Banca de DEFESA: ERICA RODRIGUES DE MATOS SILVA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : ERICA RODRIGUES DE MATOS SILVA
DATE: 28/07/2023
TIME: 14:00
LOCAL: ONLINE
TITLE:

DNA detection of Sarcocystis spp. and Neospora caninum in reproductive tissues and blood of wild domestic cats in Ilha Furtada, Mangaratiba, State of Rio de Janeiro

KEY WORDS:

Cats, Histopathology, Polymerase Chain Reaction (PCR), Neosporosis, Sarcocystosis

PAGES: 70
BIG AREA: Ciências Agrárias
AREA: Medicina Veterinária
SUBÁREA: Patologia Animal
SPECIALTY: Anatomia Patologia Animal
SUMMARY:

Sarcocystis spp. and Neospora caninum are obligate intracellular protozoa belonging to the family Sarcocystidae. They have a heteroxene cycle, requiring two hosts to complete the cycle. The sexual phase occurs in the definitive host and the asexual phase in the intermediate host, thus infecting a wide variety of animal species. Both agents cause tissue cysts in the musculature of the intermediate host. Sarcocystis spp. is the etiologic agent of intestinal sarcocystosis, a zoonosis of great economic and public health importance. Neospora caninum causes neosporosis, a disease involved in serious economic losses in Brazilian and world livestock, due to reproductive losses. The epidemiology of these coccidia in cats is poorly understood and there is a lack of studies in the literature. Thus, the main objective of this research is to identify the protozoa Sarcocystis spp. and Neospora caninum in feral cats. The samples were provided by the Pathological Anatomy Sector (SAP) of the Federal Rural University of Rio de Janeiro (UFRRJ) where all the processing and histopathological diagnosis will be carried out. Whole blood and reproductive tissues are from neutered cats from Ilha Furtada, popularly known as Ilha dos Gatos, located in the municipality of Mangaratiba, Costa Verde do Rio de Janeiro. A total of 54 tissue samples will be analyzed, such as: testes, uterus, ovaries, placenta and fetuses previously fixed in 10% formalin for histopathological analysis. Relevant information such as sex, race and life stages of the animals were also provided for data collection. The slides will be processed and stained with Hematoxylin and Eosin (HE) dyes, then they will be analyzed by optical microscopy. In cases where at least one visualized cyst is morphologically compatible with the studied protozoa, the tissues will be considered histologically positive. Whole blood and tissue fragments destined for molecular diagnosis will be kept in Eppendorf-type microtubes and frozen until the samples are processed in the laboratory. Molecular analysis by Polymerase Chain Reaction (PCR) will use Qiagen's DNeasy® kit for DNA extraction following the manufacturer's guidelines. The extracted products will be amplified in the Veretti Thermal Cyclicler (AB Applied Biosystems). For electrophoresis, agarose gel with a concentration of 0.8% (0.8 g of agarose in 100 ml of TAE buffer) will be used. For detection of the B1 gene, conventional PCR will be used with the following primers: 5'-GGA ACT GCA TCC GTT CAT GAG-3' and 5'-TCT TTA AAG CGT. For reading and capturing Nested-PCR images, a software from Carestream Molecular Imaging (Carestream Health), version 5.0 will be used. This is a pioneering study to use reproductive tissues for the diagnosis of Sarcocystis and Neospora caninum and the first to be carried out with cats from Furtada Island. It is important to highlight possible sources of infection and the participation of these animals in the coccidia cycle and a possible potential for transmission to humans and other animal species. Thus, it is expected to contribute to further studies on the subject to be carried out in the future and serve as information for pathologists and veterinarians as a differential diagnosis for other parasitic diseases.

COMMITTEE MEMBERS:
Presidente - 2932766 - ANDRESSA FERREIRA DA SILVA SPYRIDES
Interno - 1151827 - BRUNO RICARDO SOARES ALBERIGI DA SILVA
Interno - 2318733 - DANIEL DE ALMEIDA BALTHAZAR
Interno - 3701718 - DANIEL GUIMARAES UBIALI
Externo à Instituição - GIDEÃO DA SILVA GALVÃO