感染白斑综合症病毒中国对虾雄性生殖系统的PCR检测与电镜观察

通过人工投喂携带WSSV的毒饵，对性腺发育成熟的中国对虾（Fenneropenaeus chinensis）雄虾（♂）进行感染实验。采用nest-PCR（巢式PCR）技术，检测感染后的中国对虾雄性生殖系统受WSSV感染情况，同时选取感染严重的虾样进行电镜观察。巢式PCR检测结果表明，感染组中国对虾的精巢、输精管和精囊均被WSSV感染，其中精囊呈阳性的最多，输精管次之，精巢最少。通过电镜进一步观察发现，WSSV粒子只存在于精巢、输精管和精囊的结缔组织中，而在其他组织和生殖细胞中均未发现病毒粒子。其中，精巢中WSSV粒子存在于精巢内两个生精小管之间的结缔组织；输精管中WSSV粒子存在于管壁的结缔组织；精囊中WSSV粒子也只存在于精囊内膜的结缔组织和精荚膜的结缔组织中。PCR检测和电镜观察结果均表明，WSSV粒子能感染中国对虾的雄性生殖系统且对性腺感染存在着一定的组织特异性。

Infection of WSSV in the gonad of Fenneropenaeus chinensis

In shrimps,the WSSV particles infection is characterized by a wide range of target tissues,including testis,ovary,follicle cells,oogonia,oocytes and connective tissue cells,etc..Making sure of the gonad infected by WSSV is important to the vertical transmission of the white spot syndrome virus(WSSV).In this experiment,the adult males of Fenneropenaeus chinensis were artificially infected by WSSV through oral administration.To confirm the infection of WSSV in the gonads of shrimps infected by WSSV,a combine study was made using currently available tools,including polymerase chain reaction(PCR) and transmission electron microscopy(TEM).The tissue/organ samples detected included testis,vas deferens and seminal vesicle.The primary detecting results by nest PCR showed there existed certain tissue specificities on the different parts of the gonad(testis,vas deferens and seminal vesicle).The infected rate in the seminal vesicles infected by WSSV was the highest,while that in the vas deferens was less,and that in the testis was the least.The results of transmission electron microscope showed that the WSSV particles exclusively existed in the connective tissue of the testis,vas deferens and seminal vesicle,but didn't exist in the other germ cells.In addition,the WSSV particles were found in the connective tissue,which located between two testicular cords in the testis,the duct wall of the vas deferens,the inner membrane of the seminal vesicle and the spermatophores.In brief,the results of the nest PCR and the transmission electron microscope indicated that WSSV particles were able to infect the male reproductive system of F.chinensis,and there exist certain tissue specificities on the different parts of the gonad(testis,vas deferens and seminal vesicle).