USDA Funded Research Records from the Current Research Information System
(CRIS)
(1)
ACCESSION NO: 0168532 SUBFILE: CRIS
PROJ
NO: ARK01611 AGENCY: CSREES
PROJ
TYPE: HATCH PROJ STATUS: EXTENDED MULTISTATE PROJ
NO: NE-60
START:
INVESTIGATOR: Erf, G.
F.
PERFORMING INSTITUTION:
GENETIC BASES FOR RESISTANCE
AND IMMUNITY TO AVIAN DISEASES
OBJECTIVES
Identify and characterize environmental, dietary
and physiologic factors that modulate immune system development, optimal immune
function and disease resistance in poultry genetic stocks.
APPROACH: Contributing research will include the Smyth line
chickens which develop spontaneous post-hatch, autoimmune vitiligo. Three
MHC-matched lines of chickens, all homozygous for the MHC B101 haplotype will
be used in this project. Included are the autoimmune vitiliginous Smyth line
(SL), the parental Brown line (BL), and the normally pigmented Light Brown
Leghorn. Special emphasis will be placed on identifying environmental factors
required for the expression of vitiligo in genetically susceptible SL chickens
and on the immune mechanisms involved in autoimmune destruction of pigment
cells in SL vitiligo. Additionally, immunomodulatory effects of dietary supplements
on the avian immune system will be examined in broilers and in turkeys. Scientific
methods used will include in vitro culture systems and flow cytometry.
PROGRESS: 2002/01 TO 2002/12
Mutant Smyth line chickens spontaneously develop post-hatch loss of eye and
feather pigment. This loss of pigment is due to the destruction of pigment
cells by the immune system. The similarities between the autoimmune loss of
pigment cells in Smyth line chickens and the pigment loss observed in human
vitiligo have lead to the acceptance of the SL chicken as the best animal
model to study autoimmune vitiligo. During the last calendar year, we completed
a study (funded by the National Vitiligo Foundation), on the role of environmental
factors such as turkey herpesvirus (HVT) vaccine and other live virus vaccines
(
IMPACT: 2002/01 TO 2002/12
The use of an animal
model that is genetically susceptible to development of autoimmune vitiligo
provides an excellent opportunity to study the cause and effect relationship
between genetic susceptibility and the factors leading to the onset and expression
of autoimmune disease. Knowledge gained from these studies will find direct
application in the management and prevention of autoimmune disease. Additionally,
these studies on immune system dysfunction and mechanisms of pathogenesis
will yield important new knowledge regarding immune system development and
function in avian species.
PUBLICATIONS: 2002/01 TO 2002/12
1. Erf, G. F. 2002. Smyth line autoimmune vitiligo - similar to human autoimmune
vitiligo. Pages 316-332 in Modern Concepts of Immunology in Veterinary Medicine-Poultry
Immunology. Mathew, T., editor. Advances in Medical and Veterinary Virology,
Immunology and Epidemiology, Thajema Publishers,
2. Wang, W., R. F. Wideman, Jr., T. K. Bersi, and G. F. Erf. 2003. Pulmonary
and hematological immune responses to intravenous cellulose micro-particles
in broilers. Poult. Sci. in press.
3. Erf, G. F., T. K. Bersi, and H. S. Lillehoj. 2002. A role of interferon
gamma in autoimmune vitiligo of Smyth line chickens. FEMS in press.
4. Wang, W., G. F. Erf, and R. F. Wideman. 2002. Effect of cage vs floor litter
environments on the pulmonary hypertensive response to intravenous endotoxin
and on blood-gas values in broilers. Poult. Sci. 81:1728-1737.
5. Wang, W., R. F. Wideman, and G. F. Erf. 2002. Pulmonary hypertensive response
to endotoxin in cellulose-primed and unprimed broiler chickens. Poult. Sci.
81:1224-1230.
6. Wideman R. F., G. F. Erf, M. E. Chapman, W. Wang, N. B. Anthony, and L.
Xiaofang. 2002. Intravenous micro-particle injections and pulmonary hypertension
in broiler chickens: acute post-injection mortality and ascites susceptibility.
Poult. Sci. 81:1203-1217.
7. Iqbal, M., J. D. Freiburger, G. F. Erf, and W. G. Bottje. 2002. Immunohistochemical
evidence of cytochrome c oxidase subunit II involvement in pulmonary hypertension
syndrome (PHS) in broilers. Poult. Sci. 81:1231-1235.
8. Wideman, R. F., and G. F. Erf. 2002. Intravenous microparticle injection
and pulmonary hypertension in broiler chickens: Cardio-pulmonary hemodynamic
responses. Poult. Sci. 81:877-886.
PROJECT CONTACT:
Name: Erf, G. F.
Phone: 501-575-8664
Fax: 501-575-3026
Email: gferf@comp.uark.edu
(2)
ACCESSION NO: 0098242 SUBFILE: CRIS
PROJ
NO: CA-V*-PHR-4652-AH96 AGENCY: CSREES CALB
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Lam, K.
M.
PERFORMING INSTITUTION:
POPULATION HEALTH & REPRODUCTION
UNIV OF
OBJECTIVES: A. To determine that
APPROACH: A. Heterophils and macrophages will be infected with
ND V in vitro. Gel electrophoresis, electron microscopy, flow cytometry, and
in situ hybridization will be used to confirm the presence of apoptosis in
the infected cells. B. The ability of heterophils and macrophages to produce
hydrogen peroxide will be determined by the stimulation of cells with dichclorofluorescein
(DCF) and phorbol myristate acetate (PMA), and followed by flow cytometric
examination. The effect of ND V on hydrogen peroxide production will also
be determined. C. Heterophils and macrophages will be pre-treated with various
recombinant human cytokines and then determine for their oxidative burst by
DCF and PMA.
PROGRESS: 1996/10 TO 2001/09
The efforts in this year have been concentrated on chicken heterophils and
thrombocytes and the effect of
IMPACT: 1996/10 TO 2001/09
The goal of this project was to A. determine that
PUBLICATIONS: 1996/10 TO 2001/09
1. Lam KM. 1997. Myeloperoxidase activity in chicken heterophils and adherent
cells. Vet. Immunol. Immunopathol. 57:327-335.
2. Lam KM. 1997. Activation, adhesion, migration and death of chicken thrombocytes.
Comp. Haematol. Intl. 1:81-87.
(3)
ACCESSION NO: 0182013 SUBFILE: CRIS
PROJ
NO: CALV-AH-176 AGENCY: CSREES CALV
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: NEW
START:
INVESTIGATOR: Gardner,
PERFORMING INSTITUTION:
MEDICINE & EPIDEMOLOGY
UNIV OF
QUANTITATIVE METHODS TO CERTIFY
FREEDOM OF ANIMALS FROM PATHOGENS
OBJECTIVES: 1. Develop a Bayesian approach to certify disease
freedom of a country/region that incorporates uncertainty in probability estimates.
2. Compare frequentist and Bayesian approaches to certify disease freedom
using common data sets and to compare sample size requirements for surveys
with both approaches.
APPROACH: 1. The Bayesian approach will be implemented with
the Gibbs sampler, an interactive Markov-chain Monte Carlo method. The mathematical
calculations will incorporate the prior probability that a country is free
of disease, the uncertainty in sensitivity and specificity estimates and the
possible clustering of positive test results at a herd level. The output will
be a probabilistic estimate of disease freedom. 2. Frequentist and Bayesian
estimates will be compared with common published data sets on porcine reproductive
and respiratory syndrome and Newcastle Disease. The effect of selected prior
distributions for the Bayesian approach will be evaluated. Sample sites used
in frequentist calculations for surveys will be compared with estimates that
we will derive using Bayesian approaches.
NON-TECHNICAL SUMMARY: If countries and regions are able to "certify"
freedom from important animal pathogens, trade opportunities may increase
and product export costs may decrease. To develop a Bayesian statistical approach
(using Gibbs sampling) to quantification of disease freedom. The output from
the model will be probability distributions that can be used to make inferences
about the proportion of diseased herds, within-herd prevalence, and the probability
that a country is free of disease. The research will be involve collaboration
with others in the
PROGRESS: 2002/01 TO 2002/12
Quantitative approaches are needed to allow scientifically-valid inferences
about freedom of animals from important pathogens that affect animal trade.
Freedom in the context of these inferences includes a pathogen prevalence
less than a threshold (e.g. <0.2% of infected herds). We developed a hierarchical
Bayesian statistical model that uses herd-level test results from multiple
herds in a region or country or zone, and adjusts for uncertainty in the sensitivity
and specificity of tests and the prior probability of infectious agent. The
model allows inferences about the post-test probability of freedom from infection,
the proportion of infected herds, and the within-herd prevalence. Using published
survey data for porcine reproductive and respiratory syndrome and Newcastle
Disease in poultry, we have shown that inferences from our Bayesian approach
are similar to those from an alternate simulation modeling approach. The Bayesian
model is superior to previous methods because it allows inferences about the
proportion of infected herds and within-herd prevalence which are important
inputs into risk assessment models. The model has been modified to include
the possibility of different sample sizes in each of the herds, and the use
of additional tests in animals that are positive on the first screening test.
IMPACT: 2002/01 TO 2002/12
The new method has potential to be used internationally as a tool in substantiating
a country's claim of freedom from animal pathogens.
PUBLICATIONS: 2002/01 TO 2002/12
No publications reported this period
PROJECT CONTACT:
Name: Gardner, I. A.
Phone: 530-752-6992
Fax: 530-752-0414
Email: iagardner@ucdavis.edu
(4)
ACCESSION NO: 0177509 SUBFILE: CRIS
PROJ
NO: CALV-CAHFS95CDFA6601 AGENCY: CSVM CALV
PROJ
TYPE: STATE PROJ STATUS: EXTENDED
START:
INVESTIGATOR: Ardans,
A. A.
PERFORMING INSTITUTION:
ADMINISTRATION
UNIV OF
OBJECTIVES: To provide laboratory diagnostic support of the highest
quality for the surveillance and control of diseases and the enhancement of
health of livestock and poultry in
APPROACH: The CVDLS is composed of a full service, central
reference laboratory at UC Davis and four laboratories located at Turlock
(poultry), Fresno (poultry and regulatory services), Tulare (mammalian services)
and San Bernardino (general services for poultry, mammalian and regulatory).
These laboratories are linked by a computer based Management Information and
Surveillance System to function as a single entity.
PROGRESS: 2002/01 TO 2002/12
Avian influenza, H6N2, reoccurred in
IMPACT: 2002/01 TO 2002/12
The resources of the California Animal Health and Food Safety Laboratory System
(CAHFS) in concert with
PUBLICATIONS: 2002/01 TO 2002/12
1. Adaska JM,
2. Chin RP. 2002. Isolation of an unidentified, nonfermentative, gram-negative
bacterium from turkeys and chickens: 38 cases (1995-2001). Avian Diseases,
46:447-452.
3. Colagross-Schouten AM, Mazet JA, Gulland FM, Miller MA, Hietala SK. 2002.
Diagnosis and seroprevalence of leptospirosis in
4. Cramer G, Kelton D, Duffield TF, Hobson JC, Lissemore K, Hietala SK, Peregrine
AS. 2002. Neospora caninum serostatus and culling of Holstein cattle. Journal
of the American Veterinary Medical Association, 221:1165-1168.
5. Crespo R, Ghazikhanian GY, Hall CI. 2002. Avulsion of the common retinaculum
in meat turkeys. Avian Diseases, 46:245-248.
6. Crespo R, Stover SM, Shivaprasad HL, Chin RP. 2002. Microstructure and
mineral content of femora in male turkeys with and without fractures. Poultry
Science, 81:1184-1190.
7. Crespo R, Woolcock PR, Fadly AM, Hall C, Shivaprasad HL. 2002. Characterization
of T-cell lymphomas associated with an outbreak of reticuloendotheliosis in
turkeys. Avian Pathology, 31:355 -361.
8. Daft BM, Barr BC, Gardner IA, Read D, Bell W, Peyser KG, Ardans A, Kinde
H, Morrow JK. 2002. Sensitivity and specificity of western blot testing of
cerebrospinal fluid and serum for diagnosis of equine protozoal myeloencephalitis
in horses with and without neurologic abnormalities. Journal of the American
Veterinary Medical Association, 221:1007-1013.
