RESUMO
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.
Assuntos
Animais , Bovinos , Brucelose Bovina/genética , Técnicas In Vitro , Reação em Cadeia da Polimerase/métodos , Sincronização do Estro/métodos , Vacina contra Brucelose/genética , Amostras de Alimentos , Métodos , Testes SorológicosRESUMO
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.
RESUMO
ABSTRACT Serum samples from 200 vaccinated adult bovine females from two herds were analyzed by buffered antigen acidified plate test (AAT) (Rose Bengal Plate Test), indirect enzyme-linked immunosorbent assay (ELISAI), 2-mercaptoethanol test (2-ME) and complement fixation test (FC). For ELISAI, the fixed value of 45 percent positivity (PP) was used. Group A was composed of 100 animals, with description of reproductive disturbances compatible with brucellosis and reagent to the AAT. Group B was composed of 100 animals serologically free of B. abortus for at least two years. Additionally, all serum samples were tested using the AAT, the 2-ME and the FC to confirm negative status. The combination of two tests, FC and 2ME was used as the gold standard. The relative sensitivity and specificity and the Kappa were calculated for each test. The result of kappa for 2-ME in relation to the FC and of the AAT and the ELISAI in relation to the gold standard was, respectively, 0.78, 0.86 And 0.84. The relative sensitivities (Sr) were, respectively, 84.1% (53/63), 100% (53/53) and 98.1% (52/53), and the relative specificities (Er) were 93.3% (111/119), 90.1% (100/111) and 90.1% (100/111). For comparison between the ELISAI and the AAT, there was obtained a Kappa of 0.91, Sr of 93% (93/100) and Er of 98% (98/100). Conclusions: 1 The option of constituting the gold standard based on at least two tests was the most suitable for this study; 2 The ELISAI resulted in values of Sr and Er similar to the AAT. Therefore, the AAT and the ELISAI are good for screening in regard to the diagnosis of brucellosis.
RESUMO O estudo objetivou comparar o desempenho do teste mercaptoetanol (2-ME) com o teste fixação de complemento (FC) e os testes antígeno acidificado tamponado (AAT) e ELISA indireto (ELISAI) frente ao gold standard, para o sorodiagnóstico da brucelose em bovinos. Também se comparou o ELISAI com o AAT. Foram processadas 200 amostras de soro de fêmeas bovinas adultas de dois rebanhos, denominados grupos A e B. O grupo A foi composto de 100 fêmeas vacinadas contra brucelose, com histórico de problemas reprodutivos compatíveis com a doença e reatoras ao AAT. O grupo B foi composto de 100 fêmeas vacinadas e não reatoras ao AAT por mais de dois anos. Todos os soros foram submetidos aos quatro testes. Foi empregado, como gold standard, a combinação do FC com o 2ME. O índice Kappa foi usado como medida de concordância e seu resultado, para o 2-ME, o AAT e o ELISAI foi de, respectivamente, 0,78, 0,86 e 0,84. As sensibilidades relativas (Sr) foram de, respectivamente, 84,1%, 100% y 98,1% e as especificidades relativas (Er), 93,3%, 90,9% e 90,1%. Entre o ELISAI e o AAT, obteve-se Kappa de 0,91, Sr de 93% e Er de 98%. Conclusões: 1-A opção de se contituir o gold standard baseado em dois testes foi a mais adequada para este estudo; 2-Os valores de Sr e Er do ELISAI foram semelhantes aos do AAT. 3-O AAT e o ELISAI mostraram ser testes muito sensíveis e de boa especificidade e, portanto, adequados para serem usados como triagem no diagnóstico da brucelose.
RESUMO
Mycobacterium was verified in animals from a Brazilian dairy herd, a total of 42 samples from 30 cows were submitted to culture and the isolated strains were analyzed by two polymerase chain reaction (PCR), the first specific for species belonging to the Mycobacterium complex (MTBC) and the other for differentiating M. tuberculosis from M. bovis. Twenty seven samples (64.3%) from 18 animals (60%) were positive for mycobacteria by culture, including samples from 15 retrofaryngeal lymphnodes (55.5%), 9 prescapular lymphnodes (33.3%), 2 lungs (7.4%), and 1 liver (3.7%). All isolated colonies were confirmed by PCR to contain MTBC organisms, and were identified as M. bovis by the same methodology.
Assuntos
Mycobacterium bovis/isolamento & purificação , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Bovina/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Brasil , Bovinos , DNA Bacteriano/análise , Mycobacterium bovis/genética , Mycobacterium tuberculosis/genética , Reação em Cadeia da PolimeraseRESUMO
Mycobacterium was verified in animals from a Brazilian dairy herd, a total of 42 samples from 30 cows were submitted to culture and the isolated strains were analyzed by two polymerase chain reaction (PCR), the first specific for species belonging to the Mycobacterium complex (MTBC) and the other for differentiating M. tuberculosis from M. bovis. Twenty seven samples (64.3 percent) from 18 animals (60 percent) were positive for mycobacteria by culture, including samples from 15 retrofaryngeal lymphnodes (55.5 percent), 9 prescapular lymphnodes (33.3 percent), 2 lungs (7.4 percent), and 1 liver (3.7 percent). All isolated colonies were confirmed by PCR to contain MTBC organisms, and were identified as M. bovis by the same methodology.