status in white leghorn hens. 2003. Avian Diseases. 47:1196-1199. Bennett RS, Njenga MK, Nezworski J; Velayudhan BT; Nagaraja KV, DH; Dyer N; Lemire T; Lauer DC; Halvorson DA. Detection of Avian pneumovirus in wild birds and domestic turkeys from central North. Avian Dis. IN PRESS. Bliss, T. W., J.E. Dohms, M. G. Emara, and C. L. Keeler. 2004. Gene Expression Profiling of Avian Macrophage Activation. Veterinary Immunology and Immunopathology (in press). Boettger, C. M. and J. E. Dohms. Development of Methods to Separate Mycoplasma gallisepticum Field Strains from Non-pathogenic Avian Mycoplasmas. Presented at the 76th Northeastern Conference on Avian Diseases, June 9-11, State College, Pennsylvania. Bulaga, L.L., L. Garber, D.A. Senne, T.J. Myers, R. Good, S. Wainwright, S. Trock, D.L. Suarez. 2003. Epidemiologic and Surveillance Studies on Avian Influenza in Live-Bird Markets in New York and New Jersey, 2001. Avian Diseases. 47:996-1001. Bulaga, LL., L. Garber, D. Senne, T.J. Myers, R. Good, S. Wainwright, D.L. Suarez. 2003. Descriptive and Surveillance Studies of Suppliers to New York and New Jersey Retail Live Bird Markets. Avian Diseases. 47:1169-1176. Caterina, K. M., S. Frasca Jr, T. Girshick and M. I. Khan. Development of a multiplex PCR for detection of avian adenovirus, avian reovirus, infectious bursal disease virus and chicken anemia virus. Molecular and Cellular Probes. 18:293-298. 2004. Cloud, Sandra, Jack Gelb, Conrad Pope, John Rosenberger and Brian Ladman. Characterization of Avian Influenza Viruses Isolated from Delmarva Broilers. Respiratory disease virus characterization from Delmarva high mortality flocks. Proc. 39th National Meeting on Poultry Health and Processing. Ocean City, Maryland. p. 96. October 6-8, 2004. Donoghue, K., Lomniczi, B., McFerran, B., Conner, T.J., Seal, B., King, D., Banks, J., Manvell, R., White, P.S., Richmond, K., Jackson, P., Hugh-Jones, M.: Retrospective characterization of Newcastle disease virus in relation to other epidemics, past and present. Epidemiol. Infect. 2004. v. 132. p. 357-368. Gelb, J., Jr., J. K. Rosenberger, B. S. Ladman, and S. S. Cloud. Exotic Newcastle disease-Real world trials at the University of Delaware. Proc. 38th National Meeting on Poultry Health and Processing. Ocean City, Maryland. p. 17. October 22-24, 2003. Gelb, Jack, Jr. Respiratory Coronavirus of Chicken: Ongoing Challenges Posed by Infectious Bronchitis Virus. Seventh Annual Conference On Vaccine Research. National Institute for Infectious Diseases. Arlington, Virginia May 24-26, 2004. Gelb, Jack, Jr., and Mariano Salem. Avian Influenza in Delmarva. North Atlantic Regional Conference, Northeast District Veterinary Command. Fort Dix, NJ, Oct. 12, 2004. Gelb, Jack, Jr., Sandra S. Cloud, Brian S. Ladman, Mariano Salem, and John K. Rosenberger. Respiratory disease virus characterization from Delmarva high mortality flocks. Proc. 39th National Meeting on Poultry Health and Processing. Ocean City, Maryland. p. 93. October 6-8, 2004. Glanville, T. D., T. L. Richard, J. D. Harmon, D. L. Reynolds, S. S. Sadaka and S. Akinc. Environmental Impact & Biosecurity of Composting for Emergency Disposal of Livestock Mortalities. Paper No. 032262. Presented at the American Society of Agricultural Engineers 2003 Annual International Meeting. Las Vegas, Nevada. July 27-30, 2003. Glanville, T.D., T.L. Richard, J.D. Harmon, D.L. Reynolds, H. K. Ahn and S. Akinc. Environmental Impact & Bio-security of Composting for Emergency Disposal of Livestock Mortalities - Year 2 Project Update. Paper Number: 044069. ASAE/CSAE Annual International Meeting Sponsored by ASAE/CSAE, Fairmont Chateau Laurier, The Westin, Government Centre Ottawa, Ontario, Canada 1 - 4 August 2004. Hanson BA, Stallknecht DE, Swayne DE, Lewis LA, Senne DA. Avian influenza viruses in Minnesota ducks during 1998-2000. 