Periódicos Brasileiros em Medicina Veterinária e Zootecnia

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Identification of Brucella sp. isolated in Brazil from 1976 to 2013 by Bruce-Ladder PCR

Souza Lopes, EsterSilveira Luiz Machado, VerônicaTonial Simões, CristinaGarin-Bastuji, BrunoCosta, Marisa

Background: Brucella sp. are the causative agents of brucellosis, an infectious disease that affects various species of animals and can be transmitted to humans through direct contact with infected animals, indirectly by the ingestion of rawmilk products, and during the handling of strains or infected material in the laboratory. Being a zoonosis, the detection of Brucella species in animals is essential for the prevention of the disease in humans and to perform a good program of control in infected herds. This study aimed at identifying Brucella field strains isolated from 1976 to 2013 in Brazil, using the modified Bruce-Ladder method, to evaluate the performance of this technique.Materials, Methods & Results: Eighty-three strains of Brucella sp. were included in the study, i.e. 21 reference strains (nine B. abortus, one B. canis, four B. melitensis, two B. ovis and five B. suis) and 62 field strains (six B. canis, one B. suis and 55 B. abortus). For the identification of the genus and/or species of Brucella, biochemical and physiological tests, including MacConkey-agar growth, glucose fermentation, haemolysis, catalase, oxidase and urease tests, nitrate reduction, citrate utilization, H2 S production and CO2 requirement, were performed. Genomic DNA was extracted from pure cultures through heat-lysis of bacterial cultures and the genus was confirmed by a genus-specific PCR

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