Use of quantitative Real-Time PCR for differential diagnosis of Salmonella enterica subsp. enterica serovar Enteritidis, Typhimurium and Gallinarum (biovars Gallinarum and Pullorum) in domestic poultry (Gallus Gallus)

• Grant number: 14/05496-7
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: July 01, 2014
• End date: December 31, 2017
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Pathology
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Angelo Berchieri Junior
• Grantee: Marcela da Silva Rubio
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The objective of this research is the application of the technique of real-time PCR for diagnosis and indirect quantification of the bacterial load in organs and faeces of experimentally infected commercial poultry by Salmonella enterica subsp. enterica serovar Enteritidis (SE), TYPHI (STM) and Gallinarum (biovars Gallinarum and Pullorum), at different times post-infection. 2 experiments, in vitro and in vivo, will be conducted. The in vitro analyzes will be performed for differential diagnosis and specific detection of each bacterial strain (SE, STM, SG and SP), using 3 different mixes for each bacterium. The first treatment consist of a mixture of the four bacterial strains as the test sample. Then, a mixture of 3 bacterial strains shall be used, excluding the bacterium of interest, as negative control sample and in the third reaction, as positive control, will be used only bacterial strain of interest. For the in vivo experiment, will be used 36 laying birds of white commercial line, divided into 6 groups (n=6). In group A, the birds are not infected and will be kept as a negative control. The birds in groups B, C, D and E, will be infected with SE, STM, SG and SP, respectively. In group F, the birds will be infected with an inoculum containing the four bacterial strains. The inoculations are administered orally, on the first day of bird life. At 24h and 72h after challenge, 3 birds from each group will be euthanized for harvest of spleen and faeces, for the extraction of bacterial DNA and bacterial quantification by means of the technique of real-time PCR. The results will be compared by Tukey test at 5% probability. (AU)

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)

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)

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)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

RUBIO, Marcela da Silva. Use of quantitative real-time PCR for differential diagnosis between Salmonella enterica subesp. enterica serovars Enteritidis, Typhimurium and Gallinarum (biovars Gallinarum and Pullorum) in domestic birds (Gallus gallus). 2017. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Faculdade de Ciências Agrárias e Veterinárias. Jaboticabal Jaboticabal.