9. Driessen B, Zarucco L, Steffey EP, McCullough C, Del Piero F, Melton L,
Puschner B, Stover SM. 2002. Biochemical and histopathological changes associated
with prolonged sevoflurane anaesthesia in horses. Journal of Veterinary Medicine,
A 49:1-11.
10. Fosgate GT, Adesiyun AA, Hird DW,
11. Fosgate GT, Adesiyun AA, Hird DW, Johnson WO, Hietala SK, Schurig GG,
Ryan J. 2002. Comparison of serologic tests for detection of Brucella infections
in cattle and water buffalo (Bubalus bubalis). American Journal of Veterinary
Research, 63:1598-1605.
12. Fosgate GT, Hird DW, Read DH, Walker RL. 2002. Risk factors for foals
developing Clostridium piliforme infection (Tyzzer's Disease) on a California
Thoroughbred breeding farm. Journal of the Veterinary Medical Association,
220:785-790.
13. Gordus AG, Shivaprasad HL, Swift P. 2002. Salt toxicosis in ruddy ducks
that winter on an agricultural evaporation basin in
14. Haqshenas G, Huang FF, Fenaux M, Guenette DK, Pierson FW, Larsen CT, Shivaprasad
HL, Toth TE and Meng XJ. 2002. The putative capsid protein of the newly identified
avian hepatitis E virus shares antigenic epitopes with that of swine and human
hepatitis E viruses and the chicken big liver and spleen disease virus. Journal
of General Virology, 83:2201-2209.
15. Hobson JC, Duffield TF, Kelton D, Lissemore K, Hietala SK, Leslie KE,
McEwan B, Cramer G, Peregrine AS. 2002. Neospora caninum serostatus and milk
production of
16. Holstege DM, Puschner B, Whitehead G, and Galey FD. 2002. Screening and
mass spectral confirmation of beta-lactam antibiotic residues in milk using
LC-MS/MS. Journal of Agriculture Food Chemicals, 50:406-411.
17. Huang FF, Haqshenas G, Shivaprasad HL, Guenette DK, Woolcock PR, Larsen
CT, Pierson FW, Elvinger F, Toth TE and Meng XJ. 2002. Heterogeneity and Seroprevalence
of the Newly Identified Avian Hepatitis E Virus from Chickens in the
18.
19. Nieto JE, Spier S, Pipers FS,
20. Peroni DL, Stanley S, Kollias-Baker C, Robinson NE. 2002. Prednisone per
os is likely to have limited efficacy in horses. Equine Veterinary Journal,
34:283-287.
21. Puschner B, Booth MC, Tor ER, Odermatt A. 2002. Diterpenoid alkaloid toxicosis
in cattle in
22. Ridpath JF,
23. Riggs SM, Puschner B, Tell LA. 2002. Management of an ingested lead foreign
body in an Amazon Parrot. Veterinary and Human Toxicology, 44:345-348.
24. Shilton CM, Smith DA, Woods LW, Crawshaw GJ, Lehmkuhl HD. 2002. Adenoviral
infection in captive moose (Alces alces) in
25. Shivaprasad HL, Crespo, R, Puschner B, Lynch S, Wright L. 2002. Myopathy
in brown pelicans (Pelicanus occidentalis) associated with rancid feed. Veterinary
Record, 150: 307-311.
26. Shivaprasad HL, Kim TJ, Woolcock PR, Tripathy DN. 2002. Genetic and antigenic
characterizations of a poxvirus isolate from ostriches. Avian Diseases, 46:429-436.
27. Shivaprasad HL, Woolcock PR, McFarland MD, Curtis M, Karabatsos N. 2002.
Turlock-like bunyavirus associated with encephalomyelitis and myocarditis
in an ostrich chick. Journal of Veterinary Diagnostic Investigation, 14:363-370.
28. Shivaprasad HL and Droual R. 2002. Pathology of an atypical strain of
P. gallinarum in chickens. Avian Pathology, 31:399-406.
29. Stamm LV,
30. Suarez DL, Woolcock PR, Bermudez AJ, Senne D. 2002. Isolation from turkey
breeder hens of a reassortant H1N2 influenza with swine, human and avian lineage
genes. Avian Diseases, 46:111-121.
31. Taduesz FM, and Stanley SD. 2002. Improved sythesis of 13C,2H3- and 2H3-salmeterol
by Cs2CO3-mediated monoalkylation of a primary amine. Journal of Labeled Compounds
and Radiopharmaceuticals, 45:755-762.
32. Tegzes J, Puschner B. 2002. Amanita mushroom poisoning - efficacy of aggressive
treatment in 2 dogs. Veterinary and Human Toxicology, 44:96-99.
33. Tegzes JH,
34. Thurmond MC, Wesley OJ, Munoz-Zanzi CA, Chun-Lung S, Hietala SK. 2002.
A method of probability diagnostic assignment that applies Bayes theorum for
use in serologic diagnostics, using an example of Neospora caninum infection
in cattle. American Journal of Veterinary Research, 63:318-325.
35. Turay HO, Caldwell A, Barr BC, Branson KR, Cockrell MKR, Marsh AE. 2002.
Sarcocystis neurona reacting antibodies in
36. Van Hoogmoed LM, Harmon FA,
37. Walker RL, Read DH,
38. Webby RJ, Woolcock PR, Krauss SL, Webster RG. 2002. Reassortment and influenza
transmission of North American H6N2 influenza viruses. Virology, 295:44-53.
39. Woolcock PR, McFarland MD, Lai S and Chin RP. 2002. Enhanced Recovery
of Avian Influenza Virus Isolates using a Combination of Chicken Embryo Inoculation
Methods. Avian Diseases, 45:1030-1035.
40. Zakhartchouk A, Bout A, Woods LW, Lehmkuhl HD, Havenga MJE. 2002. Odocoileus
hemionus deer adenovirus is related to the members of Atadenovirus genus.
Archives of Virology, 147:841-847.
(5)
ACCESSION NO: 0181168 SUBFILE: CRIS
PROJ
NO: CONS-9802281 AGENCY: CSREES CONS
PROJ
TYPE: NRI COMPETITIVE GRANT PROJ STATUS: EXTENDED
CONTRACT/GRANT/AGREEMENT
NO: 98-35204-6954
START:
01 DEC 1998 TERM:
GRANT
AMT: $180,000
INVESTIGATOR: Sekellick,
M. J.; Marcus, P. I.
PERFORMING INSTITUTION:
MOLECULAR AND CELL BIOLOGY
UNIV OF
RECOMBINANT CHICKEN INTERFERONS
AS ANTIVIRAL AGENTS
OBJECTIVES: 9802281. Our goal is to develop chicken interferons
singly, or in synergistic mixtures, as novel biological response modifiers
for the prevention or cure of viral diseases. Specific objectives include:
(1) Determine the spectrum and degree of sensitivity of avian viruses of economic
and public health importance to the action of Types I and II recombinant chicken
interferons in vitro, in ovo, and in the chicken, acting singly, and in mixtures
that display synergy; (2) Determine the nature of the heterogeneity in avian
influenza virus sensitivity to recombinant chicken interferon; (3) Determine
the antiviral effects of recombinant chicken interferon administered orally
thorugh novel means and/or intranasally, as an effector of the humoral and
mucosal system; and (4) develop a line of chickens with genetically enhanced
sensitivity to the action of interferon.
APPROACH: The recently cloned and expressed genes of Types
I and II chicken interferons will be produced as glycosylated recombinant
molecules in stably transfected COS cells or in E. coli., purified, and tested
for their in vitro, in ovo and in vivo activity against avian viruses of economic
importance.
PROGRESS: 2000/01 TO 2000/12
Interferons (IFN) are components of the innate immune system and constitute
the first and immediate line of defense against virus infection. They are
produced rapidly by virus-infected cells, are released into the surrounding
milieu within hours, and act within minutes following binding to specific
cellular receptors on uninfected cells. Subsequent signal transduction and
activation of transcription factors result in the activation of over 100 IFN-stimulated
genes. The multiple intracellular modes of action that result from expression
of these IFN-stimulated genes, and their efficacy against a broad spectrum
of virus families, including those subject to antigenic changes that mute
the effectiveness of vaccines, make IFNs novel biological modifiers worthy
of tests to determine the range of their protective and curative properties.
Double-stranded RNA (dsRNA) is a second biological response modifier of equally
formidable activity. This class of molecules has emerged as singularly important
in both the induction and action phases of the IFN system, and as an activator
of many genes capable of producing multiple effects on cells and the immune
system. Interestingly, many viruses have evolved mechanisms to prevent activation
of cellular proteins designed to sense, and counteract, the presence of dsRNA.
These include production of viral gene products which sequester dsRNA, and
small helical RNAs. These molecules potentially prevent activation of dsRNA-dependent
pathways of interferon action, or block expression of cellular genes activated
exclusively by dsRNA that may contribute to the antiviral state. Not surprisingly,
these means have provided viruses with highly effective mechanisms against
IFN action. One of the most successful of the anti-IFN mechanisms is exemplified
by the almost absolute resistance to the action of IFN displayed by avian
reovirus (ARV). This resistance is attributed to the dsRNA-binding capacity
of the sigma alpha core protein. Thus, dsRNA could be rate limiting in an
ARV infected cell providing a means of preventing the development of an IFN-
or dsRNA-mediated antiviral state. In support of this hypothesis, we have
shown that dsRNA added exogenously to IFN-treated cells in the form of poly(rI):poly(rC)
is effective in establishing in a dose-dependent manner an antiviral state
against ARV as well as
IMPACT: 2000/01 TO 2000/12
The combined sequential application of interferon and double-stranded RNA
may be useful in overcoming the anti-interferon activity of viruses of clinical
interest, and even find relevance in other clinical conditions where interferon
by itself is marginally, if at all, effective.
PUBLICATIONS: 2000/01 TO 2000/12
1. Sekellick, M.J., Carra, S.A., Bowman, A., Hopkins, D.A. and Marcus, P.I.
2000. Transient resistance of influenza virus to interferon action attributed
to random multiple packaging and activity of NS genes. Journal of Interferon
and Cytokine Research 20:963-970.
2. Marcus, P.I. and Sekellick, M.J. 2000. Combined action of interferon and
dsRNA enhances antiviral effects. European Cytokine Network 11:186.
(6)
ACCESSION NO: 0007173 SUBFILE: CRIS
PROJ
NO: CONS00122 AGENCY: SAES CONS
PROJ
TYPE: STATE PROJ STATUS: EXTENDED
START:
INVESTIGATOR: Van Kruiningen,
H.
PERFORMING INSTITUTION:
PATHOBIOLOGY
UNIV OF
PULLORUM DISEASE CONTROL
OBJECTIVES: Pullorum-Typhoid Eradication.
APPROACH: This program involves the serologic testing of 500,000
to 700,000 avian blood samples per year for the presence of Salmonella pullorum
and S. gallinarum. Reacting birds are called to the laboratory for bacteriological
examination. IfS. pullorum or S. gallinarum is isolated, the reacting flock
is retested at 21 day intervals until two successive negative flock tests
are obtained. Control and regulatory action are administered by the Commissioner
of Agriculture through the State Veterinarian.
PROGRESS: 2002/01 TO 2002/12
This is a collaborative project with the Connecticut Department of Agriculture.
The purpose is to monitor and diagnose infectious diseases of poultry including
Salmonella pullorum, Salmonella enteritidis, Mycoplasma gallisepticum and
synoviae,
IMPACT: 2002/01 TO 2002/12
This monitoring program for infectious disease of
PUBLICATIONS: 2002/01 TO 2002/12
No publications reported this period
(7)
ACCESSION NO: 0086778 SUBFILE: CRIS
PROJ
NO: CONS00541 AGENCY: CSREES CONS
PROJ
TYPE: HATCH PROJ STATUS: TERMINATED MULTISTATE PROJ NO: NE-138
START:
INVESTIGATOR: Khan,
M. I.
PERFORMING INSTITUTION:
PATHOBIOLOGY
UNIV OF
EPIDEMIOLOGY AND CONTROL
OF EMERGING STRAINS OF POULTRY DISEASE RESPIRATORY AGENTS
OBJECTIVES: Develop and evaluate rapid diagnostic capabilities
for the identification of emerging IBV, ILTV, mycoplasmas, and IBDV.