2003. Avian Dis. 47:867-71. Hsieh, M.K.; Lin, T.L.; Wu, C.C. 2004. The effect of chicken Interleukin-2 on protection of chickens against infectious bursal disease by DNA vaccination. In: The Proceedings of The 55th North Central Avian Disease Conference. Jackwood, D. J., and S. E. Sommer. Molecular epidemiology of infectious bursal disease viruses: Distribution and genetic analysis of new variants in the United States. Abstr. 141st AVMA Mtg. 2004. Jackwood, D. J., R. E. Gough and S. E. Sommer. Nucleotide and amino acid sequence analysis of a Birnavirus isolated from penguins. Vet. Record. (in press), 2004. Jackwood, D. J., S. E. Sommer, R. E. Gough, S. E. Drury, D. de B. Weichman, J. R. Chitty and G. E. S. Summerhays. Sequence analysis of an infectious bursal disease virus isolated from penguins in the United Kingdom. Abstr. 141st AVMA Mtg. 2004. Jacobs, J.A., Njenga, M.K. Alvarez, R., Mawditt, K., Britton, P., Cavanagh, D., Seal, B.S. Subtype B avian metapneumovirus resembles subtype A more closely than subtype C or human metapneumovirus with respect to the phosphoprotein, second matrix and small hydrophobic proteins. Virus Research. 2003. v. 92. p. 171-178. Jirjis FF, Noll SL, Halvorson DA, Nagaraja KV, Martin F, Shaw DP. Effects of bacterial coinfection on the pathogenesis of avian pneumovirus infection in turkeys. Avian Dis. 2004 48:34-49. Jones Y.L., Swayne D.E. Comparative pathobiology of low and high pathogenicity H7N3 Chilean avian influenza viruses in chickens. 2004. Avian Diseases. 48:119-128. Kapczynski, D. R., Sellers, H. S. Immunization of turkeys with a DNA vaccine expressing either the F or N gene of avian metapneumovirus. 2003. Avian Dis. v. 47. p. 1376-1383. Kapczynski, D. R., Tumpey, T. M. Development of a virosome vaccine for Newcastle disease virus. 2003. Avian Dis. v. 47. p. 578-587. Khan, M. I. Avian influenza outbreak in Connecticut. . Seminar at the Institute of Animal Husbandry and Veterinary Science, Beijing Municipal Academy of Agriculture, Beijing, Peoples Republic of China, November 4, 2003. Khan, M. I. Low Path Avian Influenza outbreak in Connecticut and control measures. . Seminar at the, Institute of Animal Science, Shandong Academy of Agricultural Science, Jinan City, Shandong, Peoples Republic of China, November 7, 2003. Khan, M. I. Multiplex PCR development and its application for detection of avian pathogens. . Seminar at the, Institute of Poultry Science, Shandong Academy of Agricultural Science, Jinan City, Shandong, Peoples Republic of China, November 6, 2003. Khan, M. I. Monitoring vaccinated flocks, Low Path AI outbreak in Connecticut. North Atlantic Poultry Health & Management Conference, Harrisburg/Hershey, Pennsylvania. May 11-12, 2004. Khan, M. I., and J. J. Fabis. In- vivo expression of IBV-S gene in chicks inoculated with recombinant IBV DNA vaccine in-ovo. Proc. 4th International symposium on avian corona and pneumovirus infections. Rauischholzhausen, Germany. p 232-236. 