生物染色法在犬复孔绦虫虫种形态学鉴别中的应用  

Application of biological staining method in morphological identification of Dipylidium caninum

作　　者：邢维媚 张锐[1] 杨丽侨 张彩虹 殷慧敏 李丽花[1] 芦亚君[1] 

机构地区：[1]海南医学院热带医学与检验医学院,海南海口571199 [2]海南医学院基础医学与生命科学学院,海南海口571199

出　　处：《海南医学》2017年第22期3691-3695,共5页Hainan Medical Journal

基　　金：国家级大学生创新创业训练计划项目(编号:201611810074);海南医学院大学生创新创业训练计划项目(编号:HYCX2015019)

摘　　要：目的调查海口市犬肠道内寄生虫——犬复孔绦虫的流行情况,并研究不同生物染色剂对犬复孔绦虫虫体的染色效果,确定最佳染色方法作为虫种形态学鉴别依据。方法使用正交试验设计法,选择醋酸卡红、齐尔石碳酸复红、德氏苏木素、孔雀绿及盐酸卡红五种不同的生物染色剂对虫体节片在不同时间、不同浓度分别染色,并对这五种生物染色剂两两组合进行复合染色,光学显微镜下观察形态结构并鉴别虫种。二甲苯透明法和冬青油透明法处理虫体节片,和染色法进行效果比较。结果 20%醋酸卡红染色12 h、24 h和10%醋酸卡红染色24 h时染色成节效果更佳;齐尔石碳酸复红染液在48 h、72 h染色成节效果最佳;德氏苏木素染液在8~20 h时染色孕节效果最佳;4%的孔雀绿在6 h时染色孕节效果最佳;盐酸卡红在4~8 h时染色孕节效果最佳;复合染色时石碳酸首染,孔雀绿复染效果最佳。透明法处理孕节片效果佳。结论选择合适染色剂及确定最佳染色方案,可清晰观察到绦虫节片的形态结构,有利于虫种鉴别和病原学诊断。Objective To investigate the prevalence of canine intestinal parasites, Dipylidium caninum, in Haikou. The staining effects of different biological stains on Dipylidium caninum was studied and the best staining method was determined as the basis of morphologic identification. Methods The orthogonal design was used to design an experimental matrix to study the staining effects of five different biological stains（such as acetate carmine, carbol fuchsin,delafields hematoxylin, malachite green and hydrochloride carmine） on different parts of the body, at different time and different concentrations. Moreover, these five biological stains were combined in pairwise to stain specimens. The morphological structure was observed and identified under optical microscope, compared with Xylene transparent method and Holly oil transparent method to process proglottids. Results The staining effects of mature proglottids were better when specimens were treated by 20% acetate carmine for 12 h, 24 h, 10% acetate carmine for 24 h and carbol fuchsin for 48 h, 72 h. The best staining effects of gravid proglottids were observed when they were stained with delafields hematoxylin for 8-20 h, 4% malachite green for 6 h and hydrochloride carmine for 4-8 h. If using combined staining method, the best staining result was obtained from carbol fuchsin-malachite green pairwise staining method. Optimal staining method for gravid proglottid is transparent method. Conclusion Choosing the suitable staining agent and optimal staining method is helpful to clearly observe the structures of tapeworms and is useful for species identification and pathogen diagnosis.

关 键 词：犬复孔绦虫 流行 染色 

分 类 号：R383.3[医药卫生—医学寄生虫学]