APPROACH: Infectious bronchitis virus (IBV) specific RT-PCR.
IBV strains
PROGRESS: 1996/10 TO 2002/09
Development of recombinant DNA vaccine that expresses S1 of IBV and Immunogenicity
studies A recombinant fowlpox virus (rFPV) containing a cDNA copy of S1 gene
of IBV (rFPV-S1) was constructed and its immunogenicity and vaccine potential
were evaluated. Initially, rFPV-S1 was shown to express the S1 protein in
vitro by using indirect immunofluorescence staining and Western blot analysis.
Later, in vivo expression was demonstrated by the detection of IBV-specific
serum IgG and neutralization antibodies in the sera of chickens immunized
with rFPV-S1. That the recombinant virus elicited anti-IBV protective immunity
was indicated by the manifested, relatively mild clinical signs of disease,
decreased titers of recovered challenge virus, and less severe histological
changes of the tracheas in virulent IBV-challenged chickens previously receiving
rFPV-S1 as compared to parental FPV vaccinated control birds. In contrast,
chickens immunized with either recombinant or parental FPV were resistant
to a subsequent, virulent FPV challenge. As to a preferred method of immunization,
wing web inoculation appeared to be superior to the subcutaneous route since
a greater percentage of birds vaccinated by the former protocol exhibited
an anti-IBV humoral immune response. Thus, rFPV-S1 has potential as a poultry
vaccine against both fowlpox and infectious bronchitis.
IMPACT: 1996/10 TO 2002/09
Impact: 1. Recombinant Fowlpox virus containing S1 gene has potential for
a poultry vaccine against both fowlpox and infectious bronchitis. 2. DNA vaccine
containing whole S gene instead of S1 of IBV in pCMV plasmid vector could
provide better protection against IBV infection.
PUBLICATIONS: 1996/10 TO 2002/09
1. Wang, X., W. M. Schnitzlein, D.
2. Khan, M. I. 2002. Avian Pathogenic Mycoplasmas. PCR detection of Microbial
Pathogens. Methods in Molecular Biology. eds. J. Frey and K. Sachse. Humana
Press Inc.
(8)
ACCESSION NO: 0406071 SUBFILE: CRIS
PROJ
NO: AP-511 AGENCY: ERS MTED
PROJ
TYPE: USDA COOPERATIVE AGREEMENT PROJ STATUS: NEW
START:
INVESTIGATOR: Hahn,
W.; Salin, D.; Harvey, D.
PERFORMING INSTITUTION:
Economic Research Service
USDA/ERS
1800 M STREET NW
AP SPECIAL TOPICS: THE ECONOMIC
EFFECT OF CHANGES IN SANITARY REGULATIONS ON BROILER TRADE IN THE
OBJECTIVES: Sanitary and phytosanitary (SPS) measures are impediments
to trade and affect both the flow and the magnitude of trade. Newcastle Disease
(ND) and highly pathogenic avian influenza (HPAI), included in List A of the
International Organization for Epizootics (OIE) classification of transmissible
animal diseases, are two highly infections diseases that restrict poultry
trade. The END-free status gives the
APPROACH: Objective 1: Analyze price differentials of poultry
products between the countries in the
NON-TECHNICAL SUMMARY: This project evaluates the potential economic impact
of changes in sanitary restrictions on broiler trade in the
PROJECT CONTACT:
Name: Hahn, W.
Phone: 202-694-5175
Email: WHAHN@ers.usda.gov
(9)
ACCESSION NO: 0181427 SUBFILE: CRIS
PROJ
NO: FLA-VME-03777 AGENCY: CSREES
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: NEW
START:
04 MAR 1999 TERM:
INVESTIGATOR: Spalding,
M. G.; Forester, D. J.
PERFORMING INSTITUTION:
SURVEILLANCE FOR DISEASES
OF WILDLIFE WHICH ARE TRANSMISSIBLE TO LIVESTOCK AND POULTRY IN
OBJECTIVES: The objectives are to determine the distribution
and prevalance of wildlife diseses that are transmissible to livestock and
pountry in
APPROACH: Serologic studies and virus isolations will be conducted
on eggs, nestlings, and adult double-crested cormorants from several colonies
in
PROGRESS: 2001/10 TO 2002/10
We continue to receive specimens, especially from the Florida Fish and Wildlife
Conservation Commission and monitor them for exposure to eastern equine encephalitis
and
IMPACT: 2001/10 TO 2002/10
Pigeon paramyxovirus 1, though very similar to Newcastle Disease virus, does
not appear to be virulent in domestic poultry.
PUBLICATIONS: 2001/10 TO 2002/10
1. Forrester, D. J. and M. G. Spalding. 2001. Salmonellosis in a wild turkey
from
2. Spalding, M.G., S.A. Nesbitt, S.T. Schwidert, and R.J. Dusek. 2001. The
use of radio transmitters to monitor survival of sandhill crane chicks. Proc.
North Ammerican Crane Workshop 8: 213-215.
3.
4. Frederick, P. C., M. G. Spalding, and R. Dusek. 2002. Wading birds as bioindicators
of mercury contamination in Florida, USA: Annual and geographic variation.
Environmental Toxicology and Chemistry 21(1): 163-167.
5. Spalding, M. G., C. A. Yowell, D. S. Lindsay, E. C. Greiner, and J. B.
Dame. 2002. Sarcocystis meningoencephalitis in a northern gannet (Morus bassanus).
Journal of Wildlife Diseases 38(2): 432-437.
6. Varela, A., J.M. Kinsella, and M.G. Spalding. 2002. Presence of encysted
immature nematodes in a released whooping crane (Grus
7. Buergelt, C.D., B.L. Homer, and M.G. Spalding. 2002. Causes of mortality
in the Florida panther (Felis concolor coryi). Annals of the New Your
(10)
ACCESSION NO: 0178041 SUBFILE: CRIS
PROJ
NO: FLAV-CO-00204
AGENCY: CSREES FLAV
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START:
16 APR 1998 TERM: 30 MAR 2001 FY: 2001
INVESTIGATOR: Romero,
C. H.; Butcher, G. D.
PERFORMING INSTITUTION:
DEVELOPMENT OF RECOMBINANT
POULTRY VACCINES
OBJECTIVES: To validate existing stocks of recombinant herpes
virus of turkeys (HVT) as to gene content and expression in chicken embryo
fibroblasts (CEFs). To prepare stocks of recombinant HVT and maintain the
stocks in liquid nitrogen for future validation and preparation of vaccine.
To develop assays for identifying and testing gene expression in the recombinant
vaccines. To identify new insertion sites in the HVT genome for cloning of
immunogenic genes obtained from avian viruses of economic impact. To identify
genes of avian pathogens that code for immunogenic proteins that confer disease
resistance in vaccinated chickens. To develop new recombinant vaccines based
on HVT as a vector and to perform potency and protection tests in chickens.
APPROACH: Cultures are prepared from chicken embryos on the
11th day of incubation . Large stocks of primary cells, chicken embryo fibroblasts
(CEFs) are aliquoted and maintained frozen in liquid nitrogen tanks. Only
these cell stocks guaranteed of extraneous viral or bacterial contamination
are used in the validation or development of new assays on recombinant viruses
and DNA molecules. Recombinant and wild-type viral RNA and DNA is extracted
from infected CEFs and purified in order to locate and identify the inserted
foreign genes in the viral genome utilizing reverse transcription and polymerase
chain reaction (RT-PCR) amplification. The functionality of these genes is
determined utilizing techniques such as Western blots, immunoprecipitation
and enzymatic assays. New genomic insertion sites are found by cloning viral
fragments in bacterial plasmids and by looking for unique restriction sites
in these fragments. Then, by a process of homologous recombination in CEFs
utilizing infectious HVT DNA and recombinant plasmid DNA recombinant viruses
are screened for as proof of the non-essentiality of the insertion site. Genes
of avian pathogens known to code for immunogenic proteins are then generated
by PCR or RT-PCR and tested for protein expression before being cloned into
the HVT vector or eukaryotic expression vectors.
PROGRESS: 1998/04 TO 2001/03
Specific-pathogen-free chickens injected with 200ug of naked DNA molecules
expressing the glycoprotein B of serotypes 1 and 3 of Marek's disease virus
and the MEQ protein were not protected from challenge with the virulent RB-1B
strain. The chicken interferon gamma and interleukin-2 genes have been amplified
and cloned in order to test them as immunological adjuvants when administered
with DNA molecules that express Marek's disease virus genes. The capsid protein
VP2 of infectious bursal disease virus has been re-created and injected into
chickens in order to evaluate its expression in vivo and its fate and distribution
in lymphod tissues. Final transfer vectors for generating recombinant vaccines
based on the herpes virus of turkeys and the Rispens strain of Marek's disease
virus are now constructed and being tested in transfection experiments. Assays
based on the RT-PCR have been developed in order to test the transcription
of foreign genes expressed from recombinant vaccines based on the herpes virus
of turkeys. These assays test for the transcription of the full gB,gC and
gD genes of serotypes 1, 2 and 3 of Marek's disease virus. Similarly, primers
have been developed for the differential amplification of the complete gB,
gC and gD genes of serotypes 1, 2 and 3 of Marek's disease virus. Restriction
fragment polymorphism of the DNA products with selected enzymes add to the
specificity of the assay. Several non-essential regions in the genome of the
herpes virus of turkeys and the Rispens strain have been cloned and characterized.
The nucleotide and deduced amino acid sequence of turkey interleukin-2 has
been determined. Restriction analysis of genomic DNA extracted from fowl poxvirus
vaccines and from two fowl poxvirus recombinants used by the poultry industry
in the
IMPACT: 1998/04 TO 2001/03
The ability to generate functionally active genes of avian pathogens has been
established. The RT-PCR technique can be used for the validation and testing
of recombinant vaccines.
PUBLICATIONS: 1998/04 TO 2001/03
1. ROMERO,C.H. and CHUNG,H.Y. Restriction fragment length polymorphism of
classical and recombinant fowlpox virus vaccines used by the poultry industry
in the
2. ROMERO,C.H. and CHUNG,H.Y. Amplification and restriction analysis of a
highly conserved gene of Marek's disease virus to differentiate starins of
serotypes 1,2 and 3. American Association of Avian Pathologists Meeting,
3. ROMERO, C.H., Cai, X., Elyar, J.S. and Evans, K. Cloning, sequence, and
expression of turkey interleukin-2. Avian Diseases, 2000 (submitted).
(11)
ACCESSION NO: 0400221 SUBFILE: CRIS
PROJ
NO: 6612-32000-019-00D
AGENCY: ARS 6612
PROJ
TYPE: USDA INHOUSE
PROJ STATUS:
TERMINATED
START:
09 APR 1996 TERM: 08 APR 2001 FY: 2001
INVESTIGATOR: TUMPEY
T; MITCHELL B W; HOLT P S; SWAYNE D E
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
STIMULATION OF MUCOSAL IMMUNITY
IN CHICKENS TO PROTECT AGAINST ENTERIC AND RESPIRATORY PATHOGENS
OBJECTIVES: Examine the development of local humoral immune response
at mucosal surfaces in chickens and compare this response with systemic immunity.
Develop vaccines for mucosal immunity against intestinal and respiratory pathogens
in poultry and diagnostic tests that will predict effectiveness. Determine
the mechanisms for generation of airborne pathogens. Develop controls to improve
poultry health and enhance mucosal vaccine effectiveness by reducing airborne
pathgens and dust.
APPROACH: Birds will be orally infected with salmonella enteritidis
(SE) and serum and intestinal anti-SE antibody levels will be ascertained
over time. The birds will be re-infected to determine the development of serum
and intestinal immunological memory. Immune recognition of different components
of SE in serum and the intestinal tract will be compared. The protective role
of serum and mucosal antibodies will be ascertained by passive administration
of antibodies to naive birds and following the progression of the infection.
