Source: UNIV OF MARYLAND submitted to NRP
DEVELOPMENT OF NOVEL NEWCASTLE DISEASE VIRUS VACCINES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
ANIMAL HEALTH
Reporting Frequency
Annual
Accession No.
0231302
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Feb 18, 2016
Project End Date
Oct 30, 2019
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIV OF MARYLAND
(N/A)
COLLEGE PARK,MD 20742
Performing Department
Veterinary Medicine
Non Technical Summary
Newcastle disease (ND) causes large economic losses in poultry production worldwide. ND is a highly infectious disease caused by virulent isolates of Newcastle disease virus (vNDV). The disease is enzootic in Africa, Asia, Middle East, and many countries of Central and South America. Although vNDV is not currently found in poultry in the USA, it is a major threat to U.S. poultry industry. The last outbreak of ND in 2002-2003 required the culling of over 3 million chickens in California at a cost of over $280 million. Since vNDV is circulating in many parts of the world and in wild birds, there is a possibility that vNDV could be introduced into U.S. poultry flocks. To prevent introduction of vNDV into U.S. poultry, it is important to understand the biological and genetic characteristics of the vNDV strains circulating in other geographic areas. We plan to address these needs by performing biological and genetic characterization of vNDV strains circulating in different geographic regions of the world. There is a concern regarding the efficacy of currently used vaccines and there is a need for an improved vaccine. In this study, we will investigate whether a recombinant attenuated NDV vaccine generated from a currently circulating vNDV in our laboratory could provide better protection against prevalent vNDV strains. The research proposed here will help reduce the rise of ND in the US and protect the economy of the poultry industry.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3113210110125%
3113210117025%
3113220110125%
3113220117025%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3210 - Egg-type chicken, live animal; 3220 - Meat-type chicken, live animal;

Field Of Science
1101 - Virology; 1170 - Epidemiology;
Goals / Objectives
Poultry production in the U.S. amounts to approximately 18% of the total world poultry production. Viral disease, particularly ND, is a major threat to the U.S. poultry industry. An outbreak of ND can have a devastating effect on the economy of U.S. poultry industry. It is most likely that the next vNDV strain responsible for outbreak in the USA will be from another geographic area or from a wild bird. Therefore, it is important to know the antigenic and genetic makeup of the NDV strains circulating in different parts of the world. This information will not only help us to design better vaccines and control measures but also will improve our understanding on evolution of NDV. It is now well established that the current live attenuated vaccines B1 and LaSota are not effective against vNDV strains circulating in other parts of the world.
Project Methods
Aim 1: Determine the biological and genetic characteristics of vNDV strains circulating in different geographic locations Experimental design: Field samples of NDV collected from poultry and wild birds and live bird markets of the USA and abroad will be processed for virus isolation. Briefly, cloacal and oro-pharyngeal swabs collected will be transported in virus transport medium. The virus isolation will be performed in a USDA certified enhanced BSL-3 laboratory at the University of Maryland. The pathogenicity of the NDV isolates will be determined by the mean death time (MDT) in 9-day-old embryonated SPF eggs and the intracerebral pathogenicity index (ICPI) in 1-day-old SPF chicks. Tissue tropism and pathogenicity studies will be performed in 3-week-old chickens. Consensus primers will be used to determine the complete genome sequence of an NDV isolate. The sequence information will be used for phylogenetic analysis. Aim

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:The target audiences are scientists working on poultry diseases, poultry farmers, vaccine manufacturers, veterinarians, infectious diseases specialists/scientists and students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training of undergraduate students, graduate students, postdoctoral fellows and visiting scientists. How have the results been disseminated to communities of interest?The research outcomes were presented in conferences and scientific seminars. What do you plan to do during the next reporting period to accomplish the goals?We are planning to develop better Newcastle disease virus vaccine by codon optimization.

Impacts
What was accomplished under these goals? Newcastle disease (ND) causes severe economic loss to poultry industry worldwide. Frequent outbreaks of ND in commercial chickens vaccinated with live vaccines suggest a need to develop improved vaccines that are genetically matched against circulating Newcastle disease virus (NDV) strains. In this study, the fusion protein cleavage site (FPCS) sequence of NDV strain Banjarmasin/010 (Banj), a genotype VII NDV, was individually modified using primer mutagenesis to those of avian paramyxovirus (APMV) serotypes 2, 7 and 8 and compared with the recombinant Banjarmasin (rBanj) with avirulent NDV LaSota cleavage site (rBanj-LaSota). These FPCS mutations changed thein vitrocell-to-cell fusion activity and made rBanj FPCS mutant viruses highly attenuated in chickens. When chickens immunized with the rBanj FPCS mutant viruses and challenged with the virulent Banj, there was reduced challenge virus shedding observed compared to chickens immunized with the heterologous vaccine strain LaSota. Among the genotype VII NDV Banj vaccine candidates, rBanj-LaSota and rBanj containing FPCS of APMV-8 induced highest neutralizing antibody titers and protected chickens with reduced challenge virus shedding. These results show the effect of the F protein cleavage site sequence in generating genotype VII matched NDV vaccines.

Publications


    Progress 10/01/16 to 09/30/17

    Outputs
    Target Audience:Researchers working on poultry diseases and scientists working on paramyxoviruses. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This has provided training for graduate students and post-doctoral associates how to sequence NDV genome and compare with other published NDV sequence. How have the results been disseminated to communities of interest?A manuscript describing the NDV strain isolated in Africa is in preparation, which will be submitted for publication soon. What do you plan to do during the next reporting period to accomplish the goals?Construct two genetically engineered live-attenuated vaccines based on sequences of Newcastle disease viruses circulating in different parts of the world and evaluate the effectiveness of these vaccines in chickens.

    Impacts
    What was accomplished under these goals? The complete genome sequence of NDV strains AKO was determined. The nucleotide sequence showed high degree of divergence from the vaccine strain LaSota. The sequence determination of other NDV strains are in progress.

    Publications


      Progress 02/18/16 to 09/30/16

      Outputs
      Target Audience:Researchers working on poultry diseases and scientists working on paramyxoviruses. Changes/Problems:Under the supervision by PI, Dr. Siba Samal, one graduate student and a Postdoc (0.5 FTE) are working with him in this project. What opportunities for training and professional development has the project provided?This has provided training for graduate students and post-doctoral associates to work in BSL-3 facility, how to grow NDV, how to extract viral RNA and how to test infectivity of viral RNA. How have the results been disseminated to communities of interest?The research is in progress. It will be published after it is completed. What do you plan to do during the next reporting period to accomplish the goals?Complete the entire genome sequence of few NDV strains isolated in Africa, compare the genome sequences with those of vaccine strains LaSota, construct live-attenuated vaccines using the strains circulating in Africa.

      Impacts
      What was accomplished under these goals? Five different NDV strains isolated in Africa were grown in our BSL-3 laboratory. These virus stocks were aliquoted. The viral RNAs from these viruses were extracted and tested for infectivity. After testing, these RNAs were removed from BSL-3 lab to BSL-2 lab for sequencing.

      Publications