2004. Kim, S. J., H. W. Sung, J. H. Han, D. J. Jackwood, and H. M. Kwon. Protection against a very virulent infectious bursal disease virus in chickens immunized with DNA vaccines. Vet. Microbiol. 101:39-51. 2004. Kommers, G.D., King, D.J., Seal, B.S., Brown, C.C. Pathogenesis of chicken-passaged Newcastle disease viruses isolated from chickens and wild, and exotic birds. 2003. Avian Diseases. v. 47. p. 319-329. Kommers, G.D., King, D.J., Seal, B.S., Brown, C.C. Virulence of six heterogenous-origin Newcastle disease virus isolates before and after sequential passage in domestic chickens. 2003. Avian Pathology. v. 32. p. 81-93. Land, J.M., C. M. Boettger, and J.E. Dohms. Studies of Intracellular Survival Using Wild Type and Attenuated Mycoplasma gallisepticum strains. Presented at the 76th Northeastern Conference on Avian Diseases, June 9-11, State College, PA Lee, C.W. and D.L. Suarez. 2004. Application of real-time RT-PCR for the quantification and competitive replication study of H5 and H7 subtype avian influenza virus. Journal of Virological Investigation. 119:151-158. Lee, C.-W., D. Senne, J. A. Linares, P. Woolcock, D. Stallknecht, E. Spackman, D.E. Swayne, and D.L. Suarez. 2004. Characterization of Recent H5 Subtype Avian Influenza Viruses from U.S. Poultry. Avian Pathology. 33:288-297. Lee, C-W, D.A. Senne, and D.L. Suarez. 2003. Development of subtype-specific reference antisera by DNA vaccination of chickens. Avian Diseases. 47:1051-1056. Lopes VC, Velayudhan BT, Halvorson DA, Lauer DC, Gast RK, Nagaraja KV. Comparison of methods for differentiation of Salmonella enterica serovar Enteritidis phage type 4 isolates. Am J Vet. Res 2004;65:538-43. Lukert, P. D., and Saif, Y. M. 2003. Infectious Bursal Disease Virus. Diseases of Poultry. 11th edition, Iowa State Press, Ames, Iowa, pp. 161-179. Lwamba HCM, Alvarez R, Wise M, Yu Q, Halvorson D, Njenga MK, Seal BS. Comparison of the full-length genome sequence of Avian metapneumoviurus subtype C with other paramyxoviruses. Virus Res. IN PRESS. Maherchandani, S., Muñoz-Zanzi, C.A., Patnayak, D.P., Malik, Y.P.S., and Goyal, S.M. 2004. The effect of pooling sera on the detection of avian pneumovirus antibodies using an ELISA test. J. Vet. Diag. Investig. IN PRESS. Malik, Y. P. S., Patnayak, D.P., and Goyal, S.M. 2004. Detection of three avian respiratory viruses by single-tube multiplex reverse transcription-polymerase chain reaction assay. J. Vet. Diag. Investig, 16:244-248. Njenga, M.K., Lwumba, H.M., Seal, B.S. Metapneumovirses in birds and humans. 2003. Virus Research. v. 92. p. 163-169. Patnayak, D.P., and Goyal, S.M. 2004. Cold adapted strain of avian pneumovirus as a vaccine in one-day-old turkeys and the effect of inoculation routes. Avian Dis. 48: 155-159. Patnayak, D.P., and Goyal, S.M. 2004. Duration of immunity produced by a live attenuated vaccine against avian pneumovirus type C. Avian Pathol. IN PRESS. Patnayak, D.P., Sheikh, A.M., and Goyal, S.M. 2004. Stability of attenuation in live avian pneumovirus vaccines. J. App. Poult. Res, 13: 253-257. Perkins L.E., Swayne D.E. Comparative susceptibility of selected avian and mammalian species to a Hong Kong-origin H5N1 high-pathogenicity avian influenza virus. 