The development of immunity in the intestinal tract will be dilineated by
immunoassay of intestinal contents and elispot analysis of purified lamina
propria lymphocytes. Dust and bacterial counts will be measured in hatching
cabinets and other poultry production areas. Dust reduction techniques studied
will include lowering air velocity and using an electrostatic space charge
with a grounded collection system. Experiments will be conducted to characterize
airborne transmission of SE and to explore treatments for reducing it.
PROGRESS: 2000/10 TO 2001/09
PROTECT AGAINST ENTERIC AND RESPIRATORY PATHOGENS 1. What major problem or
issue is being resolved and how are you resolving it? The poultry industry
is still threatened by avian influenza virus (AIV) and can result in substantial
economic losses. Although the incidence of highly virulent AI strains are
relatively rare, less pathogenic strains often circulate through chicken and
turkey flocks and are responsible for significant morbidity, mortality and
production losses throughout the world. Current parenteral vaccines and vaccine
technologies provide protection against clinical signs and death from highly
pathogenic AI virus challenge. However, prevention of highly and mildly pathogenic
AI virus replication at mucosal sites is inconsistent and thus limits vaccines
in preventing transmission. This is thought to be due in part to the observation
that systemic vaccination is a poor inducer of mucosal immunity and therefore
organisms can invade the host before the systemic immunity can impede the
infection. Because AIV invade the mucosal surfaces, emphasis should be placed
on vaccines that induce strong mucosal immunity. Thus, the main objective
of this CRIS will be to develop mucosal vaccines for controlling AI in poultry
and to characterize the immune effectors mediating vaccine protection. The
development of new vaccine approaches for controlling such emerging or reemerging
pathogens can be of great importance to the profitability of the poultry industry.
In addition, effective mucosal vaccine strategies developed for AI will enable
scientists to develop vaccine approaches for controlling other respiratory
or gastrointestinal pathogens that affect poultry. 2. How serious is the problem?
Why does it matter? It has been estimated that the next outbreak of a highly
pathogenic AIV such as the 1983 outbreak in
PUBLICATIONS: 2000/10 TO 2001/09
1. Chaubal,L.H., Holt,P.S. Characterization of motility and identification
of flagella proteins in the avian pathogen Salmonella pullorum. American Journal
of Veterinary Research. 2000. v.60.p.1322-1327.
2. Katz,J.M., Lu,X., Frace,M. Morken,T., Zaki,S.R., Tumpey,T.M. Pathogenesis
of and Immunity to Avian Influenza A H5 viruses. Biomedicine Pharmacotherapy.
2000. v.54.p.178-187.
3. Tumpey,T.M., Lu,X., Morken,T., Zaki,S.R., Katz,J.M. Depletion of lymphocytes
and diminished cytokine production in mice infected with a highly virulent
influenza A virus isolated from humans. Journal of Virology. 2000. v.73.p.6105-6116.
4. Tumpey,T.M., Renshaw,M., Clements,J., Katz,J.M. Mucosal delivery of inactivated
influenza vaccine induces B cell dependent heterosubtypic protection against
a lethal influenza A H5N1 virus. Journal of Virology. 2001. v.75.p.5141-5150.
(12)
ACCESSION NO: 0402463 SUBFILE: CRIS
PROJ
NO: 6612-32000-021-00D AGENCY: ARS 6612
PROJ
TYPE: USDA INHOUSE PROJ STATUS: NEW
START:
INVESTIGATOR: KING D
J; SEAL B S; VACANT; KAPCZYNSKI D R; SWAYNE D E; MITCHELL B W
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
VIRULENCE DETERMINANTS IMPORTANT
TO PATHOGENESIS OF
OBJECTIVES: Molecular and biological characterization of new
APPROACH: Acquisition of new NDV and APV isolates from poultry
oubtreaks in the
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving
it?
PUBLICATIONS: 2000/10 TO 2001/09
1. Cameron,K., Zhang,X., Seal,B., Rodriguez,M., Njenga,M.K. Antigens to viral
capsid and non-capsid proteins are present in brain tissues and antibodies
in sera of Theiler's virus-infected mice. Journal of Virological Methods.
2001.v.91.p.11-19.
2. Gulati,B.R., Cameron,K.T., Seal,B.S., Goyal,S.M., Halvorson,D.A., Njenga,M.K.
Development of highly sensitive and specific enzyme-linked immunosorbent assay
based on recombinant matrix protein for detection of avian pneumovirus antibodies.
Journal of Clinical Microbiology. 2000.v.38. p.4010-4014.
3. Kapczynski,D.R., Koci,M., Kelley,L., Schultz-Cherry,S. Use of in vitro
expressed capsid protein from turkey astrovirus to protect poults from PEMS-associated
disease. Program of the Fifth International Congress of Veterinary Virology,
4. Kapczynski, D.R.,Smith, C. Immune response of turkeys following intranasal
vaccination with BPL-inactivated avian pneumovirus and live-virus challenge.
Program of the American Association of Avian Pathologists at the American
Veterinary Medical Association annual meeting. Boston, MA. July 14-18, 2001.(Abstract)
5. King,D.J. Efficacy of vaccine in protection against velogenic Newcastle
Disease (ND). Proceedings of the United Animal Health Association.2000. p.591-593.
6. King,D.J.,Swayne,D.E. Newcastle disease update: International ND problems.
Proceedings of the United Animal Health Association. 2000. p. 593-596.
7. King,D.J. Selection of Thermostable
8. King,D.J., Kommers,G.D., Brown,C.C., Seal,B.S. Virulence of pigeon
9. Kommers,G.D., King,D.J., Seal,B.S., Brown,C.C. Pathogenesis of five different
pigeon-origin isolates of
10. Locke,D.P., Sellers,H.S., Crawford,J.M., Schultz-Cherry,S., King,D.J.,
Meinersmann,R.J.,Seal,B.S. Newcastle disease virus phosphoprotein gene analysis
and transcriptional editing in avian cells. Virus Research.2000. v.69.p.55-68.
11. Seal,B.S., Sellers,H.S. Avian paramyxoviruses evolve independently of
their mammalian counterparts and deserve a genus designation among the subfamily
Paramyxovirinae in the family Paramyxoviridae. Program of the Ninth Annual
Meeting of the Society for Molecular Biology and Evolution, Athens, GA. July,7-10,
2001.(Abstract)
12. Seal,B.S. Molecular evolution of
13. Seal,B.S. Avian Pneumoviruses and Emergence of a New Type in the
14. Seal,B.S., King,D.J.
15. Sellers,H.S., Schultz-Cherry,S., Brown,C.C., Seal,B.S., King,D.J. Induction
of apoptosis by
16. Suarez,D.L., Swayne,D.E., King,D.J. The ongoing threat of avian influenza
and
17. Swayne,D.E., Suarez,D.L., King,D.J. Avian influenza (AI) and velogenic
(13)
ACCESSION NO: 0401966 SUBFILE: CRIS
PROJ
NO: 6612-32000-021-01T AGENCY: ARS 6612
TYPE: PROJ USDA INHOUSE PROJ STATUS:
TERMINATED
START:
INVESTIGATOR: SEAL B
S
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
CONSTRUCTION OF A
OBJECTIVES:
APPROACH: Since NDV genomic RNA is minus-sense and therefore
noninfectious, several cloned proteins must be expressed along with a positive
sense anti-genomic RNA. The nucleoprotein (NP), phosphoprotein (P) and the
polymerase (L) are currently being cloned into expression vectors. A NDV mini-genome
that contains the NP, P and L genes with the appropriate leader and trailer
sequences will be constructed. A reporter gene will be inserted between the
P and L genes. The NDV mini-genome will be coexpressed with viral NP, P and
L genes and monitored for expression of the reporter gene to validate a functional
NDV rescue system.
PROGRESS: 2000/10 TO 2001/09
1. What major problem or issue is being resolved and how are you resolving
it? 2. How serious is the problem? Why does it matter? 3. How does it relate
to the National Program(s) and National Component(s)? 4. What were the most
significant accomplishments this past year? D. Progress report. This report
serves to document research conducted under a specific cooperative agreement
between ARS and the U.S. Poultry and Egg Association. Additional details of
research can be found in the report for the parent project 6612-32000-021-00D
Paramyxovirus Infections of Poultry. Complete nucleotide sequencing of the
Newcastle disease virus (NDV) vaccine strain B1 is necessary to understand
pathogenesis and develop NDV as a vaccine vector. Using nucleotide sequencing
and mapping techniques, the full-length genomic sequence of the NDV vaccine
strain B1 was determine. The sequence has been published in Genbank for use
by all scientists. 5. Describe the major accomplishments over the life of
the project including their predicted or actual impact. 6. What do you expect
to accomplish, year by year, over the next 3 years? 7. What science and/or
technologies have been transferred and to whom? When is the science and/or
technology likely to become available to the end user (industry, farmer, other
scientists)? What are the constraints if known, to the adoption & durability
of the technology product? 8. List your most important publications in the
popular press (no abstracts) and presentations to non-scientific organizations
and articles written about your work (NOTE: this does not replace your peer-reviewed
publications which are listed below) Sellers, H.S. and Seal, B.S. The full-length
genomic sequence of NDV strain B1. GenBank accession number NC002617.
PUBLICATIONS: 2000/10 TO 2001/09
None.
(14)
ACCESSION NO: 0404751 SUBFILE: CRIS
PROJ
NO: 6612-32000-028-00D AGENCY: ARS 6612
PROJ
TYPE: USDA INHOUSE PROJ STATUS: NEW
START:
07 APR 2001 TERM:
INVESTIGATOR: TUMPEY
T; SWAYNE D E; VACANT; SUAREZ D L; MITCHELL B W
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
STIMULATION OF MUCOSAL IMMUNITY
IN CHICKENS TO PROTECT AGAINST ENTERIC AND RESPIRATORY PATHOGENS
OBJECTIVES: Examine the development of local humoral immune response
at mucosal surfaces in chickens and compare this response with systemic immunity.
Develop vaccines for mucosal immunity against intestinal and respiratory pathogens
in poultry and diagnostic tests that will predict effectiveness. Determine
the mechanisms for generation of airborne pathogens.
APPROACH: The protective role of serum and mucosal antibodies
will be ascertained by passive administration of antibodies to naive birds
and following the progression of the infection. The development of immunity
in the intestinal tract will be dilineated by immunoassay of intestinal contents
and elispot analysis of purified lamina propria lymphocytes.
(15)
ACCESSION NO: 0405248 SUBFILE: CRIS
PROJ
NO: 6612-32000-038-00D AGENCY: ARS 6612
PROJ
TYPE: USDA INHOUSE PROJ STATUS: NEW
START:
INVESTIGATOR: KING D
J; KAPCZYNSKI D R; SEAL B S; VACANT; SWAYNE D E; MITCHELL B W
PERFORMING INSTITUTION:
AGRICULTURAL RESEARCH SERVICE
IDENTIFICATION OF VIRULENCE
DETERMINANTS, PATHOGENETIC MECHANISMS, ...AVIAN PARAMYXOVIRUSES
OBJECTIVES: 1. Characterization of emergent
APPROACH: NDV and APV isolates will be acquired from outbreaks
and from surveys of North American wild bird populations. Wild waterfowl will
be surveyed for APV specific antibodies. Antigenic differentiation with monoclonal
and polyclonal antibodies and nucleotide sequence analysis of NDV and APV
genes will provide molecular characterization and epidemiologic determinations.
Virulence, persistence, and pathogenesis will be evaluated by inoculation
of chickens or turkeys. Sequence analysis in combination with results from
pathogenesis studies of isolates or infectious clones rescued from cloned
NDV will provide the basis for identification of virulence markers useful
for diagnostic development. Immunity to APV and NDV infections and newly developed
live, inactivated and subunit vaccines will be assayed for both antibody mediated
and cellular immune responses. Serum and mucosal antibody will be quantitated
and isotype determined. Cytokine regulators of the immune response will be
assayed. BSL-2 and BSL-3Ag, 8/10/01.
