Evaluation of the infection and immunity in susceptible and resistant lines of chicken (Gallus gallus) challenged with Salmonella enterica subsp. enterica sorovar Gallinarum

• Grant number: 12/24017-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: March 01, 2013
• End date: February 28, 2017
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Pathology
• Principal Investigator: Angelo Berchieri Junior
• Grantee: Rafael Antonio Casarin Penha Filho
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil
• Associated scholarship(s): 13/26122-5 - 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, BE.EP.PD

Abstract

Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (SG) belongs to serogroup D of Salmonella enterica subsp. enterica and causes fowl typhoid, a disease characterized by high morbidity and mortality in birds. The control of this disease in commercial poultry farms is accomplished through a combination of biosecurity measures including vaccination of birds. SG is a host-specific biovar that is able to infect chickens of all ages. Clinical symptoms are variable between the different commercial lines of chicken. Brown Layer-hens (semi-heavy line) are susceptible to SG infection and develop severe symptoms with high mortality rates at any age. Rather the white layer-hens (light lines) are resistant to infection and show low mortality after infection. This study will evaluate the immune response against SG in two different experiments. The first study will evaluate the innate immune response after infection of layer-hens (light and semi-heavy lines). At 4h, 2 and 6 days post-infection (dpi) birds of the two lines will be sacrificed to harvest the samples and assess the bacterial infection. Additionally, the cellular immune response will be evaluated by detecting ³´ T cells and macrophages by immunohistochemistry. RT-PCR in real time will be used to quantify the expression of receptors responsible for the detection of the bacteria and proinflammatory cytokines.The other experiment will evaluate the acquired immune response triggered by an attenuated strain of SG, with deletion of genes cobS and cbiA (SG”cobS”cbiA), with proven potential as a live vaccine against SG infection (FAPESP Proc. N. 2007/53479-0). This study seeks to identify the important elements involved in protection against challenge by pathogenic strain of SG. For this, the response will be evaluated after immunization with fifteen and 1 day before the infection (dbi) and 1, 3 and ten days after infection (dpi) with SG. The technique of RT-PCR inreal time will be used to quantify the expression of immunomodulatory cytokines and flow cytometry will be used to quantify effector T lymphocytes (CD4+CD44+ and CD8+CD44+). Additionally, the cellular immune response will be evaluated by imunohistochemistry to detect macrophages (Experiment 1) and by flow citometry to quantify effector T lymphocytes (CD4+CD44+ and CD8+CD44+). The levels of the immunoglobulins IgM and IgG will be measured by ELISA. Overall, this study was designed to generate knowledge about the different immune responses in birds vaccinated or naturally resistant or susceptible to infection by SG. (AU)

Scientific publications

(7)

(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, MAY 2016. (12/24017-7, 13/26122-5)

RUBIO, MARCELA DA SILVA; CASARIN PENHA FILHO, RAFAEL ANTONIO; DE ALMEIDA, ADRIANA MARIA; BERCHIERI JUNIOR, ANGELO. Development of a multiplex qPCR in real time for quantification and differential diagnosis of Salmonella Gallinarum and Salmonella Pullorum. AVIAN PATHOLOGY, v. 46, n. 6, p. 644-651, 2017. (14/05496-7, 12/24017-7)

RUBIO, M. S.; PENHA, F. R. A. C.; ALMEIDA, A. M.; BARBOSA, F. O.; BERCHIERI JR, A.. Duplex Real-Time PCR Using Sybr Green I for Quantification and Differential Diagnosis between Salmonella Enteritidis and Salmonella Typhimurium. Brazilian Journal of Poultry Science, v. 21, n. 1, JAN-MAR 2019. (12/24017-7, 14/05496-7)

FERREIRA, JOSEANE CRISTINA; CASARIN PENHA FILHO, RAFAEL ANTONIO; ANDRADE, LEONARDO NEVES; BERCHIERI JUNIOR, ANGELO; COSTA DARINI, ANA LUCIA. Diversity of plasmids harboring bla(CMY-2) in multidrug-resistant Escherichia coli isolated from poultry in Brazil. DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, v. 88, n. 4, p. 361-364, AUG 2017. (14/14494-8, 12/24017-7)

CASARIN PENHA FILHO, RAFAEL ANTONIO; ACELAS DIAZ, SILVIA JULIANA; FERNANDO, FILIPE SANTOS; CHANG, YUNG-FU; ANDREATTI FILHO, RAPHAEL LUCIO; BERCHIERI JUNIOR, ANGELO. Immunomodulatory activity and control of Salmonella Enteritidis colonization in the intestinal tract of chickens by Lactobacillus based probiotic. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, v. 167, n. 1-2, p. 64-69, SEP 15 2015. (13/26127-7, 12/24017-7)

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, AUG 2016. (13/26127-7, 12/24017-7, 13/26122-5)

FERREIRA, JOSEANE CRISTINA; CASARIN PENHA FILHO, RAFAEL ANTONIO; ANDRADE, LEONARDO NEVES; BERCHIERI JUNIOR, ANGELO; COSTA DARINI, ANA LUCIA. Evaluation and characterization of plasmids carrying CTX-M genes in a non-clonal population of multidrug-resistant Enterobacteriaceae isolated from poultry in Brazil. DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, v. 85, n. 4, p. 444-448, AUG 2016. (14/14494-8, 11/08892-2, 12/24017-7)