Ali Al Ahmad M Z, Fieni F, Pellerin J L, Guiguen F, Cherel Y, Chatagnon G, Bouzar A B, Chebloune Y
Department of Research into the Health Risk and Biotechnology of Reproduction ENVN, BP 40706, Nantes Cedex 03, France.
Theriogenology. 2008 Mar 1;69(4):473-80. doi: 10.1016/j.theriogenology.2007.10.017. Epub 2007 Dec 21.
The aim of this study was to determine the infectious status of semen and genital tract tissues from male goat naturally infected with the caprine lentivirus. Firstly, polymerase chain reaction (PCR) was used to detect the presence of CAEV proviral-DNA in the circulating mononuclear cells, semen (spermatozoa and non-spermatic cells), and genital tract tissues (testis, epididymis, vas deferens, and vesicular gland) of nine bucks. RT-PCR was used to detect the presence of CAEV viral RNA in seminal plasma. Secondly, in situ hybridization was performed on PCR-positive samples from the head, body, and tail of the epididymis. CAEV proviral-DNA was identified by PCR in the blood cells of 7/9 bucks and in non-spermatic cells of the seminal plasma of 3/9 bucks. No CAEV proviral-DNA was identified in the spermatozoa fraction. The presence of CAEV proviral-DNA in non-spermatic cells and the presence of CAEV in the seminal plasma was significantly higher (p<0.01) in bucks with PCR-positive blood. Two of the three bucks with positive seminal plasma cells presented with at least one PCR-positive genital tract tissue. Proviral-DNA was found in the head (3/9), body (3/9), and tail (2/9) of the epididymis. In situ hybridization confirmed the presence of viral mRNA in at least one of each of these tissues, in the periphery of the epididymal epithelium. This study clearly demonstrates the presence of viral mRNA and proviral-DNA in naturally infected male goat semen and in various tissues of the male genital tract.
本研究的目的是确定自然感染山羊慢病毒的雄性山羊精液和生殖道组织的感染状况。首先,使用聚合酶链反应(PCR)检测9只雄羊的循环单核细胞、精液(精子和非精子细胞)以及生殖道组织(睾丸、附睾、输精管和精囊)中CAEV前病毒DNA的存在情况。使用逆转录聚合酶链反应(RT-PCR)检测精浆中CAEV病毒RNA的存在情况。其次,对来自附睾头部、体部和尾部的PCR阳性样本进行原位杂交。通过PCR在7/9只雄羊的血细胞和3/9只雄羊精浆的非精子细胞中鉴定出CAEV前病毒DNA。在精子部分未鉴定出CAEV前病毒DNA。在血液PCR阳性的雄羊中,非精子细胞中CAEV前病毒DNA的存在以及精浆中CAEV的存在显著更高(p<0.01)。精浆细胞呈阳性的3只雄羊中有2只至少有一个PCR阳性的生殖道组织。在附睾的头部(3/9)、体部(3/9)和尾部(2/9)发现了前病毒DNA。原位杂交证实了在附睾上皮周围的这些组织中的每一种组织中至少有一个存在病毒mRNA。本研究清楚地证明了在自然感染的雄性山羊精液和雄性生殖道的各种组织中存在病毒mRNA和前病毒DNA。