摘要
采用差速离心和密度梯度离心,从感染虾肝肠胞虫(Enterocytozoon hepatopenaei,EHP)对虾的肝胰腺中,尝试纯化出孢子,并应用透射电镜、荧光桃红染色和血球计数板计数的方法对纯化出的孢子进行了观察和计数。结果表明,从1g的EHP载量为(4.7±2.2)×104 copies/ng DNA的肝胰腺组织中经差速离心和密度梯度离心方法分离到EHP的孢子,电镜观察孢子为大小在0.7～1.2μm的椭圆形,纯化孢子悬液用血球计数板计数显示孢子浓度为5.30×103个/μL,蔗糖密度梯度中的纯化孢子区带含有的孢子总数约1.06×106个。
Enterocytozoon hepatopenaei(EHP)spores were purified from total 1g hepatopancreas tissues with an average EHP loading at(4.7±2.2)×10^4copies/ng DNA by differential centrifugation and density gradient centrifugation.The purified spores were observed under transmission electron microscope,and counted with phloxine staining on a blood counting chamber under light microscope.Purified spores of EHP were observed under the electron microscope as ellipsoid at size of 0.7~1.2μm.The concentration of EHP spores in the suspension of purified EHP spores was counted as 5.30×10^3/μL by blood counting chamber.The total number of purified EHP spores in the band of sucrose gradients was estimated as 1.06×10^6.
作者
赵若恒
高雯
邱亮
李晨
谢国驷
黄倢
ZHAO Ruo-heng;GAO Wen;QIU Liang;LI Chen;XIE Guo-si;HUANG Jie(Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071;Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao National Laboratory for Marine Science and Technology;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306)
基金
中国-东盟海上合作基金项目(2016-2018)
现代农业产业技术体系项目(CARS-47)
青岛海洋科学与技术国家实验室主任基金项目(QNLM201706)