(16)
ACCESSION NO: 0189393 SUBFILE: CRIS
PROJ
NO: GEOV-0456 AGENCY: CSREES GEOV
PROJ
TYPE: ANIMAL HEALTH
PROJ
STATUS: NEW
START:
INVESTIGATOR: Sellers,
H. S.
PERFORMING INSTITUTION:
COLLEGE OF
110 RIVERBEND ROAD
DETECTION, ISOLATION AND
CHARACTERIZATION OF AVIAN VIRUSES
OBJECTIVES: The objectives of this proposal are to provide diagnostic
virology services for the
APPROACH: Objectives. The following objectives are broad as
this project is long-term and continuing flexibility is needed to adjust for
new situations in the field. To provide diagnostic virology services in an
accurate and reliable manner for the U.S. poultry industry Improve methods
of detection and monitoring of avian viruses Apply new monoclonal antibodies
(Mab) as they become available to diagnostic cases (i.e. monoclonal antibodies
to IBDV, J. Rosenberger) Utilize fluorescent (FITC)-labeled antibodies for
direct detection of viral antigen, as in recent subclinical Infectious Laryngotracheitis
(ILT) cases Apply PCR and nucleic acid probe technology for diagnostic applications
(i.e. real-time PCR utilizing the light cycler) Maintain contacts and working
relationships with other research and diagnostic facilities for the exchange
of data and reagents Conduct applied research on current avian virus diseases
isolated from the field in collaboration with clinicians, faculty, students
at PDRC and other poultry professionals in the field
NON-TECHNICAL SUMMARY: Despite rigorous vaccination in commercial poultry,
avian viruses continue to cause problems resulting in production losses for
the poultry industry. This project provides methods for detection, isolation
and characterization of avian viruses.
PROGRESS: 2001/10 TO 2002/09
The mission of the diagnostic virology laboratory is to provide accurate and
timely diagnostic virology services for the
IMPACT: 2001/10 TO 2002/09
Improved methods of virus detection and isolation will expedite control measures
used in the field to control and in some cases eradicate viral diseases. An
emphasis has been placed on molecular diagnostics in the past year and as
a result the time required for positive identification has been minimized
thus providing much needed information in less time.
PUBLICATIONS: 2001/10 TO 2002/09
Kapczynski, D.R., H.S. Sellers, V. Simmons, S. Schultz-Cherry. Sequence analysis
of the S3 gene from a turkey reovirus. Accepted to Virus Genes 2/2002.
PROJECT CONTACT:
Name: Sellers, H. S.
Phone: 706-542-5647
Fax: 706-542-5630
Email: hsellers@arches.uga.edu
URL: http://www.avian.uga.edu
(17)
ACCESSION NO: 0194953 SUBFILE: CRIS
PROJ
NO: GEOV-0466 AGENCY: CSREES GEOV
PROJ
TYPE: SPECIAL GRANT PROJ STATUS: NEW
CONTRACT/GRANT/AGREEMENT
NO: 2002-30001-12128 PROPOSAL NO: 2002-04435
START:
GRANT
AMT: $2,000,000
INVESTIGATOR: Prasse,
K. W.; Dickerson, H. W.; Miller, D. M.; Glisson, J. R.
PERFORMING INSTITUTION:
COLLEGE OF
110 RIVERBEND ROAD
CORE ANIMAL DIAGNOSTIC LABORATORY
OBJECTIVES: Principle objective is to develop a regional capability
to accurately and rapidly diagnose eight specific foreign animal diseases.
Secondly, to establish a secure communications network with the other designated
laboratories so that data may be shared throughout the network and with federal
authorities.
APPROACH: Personnel will be trained in diagnostic procedures
in eight foreign animal diseases. New equipment will be purchased to perform
the diagnostic tests to detect the eight diseases. Developing a computerized
reporting system in collaboration with 11 other states for reporting foreign
animal diseases.
NON-TECHNICAL SUMMARY: There is a critical need for a national animal health
reporting system to detect and report foreign animal diseases. This project
will contribute to the development of a network of detecting and reporting
foreign animal diseases nationwide.
PROJECT CONTACT:
Name: Miller, D. M.
Phone: 706-542-5568
Fax: 706-542-5977
Email: miller@vet.uga.edu
(18)
ACCESSION NO: 0152351 SUBFILE: CRIS
PROJ
NO: IND073055V AGENCY: CSREES
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START:
15 DEC 1995 TERM: 30 DEC 1998 FY: 1999
INVESTIGATOR: Guo, P.
PERFORMING INSTITUTION:
MICROBIOL PATHOLOGY & PUB HLTH
PURDUE UNIVERSITY
CONSTRUCTION OF ATTENUATED
RECOMBINANT AVIAN INFECTIOUS LARYNGOTRACHEITIS VIRUS VACCINES
OBJECTIVES: Our long term objective is to develop a polyvalent
vaccine which is both effective in stimulating a high level of mucosal immunity
against several avian respiratory tract infections, and is easy to administer,
such as via aerosal spray to facilitate vaccination on large-flock chicken
farms. Our short term goal of this proposal is to construct recombinant avian
infectious laryngotracheitis viruses (ILTV) expressing the Fusion (F) glycoprotein
of Newcastle disease virus (NDV) and the hemagglutinin (HA) of avian influenza
virus (AIV), individually or in combination. Expression of the non-ILTV proteins
will be monitored, and pathogenicity, immunogenicity and stability of the
recombinant viruses will be tested by in vivo experiments.
APPROACH: Genes coding for the F protein of NDV and HA protein
of AIV will be cloned by PCR after reverse transcription. The genes will be
introduced into the ILTV genome via homologous recombination using marker
genes for selection. Expression of the foreign genes will be monitored by
immunofluorescence and western blot. Vaccination experiments will be performed
to determine the minimum dose for 100% protection against NDV, AIV, and ILTV;
stability of the recombinant viruses will be tested through in vivo passaging.
Pathogenicity of the recombinant ILT viruses will be assessed symptomatically
as well as by screening for tracheal lesions. The best route of vaccination
to stimulate mucosal immunity will be determined by administration of the
recombinant ILT-HA virus via different inoculation routes.
PROGRESS: 1995/12 TO 2000/09
Our research has recently identified an hepatoma cell line for the cultivation
of infectious laryngotracheitis virus (ILTV), elucidated the assembly pathway
of this pathogen, constructed three dual viral promoters simultaneously recognized
by both mammalian and E. coli cells, documented the transactivation of the
early SV40 promoter by ILTV co-infection, developed a simple procedure for
ILT diagnosis and constructed recombinant ILTV with pathogenic gene deletion
and foreign gene insertion. The recombinant ILTV will be used as a vector
to develop a polyvalent vaccine for mucosal immunity against multiple avian
respiratory tract infections.
PUBLICATIONS: 1995/12 TO 2000/09
1. Guo, P., E.Scholz, B.Maloney, and E.Welniak. 1994. Construction of recombinant
avian infectious laryngotracheitis virus expressing bata-gal gene and DNA
sequencing of insertion region. Virology 202:771-781.
2. Scholz, E., R. E. Porter, and P. Guo. 1994. Differential diagnosis of infectious
laryngotracheitis from other avian respiratory disease by a simplified PCR
procedure. J Virol Meth. 50:313-322.
3. Guo,P., E.Scholz, J.Turek, R.Nordgren, and B.Maloney. 1993. Assembly pathway
of avian infectious laryngotracheitis virus. Am J Vet Res 54:2031-2039.
4. Scholz, E., C. L. Zhang, and P. Guo. 1993. Transactivation of the early
SV40 promoter by avian infectious laryngotracheitis virus in avian hepatoma
cells. J Virol Meth 45:291-301.
5. Scholz,E., E.Welniak, T.Nyholm, and P.Guo. 1993. An avian hepatoma cell
line for cultivation of infectious laryngotracheitis virus and for expression
of foreign genes with mammalian promotor. J Virol Meth 43:273-286.
6. Scholz, E. and P. Guo. 1995. Construction of Recombinant Avian Infectious
Laryngotracheitis Virus with TK Gene disrupted by Bata-gal Coding Sequence.
In Imm Viral Inf. Proc. 3rd Intl Cong Vet. Virol, 379-384..
7. Huang, Q., Y. Mat-Arip and P. Guo. 1997. Sequencing of a 5.5-kb DNA fragment
and identification of a gene for a subunit of helicase/primase complex of
avian laryngotracheitis virus. Virus Gene 15:(2): 119-121.
(19)
ACCESSION NO: 0184128 SUBFILE: CRIS
PROJ
NO: IOW03599 AGENCY: CSREES IOW
PROJ
TYPE: HATCH PROJ STATUS: NEW MULTISTATE PROJ NO: NC-228
START:
INVESTIGATOR: Reynolds,
D. L.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
AVIAN RESPIRATORY DISEASES:
PATHOGENESIS, SURVEILLANCE, DIAGNOSIS AND CONTROL
OBJECTIVES: Objective #1. Determine the pathogenesis and interactions
of specific agents. Objective #2. Surveillance, occurrence and consequences
of agents and hosts on disease susceptibility. Objective #3. Develop new and
improved methods for the diagnosis, prevention and control of avian respiratory
diseases.
APPROACH:
NON-TECHNICAL SUMMARY: Respiratory diseases afflicting poultry in modern
commercial production operations are complex entities. Numerous factors including
infectious agents, non-infectious agents and environmental factors may contribute
to the disease complex. The purpose of this project is to have a significant
impact on the diagnosis, control and prevention of poultry respiratory diseases.
PROGRESS: 2002/01 TO 2002/12
A study was initiated to evaluate biosecurity related to composting of large
amounts of animal carcasses. In order to achieve a large amount of animal
tissue, cattle carcasses were used. Twelve cattle carcasses averaging approximately
1,000 lbs. each, were delivered to the research site. The carcasses were placed
into three 20-ft long windrow segments (constructed end-to-end to produce
a single 60-ft long windrow). Each segment utilized one of three different
cover materials (silage, ground cornstalks, finished yard waste compost).
IMPACT: 2002/01 TO 2002/12
The findings indicate that composting may be a safe and environmental feasible
way to dispose of massive amounts of animal carcasses that may occur in such
catastrophic disease events as avian influenza, foot and mouth disease, etc.
PUBLICATIONS: 2002/01 TO 2002/12
No publications reported this period
PROJECT CONTACT:
Name: Good, C.
Phone: 515-294-4544
Fax: 515-294-2909
Email: cgood@iastate.edu
(20)
ACCESSION NO: 0181438 SUBFILE: CRIS
PROJ NO: IOWV-400-63-17
AGENCY: CSVM IOWV
PROJ TYPE: STATE PROJ STATUS:
TERMINATED
START:
INVESTIGATOR: Reynolds,
D. L.
PERFORMING INSTITUTION:
S. AND 16TH ELWOOD
STUDIES ON
OBJECTIVES: The objective of this project is to assess a new
recombinant vaccine for use in turkeys for the treatment of
APPROACH: Some poults will be vaccinated with rFP/NDV at the
hatchery. Vaccination of these poults will be by subcutaneous injection. Some
poults will be vaccinated orally at 3 weeks of age. Blood samples will be
collected by the medial wing vein method. Birds will be challenged with Texas
GB strain of velogenic
NON-TECHNICAL SUMMARY:
PROGRESS: 1998/05 TO 1998/12
The fowlpox vectored recombinant
IMPACT: 1998/05 TO 1998/12
The rFPNDV used by itself will not convey strong protection against challenge
with velogenic NDV.
PUBLICATIONS: 1998/05 TO 1998/12
None, 1999
PROJECT CONTACT:
Name: Reynolds, D. L.
Phone: 515-294-0914
Email: dlr@iastate.edu
(21)
ACCESSION NO: 0181440 SUBFILE: CRIS
PROJ
NO: IOWV-400-63-26 AGENCY: CSVM IOWV
PROJ
TYPE: STATE PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Reynolds,
D. L.
PERFORMING INSTITUTION:
S. AND 16TH ELWOOD
STUDIES ON OASIS IN
OBJECTIVES: The objective of this trial is to determine if the
nutritional supplement Oasis has any beneficial effect on enhancing the immune
system of young turkeys.
APPROACH: Birds will be allowed access only to Oasis for 48
hours following hatch and prior to placing them into battery brooders. Corresponding
control birds will not receive Oasis, food or water. Upon placing the birds
they will be vaccinated with a commercial adjuvanated vaccine. Blood samples
will be collected at weekly intervals for weeks. Antibody titers and total
serum immunoglobulin will be assessed on the birds and the groups will be
compared.
