PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Silva K.P.C., Takaki G.M.C., Teles J.A.A., Dantas A.F.M., Costa M.M., Felix W.P. & Mota R.A. 2014. [Partially purified malleo-proteins production for glanders diagnosis in Equidae.] Produção e purificação parcial de PPD-maleína para diagnóstico do mormo em equídeos. Pesquisa Veterinária Brasileira 34(1):57-61. Laboratório de Doenças Infecciosas dos Animais, Faculdade de Veterinária, Unidade Educacional Viçosa, Campus Arapiraca, Universidade Federal de Alagoas, Fazenda São Luiz s/n, Zona Rural de Viçosa, Viçosa, AL 57700-000, Brazil. E-mail: karla.ufal@gmail.com The objective of this study was to produce and partially purify Malleo-protein from Burkholderia mallei samples isolates from Equidae in Brazil with potential for use in the diagnosis of glanders. The strain B. mallei phenotypically characterized and proven virulent was inoculated into broth Dorset-Henley to grow and metabolize. The proteins were separated by trichloroacetic acid precipitation and amonium sulfate precipitation. The PPD mallein was concentrated 1.0mg/mL and biologically tested in guinea pigs. It was effective in the development of delayed-type hypersensitivity and consequently to identify true-positive animals and to exclude of true negatives. There is the possibility for potential use in the glanders diagnosis in Equidae.

• Mallein test glanders Burkholderia mallei maleinização mormo

Abstract in Portuguese:

RESUMO.- Silva K.P.C., Takaki G.M.C., Teles J.A.A., Dantas A.F.M., Costa M.M., Felix W.P. & Mota R.A. 2014. [Partially purified malleo-proteins production for glanders diagnosis in Equidae.] Produção e purificação parcial de PPD-maleína para diagnóstico do mormo em equídeos. Pesquisa Veterinária Brasileira 34(1):57-61. Laboratório de Doenças Infecciosas dos Animais, Faculdade de Veterinária, Unidade Educacional Viçosa, Campus Arapiraca, Universidade Federal de Alagoas, Fazenda São Luiz s/n, Zona Rural de Viçosa, Viçosa, AL 57700-000, Brazil. E-mail: karla.ufal@gmail.com Objetivou-se com este estudo produzir e purificar parcialmente a PPD-maleína a partir de amostras de Burkholderia mallei isoladas de equídeos no Brasil com potencial para uso no diagnóstico do mormo. As linhagens de B. mallei fenotipicamente caracterizadas e de virulência comprovada foram inoculadas em caldo Dorset-Henley para crescer e metabolizar. Em seguida, as proteínas foram separadas por precipitação com ácido tricloroacético e precipitadas com sulfato de amônia. As PPDs-maleínas foram concentradas em 1,0mg/mL e na avaliação realizada em cobaios foi eficaz no desenvolvimento da hipersensibilidade do tipo tardia e consequentemente na identificação de animais verdadeiro positivos e exclusão dos verdadeiro negativos, sendo uma possibilidade em potencial para utilização no diagnóstico do mormo.

Abstract in English:

ABSTRACT.- Teles J.A.A., Campos A.C., Silva K.P.C., Santos A.S., Santana V.L.A., Castro R.S. & Mota R.A. 2012. [Standardization and evaluation of an indirect ELISA for the serological diagnosis of glanders in horses.] Desenvolvimento e avaliação de um teste ELISA indireto para o diagnóstico sorológico do mormo em equídeos. Pesquisa Veterinária Brasileira 32(9):838-842. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Av. Dom Manuel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com Glanders is an infectious-contagious disease of acute or chronic character which principally affects horses, causing enormous losses in the productive chain of this animal. To control the disease, the Ministry of Agriculture, Husbandry and Supply instituted mandatory sanitation measures in the entire national territory which include an official diagnosis through the complement fixation (CF) test, maleinization and sacrifice of the animals that are positive. Nowadays the kits used for the diagnosis of the disease are imported, making their routine application difficult and more expensive. The objective of this study was to standardize an indirect ELISA test, using the proteic extract of Burkholderia mallei isolated from a carrier horse in the state of Pernambuco. The samples were cultivated in 10% blood agar and incubated for 48h at 37°C; later, one of the isolated colonies was characterized phenotypically and genotypically and immediately cultivated in brain heart infusion (BHI) for enrichment; then it was peaked (repicada) for the Dor-set Henley medium which was incubated at 37ºC under 60rpm for eight weeks. To standardize the test the Protein G Peroxidase Sigma Conjugate was used in the dilution of 1:90.000, with serums diluted in 1:100 and the antigen in 1:400. Sixty serums were used as negative controls, tested before the CF to determine the cutting point which was 0.042nm. After establishing the standardization, 300 samples were tested, of which 99% (297) were in agreement with the results obtained in the CF. At the end, of assay presented 100% sensibility and 98.2% specificity, with predictive (preditivo) positive and negative values of 97.7% and 100% respectively. The Kappa concordance test was 0.98 and the intra and interplac repeatability were 8.8% and 10.3% respectively. From the results obtained, it is possible to affirm that the indirect ELISA test can be used as an efficient diagnosis tool. However, more essays must be carried out to consolidate the reliability of this test.

• Burkholderia mallei glanders mormo

Abstract in Portuguese:

RESUMO.- Teles J.A.A., Campos A.C., Silva K.P.C., Santos A.S., Santana V.L.A., Castro R.S. & Mota R.A. 2012. [Standardization and evaluation of an indirect ELISA for the serological diagnosis of glanders in horses.] Desenvolvimento e avaliação de um teste ELISA indireto para o diagnóstico sorológico do mormo em equídeos. Pesquisa Veterinária Brasileira 32(9):838-842. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Av. Dom Manuel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com O mormo é uma enfermidade infecto-contagiosa de caráter agudo ou crônico que acomete principalmente os equídeos, causando enormes prejuízos na cadeia produtiva do cavalo. Para controlar a enfermidade o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) instituiu medidas sanitárias obrigatórias em todo território nacional que incluem o diagnóstico oficial pela fixação do complemento (FC), maleinização e sacrifício dos animais positivos. Os kits atuais utilizados no diagnóstico da doença são importados, dificultando e encarecendo sua aplicação na rotina. Objetivou-se com este estudo padronizar um teste de ELISA indireto utilizando o extrato protéico de Burkholderia mallei isolada a partir de equídeo portador no estado de Pernambuco. As amostras foram cultivadas em ágar sangue 10%, incubada por 48h a 37°C; posteriormente caracterizou-se fenotípica e genotipicamente uma das colônias isoladas, e em seguida a cultivou em BHI para enriquecimento; logo após, esta foi repicada para o meio Dor-set Henley o qual foi incubado a 37ºC sob 60rpm por oito semanas. Para padronização do teste utilizou-se o Conjugado Proteína G Peroxidase Sigma na diluição de 1:90.000, com soros diluídos em 1:100 e o antígeno em 1:400. Utilizou-se 60 soros como controle negativo testados frente à FC para determinação do ponto de corte o qual ficou em 0,042nm. Feitas as padronizações, foram testadas 300 amostras, onde 99% (297) foram concordantes com os resultados obtidos na FC. Ao final, o ensaio apresentou 100% de sensibilidade e 98,2% de especificidade com valores preditivo positivo e negativo de 97,7% e 100%, respectivamente. O teste de concordância kappa foi 0,98 e a repetibilidade intra e interplaca ficaram em 8,8 e 10,3%, respectivamente. Diante dos resultados obtidos durante os ensaios, conclui-se que o teste de ELISA indireto pode ser utilizado como uma ferramenta de diagnóstico eficiente. Entretanto, mais ensaios devem ser realizados visando consolidar a confiabilidade do referido teste.

