中国实验动物中结肠小袋纤毛虫快速鉴定及感染调查

Rapid Identification and Prevalence of Balantidium Coli Infection among Laboratory Animals in China

目的建立结肠小袋纤毛虫快速鉴定方法,进行国内实验动物感染调查,为国家标准的修订提供参考依据。方法选择2012年12月～2017年12月期间全国60个不同厂家的14 964只实验动物(包括:猴382只,小型猪725只,犬806只,兔1 517只,地鼠422只,豚鼠1 545只,大鼠1 489只,小鼠8 053只,鸡25只)作为调查对象。应用实时动态显微技术识别结肠小袋纤毛虫病原体。利用PCR和测序技术鉴定结肠小袋纤毛虫ITS,18SrRNA和SSU RNA基因。结果通过实时动态显微技术,在这些样本中发现了少量的结肠小袋纤毛虫滋养体和包囊。通过PCR扩增和测序鉴定ITS,18S rRNA和SSU RNA基因的分子特性证明了样本中结肠小袋纤毛虫的存在。2012年12月～2017年12月期间,对14 964只实验动物调查结果显示,结肠小袋纤毛虫感染率为0.03%(4/14 964)。结论实时动态显微技术、PCR和测序技术相结合可完成实验动物的结肠小袋纤毛虫检验工作。国内实验动物中存在结肠小袋纤毛虫感染。因此,建议设计合理的筛查程序用于防止动物传播的结肠小袋纤毛虫病。有必要开发新的诊断工具,特别是快检技术用于这一重要人兽共患病的未来研究。

Objective To establish a rapid identifieation method for Balantidium coli （B. coli）, and acquire the prevalence data on B. coti infestation in laboratory animals in China, and provide reference for the revision of national standard. Methods A total of 14 964 laboratory animals （including 382 monkeys, 725 mini-pigs, 806 dogs, 1 517 rabbits, 422 hamsters, 1 545 guinea pigs,1 489 rats,8 053 mice and 25 chicken） came from 60 different manufactures in China ftom December 2012 to December 2017 were chosen as respondents. Real-time dynamic microscopy technique was performed to identify the B. coli pathogen. Polymerase chain reaction （PCR） and sequencing techniques were used to detect ITS, 18S rRNA and SSU RNA genes of B. coll. Results A small number of trophozoites and cysts of B. coli were detected in these animals by using realtime dynamic microscopy technique. Molecular characterization by PCR amplification and sequencing of ITS, 18S rRNA and SSU RNA genes demonstrated the presence of B. coli in these animals. Among the total 14 964 laboratory animals investigated ftom December 2012 to December 2017, the prevalence of B. coli infection was 0.03f （4/14 964）. Conclusion Realtime dynamic microscopy technique combined with PCR and sequencing technique can be used for the detection of B. coli. g coli infection was present among laboratory animals in China. Thus, it is suggested to design reasonable screening programs for preventing animal-transmitted Balantidiosis. New diagnostic tools, especially rapid detection technologies, which is necessary to be developed in the future studies on this important zoonotic disease.