NON-TECHNICAL SUMMARY: Treatment/vaccine for
PROGRESS: 1998/10 TO 1999/09
Oasis is a commercial product that is used when transporting turkey poults
or chick from the hatchery to remote locations. The product is a hydrated
gel that is placed in the transport box. The hatchling birds pick and eat
the material and is supplies nutrition and hydration during the transport
period. The objective of this trial was to determine if Oasis had a positive
influence on the immune response of the bird. Birds were hatched and one grup
was placed on oasis for 48 hours while a control group received nothing. The
birds were also vaccinated with IBD immediately following hatch to determine
if birds receiving Oasis would mount a higher antibody response. The results
indicated there was no difference in the immune response to vaccination between
those birds receiving oasis and those that did not.
IMPACT: 1998/10 TO 1999/09
Hydrated gel material will not act to stimulate a better immune response in
commercial poultry.
PUBLICATIONS: 1998/10 TO 1999/09
None, 1999
PROJECT CONTACT:
Name: Reynolds, D. L.
Phone: 515-294-0914
Email: dlr@iastate.edu
(22)
ACCESSION NO: 0181442 SUBFILE: CRIS
PROJ
NO: IOWV-400-63-88 AGENCY: CSVM IOWV
PROJ
TYPE: STATE PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Reynolds,
D. L.
PERFORMING INSTITUTION:
S. AND 16TH ELWOOD
STUDIES ON
OBJECTIVES: The objective of this proposal is to determine if
intrayolk-sac (IYS) injection could be used to efficaciously vaccinate birds
against
APPROACH: Groups of birds were vaccinated by the IYS route
with commercial licensed
NON-TECHNICAL SUMMARY:
PROGRESS: 1997/09 TO 1998/08
Immunoglobulins purified from the egg yolks of chickens vaccinated against
IMPACT: 1997/09 TO 1998/08
Passive immunization may be an important strategy in controlling certain avian
pathogens. The results of this study that passive immunity will require the
entire immunoglobulin molecule.
PUBLICATIONS: 1997/09 TO 1998/08
Reynolds, D. L., S. Akinc and A. Ali. Studies on the passive immunity employing
the Fab' fragment of chicken immunoglobulin. Oral presentation. AAAP/AVMA
annual convention meeting. New Orleans, LA. July 10 - 14, 1999.
PROJECT CONTACT:
Name: Reynolds, D. L.
Phone: 515-294-0914
Email: dlr@iastate.edu
(23)
ACCESSION NO: 0132087 SUBFILE: CRIS
PROJ
NO: IOWV-701-23-10-0005 AGENCY: CSVM IOWV
PROJ
TYPE: STATE PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Reynolds,
D. L.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
RESPIRATORY DISEASES OF POULTRY
OBJECTIVES: To develop diagnostic techniques to aid in the control
of poultry respiratory diseases of economic importance. To improve current
methods of management and prevention of poultry respiratory diseases.
APPROACH: To apply modern laboratory techniques and develop
new products that relate to the above objectives.
PROGRESS: 1986/07 TO 2001/06
Newcastle disease (ND) is a highly contagious viral disease of poultry capable
of causing high morbidity and mortality. The traditional strategy for controlling
outbreaks of highly pathogenic ND is to eradicate exposed, or potentially
exposed, flocks of birds. Although this strategy has proved successful, it
typically results in large numbers of birds being euthanized. The environmental,
economic and animal ethical issues of this strategy are of increasing concern.
The objective of this study was to examine the potential for using passive
immunization as an alternative strategy for controlling highly pathogenic
outbreaks of ND. Here we determine the time interval between exposure and
providing protection by administering anti-Newcastle disease virus (NDV) specific
immunoglobulin subsequent to virulent NDV challenge. Different groups of chickens
were passively immunized (i.e. received Anti-NDV antibody) at various times
with respect to challenge corresponding to 24 hrs. prechallenge, day of challenge,
1, 2, 3, 4, 5, 6, 8 and 9 days post challenge. HI titers were evaluated prechallenge
prior to antibody injection and 24 hours post passive immunization. Serologic
results indicated that all birds passively immunized had titers between 10
and 12 log2. The results of the challenge indicated that all birds that received
passive immunization by 3 days following challenge were protected. Protection
began to wane by 4 days post challenge and little (if any) protection was
afforded by 8 days post challenge. In general, if birds were administered
immunoglobulins prior to clinical signs of ND, they were afforded protection.
IMPACT: 1986/07 TO 2001/06
The results of the passive immunity studies indicate that providing passive
protection to birds at the time of, or subsequent to, challenge can provide
protection. This method of protection may be an alternative to eradication.
PUBLICATIONS: 1986/07 TO 2001/06
Reynolds, Donald and Sevinc Akinc. Passive immunization protects birds following
challenge with virulent NDV. Oral presentation. AVMA / AAAP annual meeting.
PROJECT CONTACT:
Name: Reynolds, D. L.
Phone: 515-294-0914
Fax: 515-294-1401
Email: dlr@iastate.edu
(24)
ACCESSION NO: 0175014 SUBFILE: CRIS
PROJ
NO: MD-D-115 AGENCY: CSREES MD.
PROJ
TYPE: HATCH PROJ STATUS: TERMINATED MULTISTATE PROJ NO: NE-138
START:
INVESTIGATOR: Vakharia,
V. N.; Heckert, R. A.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
UNIV OF
COLLEGE PARK
EPIDEMIOLOGY AND CONTROL
OF EMERGING STRAINS OF POULTRY RESPITORY DISEASE AGENTS
OBJECTIVES: Design and implement novel immune and genetic prophylactic
strategies for effective control of respitory diseases caused by emerging
IBV, ILTV, mycoplasmas, IBDV, and CAV.
APPROACH: Approach: The genomic DNA of chicken anemia vius
(CAV) will be cloned and complete nucleotide sequence determined. The VPI
and VP2 genes will be subcloned and expressed in a baculovirus expression
system. The synthesized proteins will be characterized with monoclonal antibodies
and then evaluated as immunogens serologically and via challenge.
PROGRESS: 1996/10 TO 2002/09
To develop an attenuated, multi-spectrum vaccine candidate that can protect
against classical and variant strains of infectious bursal disease virus (IBDV),
we constructed several full-length and chimeric cDNA clones of segments A
and B of the D78 and GLS strains. Using the cRNA-based reverse genetics system
developed for IBDV, we generated recombinant viruses after transfection in
Vero cells. A panel of IBDV-specific monoclonal antibodies (MAbs) was used
to characterize the recovered viruses and their replication kinetics was compared
with that of the parental D78 strain in vitro. Viruses deficient in the expression
of VP5 nonstructural protein (NS) grew to slightly lower titers than D78 virus
and exhibited decreased cytotoxic and apoptotic effects in cell culture. To
evaluate the efficacy of the recombinant IBDV vaccine, we inoculated 3-week-old
chickens with this virus and challenged them with STC and variant GLS viruses.
Based on histopathology and serology tests, we observed that the chickens
inoculated with recombinant IBDV vaccine (containing a GLS-specific epitope)
failed to induce any pathological lesions or clinical signs of disease, and
were completely protected against both classic and variant IBDV strains. In
another study, we show that dimethylsulfoxide (DMSO) enhances liposome-mediated
transfection of nucleic acid in chicken macrophage cells and that this could
be exploited for transcutaneous delivery of naked DNA through the intact skin
of chickens. We found that DMSO enhanced transfection efficiencies of lipofectamine
and polyethyleneimine in HD-11 chicken macrophage cells. Based on this principle,
we showed that transcutaneous delivery of a DNA plasmid-dimethylsulfoxide
mixture (1:1) to untreated skin of chickens result in a wide distribution
of the plasmid in the body. Distribution studies were done using plasmids
encoding enhanced green fluorescent protein (EGFP) reporter gene and a bivalent
DNA vaccine coding for IBDV and
IMPACT: 1996/10 TO 2002/09
We have developed an attenuated, multivalent IBDV vaccine that can protect
against classic and variant strains, and have developed methods for delivery
of a DNA vaccine. These findings will aid in the development of cost-effective
vaccines for viral pathogens and better equip the chicken farmers with tool
necessary to combat viral diseases.
PUBLICATIONS: 1996/10 TO 2002/09
1. Elankumaran, S., Heckert, R.A., and Moura, L. 2002. Persistence and tissue
distribution of a variant strain of infectious bursal disease virus in commercial
broiler chickens. Avian Dis. 46:169-176.
2. Heckert, R.A., Elankumaran S, Oshop G, and Vakharia V.N. 2002. A novel
transcutaneous plasmid-dimethylsulfoxide delivery technique for avian nucleic
acid immunization. Vet Immunol Immunopathol. 89:67-81.
3. Oshop G, Elankumaran S, Heckert R.A. 2002. DNA vaccination in the avian.
Vet Immunol Immunopathol. 89:1-12. Oshop, G.L., Elankumaran, S., Vakharia,
V.N., and Heckert, R.A. 2002. In ovo delivery of DNA to the avian embryo.
Vaccine (in press).
4. Vakharia, V.N. 2002. Molecular determinants of virulence in infectious
bursal disease virus. 3rd European Concerted Research Action (COST 839) Meeting
on Immunosuppressive Viral Diseases in Poultry, April 25-28,
5. Liu, M., Brandt, M., Liu, Y., Edwards, G.H., and Vakharia, V.N. 2002. Recombinant
attenuated IBDV vaccine that protects against classic and variant strains.
138th American Veterinary Medical Association Annual Convention,
6. Liu, M., and Vakharia, V.N. 2002. Two amino acid residues in VP2 protein
of IBDV are involved in cell entry and efficient replication in vivo. 21st
Annual Meeting of American Society for Virology, July 20-24,
(25)
ACCESSION NO: 0190633 SUBFILE: CRIS
PROJ
NO: MD-VTMD-9201 AGENCY: CSREES MD.
PROJ
TYPE: NRI COMPETITIVE GRANT PROJ STATUS: NEW
CONTRACT/GRANT/AGREEMENT
NO: 2002-35204-11601 PROPOSAL NO: 2001-02372
START:
15 DEC 2001 TERM: 31 DEC 2003 GRANT YR: 2002
GRANT
AMT: $228,000
INVESTIGATOR: Samal,
S.; Gelb, J.
PERFORMING INSTITUTION:
VETERINARY MEDICINE
UNIV OF
COLLEGE PARK
RECOMBINANT
OBJECTIVES: A recombinant Newcastle Disease Virus (NDV) containing
the spike S1 glycoprotein gene of avian infectious bronchitis virus (IBV)
will be recovered from cloned cDNAs. The level of expression, intracellular
transport, and processing of the IBV S1 protein expressed from the recombinant
NDV will be examined. The recombinant NDV expressing the S1 protein will be
evaluated as a vaccine to control both of these economically important diseases.
APPROACH: A reverse genetic system developed by our laboratory
will be used to recover a recombinant
NON-TECHNICAL SUMMARY: IBV is the most common respiratory disease in poultry
in the
PROGRESS: 2002/01 TO 2003/01
We have constructed two NDV cDNA constructs containing the S1 gene of IBV
strain Mass 41. In one construct, the S1 gene was introduced into the first
position, before the NP gene, in the full-length cDNA of NDV strain Beaudette
C. In the other construct, the S1 gene of Mass 41 was introduced into the
third position, after P gene, in the full-length cDNA of NDV strain LaSota.
Recombinant NDV strains Beaudette C and LaSota containing the S1 gene were
recovered after transfection of HEp-2 cells following our standard recovery
protocol. Recombinant NDV strains containing the S1 gene of IBV were first
examined by RT-PCR to confirm the presence of the S1 gene in the genome of
recombinant NDV. To determine whether the recombinant NDV strains correctly
expressed the IBV S1 protein in infected cells, Western blot analysis was
performed. Our results showed that correct size S1 protein was expressed from
recombinant NDV strains. Pathogenicity studies of these recombinant NDV strains
in 10-day-old chicks showed that the ICPI values were slightly lowered. Studies
are underway to determine whether the S1 protein is incorporated into NDV
particles.