2003. Avian Diseases 47:956-967. Peters, M, Lin, T.L., and Wu, C.C. 2004. Infectious bursal disease virus polyprotein expression arrests growth and mitogenic Stimulation of B lymphocytes. Archives of Virology, in press. Reynolds, D. L., S. Akinc and T. Glanville. Biosecurity of Large-Scale Composting. Poster presentation. AAAP / AVMA annual meeting. Denver Colorado. July 19 23, 2003. Reynolds, D. L., S. Akinc and T. Glanville. Biosecurity of Large-Scale Composting. Poster presentation. Conference of Research Workers in Animal Diseases. Chicago, IL. November 9 -11, 2003. Reynolds, Don, Sevinc Akinc, Thomas Glanville, Jay Harmon, Tom Richards, H. K. Ahn, Samy Sadaka. Biosecurity of Large Scale Animal Mortality Composting. Conference of Research Workers in Animal Diseases. Chicago, IL. November 12 -14, 2003. Reynolds, Don. Biosecurity Issues Related to Disposal of Catastrophic Mortality. Oral presentation. Proceedings of the North Atlantic Poultry Health & Management Conference. May 11-12, 2004. Harrisburg / Hershey Pennsylvania. Saif, Y. M. 2003. Introduction of Emerging Diseases and Diseases of Complex or Unknown Etiology. Diseases of Poultry. 11th edition, Iowa State Press, Ames, Iowa, p. 1163. Saif, Y.M. 2004. Control of infectious bursal disease by vaccination. Proc. International Conference on the Control of Infectious Animal Diseases by Vaccination, Abstract, p. 25, Buenos Aires, Argentina, April 13-16, 2004. Saif, Y.M. 2004. Virulent infectious bursal disease virus. Symp. On Emerging and re-emerging diseases. Co-sponsored by the AAAP/AVMA, Philadelphia, PA, July 25, 2004. Schultz-Cherry S, Koci M, Thompson E, Tumpey TM. Examining the cellular pathways involved in influenza virus induced apoptosis. 2003. Avian Dis. 47:968-71. Senne,D.A. D. L. Suarez, J. C. Pedersen and B. Panigrahy. 2003. Molecular and Biological Characteristics of H5 and H7 Avian Influenza Viruses Found in Live-bird Markets of Northeastern United States During 1994-2001. Avian Diseases. 47:898-904. Spackman, E., D. A. Senne, S. Davison and D. L. Suarez. 2003. Sequence Analysis of Recent H7 Avian Influenza Viruses Associated with Three Different Outbreaks in Commercial Poultry in the United States. Journal of Virology. 77:13399-13402. Suarez, D.L. Erica Spackman, and Dennis Senne. 2003. Update on Molecular Epidemiology of H1, H5 and H7 influenza infections in poultry in North America. Avian Diseases. 47:888-897. Suarez,D.L. D.A. Senne, J. Banks , I.H. Brown, S.C. Essen, C.W. Lee, R.J. Manvell, , C. Mathieu-Benson V. Moreno, J. Pedersen, B. Panigrahy, H. Rojas, E. Spackman, and D.J. Alexander. 2004. Recombination Resulting in Virulence Shift on Avian Influenza Outbreak, Chile. Emerging and Infectious Diseases. 10:693-699. Subler, K. A., C. O. Silva and D. J. Jackwood. Analysis of IBDV-induced immunosuppression and C. jejuni colonization and shedding in chickens. Abstr. 141st AVMA Mtg. 2004. Swayne, D.E., D.L. Suarez, E. Spackman, T.M. Tumpey, J.R. Beck, D. Erdman, P. E. Rollin and T. G. Ksiazek. 2004. SARS-Coronavirus Does Not Cause Disease or Infection in Experimentally Inoculated Domestic Poultry. Emerging and Infectious Diseases. 10:914-916. Swayne, D.E., King, D.J.. Zoonosis update: avian influenza and Newcastle disease. 2003. Journal of the American Veterinary Medical Association. v. 222. p. 1534-1540. Tumpey T.M., Garcia-Sastre A., Taubenberger J.K., Palese P., Swayne D.E.Basler C.F. Pathogenicity and immunogenicity of influenza viruses with genes from the 1918 pandemic virus. 