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Identification of homologous protein antigen in Salmonella enterica subsp. enterica serovars Typhimurium, Enteritidis, Heidelberg and Schwarzengrund for production of recombinant protein and evaluation as a vaccine candidate in chickens

Grant number: 13/26122-5
Support Opportunities:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): June 10, 2014
Effective date (End): June 09, 2015
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Pathology
Principal Investigator:Angelo Berchieri Junior
Grantee:Rafael Antonio Casarin Penha Filho
Supervisor: Yung-Fu Chang
Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil
Research place: Cornell University, United States  
Associated to the scholarship:12/24017-7 - Evaluation of the infection and immunity in susceptible and resistant lines of chicken (Gallus gallus) challenged with Salmonella enterica subsp. enterica sorovar Gallinarum, BP.PD


Salmonellosis are diseases frequently ocurring in commercial flocks of chickens with failures in the biosafety program. During the last few years, vaccination has been applied in commercial poultry flocks throughout Brazil in efforts to control Salmonella spp. infections. Live vaccines against S. Gallinarum, S. Typhimurium and S. Enteritidis in poultry are available. However, live or killed vaccines against non-typhoidal salmonellae (serovars with flagella; e.g. S. Enteritidis) generates a partial protection in chickens, mainly for controlling the intestinal colonization and fecal excretion by these motile serovars. Emerging Salmonella enterica serovars (e.g. S. Heidelberg and S. Schwarzengrund) have demonstrated increasing prevalence in commercial poultry flocks worldwide, including in Brazil. As a consequence, the contamination of poultry products (e.g. eggs and meat) and the risk of infection of humans by these non-typhoidal salmonellae serovars also increased. Thus, it is important to develop new methods and generate tools to control new challenges, in an effective and fast manner. The use of recombinant proteins in vaccine formulations has been studied for controlling various pathogens. However, the use of this approach to produce immunogenic antigens, capable to stimulate cross-protection against different Salmonella serovars, is a method poorly developed for use in poultry.In the proposed project, bioinformatics tools will be used to compare the amino acid sequences of different immunogenic proteins of S. Typhimurium, which are also present in serovars S. Enteritidis, S. Heidelberg and S. Schwarzengrund. The protein with the highest homology among these four serovars will be chosen for cloning and expression in E. coli for the production as a recombinant protein.After producing and purifying, the recombinant protein will be evaluated as a potential vaccine candidate in commercial chickens to protect against an experimental challenge with S. Typhimurium. The birds will be divided into two groups A and B. In group A, the birds will not be vaccinated. In group B , the birds will be vaccinated with 100 µg of recombinant protein, eluted in adjuvant (EMULSIGEN) at 5 and 25 days of age. All birds will be challenged with S. Typhimurium at 30 days of age. At 2 and 5 days post-challenge, 3 birds from each group will be euthanized to analyse the infection by the challenge strain, evaluating the bacterial numbers in spleen, liver and cecal contents. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
CASARIN PENHA FILHO, RAFAEL ANTONIO; FERREIRA, JOSEANE CRISTINA; IBA KANASHIRO, ANA MARIA; DA COSTA DARINI, ANA LUCIA; BERCHIERI JUNIOR, ANGELO. Suscetibilidade a antimicrobianos de Salmonella Gallinarum e Salmonella Pullorum isolados de aves doentes no Brasil. Ciência Rural, v. 46, n. 3, p. 513-518, . (12/24017-7, 13/26122-5)
CASARIN PENHA FILHO, RAFAEL ANTONIO; ACELAS DIAZ, SILVIA JULIANA; MEDINA, TIAGO DA SILVA; CHANG, YUNG-FU; DA SILVA, JOAO SANTANA; BERCHIERI, JR., ANGELO. Evaluation of protective immune response against fowl typhoid in chickens vaccinated with the attenuated strain Salmonella Gallinarum Delta cobS Delta cbiA. Research in Veterinary Science, v. 107, p. 220-227, . (13/26127-7, 12/24017-7, 13/26122-5)

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