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A brucelose e uma doenca infecto-contagiosa, de carater zoonotico e de grande importância tanto em producao animal, quanto na saude publica. E causada por bacterias do genero Brucella , que provocam orquites e epididimites uni ou bilaterais em touros, podendo levar a sub-fertilidade e a esterilidade. Machos bovinos assintomaticos tornam-se importantes disseminadores desta doenca nos rebanhos. Desta forma objetivou-se avaliar o protocolo de extracao de DNA de Brucella spp em semen in natura bovino, submetendo-os aos testes diagnosticos PCR (protocolo Lise Enzimatica por Proteinase e Fenol: Cloroformio – PK), Antigeno Acidificado Tamponado (AAT) e Semem Plasma Aglutinacao (SPA), de 33 touros utilizados em monta natural. O protocolo PK foi eficaz na extracao de DNA de Brucella spp. Dos 33 touros testados, oito foram reagentes na PCR (24,24%), seis (18,18%) reagentes na SPA, e dos 23 animais com amostra de soro sanguineo, um (4%) foi reagente para o AAT. Apenas um animal foi reagente na SPA e no AAT, e nao reagente na PCR. Por outro lado sete animais apresentaram-se com semem reagente na PCR, e negativos nos testes de SPA e AAT. Portanto os resultados indicam a necessidade de associacao de testes laboratoriais diretos, como a PCR, com os testes indiretos AAT e SPA, para um diagnostico mais adequado de brucelose, quando forem utilizadas amostras de semen bovino. SUMMARY Brucellosis is an infectious disease of zoonotic character and great importance in animal production, as well as a public health matter. It is caused by bacteria of the genus Brucella , which cause unilateral or bilateral epididymitis and orchitis in bulls and can lead to sub-fertility and sterility. Asymptomatic bulls become important disseminators of the disease in herds. Therefore, this study evaluates the extraction protocol of DNA of Brucella spp. from bovine semen in natura . The semen samples of 33 bulls used for natural mating underwent PCR diagnostic tests (Lise Enzyme protocol by Proteinase and Phenol:Chloroform-PK), Buffered Acidified Antigen (AAT) and Semen plasma agglutination (SPA). The PK protocol was effective in the extraction of Brucella spp. Of the 33 bulls tested, eight were positive to PCR (24.24%), six (18.18%) to the SPA reagent, and from the 23 animals with blood serum sample, one (4%) was positive for AAT. Only one animal was positive in SPA and AAT, and not reactive in PCR. On the other hand, seven animals had semen reactive in PCR, and negative tests for SPA and AAT. Therefore, the results indicate the need for association of direct laboratory tests, such as PCR, with indirect tests AAT and SPA, for a more accurate diagnosis of brucellosis, when using samples of bovine semen.

AVALIAÇÃO DE PROTOCOLO DE EXTRAÇÃO DE DNA DE BRUCELLA SPP. EM SÊMEN BOVINO / EVALUATION PROTOCOL FOR DNA EXTRACTION OF Brucella spp. IN BOVINE SEMEN