2004. PNAS. 101:3166-3171. Tumpey, T.M., D.L. Suarez, L.E.L. Perkins, D.A. Senne, J. Lee, Y. Lee, I. Mo, H. Sung, D.E. Swayne. 2003. Evaluation of highly pathogenic H5N1 avian influenza A virus isolated in duck meat. Avian Diseases. 47:951-955. Turpin, E.A., Lauer, D.C. and Swayne, D.E. Development and evaluation of a blocking ELISA for the detection of type C avian metapneumovirus antibodies in multiple domestic avian species. Journal of Clinical Microbiology. 2003. v.41. p.3579-3583. Wise, M.G., Suarez, D.L., Seal, B.S., Pedersen, J.C., Senne, D.A., King, D.J., Kapczynski, D.R., Spackman, E.: Development of a real-time, reverse-transcriptase PCR for detection of Newcastle disease virus RNA in clinical samples. J. Clin. Micro. 2004. v. 42. p. 329-338. Woolcock, P.R., D.L. Suarez, and D. Kuney. 2003. Low pathogenicity avian influenza virus (H6N2) in chickens in California, 2000-2001. Avian Diseases. 47:872-881. Wu, C.C., Hsieh, M.K., and Lin, T.L. 2004. Co-administration of Chicken IFN-³ for protection of chickens against infectious bursal disease by DNA vaccination. In: The Proceedings of The 141st Annual Meeting of the American Veterinary Medical Association. Wu, C.C., Peters, M., and Lin, T.L. 2004. Infectious bursal disease virus polyprotein induces immunosuppression independently of virus infection. In: The Proceedings of The 141st Annual Meeting of the American Veterinary Medical Association. Yamnikova, S.S., A.S. Gambaryan, A.B. Tuzikov, N.V. Bovin, M.N. Matrosovich, I.T. Fedyakina, A.A. Grinev, V.M. Blinov, D.K. Lvov, D.L. Suarez, and D.E. Swayne. 2003. Differences between HA receptor-binding sites of avian influenza viruses isolated from Laridae and Anatidae. Avian Diseases. 47:1164-1168. Zanetti, F., Rodriquez, M., King, D.J., Capua, I., Carillo, E., Seal, B.S. Berinstein, A. Matrix protein gene sequence analysis of avian paramyxovirus 1 isolates obtained from pigeons. 2003. Virus Genes. v. 26. p. 199-206.

Impact Statements

Objective 1. Determine the pathogenesis and interactions of specific agents. AVIAN FOWLPOX: The hemagglutinin gene homologue site can be used effectively for insertion of a foreign gene to generate recombinant fowlpox virus vaccines (ILLINOIS).
AVIAN INFLUENZA VIRUS (AIV): Establishing the pathogenicity of AIV from Delmarva broilers will be needed for future studies that will examine interactions of AIV and common respiratory viruses and immunosuppressive viruses to define their role in identifying and diagnosing low path AIV infections in chickens and turkeys. (DELAWARE). Pathogenesis, sequence analysis and vaccination studies using the highly pathogenic H5N1 AIV from Asia was performed to assess what vaccines will be effective if t
AVIAN METAPNEUMOVIRUS (APV): Studies suggested that secondary bacterial infection of APV-infected turkeys aggravates disease (MINNESOTA).
INFECTIOUS BURSAL DISEASE VIRUS (IBDV): A reverse genetically engineered IBDV is important for understanding factors that will lead to improved disease control (INDIANA). Interference in the replication of a virulent strain of IBDV by a vaccine strain may be important for designing control strategies (OHIO). Control of IBDV will lower the risk of human exposure to food-borne pathogens such as C. jejuni (OHIO).
