This study was conducted in order to evaluate the transmission of caprine lentivirus to sheep using different experimental groups. The first one (colostrum group) was formed by nine lambs receiving colostrum from goats positive for small ruminant lentiviruses (SRLV). The second group (milk group) was established by nine lambs that received milk of these goats. Third was a control group, consisting of lambs that suckled colostrum and milk of negative mothers. Another experimental group (contact group) was formed by eight adult sheep, confined with two naturally infected goats. The groups were monitored by immunoblotting (IB), enzyme-linked immunosorbent assay (ELISA), agar gel immunodiffusion (AGID) and nested polymerase chain reaction (nPCR). All lambs that suckled colostrum and milk of infected goats and six sheep of the contact group had positive results in the nPCR, although seroconversion was detected only in three of the exposed animals, with no clinical lentiviruses manifestation, in 720 days of observation. There was a close relationship between viral sequences obtained from infected animals and the prototype CAEV-Cork. Thus, it was concluded that SRLV can be transmitted from goats to sheep, however, the degree of adaptation of the virus strain to the host species probably interferes with the infection persistence and seroconversion rate.
RESUMOA artrite-encefalite caprina (CAE) é diagnosticada rotineiramente pela técnica de imunodifusão em gel de agarose (IDGA), que é considerada pouco sensível. Objetivou-se com este estudo padronizar testes de Elisa-i e Western Blot (WB) para diagnóstico precoce de anticorpos em caprinos contra CAEV e comparar os resultados obtidos nesses testes com a prova de IDGA. Para a padronização dos testes Elisa-i e WB, utilizaram-se diferentes concentrações e diluições de antígeno, soros e conjugado. No Elisa-i, adotaram-se microplacas rígidas com 96 poços, sendo a combinação de concentração de 0,5µg/poço de antígeno e diluições de 1:100 de soro e 1:1500 de conjugado a que apresentou melhor resultado. No WB foram utilizadas membranas de nitrocelulose, definindo-se as diluições de 1:50 de soro e 1:15000 de conjugado. Para avaliar o desempenho das técnicas, 222 amostras de soro caprino foram testadas e os dados obtidos foram comparados com o IDGA. A sensibilidade e a especificidade do Elisa-i/IDGA, WB/IDGA e WB/Elisa-i foram de 70% e 91%, 100% e 72,6%, 84,6% e 76,5%, concomitantemente. O índice Kappa desses testes foi de 0,35, 0,2 e 0,36, respectivamente. As técnicas de Elisa-i e WB apresentaram-se mais sensíveis que a IDGA, podendo ser utilizadas como ferramentas para o diagnóstico precoce da CAE.Palavras-chave: especificidade, infecção, LVPR, sensibilidade, sorologia ABSTRACT Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and
Resumo -O objetivo deste trabalho foi avaliar a presença do DNA pró-viral do lentivírus caprino (LVC) em ejaculados de machos infectados naturalmente, e verificar a influência da lavagem do sêmen e da presença de inflamação testicular sobre a carga viral. Foram realizadas oito coletas de sêmen de sete reprodutores soropositivos para o LVC: quatro antes dos animais sofrerem dano testicular e quatro depois. Entre as coletas realizadas na mesma semana, em uma, o ejaculado era lavado, para retirada do plasma seminal, e na outra, não. O DNA pró-viral do LVC foi identificado pela reação em cadeia da polimerase Nested (PCR Nested), e pelo isolamento viral. O vírus foi isolado em 7,1% das amostras. A PCR identificou o DNA pró-viral em 35,7% do total das amostras: 17,9% nas amostras lavadas e 53,6% das amostras de sêmen integrais. O dano ao testículo permite maior fluxo do vírus para o sêmen, pois antes do dano, 21,4% das amostras foram positivas e pós-dano, 50%. A transmissão do LVC pelo sêmen de reprodutores caprinos é potencializada pela presença de inflamações testiculares e pelo fato de o sêmen criopreservado conter o LVC na forma infectante.Termos para indexação: reprodução, PCR, artrite-encefalite caprina, transmissão. Risk factors in caprine lentivirus transmission through semenAbstract -The objective of this work was to evaluate the presence of the DNA provirus of the caprine lentivirus (LVC) in ejaculates of naturally infected males, and to verify the influence of the wash of the semen as well as the presence of testicle inflammation on the viral load. Eight semen collections of seven soropositive reproducers were accomplished, four before testicle injury and four after injury. Amongst the collections carried out at the same week, in one the ejaculate was washed, to withdraw the plasma seminal, and in the other it was not. The provirus DNA was identified both by Nested polymerase chain reaction technique (Nested PCR) and by the viral isolation. The virus was isolated in 7.1% of the samples. The PCR identified the provirus DNA in 35.7% of all samples, 17.9% in the washed samples and 53.6% of the integral semen samples. The injury of the testicle tends to greater flow of virus for the semen, therefore, before injury, 21.4% of the samples were positive and after-injury, 50%. Risk of transmission of the LVC by semen of goat reproducers is strengthened by the presence of testicle inflammations and the fact that the criopreserved semen contains the LVC in infecting form.Index terms: reproduction, PCR, caprine arthritis encephalitis, transmission. IntroduçãoO lentivírus caprino (LVC) é o agente etiológico da artrite-encefalite caprina (AEC), uma enfermidade crônica, incurável, de alta prevalência em rebanhos leiteiros nacionais e associada à perdas econômicas. Há grande necessidade de pesquisa dos modos de transmissão desse vírus para o delineamento eficiente de medidas de controle e possível erradicação.O avanço das técnicas de conservação de germoplasma, aliado ao aumento da importância econômica da caprinocultura, ...
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