Banca de QUALIFICAÇÃO: LAURA RIBEIRO

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : LAURA RIBEIRO
DATA : 16/12/2018
HORA: 00:00
LOCAL: PPGMV / IV / UFRRJ
TÍTULO:

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

PALAVRAS-CHAVES:

trichomoniasis, diagnostic, semen

PÁGINAS: 13
GRANDE ÁREA: Ciências Agrárias
ÁREA: Medicina Veterinária
RESUMO:

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. Their presence in the herd is shown onto 57% of non-pregnant female at the mounting season’s end, being configured by huge financial damages to the farmer. 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 diagnosis is made mainly through clinical signs and reproductive history of animals and property, and is confirmed by laboratory tests. The identification and detection of 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 is the current way to prevent BGT in Brazil. T. foetus has already been found in semen samples and is also able to remain viable frozen, raising artificial insemination to a risk factor when it comes to a fail sanitary or preventive measure. Thus, this study is aimed to detect the presence of T. foetus in samples of commercial bovine semen, through microbiological culture and Polymerase Chain Reaction (PCR). The samples will be purchased from artificial insemination centers and thawed at 37ºC and visualized under a microscope for the analysis of the main sperm caracteristics and a possible microorganism’s presence. The samples will be inoculated in Diamond medium, incubated at 35ºC with consequent growth evaluation 24 hours later by direct examination under a microscope. Samples will be positive when containing protozoan with characteristic and motile prototypes of T. foetus and negative if no growth of the agent is observed after 10 days of incubation. The DNA extraction from the samples will be performed for molecular detection. It will be used a set of primers based on the 5.8S rRNA sequence (TFR3 and TFR4) for the reading, a positive control of the extracted K strain of T. foetus DNA of the urogenital tract of a positive bull (GENBANK AY485677.1) and a negative control containing the same reagents used in the reaction of the samples, however, without inoculation of semen or standard strain. The PCR results will be compared with the data by direct microscopy and culture of each sample and correlated by the Chi Square test method for statistical analysis.

MEMBROS DA BANCA:
Interno - 2202309 - JULIO CESAR FERRAZ JACOB
Interno - 1548043 - MARCO ROBERTO BOURG DE MELLO
Interno - 6387102 - MARILENE DE FARIAS BRITO QUEIROZ
Externo ao Programa - 387340 - CARLOS WILSON GOMES LOPES
Externo ao Programa - 1354903 - DOUGLAS MCINTOSH