Banca de DEFESA: LAURA RIBEIRO

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : LAURA RIBEIRO
DATE: 15/12/2020
TIME: 14:00
LOCAL: meet.google.com/boa-eupu-shs
TITLE:

Tritrichomonas foetus Detection in Cryopreserverd Bovine Semen by Isolation and Polymerase Chain Reaction

KEY WORDS:

Trichomoniasis, molecular biology, frozen semen

PAGES: 25
BIG AREA: Ciências Agrárias
AREA: Medicina Veterinária
SUBÁREA: Reprodução Animal
SPECIALTY: Fisiopatologia da Reprodução Animal
SUMMARY:

The Bovine Genital Trichomoniasis (BGT) is an infectious disease triggered by a protozoan called Tritrichomonas foetus, which is transmitted by venereal route and also by contaminated fomites. The disease is economically relevant, since its main consequence is abortion, besides producing temporary infertility. In males the disease is asymptomatic and permanent, characterizing the bulls as silent nurses and emphasizing the direction of diagnosis in this category of animals. The identification and detection of the protozoan by microbiological culture are considered as "gold standard" among diagnostic techniques in Brazilian artificial insemination centers. However, the possibility of a false negative is a reality and is needed more sensitive and specific techniques of diagnose, since the Artificial Insemination (AI) is the current way to prevent BGT. T. foetus has already been found in semen samples and is also able to remain viable when frozen, raising AI to a risk factor when sanitary measures are not applied or the diagnostic method is insufficient Thus, this study is aimed to detect the presence of T. foetus in samples of cryopreserved bovine semen, through culture and Polymerase Chain Reaction (PCR). The semen straws will be purchased from artificial insemination centers. A total of 27 samples were analyzed. Once in the laboratory, they were thawed in water at 37ºC, part was inoculated in a test tube containing Diamond medium, incubated at 35ºC with consequent growth evaluation and evaluated every 24 hours via direct microscope examination and the other part diluted in PBS for subsequent molecular analysis. After 10 days of cultivation, all samples were negative. For molecular detection, the Quick-DNA Miniprep Kit (Zymo Research) without proteinase K was used for DNA extraction. The primers used in PCR were: TRF1 and TRF2 (first set) and TRF3 and TRF4 (second set). The PCR result was negative for all samples, corroborating with that obtained in the culture. It was concluded that the samples used were really negative for the presence of T. foetus in both diagnostic methods, proving the innocuousness of the commercial semen tested

BANKING MEMBERS:
Presidente - 1201559 - VERA LUCIA TEIXEIRA DE JESUS
Interno - 1548043 - MARCO ROBERTO BOURG DE MELLO
Interna - 6387102 - MARILENE DE FARIAS BRITO QUEIROZ
Externa à Instituição - MÁRCIA CRISTINA MENDES
Externo à Instituição - JOAQUIM ESQUERDO FERREIRA - OUTRO