Abstract in English:

ABSTRACT.- Andrade J.S.L., Azevedo S.S., Teles J.A.A., Higino S.S.S. & Azevedo E.O. 2012. [Occurrence and risk factors associated with the Corynebacterium pseudotuberculosis infection in sheep and goats from the semiarid region of the Paraiba state, Northeastern Brazil.] Ocorrência e fatores de risco associados à infecção por Corynebacterium pseudotuberculosis em caprinos e ovinos do semiárido paraibano. Pesquisa Veterinária Brasileira 32(2):116-120. Unidade Acadêmica de Medicina Veterinária, Centro de Saúde e Tecnologia Rural, Universidade Federal de Campina Grande, Av. Universitária s/n, Patos, PB 58700-970, Brazil. E-mail: edisio@cstr.ufcg.edu.br This study aimed to determine the occurrence and risk factors associated with Corynebacterium pseudotuberculosis infection in goats and sheep in semiarid region of Paraiba State, Northeastern Brazil. Of the 640 animals examined, 7.7% (49/640) had clinical evidence of caseous lymphadenitis. In 59.2% (29/49) of these animals there was only the scars of previously ruptured abscesses and in 40.8% (20/49) of the animals abscesses were intact. Of these 20 animals 13 (65%) goats had 14 abscesses, whereas seven (35%) sheep had eight abscesses. In both species, pre-scapular lymph node was the most involved. C. pseudotuberculosis was the agent most frequently isolated, in 15 (68.2%) samples, and in one (4.5%) coagulase-negative Staphylococcus was isolated, one (4.5%) Enterococcus sp., one (4.5%) Proteus mirabilis and Pseudomonas aeruginosa, and in four (18.2%) samples there was no bacterial growth. The final logistic regression model showed that animals from herds where their owners let the abscesses break naturally presented larger odds of caseous lymphadenitis (odds ratio = 8.19, 95% CI = 1.75 - 38.25, p = 0.008). We conclude that goat/sheep owners of the region should adopt preventive measures in their herds, such as early opening and drainage of superficial abscesses, and appropriate destination of the content. Such measures, in addition to regular inspection of the flock, disposal of diseased animals and introduction of non-infected animals will contribute to the control of this infection.

• Corynebacterium pseudotuberculosis lymphadenitis linfadenite caseosa

Abstract in Portuguese:

RESUMO.- Andrade J.S.L., Azevedo S.S., Teles J.A.A., Higino S.S.S. & Azevedo E.O. 2012. [Occurrence and risk factors associated with the Corynebacterium pseudotuberculosis infection in sheep and goats from the semiarid region of the Paraiba state, Northeastern Brazil.] Ocorrência e fatores de risco associados à infecção por Corynebacterium pseudotuberculosis em caprinos e ovinos do semiárido paraibano. Pesquisa Veterinária Brasileira 32(2):116-120. Unidade Acadêmica de Medicina Veterinária, Centro de Saúde e Tecnologia Rural, Universidade Federal de Campina Grande, Av. Universitária s/n, Patos, PB 58700-970, Brazil. E-mail: edisio@cstr.ufcg.edu.br Este trabalho teve como objetivo determinar a ocorrência e os fatores de risco associados à infecção por Corynebacterium pseudotuberculosis em caprinos e ovinos do semiárido da Paraiba. De 640 animais examinados, 7,7% (49/640) apresentavam evidências clínicas de linfadenite caseosa. Em 59,2% (29/49) destes animais havia apenas as cicatrizes de abscessos anteriormente rompidos; em 40,8% (20/49) dos animais, os abscessos estavam intactos. Desses 20 animais, 13 (65%) caprinos apresentaram 14 abscessos, enquanto que sete (35%) ovinos apresentaram oito abscessos. Em ambas as espécies, o linfonodo pré-escapular foi o mais acometido. No exame microbiológico, constatou-se que C. pseudotuberculosis foi o agente mais frequentemente isolado, em 15 (68,2%) amostras; em uma (4,5%) foi isolado Staphylococcus coagulase negativa; uma (4,5%) Enterococcus sp.; uma (4,5%) o Proteus mirabilis e Pseudomonas aeruginosa; e em quatro (18,2%) amostras não houve crescimento bacteriano. O modelo final de regressão logística mostrou que animais provenientes de rebanhos em que seus proprietários deixavam os abscessos romperem naturalmente tiveram maior chance de apresentar linfadenite caseosa (odds ratio =8,19; IC 95% =1,75-38,25; p=0,008). Conclui-se que os caprinovinocultores da região devem adotar medidas profiláticas em seus rebanhos, como abertura e drenagem precoce dos abscessos superficiais e destino adequado do conteúdo. Tais medidas, associadas à inspeção periódica do rebanho, descarte de animais doentes e não introdução de animais infectados, contribuirão significativamente para o controle desta infecção.

