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Molecular characterization of Cryptosporidium spp. in rabbits domiciled in the cities of Araçatuba (SP) and São José do Rio Preto (SP)

Grant number: 19/06274-1
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): August 01, 2019
Effective date (End): July 31, 2020
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Katia Denise Saraiva Bresciani
Grantee:Bruno Criado de Araújo Mendes
Home Institution: Faculdade de Medicina Veterinária (FMVA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil

Abstract

This research project will be carried out with the aim of characterizing molecularly the species of Cryptosporidium spp. in rabbits kept in pets. The faeces will be collected from 25 animals from Araçatuba and 40 from São José do Rio Preto, totaling 65 animals. For the detection of Cryptosporidium spp. Oocysts, the malachite green negative staining technique will be performed. Afterwards, all fecal samples will be subjected to DNA extraction from this coccidia with subsequent molecular characterization by the polymerase chain reaction (PCR) to amplify 18S rRNA subunit fragments and sequencing the amplified fragments. In case of positivity for Cryptosporidium parvum, Cryptosporidium hominis or Cryptosporidium cuniculus, subtyping will be carried out by means of PCR (gp60 gene) and sequencing of the amplified fragments. The Kappa correlation coefficient will be calculated to analyze the concordance index between the results obtained in the diagnostic methods employed, and the McNemar Test to compare the proportion of positive results of the performed techniques.