Identification of gender and species of Cryptosporidium in dogs and cats

Author(s)

Camila Guariz Homem

• Document type: Doctoral Thesis
• Press: Araçatuba. 2016-12-02.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Medicina Veterinária. Araçatuba
• Defense date: 2016-10-13
• Advisor: Marcelo Vasconcelos Meireles

Abstract

This study aimed the identification and molecular characterization of Cryptosporidium isolates in stool samples from dogs and cats, as well as the development of a polymerase chain reaction (PCR) in real time to specific detection of Cryptosporidium canis in fecal samples from dogs. Three hundred fecal samples from cats and 367 fecal samples from dogs were collected in the cities of Araçatuba and Jaboticabal (SP) and subjected to the processes of purification and DNA extraction. Nested PCR (nPCR) for the 18S rRNA gene was performed for identification of Cryptosporidium spp., followed by sequencing of the amplified fragments. A real time PCR to a partial fragment of the HSP70 gene was standardized for detection of Cryptosporidium canis in dogs fecal samples. The positivity for Cryptosporidium spp. by nPCR in cats samples was 11.33% (34/300), and in dog samples was 10.4% (38/367). The real-time PCR resulted in 15.3% (58/367) of samples positive for C. canis. The sequencing of the amplified fragments allowed the identification of three samples positive for Cryptosporidium felis and six samples positive for C. canis. A higher positivity was observed in puppies and kittens when compared to adult animals. The analytical sensitivity of real-time PCR was 1 copy of DNA of C. canis and was not observed DNA amplification for other species of Cryptosporidium.We concluded that dogs and catsn of the investigated regions can present infected for zoonotic species of Cryptosporidium and the real-time PCR developed in this work is a sensitive and specific method for detection of C. canis in fecal samples from dogs. (AU)

• FAPESP's process: 11/09614-6 - Diagnosis of cryptosporidiosis in dogs and cats using the real-time polymerase chain reaction
• Grantee: Camila Guariz Homem
• Support Opportunities: Scholarships in Brazil - Doctorate