Abstract in English:

ABSTRACT.- Silva K.P.C., Mota R.A., Cunha A.P., Silva L.B.G., Leal N.C., Cavalcante Y.V.N., Teles J.A.A., Pereira M.C.C. & Freitas N.S. 2009. [Phenotypic and molecular characterization of Burkholderia mallei isolated in northeastern Brazil.] Caracterização fenotípica e molecular de amostras de Burkholderia mallei isoladas na Região Nordeste do Brasil. Pesquisa Veterinária Brasileira 29(5):439-444. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The objective of this paper was to study the molecular performance and phenotypic characterization of Burkholderia mallei isolated from horses with clinical and serological diagnosis of glanders, originating from the Metropolitan District of Recife and Zona da Mata of Pernambuco and Alagoas. The isolation and biochemical identification of B. mallei was carried out by microbiological and molecular techniques of PCR-fingerprinting and RAPD-PCR. From the eight samples studied, four showed little phenotype variations. In the molecular tests, the samples formed 4 groups of different ribotype profiles and 4 genotype profiles. There was some association of PCR-fingerprinting with RAPD-PCR results. It was concluded that the slight biochemical variations were not associated with different molecular profiles. They also indicated that these differences show heterogeneity associated with the origin of the sample, indicating that the infection was caused by clones of different strains and that the polymorphism of DNA observed could make it

• Burkholderia mallei glanders mormo doenças de eqüdeos

Abstract in Portuguese:

ABSTRACT.- Silva K.P.C., Mota R.A., Cunha A.P., Silva L.B.G., Leal N.C., Cavalcante Y.V.N., Teles J.A.A., Pereira M.C.C. & Freitas N.S. 2009. [Phenotypic and molecular characterization of Burkholderia mallei isolated in northeastern Brazil.] Caracterização fenotípica e molecular de amostras de Burkholderia mallei isoladas na Região Nordeste do Brasil. Pesquisa Veterinária Brasileira 29(5):439-444. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The objective of this paper was to study the molecular performance and phenotypic characterization of Burkholderia mallei isolated from horses with clinical and serological diagnosis of glanders, originating from the Metropolitan District of Recife and Zona da Mata of Pernambuco and Alagoas. The isolation and biochemical identification of B. mallei was carried out by microbiological and molecular techniques of PCR-fingerprinting and RAPD-PCR. From the eight samples studied, four showed little phenotype variations. In the molecular tests, the samples formed 4 groups of different ribotype profiles and 4 genotype profiles. There was some association of PCR-fingerprinting with RAPD-PCR results. It was concluded that the slight biochemical variations were not associated with different molecular profiles. They also indicated that these differences show heterogeneity associated with the origin of the sample, indicating that the infection was caused by clones of different strains and that the polymorphism of DNA observed could make it

Abstract in English:

ABSTRACT.- Medeiros E.S., Santos M.V., Pinheiro Jr J.W., Faria E.B., Wanderley G.G., Teles J.A.A. & Mota R.A. 2009. [In vitro evaluation of the efficacy of commercial disinfectants used in pre and post-dipping against Staphylococcus spp. isolated from bovine mastitis.] Avaliação in vitro da eficácia de desinfetantes comerciais utilizados no pré e pós-dipping frente amostras de Staphylococcus spp. isoladas de mastite bovina. Pesquisa Veterinária Brasileira 29(1):71-75. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The objective of this investigation was to evaluate the in vitro sensibility of Staphylococcus spp. to several commercially available disinfectants used for pre and post dipping. A total of 60 isolates of Staphylococcus spp., identified as Staphylococcus aureus (50) and Positive coagulase Staphylococcus (10) were obtained from the mammary glands of dairy cows with subclinical mastitis in the regions of Metropolitan Recife, the Agreste and the Zona da Mata of the State of Pernambuco, Brazil. As active ingredients were used a chlorine base (25%), iodine (0.6%), chlorhexidine (2.0%), quaternary ammonium (4.0%), and lactic acid (2.0%) at four specific intervals (15", 30", 60", and 300"). One hundred percent of S. aureus was found to be sensitive to iodine, 93.3% to chlorhexidine, 80% to ammonia, 35.6% to lactic acid, and 97.8% were resistant to chlorine at a 60-minute interval. With respect to the Positive coagulase Staphylococcus (SCP), 100.0% of the isolates were sensitive to iodine, 81.8% to quaternary ammonium, 99.9% to lactic acid, 72.7% to chlorhexidine, and 100% was resistant to chlorine at an interval of 60 minutes. It can be concluded that the highest disinfectant activity in vitro was found to be with iodine and chlorhexidine for S. aureus, and with iodine and lactic acid for SCP. A further conclusion was that it is important to undertake a periodic evaluation of the disinfectants used on the dairy properties in the regions studied, given the variety of sensibilities and resistance to disinfectants used, which may prejudice the control of bovine mastitis caused by Staphylococcus spp.

• Disinfectants mastitis Staphylococcus spp. dairy cows

Abstract in Portuguese:

ABSTRACT.- Medeiros E.S., Santos M.V., Pinheiro Jr J.W., Faria E.B., Wanderley G.G., Teles J.A.A. & Mota R.A. 2009. [In vitro evaluation of the efficacy of commercial disinfectants used in pre and post-dipping against Staphylococcus spp. isolated from bovine mastitis.] Avaliação in vitro da eficácia de desinfetantes comerciais utilizados no pré e pós-dipping frente amostras de Staphylococcus spp. isoladas de mastite bovina. Pesquisa Veterinária Brasileira 29(1):71-75. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The objective of this investigation was to evaluate the in vitro sensibility of Staphylococcus spp. to several commercially available disinfectants used for pre and post dipping. A total of 60 isolates of Staphylococcus spp., identified as Staphylococcus aureus (50) and Positive coagulase Staphylococcus (10) were obtained from the mammary glands of dairy cows with subclinical mastitis in the regions of Metropolitan Recife, the Agreste and the Zona da Mata of the State of Pernambuco, Brazil. As active ingredients were used a chlorine base (25%), iodine (0.6%), chlorhexidine (2.0%), quaternary ammonium (4.0%), and lactic acid (2.0%) at four specific intervals (15", 30", 60", and 300"). One hundred percent of S. aureus was found to be sensitive to iodine, 93.3% to chlorhexidine, 80% to ammonia, 35.6% to lactic acid, and 97.8% were resistant to chlorine at a 60-minute interval. With respect to the Positive coagulase Staphylococcus (SCP), 100.0% of the isolates were sensitive to iodine, 81.8% to quaternary ammonium, 99.9% to lactic acid, 72.7% to chlorhexidine, and 100% was resistant to chlorine at an interval of 60 minutes. It can be concluded that the highest disinfectant activity in vitro was found to be with iodine and chlorhexidine for S. aureus, and with iodine and lactic acid for SCP. A further conclusion was that it is important to undertake a periodic evaluation of the disinfectants used on the dairy properties in the regions studied, given the variety of sensibilities and resistance to disinfectants used, which may prejudice the control of bovine mastitis caused by Staphylococcus spp.