IMPACT: 2002/01 TO 2003/01
The use of the recombinant NDV vector to deliver the S protein of IBV should
provide immunity against IBV and NDV strains. Unlike currently available IBV
attenuated vaccines and field strains, NDV is highly stable. Thus, the recombinant
NDV/IBV vaccine should not contribute to the evolution of new IBV variants
that continue to plague the poultry industry. Our proposed vaccine will be
highly beneficial to the poultry industry.
PUBLICATIONS: 2002/01 TO 2003/01
No publications reported this period
PROJECT CONTACT:
Name: Samal, S.
Phone: 301-314-6813
Fax: 301-314-6855
Email: ss5@umail.umd.edu
(26)
ACCESSION NO: 0180471 SUBFILE: CRIS
PROJ
NO: MDR-9802290 AGENCY: CSREES MD.R
PROJ
TYPE: NRI COMPETITIVE GRANT PROJ STATUS: EXTENDED
CONTRACT/GRANT/AGREEMENT
NO: 98-35204-6427
START:
GRANT
AMT: $140,000
INVESTIGATOR: Samal,
S. K.
PERFORMING INSTITUTION:
UNIV OF
COLLEGE PARK
PRODUCTION OF INFECTIOUS
OBJECTIVES: 9802290. 1. Construction of a full-length NDV cDNA
clone. 2. Recovery and characterization of infectious NDV from cDNA. 3. Construction
of NDV with mutations in the fusion protein cleavage site.
APPROACH: A full-length cDNA of the genomic RNA of NDV will
be constructed using reverse-transcriptase polymerase chain reaction. The
cDNA will be cloned into a plasmid flanked by a T7 promoter and ribozyme sequences.
Infectious NDV will be produced by the intercelluler coexpression of T7 based
plasmid cDNAs. One cDNA will encode the complete NDV genome and the other
cDNAs will encode NDV proteins required for first round of virus specific
mRNA synthesis. T7 polymerase will be supplied by a replication-deficient
recombinant vaccinia virus. The recovered virus will be characterized using
several invitro methods. Mutations will be introduced into NDV cDNA by site
directed mutagenesis.
PROGRESS: 1999/10 TO 2000/09
A recombinant NDV strain, Beaudette C, was generated from cloned cDNAs. Characterization
of the recombinant NDV showed similarities in growth and pathogenicity to
that of the parental wild-type virus. The sequence of the cleavage site of
the fusion protein of the recombinant NDV were altered by site-directed mutagenesis.
Recombinant NDV containing the mutation required trypsin activation for fusion
and infectivity in cell culture. The virulence of the recombinant NDV with
altered fusion protein cleavage site was also lowered. This result showed
that the cleavage of the fusion protein plays an important role in the pathogenesis
of NDV.
IMPACT: 1999/10 TO 2000/09
Recovery of infectious recombinant Newcastle Disease Virus from cloned DNA
can produce better vaccines against Newcastle Disease in poultry. This new
technology will also enable recombinant Newcastle Disease Virus to be used
as vaccine vector for other avian pathogens. Thus, production of recombinant
Newcastle Disease Virus has a great potential for the development of vaccines
against avian pathogens and will significantly benefit the poultry industry.
PUBLICATIONS: 1999/10 TO 2000/09
Krishnamurthy, S., Huang, Z. and Samal, S.K. 2000. Recovery of a virulent
strain of Newcastle Disease Virus from cloned cDNA: epression of a foreign
gene results in growth retardation and attenuation. Virology 278: 168-182.
(27)
ACCESSION NO: 0180895 SUBFILE: CRIS
PROJ
NO: MICK-9803219 AGENCY: CSREES MICK
PROJ
TYPE: SMALL BUSINESS GRANT PROJ STATUS: EXTENDED
CONTRACT/GRANT/AGREEMENT
NO: 98-33610-6303
START:
GRANT
AMT: $225,000
INVESTIGATOR: Reilly,
J. D.
PERFORMING INSTITUTION:
ORIGEN, INC.
NATURA, INC.
IMMORTAL CELL LINE FOR POULTRY
VACCINE PRODUCTION & DIAGNOSTICS
OBJECTIVES: 9803219. There are two technical objectives for this
SBIR Phase II proposal. The first technical objective is to develop methods
for using OCLTM cells to make vaccines that are at least comparable to traditional
methods in yield and efficacy. The viruses that will be used are SB1-OCLTM,
CVI988-OCLTM, avian influenza virus, and vaccine strains of avian reovirus,
APPROACH: The components of the first objective are: 1) One-step
growth curves to determine yield of virus in OCLTM cells compared to traditional
methods, 2) Modify various growth parameters including initial cell density,
MOI, temperature, media composition to maximize the yield of each virus, 3)
Compare immunogenicity of virus produced on OCLTM cells to virus produced
by traditional methods, 4) Compare safety of virus produced on OCLTM cells
to virus produced by traditional methods, and 5) Scale growth of each virus
up to production levels. Specifically, the components of the second objectives
are: 1) Compare sensitivity of OCLTM-based VI assays to traditional methods,
and 2) Compare susceptibility of a panel of virus strains and field isolates
for each virus tested to traditional methods of detection. The Immunogenicity
and safety tests will be performed as described for each virus in 9 CFR PP113.
(28)
ACCESSION NO: 0172971 SUBFILE: CRIS
PROJ
NO: MISV-322090 AGENCY: CSREES MISV
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Wang,
C.; Magee, D.; Keirs, R. W.
PERFORMING INSTITUTION:
COLLEGE OF
MISSISSIPPI STATE UNIV
MISSISSIPPI STATE
INFECTIOUS BRONCHITIS VIRUS
INFECTION IN COMMERCIAL BROILERSAND CHICKEN EMBRYOS
OBJECTIVES: Serotype IBV from selected
APPROACH: Trachea, cecal tonsils and cloacal contents will
be collected from birds suffering from respiratory problems within
NON-TECHNICAL SUMMARY: The project seeks to understand how infectious bronchitis
virus infection occurs in and affects commercial broilers and chicken embryos.
PROGRESS: 1996/10 TO 2002/06
Infectious bronchitis (IB) is an acute, highly contagious viral respiratory
disease and one of the most common and economically important diseases in
the poultry industry. The objectives of this study were to 1) study the epidemiology
of IB in
IMPACT: 1996/10 TO 2002/06
This study help to understand the pathogenesis of infectious bronchitis virus
and to utilize the best method to diagnose the disease when there is an IB
outbreak. The resulting information provided prevention and control strategies
to save millions of dollars lost annually due the virus, consequently significantly
enhancing profitability.
PUBLICATIONS: 1996/10 TO 2002/06
1. Wang C, Miguel B, Hong C, Austin FW, and Keirs RW. Comparision of the immunofluorescent
assay and reverse transcription-polymerase chain reaction to detect and type
infectious bronchitis virus. Avian Dis, 1999, 43:590-596.
2. Shi Q and Wang C. Genetic relationships of infectious bronchitis virus
isolates from
3. Miguel B,
PROJECT CONTACT:
Name: Wang, C. L.
Phone: 662-325-1205
Fax: 662-325-1031
Email: wang@cvm.msstate.edu
(29)
ACCESSION NO: 0172972 SUBFILE: CRIS
PROJ
NO: MISV-329040 AGENCY: CSREES MISV
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Montgomery,
R. D.; Maslin, W.; Boyle, C. R.
PERFORMING INSTITUTION:
COLLEGE OF
MISSISSIPPI STATE UNIV
MISSISSIPPI STATE
THE HEAD-ASSOCIATED LYMPHOID
OF CHICKENS AND ITS IMMUNOLOGICAL ROLE
OBJECTIVES: Improve the methodology used to evaluate the HALT;
Effects of various stressors on the HALT; and (3) Effect of GH on pathogenesis
of various pathogens.
APPROACH: An ELISA will be developed to quantitate the various
classes of immunoglobulin (Ig) present in tears (IgA, IgG, IgM) of chickens
exposed to a test antigen (B. abortus). Once developed, the ELISA will be
used to re-evaluate the default GH B. abortus assay to determine which parameters
can be modified to increase the sensitivity and specificity of the assay;
Stressors known to have an impact on lymphoid systems and/or the respiratory
tract will be given to various ages of SPF chicks. Following exposure, these
chicks will be subjected to both structural and functional analysis to determine
the effect of these agents on the HALT; and Respiratory tract-oriented microbial
pathogens, including viruses and bacteria will be given to both GH-intact
and GHx chicks of various ages. The pathogenesis of these stressors will be
monitored in both types of subjects by conducting sequential microbial recoveries
and histological analyses of targtet sites specific for the particular agent
used. Optionally, if the pathogenesis of vaccine viruses are studied, the
GH-intact and GHx chicks may be challenged at the end of the trial to determine
the degree of protection in both.
NON-TECHNICAL SUMMARY: Defining the immunological role of the head-associated
lymphoid tissues of chickens is the focus of this project. This work will
aid us in understanding how to utilize the tissue to enhance immune responses
in the chicken.
PROGRESS: 1996/10 TO 2002/06
Thirty-six modified-live virus vaccines, including 16 infectious bronchitis
virus (IBV), 10 Newcastle Disease virus (NDV), and 10 NDV/IBV vaccines were
evaluated for their effects on the gland of Harder (GH) and other head-associated
lymphoid tissue (HALT) sites. Some of the IBV vaccines, either alone or in
combination with NDV, were found to interfere with the GH/HALT's ability to
respond to antigenic stimulation and to alter specific histological attributes.
Two hundred and eight E. coli were collected from various lesions (respiratory,
intestinal, yolk, joint, etc.). These isolates were characterized biochemically,
analyzed for their plasmid content(s), analyzed for their sensitivity to antibiotics,
and evaluated for their lethality in embryonated chicken eggs, which reputedly
correlates with in vivo pathogenicity in young chickens. One of the E. coli,
which repeatedly spared embryonated eggs was adapted to grow in the presence
of nalidixic acid and evaluated in the chicks that hatched. Although overall
hatchability was reduced, a number of infected embryos did hatch into viable
and healthy-appearing chicks. However, those chicks had lowered body weights
and increased early mortality. Consistently high levels of the nalidixic acid
resistant-E. coli were recovered from the yolk of these chicks and moderate
levels were detected in their lung and trachea. E. coli was also recovered
from the respiratory tract of non-infected chicks that were hatched in contact
with the E. coli chicks, indicating that E. coli can be transmitted vertically
through the embryo and amplified horizontally to susceptible neonates at the
time of hatching. Data from an extensive respiratory outbreak that occurred
in Mississippi broilers during 1998-1999 was collected and analyzed. Arkansas
and, to some extent, Connecticut IBV, were the principal agents detected in
this outbreak. Epidemiological factors collected with these cases included
date received, identity of growout company involved, age of birds, strain(s)
of IBV in vaccination program, infectious bursal disease (IBD) vaccination
status, condition of samples received, any respiratory lesions noted, and
the geographic location of farm. In general, the number and distribution of
1) cases received, 2) cases positive for virus, and 3) viruses detected were
proportional to the epidemiological factors collected.
IMPACT: 1996/10 TO 2002/06
This research indicated that by our use of 36 modified live virus vaccines
which included 16 infectious bronchitis virus, 10 Newcastle Disease viruses,
and 10 NDV/IBV vaccines, either alone or in combination, we were successful
in interfering with the GH/HALT's ability to respond to antigenic stimulation
and to alter specific histological attributes.
PUBLICATIONS: 1996/10 TO 2002/06
1. Montgomery, R. D., C. R. Boyle, W. R. Maslin, and D. L. Magee. Attempts
to reproduce a runting/stunting-type syndrome using infectious agents isolated
from affected
2. Montgomery, R. D., W. R. Maslin, and C. R. Boyle Effects of Newcastle disease
vaccines and Newcastle disease/infectious bronchitis virus combination vaccines
on the head-associated lymphoid tissues of chickens. Avian Dis. 41:399-406.
1997.
3. Montgomery, R. D., C. R. Boyle, T. A. Lenarduzzi, and L. S. Jones. Chicks
Hatched from Escherichia coli-infected Embryos. Avian Diseases. 43:553-563.
