Detecção do vírus da Artrite Encefalite Caprina por nested PCR e nested RT-PCR em ovócitos e fluido uterino

Abstract

Caprine arthritis-encephalitis (CAE) is aninfectious disease caused by the caprine arthritis-encephalitis virus (CAEV), belonging to the lentivirus genus. The presence of the virus has been observed in the nervous system, respiratory tract and mammary gland, and also in the male and female genital tract. The objective of this study is to identify the virus in oocyte and uterine fluid of infected goats by molecular diagnostic techniques, in order to assess the possibility of CAEV transmission with reproduction. Thirteen infected goats were selected and submitted to euthanasia for the collection of the reproductive system, aspiration of the uterine fluid and dissection of ovaries for oocyte collection. In order to identify the CAEV in the collected material, in the protovirus and free forms, it was submitted to the nRT-PCR and nPCR techniques, respectively. As a result, it was observed that 53.8% of oocytes were positive to nRT-PCR, while only 9.1% were positive to nPCR. The nRT-PCR also identified the virus in the uterine fluid of 46.1% of the tested females. Even though the 13 goats had CAEV, 30.8% presented negative results in nPCR and nRT-PCR in all of the analyzed samples (oocyte and uterine fluid). This work concludes that nRT-PCR and nPCR can be used in the diagnosis of CAE for the analysis with oocytes and uterine fluid, and that the presence of CAEV in these materials points out to the risk of CAEV transmission through reproductive technologies used in females.

Evaluation of caprine arthritis-encephalitis virus transmission in newborn goat kids

“…ANDRIOLI et al (2002) made viral isolation followed by nested-PCR and found that CAEV was present in the uterine fluid of positive goat nannies, corroborating with the findings of FIENI et al (2003), who also identified the presence of this virus in embryo washings. CAVALCANTE et al (2013) observed with reverse transcription polimerase chain reaction (RT-PCR) that six goats presented free viral particles in the uterine fluid, demonstrating the potential to transmit this pathogen, which explains the production of antibodies detected by WB in the newborn goat kids in this study.…”

“…However, viral particles may eventually pass through to the fetus, considering that they may be found in the reproductive tract (ANDRIOLI et al, 2002;CAVALCANTE et al, 2013). ANDRIOLI et al (2002) made viral isolation followed by nested-PCR and found that CAEV was present in the uterine fluid of positive goat nannies, corroborating with the findings of FIENI et al (2003), who also identified the presence of this virus in embryo washings.…”

“…CUTLIP et al (1981) isolated MVV from a fetus, which was removed via cesarean section from a seronegative sheep that was in contact with seropositive individuals. CAVALCANTE et al (2013) studied experimentally 13 female goats inoculated with CAEV-Cork viral strain and all of them were positive in AGID and WB with 30 to 60 days post-infection. After 24 months, all animals presented negative AGID results and only three were positive in WB.…”

Transmissibilidade de Lentivírus de Pequenos Ruminantes para cabritos e cabras adultas por meio de sêmen infectado experimentalmente

“…A fêmea "E" apresentou soroconversão a partir dos 30 dias pós-infecção à técnica de cELI-SA, e a partir dos 60 dias pós-infecção para IDGA. A fêmea "D" não apresentou nenhum resultado positivo durante o período monitorado, para as três técnicas diagnósticas, indicando que, naquele momento, a infecção estivesse localizada no sistema reprodutor conforme foi observado por Cavalcante et al (2013) ou uma possível transmissão para o embrião diretamente pelo sêmen infectado como sugerido por Ricarte et al 2010. As provas de imunodiagnóstico são dependentes da quantidade de anticorpos no organismo, que será afetada de acordo com o nível de replicação viral em cada animal (Tigre et al 2006), sendo observado menor tempo médio de soroconversão em rebanhos com maior prevalência (Peterhans et al 2004).…”

Nonsurgical embryo recovery and transfer in sheep and goats

“…Recent data pointed that the nonsurgical uterine flushing seems to have an important role in the caprine arthritis encephalitis virus (CAEV) control programs [55]. Many studies revealed the presence of the virus in uterine flushing media after surgical technique [56][57][58][59][60][61]. However, no positive samples for CAEV were reported from flushing media and embryos recovered by nonsurgical procedure in the first centrifugation.…”