Laboratory of Animal Virology, College of Veterinary Medicine, UNESP-São Paulo State University, São Paulo, Brazil.
Theriogenology. 2010 Feb;73(3):324-31. doi: 10.1016/j.theriogenology.2009.09.016. Epub 2009 Nov 6.
Bovine (Bos indicus) herpesviruses have been associated with reproductive disease. Type 1, the most studied species, is best known for its reproductive and respiratory effects. Type 5 (BoHV-5) has been detected in bull semen and aborted fetuses but not in oocytes and embryos. This study consisted of three experiments that evaluated (1) BoHV-5-infected oocytes matured in medium with fetal bovine serum (BoHV-FBS) or polyvinyl alcohol (BoHV-PVA) and fertilized by noninfected sperm; (2) noninfected oocytes fertilized by BoHV-5-infected sperm; and (3) infection of presumptive zygotes by BoHV-5. Each treatment involved nine drops of 15 to 20 oocytes. Infection with BoHV-5 was detected by polymerase chain reaction and in situ hybridization assay, and fertilization capacity and embryonic development were assessed using in vitro culture. Experimentally induced infection was obtained in all experiments, and vertical transmission of BoHV-5 by gametes was confirmed. The cleavage rate was reduced (P=0.0201) in BoHV-FBS (80.4+/-8.9%; mean+/-SD) compared with that of noninfected oocytes (89.9+/-6.5%); neither differed from BoHV-PVA (87.3+/-7.1%), and the resulting embryo production rate was not significantly different among groups. Rates of cleavage (87.5+/-7.5% vs. 92.2+/-5.5%, control vs. infected) and development of embryos (41.7+/-9.9% vs. 44.3+/-7.7% to morula/blastocyst/expanded blastocyst [M/B/EB] and 39.6+/-10.3% vs. 40.8+/-9.2% to blastocyst/expanded blastocyst/hatching blastocyst [B/EB/HB] stages) were not compromised by infected sperm (P=0.1462, P=0.5402, and P=0.8074, respectively). However, presumptive zygotes directly infected 1 d after fertilization produced a lower number (P=0.0140 to M/B/EB and P=0.002 to B/EB/HB stages) of in vitro-produced embryos (31.6+/-4.6 vs. 25.0+/-5.5 and 31.6+/-4.6 vs. 20.2+/-5.4; control vs. infected). In conclusion, BoHV-5 infected gametes and was transmissible to the embryo during in vitro development. As zygotes infected 1 d after fertilization had compromised development, BoHV-5 has the potential to be a pathogen with economic consequences.
牛疱疹病毒与生殖疾病有关。研究最多的 1 型,以其对生殖和呼吸的影响而闻名。5 型(BoHV-5)已在公牛精液和流产胎儿中检测到,但未在卵母细胞和胚胎中检测到。本研究包括三个实验,评估了(1)在含有胎牛血清(BoHV-FBS)或聚乙烯醇(BoHV-PVA)的培养基中成熟的 BoHV-5 感染的卵母细胞,并与未感染的精子受精;(2)BoHV-5 感染的精子受精的未感染卵母细胞;(3)BoHV-5 对假定受精卵的感染。每个处理涉及 15 到 20 个卵母细胞的 9 滴。通过聚合酶链反应和原位杂交检测 BoHV-5 感染,通过体外培养评估受精能力和胚胎发育。在所有实验中均获得了实验诱导的感染,并证实了 BoHV-5 通过配子的垂直传播。与未感染的卵母细胞(89.9+/-6.5%)相比,BoHV-FBS(80.4+/-8.9%;平均值+/-标准差)中的卵裂率降低(P=0.0201);与 BoHV-PVA(87.3+/-7.1%)均无差异,各组胚胎产生率无显著差异。卵裂率(87.5+/-7.5% vs. 92.2+/-5.5%,对照 vs. 感染)和胚胎发育率(41.7+/-9.9% vs. 44.3+/-7.7%至桑葚胚/囊胚/扩张囊胚[M/B/EB]和 39.6+/-10.3% vs. 40.8+/-9.2%至囊胚/扩张囊胚/孵化囊胚[B/EB/HB]阶段)不受感染精子的影响(P=0.1462,P=0.5402,P=0.8074)。然而,直接在受精后 1 天感染的假定受精卵产生的体外产生的胚胎数量较少(在 M/B/EB 和 B/EB/HB 阶段,P=0.0140 至 P=0.002)(31.6+/-4.6 vs. 25.0+/-5.5 和 31.6+/-4.6 vs. 20.2+/-5.4;对照 vs. 感染)。总之,BoHV-5 感染了配子,并在体外发育过程中传播给胚胎。由于受精后 1 天感染的受精卵发育受损,BoHV-5 有可能成为一种具有经济后果的病原体。