1999.
PROJECT CONTACT:
Name: Montgomery, R.
D.
Phone: 662-325-1205
Fax: 662-325-1031
Email: montgomery@cvm.mssstate.edu
(30)
ACCESSION NO: 0187676 SUBFILE: CRIS
PROJ
NO: MO-ASAH0594 AGENCY:
PROJ
TYPE: ANIMAL HEALTH PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Ledoux,
D. R.; Bermudez, A. J.; Rottinghaus, G. E.
PERFORMING INSTITUTION:
CHARACTERIZATION OF TOXICOLOGICAL
EFFECTS OF MULTIPLE MYCOTOXINS IN POULTRY
OBJECTIVES: Determine the additive, synergistic, or antagonistic
effects of low levels of multiple mycotoxins on performance, organ weights,
hematology, serum chemistry, and immune function of poultry.
APPROACH: Chicks and poults will be fed combinations of low
levels of mycotoxins (naturally occurring levels) for three to four weeks
and the individual and combined effects of the mycotoxins will be assessed
based on growth performance, organ weights, hematology, serum chemistry, and
immune function. The primary and secondary antibody response to inactivated
NON-TECHNICAL SUMMARY:
PROGRESS: 2001/01 TO 2001/12
A 21-day experiment was conducted to determine if the turkey could be used
as a model for evaluating the efficacy of adsorbents to ameliorate the toxic
effects of aflatoxin (AF). Dietary treatments fed from day of hatch included:
0 ppb AF, 100 ppb AF, 200 ppb AF, 300 ppb AF, 400 ppb AF, 500 ppb AF, and
600 ppb AF. AF was supplied by A. parasiticus culture material that contained
986 ppm AFB1, 29 ppm AFB2, 464 ppm AFG1, and 9 ppm AFG2. Compared with controls,
poults fed 200 ppb AF or higher had reduced (P < .001) feed intake and
lower (P < .001) body weight gains. Significant mortality (14/20) occurred
in poults fed 600 ppb AF. Compared with controls, poults fed 100 ppb AF or
higher had lower (P < .001) relative liver weights, whereas poults fed
200 ppb AF or higher had increased (P < .001) relative kidney weights.
Histopathological analysis indicated the presence of liver lesions in poults
fed 100 ppb AF or higher. The primary hepatic lesions were biliary hyperplasia,
hepatocellular hyperplasia, and hepatic necrosis with the severity of lesions
increasing with increasing AF dose. Kidney lesions were noted in poults fed
diets containing 400 ppb AF or higher with a mild to moderate membranous thickening
of the glomerular capillary basement membrane noted in most specimens. Results
confirm previous reports that suggest turkeys are very sensitive to the toxic
effects of AF. The lowest level of AF (100 ppb) that caused toxic effects
in poults in this study is 20 fold lower than the levels (2 ppm or higher)
reported to cause toxic effects in broilers under laboratory conditions.
IMPACT: 2001/01 TO 2001/12
Results suggest that the turkey could be used as a more sensitive model to
evaluate the efficacy of adsorbents to ameliorate the toxic effects of AF
at levels that have been reported to cause toxicity under field conditions.
PUBLICATIONS: 2001/01 TO 2001/12
1. Broomhead, J. N., D. R. Ledoux, A. J. Bermudez, and G. E. Rottinghaus,
2002. Chronic effects of fumonisin B1 in broilers and turkeys fed dietary
treatments to market age. Poultry Science 81:56-61.
2. Ledoux, D. R., G. E. Rottinghaus, and A. J. Bermudez, 2001. In vitro binding
of mycotoxins by adsorbents does not always translate into in vivo efficacy.
Pp. 279-287 In: Mycotoxins And Phycotoxins In Perspective At The Turn Of The
Millenium. Proceedings of the Xth International IUPAC symposium on Mycotoxins
and Phycotoxins.
3. Butkeraitis, P., J. N. Broomhead, E. A. Guaiume, D. R. Ledoux, A. J. Bermudez,
and G. E. Rottinghaus, 2002. A turkey model for evaluating the efficacy of
adsorbents to ameliorate the toxic effects of aflatoxin. Abstracts International
Poultry Scientific Forum, January 14-15,
4. Ledoux, D. R., J. Broomhead, A. J. Bermudez, and G. E. Rottinghaus, 2001.
Mycotoxin-Nutrient interactions. pp. 82-94. Proceedings 62nd
5. Ledoux, D. R., G. E. Rottinghaus, A. J. Bermudez, and J. N. Broomhead,
2001. Is fumonisin B1 a threat to the poultry industry? Abstracts The World
Mycotoxin Forum, 14-15 May 2001, Noordwijk, The
PROJECT CONTACT:
Name: Ledoux, D. R.
Phone: 573-882-1140
Email: ledouxd@missouri.edu
(31)
ACCESSION NO: 0088380 SUBFILE: CRIS
PROJ
NO: OHO00740 AGENCY: CSREES OHO
PROJ
TYPE: HATCH PROJ STATUS: TERMINATED MULTISTATE PROJ
NO: NC-168
START:
INVESTIGATOR: Nestor,
K. E.; Velleman, S. G.
PERFORMING INSTITUTION:
ADVANCED TECHNOLOGIES FOR
THE GENETIC IMPROVEMENT OF POULTRY
OBJECTIVES: Develop, compare, and integrate emerging technologies
with classical quantitative genetics for improvement of economic traits in
poultry.
APPROACH: Long-term lines of turkeys selected for increased
egg production and increased 16-week body weight and their corresponding randombred
controls will be maintained. Also, Japanese quail lines divergently selected
for 4-week body weight (HW, LW) and plasma yolk precursor (HP, LP) and lines
(HW-HP, HW-LP) selected for a combination of these traits will be maintained.
A randombred (R1) control population of Japanese quail will be maintained
without selection. Attempts will be made to study genetic variation among
the experimental turkey and Japanese quail lines by DNA fingerprinting and
study of MHC haplotypes. The turkey lines will be studied for resistance to
certain diseases including fowl cholera and
PROGRESS: 1997/10 TO 2003/09
Beltsville Small White (BSW) turkeys have been utilized as an experimental
model in the study of bacterial, parasitic, and fungal diseases. Given the
critical role of the major histocompatibility comple (MHC) antigens in the
initial steps of immune response to specific pathogens, the MHC Class II of
BSW turkeys was characterized. Southern blot analysis of PvuII-digested turkey
DNA that was hybridized with a chicken Class II beta gene genomic clone revealed
two restriction fragment length polymorphism profiles not previously identified
in experimental or commercial breeder lines of turkeys. These fingerprint
profiles differed in a single 6.0-kb gand that was present in approximately
38% of the birds examined. The DNA fragments of 5.0, 4.1, 3.3, and 3.1 were
present in both profiles. Furthermore, no mixed lymphocyte reaction was observed
between individuals within the BSW turkey line. The present results indicate
that BSW turkeys represent a unique source of genetic diversity for MHC Class
II haplotypes. Candidate male and female breeders from a number of genetic
lines of turkeys that were reared intermingled, with the sexes housed in different
buildings on the same farm, were vaccinated with a live Newcastle disease
virus vaccine (type B1, strain B1, Lasota) just prior to the commencement
of egg production. In 1999, an average mortality of 5.8 % occurred immediately
following vaccination and the level of mortality varied among lines. Mortality
was greater in large-bodied lines than in small-bodied lines. Affected birds
exhibited multiple areas of focal necrosis in the liver and spleen and congestion
of the heart and lungs. The percentage mortality occurring following similar
vaccination in 2000 averaged 2.6 and mortality was greater in one line (F
line) than the other genetic groups and higher in females than in males. Mortality
in the F line, selected for increased body weight and known to be susceptible
to various diseases, averaged 15.1% for both years. Attempts failed in both
years to isolate Pasteurella multocida or other bacteria. There was a positive
correlation between increased body weight and increased mortality following
vaccination with the live LaSota vaccine.
IMPACT: 1997/10 TO 2003/09
The MHC has been shown to be involved in disease resistance in turkeys. Knowledge
of variation in MHC haplotypes is important for turkey breeders when they
select for disease resistance. Selection for increased growth rate reduces
disease resistance in turkeys so commercial turkey breeders should include
selection for disease resistance in their program.
PUBLICATIONS: 1997/10 TO 2003/09
1. Sacco, R. E., K. E. Nestor, and R. A. Kunkle, 2000. Genetic variation in
response of turkeys to experimental infection with Bordetella avium. Avian
Dis. 44:197-200.
2. Sacco, R. E., R. B. Rimler, X. Ye, and K. E. Nestor, 2001. Identification
of new major histocompatibility complex Class II restriction fragment length
polymorphisms in a closed experimental line of Beltsville Small White turkeys.
Poultry Sci. 80:1109-1111.
(32)
ACCESSION NO: 0166758 SUBFILE: CRIS
PROJ
NO: VA-135458 AGENCY:
PROJ
TYPE: HATCH PROJ STATUS: TERMINATED
START:
INVESTIGATOR: Lee, J.
C.
PERFORMING INSTITUTION:
DISEASE, EPIDEMIOLOGIC AND
TOXICOLOGIC INVESTIGATIONS IN
OBJECTIVES: To identify and respond in a timely manner to sudden
disease outbreaks and toxicoses affecting
APPROACH: Funds will be allocated to individual investigators
or interdisciplinary teams for 3 purposes: To investigate disease entities
of unknown etiology and with potential risk of significant economic loss to
agricultural animals in
NON-TECHNICAL SUMMARY: Sudden disease outbreaks and toxicoses impact
PROGRESS: 1995/01 TO 1998/12
This project was designed to provide a mechanism for quick, initial response
to disease and toxicity, particularly in
IMPACT: 1995/01 TO 1998/12
This project allows quick response to conditions of animal and related human
health in the
PUBLICATIONS: 1995/01 TO 1998/12
No publications reported this period
PROJECT CONTACT:
Name: Lee, J. C.
Phone: 540-231-4807
Fax: 540-231-7367
Email: jclee@vt.edu
(33)
ACCESSION NO: 7000416 SUBFILE: ICAR
PROJ
NO: W9601 AGENCY: OTHER
FEDERAL
START:
01 APR 1997 TERM: 31 MAR 2000 FY: 1997
INVESTIGATOR: WRIGHT
P
PERFORMING INSTITUTION:
CANADIAN FOOD INSPECTION AGENCY NATIONAL CENTRE FOR FOREIGN ANIMAL DISEASES
DEVELOPMENT, STANDARDIZATION
AND VALIDATION OF SEROLOGICAL TESTS FOR THE DIAGNOSIS OF FOREIGN ANIMAL DISEASES.
NARRATIVE: IMPACT: Oct 1996 Enhanced capability and confidence
in the detection of: - E/WEE (Eastern/Western equine encephalitis), ND (
OBJECTIVES: Oct 1996 To develop, monitor and improve, on an ongoing
basis, diagnostic reagents and protocols for the diagnosis of foreign animal
disease, and ensure that they meet or exceed the standards for prescribed
tests as set by the OIE Standards Commission.
PROGRESS:
Oct 1996 new project
NOTES: CONTACT: WRIGHT P; PHONE: 204-984-1007; FAX: 204-275-0402
(34)
ACCESSION NO: 7000548 SUBFILE: ICAR
PROJ
NO: NV9401 AGENCY: OTHER FEDERAL
START:
01 APR 1997 TERM: 31 MAR 1998 FY: 1997
INVESTIGATOR: VYDELINGUM
S; TECH
PERFORMING INSTITUTION:
CANADIAN FOOD INSPECTION AGENCY CENTRE OF EXPERTISE FOR PLANT QUARANTINE PESTS
VALIDATION OF PCR METHODS
FOR THE DETECTION OF FAD VIRUSES
NARRATIVE: IMPACT: Oct 1996 Reliable, accurate, sensitive, rapid
and easy to perform FAD tests is the anticipated benefit.
OBJECTIVES: Oct 1996 - Complete the validation of the PCR method
for the detection of Hog cholera, Bluetongue and pseudorabies viruses. - Validate
the PCR method for African swine fever,
PROGRESS:
Oct 1996 new project
Return to:
Contents