Brief Summary of Minutes of Annual Meeting

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.

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).

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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. 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