Banca de QUALIFICAÇÃO: ANA PAULA MARTINEZ DE ABREU

Uma banca de QUALIFICAÇÃO de DOUTORADO foi cadastrada pelo programa.
DISCENTE : ANA PAULA MARTINEZ DE ABREU
DATA : 25/09/2019
HORA: 09:00
LOCAL: Sala 03
TÍTULO:

Molecular characterization of Trypanosoma sp from cattle from the State of Rio de Janeiro based on 18S and cathepsin L-like gene

PALAVRAS-CHAVES:

Trypanosoma vivax; cattle; molecular diagnosis;

PÁGINAS: 52
GRANDE ÁREA: Ciências Agrárias
ÁREA: Medicina Veterinária
RESUMO:

The objective of this work was to characterize the Trypanosoma vivax and Trypanosoma theileri through 18S and CatL-like genes from naturally infected cattle and to evaluate the distribution of these agents in the state of Rio de Janeiro, to analyze the phylogeny and virulence of T. vivax and T. theileri in the state of Rio de Janeiro comparing the positivity of these agents to biochemical changes in whole blood of infected and uninfected cattle. To do this, during the period from 2016 to 2018, in 15 municipalities in the state of Rio de Janeiro, whole blood samples were collected from 391 bovines through venipuncture with a volume of 3 ml, which were packed in 10% EDTA tubes to hematocrit analysis, hemoparasite screening and conventional PCR by 18S genes for identification of Trypanosoma sp, CatL-like (Cathepsin L-like) for T. theileri identification and PRAC (Proline Recemase) for T. vivax, from the samples of DNA extracted from bovine blood. After analysis of prevalence, some samples from each municipality were selected for sequencing from the 18S and CatL-like genes PCR products. In addition, 5 ml of whole blood were collected and collected in tubes without anticoagulant for biochemical analysis of the following analytes: urea, creatinine, PTS and fractions, ALT, AST, GGT, alkaline phosphatase, LDH and bilirubin. 4.6% (18/391) of the samples were positive for Trypanosoma vivax in blood smear, and no positive samples were seen for T. theileri. The same samples were tested for the 18S gene, where 15.3% (60/391) were positive for Trypanosoma sp, significantly more than detected in hemoparasite screening.
The conventional PCR technique was more specific than the blood smear, due to the false negative results obtained by optical microscopy, which has low sensitivity when compared to conventional PCR. The prevalence of T. vivax in cattle in the municipalities of Piraí (25%), Areal (15.7%), Santo Antônio de Pádua (18.4%), Barra do Piraí (55.5%), Rio Claro %), Cantagalo (25%) and Trajano de Moraes (50%) were above average. All analyzed sequences of the 18S rDNA and CatL-like genes for T. vivax, respectively, showed 99-100% identity with T.
vivax sequences reported in other countries. The same occurred for the sequences analyzed for T. theileri of the 18S and CatL-like genes for T. theileri. Phylogenetic analysis showed xxxxx clusters defined for the 18S rDNA gene and xxx defined clusters for the CatL-like gene. Based on the 18S gene, the deduced amino acid sequence demonstrated xxx non-synonymous mutation points at positions xx and xxx compared to reference sequence xxxxx. The present study in addition to reporting the first evidence of T. vivax in the state of Rio de Janeiro that
contributes to the tracing of T. vivax and T. theileri in the municipalities, as well as in the correlation of the biochemical alterations with the infection and not infection by these agents.

MEMBROS DA BANCA:
Externo ao Programa - 3103478 - MARISTELA PECKLE PEIXOTO
Externo ao Programa - 3103528 - CLAUDIA BEZERRA DA SILVA
Externo à Instituição - MARCUS SANDES PIRES - UFF
Externo à Instituição - DANIEL DA SILVA GUEDES JUNIOR - FIOCRUZ