【作者】 范希萍；

【导师】 王雯慧；

【作者基本信息】 甘肃农业大学 ， 基础兽医学， 2013， 博士

【摘要】 兔豆状囊尾蚴病呈世界性分布，严重威胁着养兔业的发展，对该病的研究有重要的兽医公共卫生学意义。为明确豆状带绦虫不同发育阶段的结构特点及其致病机理，本研究通过人工感染的方法成功建立了犬豆状带绦虫感染模型和家兔豆状囊尾蚴感染模型,并采用电子显微技术和病理组织学技术等研究方法对豆状带绦虫、六钩蚴及豆状囊尾蚴的超微结构和组织学结构进行了系统观察；同时对家兔感染豆状囊尾蚴后的病理学变化规律进行了研究与探讨。1）驱虫后的犬人工感染豆状囊尾蚴，发现感染后42d开始排出孕卵节片，收集孕卵节片。感染70d后剖杀犬，收集完整成虫，取头节、颈节、成节和孕节分别制备电镜样品，透射电镜观察；取头节、颈节、成节和孕节制备石蜡切片，分别采用HE常规染色法、Masson氏特殊染色法和Langhan氏特殊染色法染色，光镜观察。结果显示豆状带绦虫的基本结构与猪带绦虫结构相似，而豆状带绦虫小钩芯髓不均质，与猪带绦虫存在差异。2）分离六钩蚴，用人工肠液激活，制备电镜样品，透射电镜观察。结果显示豆状带绦虫六钩蚴可分为皮质层、皮下层和实质层几部分，基本结构与猪带绦虫六钩蚴结构相似，但小钩基部直径小于端部，芯髓不均质，与猪带绦虫六钩蚴存在差异；在实质层观察到含有9个核的合胞体，其周围的结构与合胞体结构相同，证实了六钩蚴的基本结构由合胞体组成。3）血清学检测豆状囊尾蚴抗体阴性的家兔，人工感染六钩蚴并隔离饲养两个月，取成熟豆状囊尾蚴，采用上述相同方法，观察其组织结构和超微结构。结果显示豆状囊尾蚴与猪囊尾蚴、羊脑多头蚴结构基本相似，但豆状囊尾蚴主要寄生于大网膜和肠系膜表面，肌肉中未发现；另外，在豆状囊尾蚴囊壁上未见微绒毛。表明带科绦虫囊尾蚴的基本结构相似，但存在种间差异。4）血清学检测豆状囊尾蚴抗体阴性的家兔人工感染六钩蚴，根据感染数量将试验动物分为高剂量组（6只）和低剂量组（6只），设对照组（6只）。于感染后第7d、30d和60d分别采取试验组和对照组家兔的肝脏、脾脏、肾脏，制作石蜡切片，HE染色，光镜观察；同时，每周耳缘静脉采血，进行血液学检查。结果显示：感染初期，不同试验组家兔的肝脏、脾脏和肾脏均呈急性出血性炎症变化，肝细胞空泡变性，间质可见六钩蚴，肉芽肿初步形成。感染一个月后，可见囊尾蚴移行道，肝细胞严重空泡变性，间质中有未成熟囊尾蚴。感染后期，肝细胞空泡变性和颗粒变性，可见寄生虫性特殊肉芽肿。血液学检查结果显示，试验组家兔外周血中的中性粒细胞和淋巴细胞在感染一周后明显高于对照组，一个月后开始下降，感染两个月时基本接近对照组。同对照组相比，试验组单核细胞和嗜酸性粒细胞的变化规律为感染一周后明显增多，一个月后继续上升，感染两个月时下降，趋于稳定。更多还原

【Abstract】 Rabbit cysticercus pisiformis poses a worldwide distribution and serious threat to thedevelopment of rabbit and canine keeping. It is important value on veterinary public health tostudy the disease. To clear structural characteristics and pathogenesis at different developmentalstages of taenia pisiformis, canine and rabbits models respectively artificial infected taeniapisiformis and cysticercus pisiformis were successfully developed to observe ultrastructure andhistological structure of taenia pisiformis, hexacanth and cysticercus pisiformis with electronmicroscope and histopathological techniques. Moreover, pathological changes of rabbit infectedcysticercus pisiformis was studied and discussed.1）Canine was dewormed before artificially infection with cysticercus pisiformis, andgravid proglottid was exerted and collected after42days. The infected canine was killed tocollect whole adult worm after70days, the scolex, collum segment, mature and gravid proglottidof adult worm were prepared for transmission electron microscopic eximination, and wereprepared as paraffin section, then stained with HE, Masson’s special staining and Langhan’sspecial staining respectiveluy for light microscopic eximination. The results showed that thebasic structure of taenia pisiformis was as simialr as that of taenia solium, but the hamulus coreof taenia pisiformis was heterogeneous, that was different from taenia solium.2）oncosphere were separated,activated in artificial intestinal juice and prepared fortransmission electron microscopic eximination. The results showed that oncosphere of taeniapisiformis included cortical, subcutaneous and stroma layers, the basic structure of taeniapisiformis oncosphere was as simialr as that of taenia solium oncosphere, but the diameter ofhamulus basal partof was smaller than that of its end part, and hamulus core was heterogeneousin taenia pisiformis oncosphere, that was different in taenia solium oncosphere. Syncytium with9nuclear was observed in stroma layer and its structure was as same as the aruond structure,which confirmed that the basic structure of taenia pisiformis oncosphere was comprised bysyncytium.3）Rabbits, identified serologically without cysticercus pisiformis antibody, were artificiallyinfected oncosphere and isolated for2months to collcet adult cysticercus pisiformis, of whichultrastructure and histological structure were observed with the above method. The results showed that the structure of cysticercus pisiformis was almost same as that of cysticercuscellulosa and sheep coenosis. However, cysticercus pisiformis parasitized the surface ofomentum majus and mesentery, and were not checked out in muscle. Microvillus was notobserved on cyst wall of cysticercus pisiformis. This indicated that the basic structure wasalomost same among cysticercus of taeniidae taenia.4) Rabbits, identified serologically without cysticercus pisiformis antibody, were artificiallyinfected oncosphere and divided into high dose group(6) and low dose group(6) according to theinfectious amount, the control group was arranged6rabbits. At7,30,and60days after infection,the liver, spleen and kidney of all groups were collected to prepared paraffin section and stainwith HE for light microscopic observation. At the same time, the blood was taken from ear veinper week for hematologic examination. The light microscopic results showed that the liver,spleen and kidney of experimetal groups presented a pathological change of acute hemorrhagicinflammation, hepatocellular showed vacuolar degeneration, oncosphere can be found onhepatocellular mesenchyme and granuloma had preliminarily formed at early stage; that thecysticercus migrating channels can be observed, hepatocellular showed serven vacuolardegeneration, immature cysticercus can be found on hepatocellular mesenchyme afer one months;and that hepatocellular showed vacuolar and flocular degeneration, and parasitic specialgranuloma can be observed at late stage. The results of hematologic examination showed thatperipheral blood neutrophilic granulocyte and lymphocyte of experimetal groups were obvioushigher than that of control group after one week, but began to decrease after one month and werealmost close to that of control group after two months. Compared with control group, monocyteand eosinophilic granulocyte of experimental groups increase obviously after one week, andcontinued to increase after one month, but decrease to the near normal level after two months.更多还原