ESCHERICHIA COLI: Understanding the gene expression of avian macrophage activation during phagocytosis of Escherichia coli is important in modulating the immune response and ultimately controlling infection (DELAWARE).
NEWCASTLE DISEASE VIRUS (NDV): Establishing poultry host range susceptibility by exotic Newcastle disease (END) virus is important in recognizing the variability of lesions and may impact diagnostics (USDA ARS SEPRL).
SARS CORONAVIRUS: The findings that the five common poultry species could not be easily infected with the SARS virus suggested that poultry likely played no role in either the origin or spread of the virus (USDA ARS SEPRL in collaboration with the Centers for Disease Control).
Objective 2. Surveillance, occurrence and consequences of agents and host variation on disease susceptibility. AVIAN INFLUENZA VIRUS: The identification of a H3N2 in turkeys with a matrix gene sharing a high homology with A/Swine/Illinois/100084/01 (H1N2) emphasizes the need for ongoing influenza surveillance as well as understanding the events associated with crossing the species barrier. Identification of new strains may require new vaccination strategies (OHIO collaborating with USDA Sout
INFECTIOUS BRONCHITIS VIRUS: In Delmarva broilers, IBV continues to be the most frequent virus recovered from the respiratory tract. Arkansas S1 genotype type IBVs were much more often isolated than were genotypes Massachusetts and Connecticut. Establishing the frequency of IBV isolations from commercial broiler chickens is important for monitoring pathogenicity of field strains and the effectiveness of vaccination programs (DELAWARE).
INFECTIOUS BURSAL DISEASE VIRUS: The presence of a serotype 2 IBDV strain in penguins from a zoological park in the United Kingdom demonstrates the disease may occur in other species (OHIO). Identification of multiple mutations across the hydrophilic B epitope region of VP2 among field isolates of IBDV suggests that antigenic diversity due to genetic drift may be responsible for continuing problems with infections in the U.S. Studies on the type and distribution of IBDV in the field will help
MYCOPLASMOSIS: The isolation of an atypical strain of M. synoviae from broilers emphasizes the need for surveillance (DELAWARE).
NEWCASTLE DISEASE VIRUS: Establishing the frequency of NDV isolations from commercial broiler chickens is important for monitoring pathogenicity of field strains and the effectiveness of vaccination programs (DELAWARE). Comparison of v-ND isolates from South and Central America Mexico and Honduras 1996/2000 and the ENDin southern California, Nevada, Arizona and Texas indicated a close phylogenetic relationship. Continued aggressive surveillance is necessary to prevent further introductions of
Objective 3. Develop new and improved methods for the diagnosis, prevention, and control of avian respiratory diseases. AVIAN INFLUENZA VIRUS: Studies using molecular epidemiology methods provided detailed information on the origins of outbreaks of avian influenza in the U.S., and helped to identify risk factors for how a virus was introduced to poultry farms (USDA ARS SEPRL). Surveillance in layers may be more easily facilitated by an egg yolk antibody test that was developed (USDA ARS SEPRL)
AVIAN METAPNEUMOVIRUS: Sequencing of the APV genome was completed thus paving way for generation of reverse genetic system for vaccine development and further characterization of the virus. (MINNESOTA and USDA ARS SEPRL). A real-time RT PCR test capable of detecting the major APV subtypes has been developed for use in surveillance against introduction of European subtypes into the U.S. turkey population (MINNESOTA). An APV isolate from wild geese was determined to be avirulent for turkeys, thu
INFECTIOUS BURSAL DISEASE VIRUS: Plasmids vaccines for IBDV encoding the chicken IL-2 gene enhanced protection of chickens and may offer another approach to control (INDIANA).
NEWCASTLE DISEASE VIRUS: Composting studies of contaminated animal and poultry carcasses resulted in the inactivation of enveloped (NDV) and non-enveloped (avian encephalomyelitis virus) avian viruses and suggest that composting is a biosecure, safe, environmentally friendly and feasible way to deal with disease events such as avian influenza and exotic Newcastle disease (IOWA).