This paper aimed to determine the excretion period of B19 vaccine strain during a complete reproductive cycle (from estrus synchronization, artificial insemination, pregnancy and until 30 days after parturition) of dairy cows from 3 to 9 years old that were previously vaccinated from 3 to 8 months. Three groups were monitored with monthly milk and urine collection during 12 months: G1 with seven cows from 3 to 4 years old; G2 with three cows from 5 to 6 years old; and G3 with four cows from 7 to 9 years old. Urine and milk samples were submitted to bacteriological culture and urine and PCR reactions for detection of Brucella spp. and PCR-multiplex for B19 strain identification. Ring test (RT) was also performed in the milk samples, and serum samples were tested by buffered acidified plate antigen test (BAPA). All animals were serologically negative at BAPA and Brucella spp. was not isolated from both urine and milk samples. RT revealed 13/210 (6.2%) positive milk samples. PCR reactions detected DNA of Brucella spp. in 86/420 (20.5%) samples. In urine it was found a significantly higher frequency (35.2%; 74/210) than in milk (5.7%; 12/210), more frequently from the estrus to 150 days of pregnancy and after parturition (6.7%; 10/150), and from 150 days of pregnancy to parturition (3.4%; 2/60), and they were all identified as B19 strain. In three groups, intermittent excretion of B19 strain was detected mainly in urine samples, which confirmed its multiplication and persistence in cows for until 9 years.

A polymerase chain reaction (PCR) assay to detect Clostridium perfringens alpha toxin gene (cpa) was used to identify eighty-nine C. perfringens strains obtained from bovine clinical material. The strains were biochemically characterized as C. perfringens. The isolated strains were cultured on plates containing brain heart infusion agar with 5% sheep blood under anaerobic conditions. DNA extraction was performed by boiling. The 324 bp amplification product of cpa was observed in all isolates. C. sordellii, C. botulinum, C. novyi, and C. septicum were also tested but did not produce any alpha toxin gene amplification. These findings suggest that PCR is a useful assay in identifying C. perfringens toxin types.