巨型艾美耳球虫裂殖子的分离和纯化  

Isolation and purification of merozoites of Eimeria maxima

作　　者：张榕珍 王黎霞[2] 郭许情 张健 张天宇 张建军[1] 安健[1] ZHANG Rongzhen;WANG Lixia;GUO Xuqing;ZHANG Jian;ZHANG Tianyu;ZHANG Jianjun;AN Jian(College of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China;College of Animal Science and Veterinary,Beijing Vocational College of Agriculture,Beijing 102206,China)

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

出　　处：《北京农学院学报》2021年第3期67-71,共5页Journal of Beijing University of Agriculture

基　　金：北京市特色高水平骨干专业群项目-动物医学专业群项目-技术平台与社会服务建设项目(PXM2020-157102-000060-15);北京农业职业学院院级科研项目(XY-JT-03)。

摘　　要：[目的]为了获得纯净的巨型艾美耳球虫(Eimeria maxima)裂殖子。[方法]对25日龄白羽肉鸡口服接种3×105巨型艾美耳球虫卵囊,在其感染后48~124 h取十二指肠末端到卵黄蒂后5 cm处肠段,收集裂殖子并计数,确定分离裂殖子的最佳时间;将所取肠段均分为5段,分别对裂殖子进行计数,筛选分离裂殖子的最佳肠段;通过DEAE-52纤维素柱层析对裂殖子进行纯化并分管收集,确定收集较高纯度裂殖子的最佳时间。[结果]在感染后84 h所收集的裂殖子最多;此外,从空肠中后段及空肠末端肠段中分离出较多数量的裂殖子,占总数的62%以上;通过DEAE-52纤维素柱层析获得44 mL纯度较高的裂殖子。[结论]分离巨型艾美耳球虫裂殖子的最佳时间为感染后84 h,最佳肠段为空肠后段,收集1~44 mL洗脱液能够获得纯度较高的裂殖子。[Objective] The study aims to isolate merozoites of Eimeria maxima from specific location of small intestine at appropriate time and obtain the merozoites with high purity. [Method] 25-day-old white feather broilers were inoculated with 3×105 sporulated oocysts of Eimeria maxima. Merozoites were harvested from midintestines(the end of duodenum to 5 cm behind the Meckel’s diverticulum) during 48-124 h postinfection and the number of merozoites were counted. In addition, the intestines were equally divided into 5 segments and numbered, and the merozoites in different segments were counted respectively. The harvested merozoites were further purified by DEAE-cellulose 52 anion exchange chromatography. [Result] The number of merozoites harvested at 84 h postinfection was the largest;In addition, the majority number of merozoites were isolated from the middle and distal segments of jejunum, which accounts for 62% of the total. 44 mL merozoites in purity were collected through DEAE-52 chromatography. [Conclusion] The appropriate time to isolate merozoites of Eimeria maxima is 84 h postinfection. The location to separate the merozoites is the distal jejunum. The merozoites with high purity can be obtained by collecting 1-44 mL liquid.

关 键 词：巨型艾美耳球虫 裂殖子 分离 纯化 

分 类 号：S853.723[农业科学—临床兽医学]