Phenotypic and molecular characterization of bovine Campylobacter fetus strains isolated in Brazil

Vargas, Agueda Castagna de; Costa, Mateus Matiuzzi da; Vainstein, Marilene Henning; Kreutz, Luiz Carlos; Neves, Jennifer

doi:10.1016/s0378-1135(03)00018-x

Cited by 34 publications

(55 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In a second PCR, C. fetus subsp. venerealis was detected in all tested samples confirming previous reports of the occurrence of these subspecies in Brazilian herds (Vargas et al, 2003) and indicating the technical potential to subspecies differentiation although more detailed studies are until necessary to prove this hypotheses. Therefore, a multiplex PCR to direct detection of microorganisms may produce mistakes due to DNA quality and PCR reagents standardization (McPherson & Moller 2000).…”

Section: Discussionsupporting

confidence: 87%

“…The primers used in the present did not amplify DNA of other bacteria that could be present in the genital tract of bovines, indicating the accuracy of PCR in obtaining a diagnosis of infection caused by C. fetus. The specificity of these primers was confirmed by previous studies (Hum et al 1997, Muller et al 2003, Vargas et al 2003.…”

Section: Discussionsupporting

confidence: 76%

“…The amplification conditions were as previously described (Lander 1990, Hum et al 1997) and modified (Vargas et al 2003) The species-specific primers used were MG3F (5'-GGTAGCCGCAGCTGCTAAGAT-3') and MG4R (5'-TAG CTA CAA TAA CGA CAA CT-3'), with amplicon of 750 bp and also for positive samples a subspecies venerealis specific PCR was used , being VenSF (5'-CTT AGC AGT TTG CGA TAT TGC CAT T-3') and VenSR (5'-GCT TTT GAG ATA ACA ATA AGA GCT T-3') with amplicon for 142 bp (Hum et al 1997). DNA template (5ml) was mixed with 45ml of PCR mix: 200mmol of each deoxyribonucleotide triphosphate (dNTP), 10x PCR buffer containing 10mmol Tris-HCl (pH 8.4), 50mmol KCl, 2.5mmol MgCl 2 , 5 U of Taq DNA polymerase (Ludwig Biotechnology Ltd), and 30pmol of primers.…”

Section: Pcr Techniquementioning

confidence: 99%

See 2 more Smart Citations

Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis

et al. 2010

View full text Add to dashboard Cite

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 conteú-do 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. 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. 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.

show abstract

Section: Discussionsupporting

confidence: 87%

Section: Discussionsupporting

confidence: 76%

Section: Pcr Techniquementioning

confidence: 99%

See 1 more Smart Citation

Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis

et al. 2010

View full text Add to dashboard Cite

show abstract

“…venerealis (ATCC 19438), e 21 amostras de C. fetus subsp. venerealis previamente isoladas e caracterizadas fenotípica e molecularmente por Vargas et al (2003) foram utilizadas.…”

Section: Methodsunclassified

Susceptibilidade antimicrobiana de Campylobacter fetus subsp. venerealis isolado de bovinos

et al. 2005

View full text Add to dashboard Cite

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 KirbyBauer 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.INDEX TERMS: Campylobacteriosis, cattle, semen, antibiomicrobials.

show abstract

“…Quando tipificados fenotípica e molecularmente, a maioria dos isolados de C. fetus de abortos ou problemas reprodutivos em bovinos foi classificada como C. fetus subsp. venerealis (Vargas et al 2003). A prevalência da CGB em bovinos de corte foi estimada em 2000, nos 12 estados com a maior produção de bovinos do país, em 19,7% (IC 95%: 13,3%; 25,1%) de animais infectados e em 50,8% (IC 95%: 41,6%; 60,1%) de propriedades contaminadas (Miranda 2005).…”

Section: Distribuiçãounclassified

Campilobacteriose genital bovina e tricomonose genital bovina: epidemiologia, diagnóstico e controle

et al. 2011

View full text Add to dashboard Cite

Pesq. Vet. Bras. 31(4): 336-344, abril 2011 RESUMO.-A presente atualização trata de duas das mais importantes doenças sexualmente transmitidas de bovinos, a campilobacteriose genital bovina e a tricomonose genital bovina. São abordados aspectos relacionados à epidemiologia destas doenças, principalmente em relação a sua distribuição no Brasil. Também são revisados aspectos importantes de diagnóstico, incluindo as técnicas e interpretação dos resultados, além de medidas de controle para ambas as doenças. INTRODUÇÃOA campilobacteriose genital bovina (CGB) e a tricomonose genital bovina (TGB) são doenças infecto-contagiosas de transmissão sexual, que acometem bovinos em idade reprodutiva e causam grandes perdas econômicas em decorrência dos problemas reprodutivos desencadeados (Dekeyser 1984, Goodger & Skirrow 1986 The present update deals with two of the most important sexually transmitted diseases of cattle: bovine genital campylobacteriosis and bovine genital trichomonosis. Epidemiological aspects, mainly their distribution in Brazil, alongside with their diagnosis in cattle are presented and commented. The main points in their diagnoses, including the description of the techniques and the interpretation of the results are also reviewed. Finally the control and prevention of both diseases are discussed.

show abstract

Phenotypic and molecular characterization of bovine Campylobacter fetus strains isolated in Brazil

Cited by 34 publications

References 27 publications

Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis

Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis

Susceptibilidade antimicrobiana de Campylobacter fetus subsp. venerealis isolado de bovinos

Campilobacteriose genital bovina e tricomonose genital bovina: epidemiologia, diagnóstico e controle

Contact Info

Product

Resources

About