LUNAM University, Oniris, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Sanitary Security of Reproduction Biotechnology Unit, Nantes, F-44307, France.
Theriogenology. 2012 May;77(8):1673-8. doi: 10.1016/j.theriogenology.2011.12.012. Epub 2012 Feb 14.
The aim of this study was to determine, in vivo, whether in vitro infected cryopreserved caprine sperm is capable of transmitting caprine arthritis-encephalitis virus (CAEV) vertically to early embryo development stages via artificial insemination with in vitro infected semen. Sperm was collected from CAEV-free bucks by electroejaculation. Half of each ejaculate was inoculated with CAEV-pBSCA at a viral concentration of 10(4) TCID(50)/mL. The second half of each ejaculate was used as a negative control. The semen was then frozen. On Day 13 of superovulation treatment, 14 CAEV-free does were inseminated directly into the uterus under endoscopic control with thawed infected semen. Six CAEV-free does, used as a negative control, were inseminated intrauterine with thawed CAEV-free sperm, and eight CAEV-free does were mated with naturally infected bucks. Polymerase chain reaction (PCR) was used to detect CAEV proviral-DNA in the embryos at the D7 stage, in the embryo washing media, and in the uterine secretions of recipient does. At Day 7, all the harvested embryos were PCR-negative for CAEV proviral-DNA; however, CAEV proviral-DNA was detected in 8/14 uterine smears, and 9/14 flushing media taken from does inseminated with infected sperm, and in 1/8 uterine swabs taken from the does mated with infected bucks. The results of this study confirm that (i) artificial insemination with infected semen or mating with infected bucks may result in the transmission of CAEV to the does genital tack seven days after insemination, and (ii) irrespective of the medical status of the semen or the recipient doe, it is possible to obtain CAEV-free early embryos usable for embryo transfer.
本研究旨在通过用体外感染的精液进行人工授精，从活体上确定体外感染的冷冻保存山羊精子是否能够将山羊关节炎-脑炎病毒（CAEV）垂直传播给早期胚胎发育阶段。通过电刺激法从 CAEV 阴性种羊中采集精子。每份精液的一半用 CAEV-pBSCA 接种，病毒浓度为 10(4)TCID(50)/mL。另一半作为阴性对照。然后对精液进行冷冻。在超数排卵处理的第 13 天，在内窥镜控制下，用解冻的感染精液直接将 14 只 CAEV 阴性母羊输精到子宫内。6 只 CAEV 阴性母羊作为阴性对照，用解冻的 CAEV 阴性精子进行宫内授精，8 只 CAEV 阴性母羊与自然感染的种羊交配。用聚合酶链反应（PCR）检测胚胎在第 7 天的胚胎洗涤液和受体母羊子宫分泌物中的 CAEV 前病毒-DNA。在第 7 天，所有收获的胚胎均为 CAEV 前病毒-DNA 阴性；然而，在 14 只用感染精子授精的母羊中，有 8/14 子宫涂片、9/14 冲洗液和 1/8 子宫拭子中检测到 CAEV 前病毒-DNA。本研究结果证实：（i）用感染精液人工授精或与感染种羊交配可能导致 CAEV 在输精后 7 天传播到母羊生殖道，（ii）无论精液的医疗状况如何或受体母羊的状况如何，都有可能获得可用于胚胎移植的 CAEV 阴性早期胚胎。
