Project Number: 6040-32000-071-07-T
Project Type: Trust Fund Cooperative Agreement
Start Date: Jun 1, 2018
End Date: May 31, 2020
This proposal addresses the Foundation's Specific Research Priority "Identify and validate interventions to reduce pathogen contamination of poultry parts." Our overall project goals are to identify the Salmonella protein antigens that are able to induce humoral immune response in broilers, and consequently these antibodies can prevent Salmonella colonization in the broiler gastrointestinal tracts (GI). The specific objectives of this proposal are to: (1) Purify recombinant Salmonella proteins from recombinant E. coli cell lysates. (2) To assess the immune response and protection of broilers after immunization with these recombinant Salmonella proteins. (3) To evaluate the GI microbiota of broilers following immunization with these recombinant Salmonella proteins.
Due to advances in molecular biology technologies, we have applied these to perform bioinformatic searches for the virulence factor genes that are conservative among Salmonella serotypes important to the U.S. poultry, and have cloned and expressed a battery of these Salmonella virulence genes in the E. coli expression system. Next, our plans are to generate and purify these recombinant proteins by affinity chromatography and size exclusion chromatography. These recombinant proteins will be emulsified in a Montanide adjuvant, and the mixtures will be administered subcutaneously in broilers. The schedules for immunization and Salmonella challenges are very crucial and will be determined experimentally. To determine the efficacies of these recombinant proteins in inducing antibody production and protecting colonization, the Salmonella counts in the broiler cecal contents will be carried out. Immunoblot assays will also be performed to determine the specificity of the antibody reactivity. Because of the importance of the microflora in the gut health, the bacterial population in cecae will be examined to determine the effect of the vaccines on gut mircobiome.