堆型艾美耳球虫体外培养细胞的筛选及其发育研究  

Study on the cell selection for Eimeria acervulina culture and development in vitro

作　　者：王黎霞[1] 黄芳[2] 张建军[2] 安健[2] WANG Lixia;HUANG Fang;ZHANG Jianjun;AN Jian(College of Animal Science and Veterinary,Beijing Vocational College of Agriculture,Fangshan 102442,China;College of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农业职业学院牧医系,北京102442 [2]北京农学院动物科学技术学院,北京102206

出　　处：《北京农学院学报》2020年第3期81-85,共5页Journal of Beijing University of Agriculture

基　　金：北京市“菜篮子”新型生产经营主体科技能力提升工程项目(201615,201418)。

摘　　要：【目的】为了选择适宜堆型艾美耳球虫(Eimeria acervulina)培养的细胞。【方法】利用纯化的E.acervulina子孢子与不同细胞(非洲绿猴肾细胞、牛肾传代细胞、兔肾传代细胞-15、鸡胚成纤维细胞及鸡肾细胞)共培养,比较E.acervulina在原代细胞和传代细胞中的入侵及发育情况。【结果】37℃比41℃适宜进行E.acervulina的体外培养;5种细胞中,牛肾传代细胞入侵率最高;继续培养,牛肾传代细胞中E.acervulina可以形成第二代裂殖体,而非洲绿猴肾细胞、兔肾传代细胞-15只能发育到第一代裂殖体阶段,鸡胚成纤维原代细胞和鸡肾原代细胞中E.acervulina有第一代裂殖子的释放,但未再次入侵,并且鸡胚成纤维原代细胞和鸡肾原代细胞不易传代,5种细胞中,牛肾传代细胞最适宜E.acervulina培养;与牛肾传代细胞共培养1 h子孢子附着于细胞表面,2 h子孢子入侵,子孢子入侵后24 h形成第一代滋养体,52 h后形成第一代成熟裂殖体,60 h第一代裂殖子释放出细胞,72 h后形成第二代裂殖体。【结论】在非洲绿猴肾传代细胞、牛肾传代细胞、兔肾传代细胞-15、鸡胚成纤维原代细胞及鸡原代细胞这5种细胞中,牛肾传代细胞是培养E.acervulina的最适宜细胞,在其中可以发育到第二代成熟裂殖体。【Objective】 The aim of the study is to select the cell for E. acervulina culture in vitro from five kind of cells(Madin-Darby bovine kidney cell, African green monkey kidney cell, Porcine kidney cell-15, chicken embryo fibroblasts cell, Chicken kidney cell). 【Methods】 Sporozoites of E. acervulina were co-cultured with the five kind of cells at 37 ℃ and 41 ℃,respectively. The invasion and development were observed under microscope by hematoxylin-eosin staining. 【Results】 The results indicated that E. acervulina could invade all of the five kind of cells in vitro. 37 ℃ was more suitbale to E. acervulina culture and development than 41 ℃. The invasion rate and development in Madin-Darby bovine kidney cell was the best in the five kind of cells. The feeding trophozoites were found within membrane bound parasitophorous vacuole at 24 h post-inoculation. The first generation mature schizonts were found in the host cytoplasm at 52 h post-inoculation. The first generation merozoites released from the host cell at 60 h post-inoculation. The second generation mature schizonts were found at 72 h. 【Conclusion】 The results suggested that the Madin-Darby bovine kidney cell is the best cell for E. acervulina culture in the five kind of cells(Madin-Darby bovine kidney cell, African green monkey kidney cell, Porcine kidney cell-15, chicken embryo fibroblasts cell, Chicken kidney cell), in which E. acervulina could develop to the second generation mature schizonts.

关 键 词：堆型艾美耳球虫 子孢子 体外培养 细胞 筛选 

分 类 号：S851.34[农业科学—预防兽医学]