PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Leal D.C., Madruga C.R., Matos P.F., Souza B.M.P.S. & Franke C.R. 2011. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi. Pesquisa Veterinária Brasileira 31(7):575-578. Departamento de Produção Animal, Escola de Medicina Veterinária, Universidade Federal da Bahia, Av. Adhemar de Barros 500, Ondina, Salvador, BA 40110170, Brazil. E-mail: franke@ufba.br Conventional PCR (PCRTeq) for diagnosing Theileria equi and multiplex PCR (M/PCRTeq-Bc) for diagnosing T. equi and Babesia caballi were comparatively evaluated with nested PCR (N/PCR-Teq) for diagnosing equine piroplasmosis. In DNA sensitivity determinations, in multiple dilutions of equine blood that had tested positive for T. equi, PCR-Teq and N/PCR-Teq detected hemoparasite DNA in the larger dilutions (1:128), but did not differ significantly from the M/PCRTeq-Bc (1:64). In analyses on equine serum tested by ELISA, there was high agreement between this serological test and PCR-Teq (k = 0.780) and moderate agreement with N/PCR-Teq (k = 0.562) and M/PCRTeq-Bc (k = 0.488). PCR-Teq found a higher frequency of T. equi both in extensively and intensively reared horses, but this was not significant in relation to N/PCR-Teq (P>0.05), and both PCRs indicated that there was an endemic situation regarding T. equi in the population of horses of this sample. PCR-Teq was only significantly different from M/PCR-Teq-Bc (P<0.05). PCR-Teq presented high sensitivity and specificity, comparable to N/PCR-Teq, but with the advantage of higher speed in obtaining results and lower costs and risks of laboratory contamination. This accredits PCR-Teq for epidemiological studies and for determinations on affected horses.

• Theileria equi

Abstract in Portuguese:

RESUMO.- Leal D.C., Madruga C.R., Matos P.F., Souza B.M.P.S. & Franke C.R. 2011. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi. Pesquisa Veterinária Brasileira 31(7):575-578. Departamento de Produção Animal, Escola de Medicina Veterinária, Universidade Federal da Bahia, Av. Adhemar de Barros 500, Ondina, Salvador, BA 40110170, Brazil. E-mail: franke@ufba.br Uma PCR convencional (PCRTeq) para diagnóstico de Theileria equi e PCR multiplex (M/PCRTeq-Bc) para diagnóstico T. equi e Babesia caballi foram avaliadas comparativamente a nested PCR (N/PCR-Teq) no diagnóstico de piroplasmose equina. Na determinação da sensibilidade com DNA em diluições múltiplas de sangue de equino positivo para T. equi, as PCR-Teq e N/PCR-Teq detectaram DNA do hemoparasito nas diluições maiores (1:128), mas não diferiu significativamente da M/PCRTeq-Bc (1:64). Na análise com soros de equinos testados por ELISA houve uma concordância elevada entre esse teste sorológico e a PCR-Teq (k =0,780) e moderada com N/PCR-Teq (k = 0.562) e M/PCRTeq-Bc (k = 0.488). A PCR-Teq determinou freqüência maior de T. equi tanto nos equinos de criação extensiva como na intensiva, entretanto essa não foi significativa com relação N/PCR-Teq (P>0.05), e ambas PCRs indicaram que há uma situação endêmica para T. equi na população de equinos que constaram da amostragem. A PCR-Teq diferiu significativamente apenas para a M/PCRTeq-Bc (P<0.05). A PCR-Teq apresentou alta sensibilidade e especificidade comparável a N/PCR-Teq, mas com a vantagem de maior rapidez na obtenção dos resultados e menor custo e risco de contaminação no laboratório. Isso credencia a PCR-Teq para estudos epidemiológicos e para determinação de equinos portadores.

Abstract in English:

ABSTRACT. Riet-Correa F., Haraguchi M., Dantas A.F., Burakovas R.G., Yokosuka A., Mimaki Y., Medeiros R.M.T. & Matos P.F. 2009. Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil. Pesquisa Veterinária Brasileira 29(1):94-98. Hospital Veterinário, Centro de Saúde e Tecnologia Rural, Universidades Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Different species of Panicum, including P. dichotomiflorum, have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to 1H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

• Lithogenic steroidal saponins Panicum dichotomiflorum photosensitization protodioscin

Abstract in Portuguese:

ABSTRACT. Riet-Correa F., Haraguchi M., Dantas A.F., Burakovas R.G., Yokosuka A., Mimaki Y., Medeiros R.M.T. & Matos P.F. 2009. Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil. Pesquisa Veterinária Brasileira 29(1):94-98. Hospital Veterinário, Centro de Saúde e Tecnologia Rural, Universidades Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Different species of Panicum, including P. dichotomiflorum, have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to 1H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.