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作 者:陈洁[1] 廖国礼[2] 吴玉娇[1] 张庆[1] 杨小娟 范晓东[1,3] 龙梦娴[1] 潘国庆[1] 周泽扬[1,3] CHEN Jie;LIAO Guo-li;WU Yu-jiao;ZHANG Qing;YANG Xiao-juan;FAN Xiao-dong;LONG Meng-xian;PAN Guo-qing;ZHOU Ze-yang(State Key Laboratory of Silkworm Genome Biology/Chongqing Key Laboratory of Microsporidia Infection and Control,Southwest University,Chongqing,400715;Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou,510380;College of Life Science,Chongqing Normal University,Chongqing,401331)
机构地区:[1]西南大学家蚕基因组生物学国家重点实验室/微孢子虫感染与防控重庆市重点实验室,重庆400715 [2]中国水产科学院珠江水产研究所,广州510380 [3]重庆师范大学生命科学学院,重庆401331
出 处:《西南大学学报(自然科学版)》2021年第3期17-23,共7页Journal of Southwest University(Natural Science Edition)
基 金:国家自然科学基金项目(31602012);重庆市教委科学技术研究项目(KJ1703053);家蚕基因组生物学国家重点实验室开放课题(sklsgb181820-11).
摘 要:虾肝肠胞虫Enterocytozoon hepatopenaei是近年来备受关注的病原性微孢子虫,严重影响对虾生长,并在东南亚及我国大部分对虾主产区迅速蔓延,极大地制约对虾养殖业的健康发展.特异、快速、便利地确诊EHP感染,在对虾育种、育苗及养殖过程中具有重要意义.本研究基于微孢子虫的孢子具有高度折光性,孢壁结构含较厚几丁质层及碘化丙啶可穿透孢子活细胞膜的特性,开展虾肝肠胞虫的光学显微成像研究,展示普通光镜法、微分干涉差显微成像法、荧光增白剂(fluorescent brightener 28)染色法及与碘化丙啶(Propidium Iodide)共染的双色荧光法在虾肝肠胞虫快速光镜检测诊断中的应用.实验建立的双色荧光镜检法无需固定和洗涤,具有操作简便,无专业技术要求的特点,可在10 min内完成样品的初筛检测,为虾肝肠胞虫的早期筛查及养殖塘样品快速诊断提供价廉、省时且易操作的检测手段.Enterocytozoon hepatopenaei(EHP)is a pathogenic microsporidia that has attracted much attention in recent years.It affects the growth of shrimp and spreads rapidly in Southeast Asia and most of the shrimp farm region in China,seriously restricts healthy development of shrimp farming.Specific,rapid and convenient diagnosis of EHP is significant in breeding,seedling and culturing of shrimp.Herein,light microscopy of EHP spores was studied based on the characteristics of highly refractive spores,a thick chitin layer within the spore wall and the specific permeability of Propidium Iodide to sporoplasm membrane.We exhibited the application of routine light microscopy,differential interference contrast microscopy,fluorescent brightener 28 staining method and dual-color fluorescence staining method in diagnose of EHP.The dual-color fluorescence microscopy method showed in this study has the characteristics of strong specificity,easy operation,and no professional technical requirements without fixing and washing.The detection of samples can be completed in 10 minutes,which lead it to be an effective method for rapid screening of EHP.
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