PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Groff A.C.M., Kirinus J.K., Sá e Silva M., Machado G., Costa M.M. & Vargas A.P.C. 2010. Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis. Pesquisa Veterinária Brasileira 30(12):1031-1035. Laboratório de Bacteriologia, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Av. Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail: agueda.vargas@gmail.com Bovine genital campylobacteriosis is a common venereal disease of cattle; the prevalence of this disease can be underestimated mostly because of the nature of the etiological agent, the microaerobic Campylobacter fetus subspecies venerealis. The purpose of the current study was to evaluate the utilization of polymerase chain reaction (PCR) in the diagnosis of genital campylobacteriosis in samples obtained from bull prepuce aspirate, cow cervical mucus, and abomasum contents of aborted fetuses, collected into enrichment medium. Five different DNA extraction protocols were tested: thermal extraction, lysis with proteinase K, lysis with guanidine isothiocyanate, lysis with DNAzol, and lysis with hexadecyltrimethylammonium bromide (CTAB). The specificity, sensitivity, and technical application of the PCR assay were also evaluated with clinical samples and compared to bacterial isolation by standard culture. DNA extraction by the CTAB protocol provided better results in PCR, and it was able to detect 63 colony-forming units per ml of C. fetus. Out of 277 clinical samples tested, 68 (24%) were positive for Campylobacter fetus using PCR, while only 8 (2.8%) of the samples were positive by bacterial isolation in solid medium, proving the superiority of the PCR technique when compared to the standard isolation method, and providing evidence for its usefulness as a better screening test in cattle for the diagnosis of bovine genital campylobacteriosis.

• Bovine genital campylobacteriosis campylobacteriosis Campylobacter fetus campilobacteriose genital bovina

Abstract in Portuguese:

RESUMO.- Groff A.C.M., Kirinus J.K., Sá e Silva M., Machado G., Costa M.M. & Vargas A.P.C. 2010. Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis. [Reação em cadeia da polimerase para o diagnóstico de campilobacteriose genital bovina.] Pesquisa Veterinária Brasileira 30(12):1031-1035. Laboratório de Bacteriologia, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Av. Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail: agueda.vargas@gmail.com RESUMO.- [Reação em cadeia da polimerase para o diagnóstico de campilobacteriose genital bovina.] Campilobacteriose genital bovina é uma doença venérea comum em bovinos. A prevalência desta doença pode ser subestimada na maioria das vezes pela natureza microaeróbica do agente etiológico, Campylobacter fetus subspecies venerealis. O propósito do presente estudo foi avaliar a utilização da reação de polimerase em cadeia (PCR) no diagnóstico de campilobacteriose genital em amostras obtidas de aspirado prepucial de touros, muco cervical de vacas e conteúdo abomasal de fetos abortados, coletados em meio enriquecido. Cinco protocolos diferentes de extração de DNA foram testados: termo extração, lise com proteinase K, lise com guanidine isothiocyanate, lise com DNAzol e lise com hexadeciltrimetilamônio brometo (CTAB). A especificidade, sensibilidade e a aplicação da técnica da PCR foram também avaliadas com amostras clínicas e comparadas com bactérias isoladas por cultura padrão. DNA extraído pelo protocolo de CTAB demonstrou os melhores resultados na PCR, e foi capaz de detectar 63 unidades formadoras de colônias de C. fetus por ml de meio. Das 277 amostras clínicas testadas, 68 (24%) foram positivas para Campylobacter fetus pela PCR, enquanto 8 (2,8%) das amostras foram positivas por isolamento bacteriológico, provando a superioridade da técnica de PCR quando comparada com métodos padrão de isolamento, e fornecendo evidências de sua utilização como um teste de melhor projeção para diagnóstico em campilobacteriose genital bovina.

Abstract in English:

Vargas A.C., Costa M.M., Groff A.C.M., Viana L.R., Krewer C.C., Spricigo D.A. & Kirinus J.K. 2005. [Antimicrobial susceptibility of Campylobacter fetus subsp. venerealis isolated from cattle.] Susceptibilidade antimicrobiana de Campylobacter fetus subsp. venerealis isolado de bovinos. Pesquisa Veterinária Brasileira 25(1):1-3. Laboratório de Bacteriologia do Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: agueda@ccr.ufsm.br Venereal campylobacteriosis is associated with infection of Campylobacter fetus subsp. fetus and Campylobacter fetus subsp. venerealis. The etiological agent is transmitted by natural bull breeding or artificial insemination using contaminated semen. The present study aimed to determine the in vitro susceptibility of C. fetus subsp. venerealis isolates to antimicrobial drugs generally used in clinical and semen treatment. Reference strains of C. fetus subsp. fetus and C. fetus subsp. venerealis and 21 C. fetus isolates were tested. The susceptibility test was performed by using the modified Kirby-Bauer diffusion disc method. C. fetus subsp. fetus reference strain was resistant to lincomycin and penicillin. C. fetus subsp. venerealis was susceptible to all antimicrobial tested, with exception to C.C. KrewerDnalidixic acid. C. fetus subsp. venerealis samples were sensible to amikacin, ampicillin, cefalotin, streptomycin, gentamycin, penicillin and tetracycline. Drug resistance was observed on 42.86% of lincomycin, 4,76 % of enrofloxacin, and 100% to nalidixic acid. In addition 4.76% of the isolates showed intermediate susceptibility to enrofloxacin, neomycin, polimixin B and 9.52% to lincomycin. The susceptibility of C. fetus isolates to antimicrobial drugs commonly used in clinical and semen treatment was demonstrated.

• Campylobacteriosis cattle semen antibiomicrobials.

Abstract in Portuguese:

Vargas A.C., Costa M.M., Groff A.C.M., Viana L.R., Krewer C.C., Spricigo D.A. & Kirinus J.K. 2005. [Antimicrobial susceptibility of Campylobacter fetus subsp. venerealis isolated from cattle.] Susceptibilidade antimicrobiana de Campylobacter fetus subsp. venerealis isolado de bovinos. Pesquisa Veterinária Brasileira 25(1):1-3. Laboratório de Bacteriologia do Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: agueda@ccr.ufsm.br Venereal campylobacteriosis is associated with infection of Campylobacter fetus subsp. fetus and Campylobacter fetus subsp. venerealis. The etiological agent is transmitted by natural bull breeding or artificial insemination using contaminated semen. The present study aimed to determine the in vitro susceptibility of C. fetus subsp. venerealis isolates to antimicrobial drugs generally used in clinical and semen treatment. Reference strains of C. fetus subsp. fetus and C. fetus subsp. venerealis and 21 C. fetus isolates were tested. The susceptibility test was performed by using the modified Kirby-Bauer diffusion disc method. C. fetus subsp. fetus reference strain was resistant to lincomycin and penicillin. C. fetus subsp. venerealis was susceptible to all antimicrobial tested, with exception to C.C. KrewerDnalidixic acid. C. fetus subsp. venerealis samples were sensible to amikacin, ampicillin, cefalotin, streptomycin, gentamycin, penicillin and tetracycline. Drug resistance was observed on 42.86% of lincomycin, 4,76 % of enrofloxacin, and 100% to nalidixic acid. In addition 4.76% of the isolates showed intermediate susceptibility to enrofloxacin, neomycin, polimixin B and 9.52% to lincomycin. The susceptibility of C. fetus isolates to antimicrobial drugs commonly used in clinical and semen treatment was demonstrated.