本文選題：椎實螺 + 片形吸蟲��； 參考：《青海大學(xué)》2016年碩士論文

【摘要】：本研究以青海藏羊體內(nèi)片形吸蟲和片形吸蟲中間宿主椎實螺為研究對象,結(jié)合形態(tài)學(xué)和分子分類學(xué)方法進行了螺體內(nèi)蚴蟲以及成蟲的蟲種分類、鑒定。采用壓片鏡檢的方法對青海省部分地區(qū)中間宿主螺、螺體內(nèi)蚴蟲感染率進行了調(diào)查,同時,用大蒜素和苦參堿兩種植物提取物對椎實螺進行了急性毒性試驗,以在實驗室條件下,篩選出殺死中間宿主螺的有效藥物,結(jié)果如下:1.在椎實螺中收集到雷蚴、尾蚴并在培養(yǎng)青海蘿卜螺的水中收集到囊蚴。經(jīng)形態(tài)學(xué)初步鑒定三種蚴蟲均為肝片吸蟲蚴蟲,進一步提取寄生于蚴蟲基因組DNA,對18S r RNA基因片段進行克隆與測序,對收集到的蚴蟲進行分子分類學(xué)鑒定,所測片段長度為1805bp,用DNAMAN進行序列比對分析,顯示與Gen Bank中的大片吸蟲(Fasciola gigantica)18S r RNA基因序列相似性為92.03%,與肝片吸蟲(Fasciola hepatica)的相似性為99.12%。綜合同源進化樹圖譜可以進一步確定所收集的蚴蟲為肝片吸蟲的蚴蟲。2.傳統(tǒng)的形態(tài)學(xué)鑒定方法無法對肝片吸蟲及大片吸蟲的中間種類進行分類鑒定,本試驗對其成蟲采用18S r RNA基因克隆及序列分析的方法,測出目的片段大小為1834bp。并將測得序列用DNAMAN軟件與Gen Bank中已發(fā)表的大片吸蟲的18S r RNA基因序列比對,其相似性為92.92%,與肝片吸蟲的18S r RNA基因序列相似性為99.78%,結(jié)合同源進化樹圖譜可以確定所采成蟲樣品為肝片吸蟲。3.試驗中發(fā)現(xiàn)所調(diào)查地區(qū)的椎實螺主要有三個種,分別為青海蘿卜螺(Radix cucnnorica)、橢圓蘿卜螺(Radix swinhoei)和狹蘿卜螺(Radix lagotis)。通過對螺體進行壓片鏡檢及在實驗室條件下培養(yǎng)椎實螺,結(jié)果發(fā)現(xiàn):橢圓蘿卜螺體內(nèi)未發(fā)現(xiàn)肝片吸蟲蚴蟲寄生,狹蘿卜螺體內(nèi)發(fā)現(xiàn)雷蚴寄生,感染率較低,10%。青海蘿卜螺體內(nèi)發(fā)現(xiàn)有雷蚴和尾蚴寄生,感染率較高,達41.33%,隨后在培養(yǎng)青海蘿卜螺的器皿中成功收集到囊蚴,而在狹蘿卜螺和橢圓螺的培養(yǎng)過程中并未收集到囊蚴。表明青海藏羊肝片吸蟲的主要中間宿主螺為青海蘿卜螺。4.利用大蒜素和苦參堿兩種植物提取物對椎實螺在實驗室條件下浸殺滅螺效果進行了試驗,研究發(fā)現(xiàn),大蒜素經(jīng)48h,濃度為8mg/L的條件下,死亡率為70%;苦參堿經(jīng)96h,濃度為24mg/L的條件下,死亡率為100%。這兩種提取物的48h的半數(shù)致死濃度(LC50)分別為6.02mg/L和45.55mg/L。表明大蒜提取物對椎實螺的滅殺效果較苦參提取物的滅殺效果好,即椎實螺對大蒜提取物的抵抗能力比對苦參提取物的抵抗能力弱。
[Abstract]:In this study, the parasites of cercariae and adults in Qinghai Tibetan sheep were classified and identified by means of morphology and molecular taxonomy. The infection rate of the intermediate host snail and the parasitic parasite in some parts of Qinghai Province was investigated by the method of pressing microscopic examination. At the same time, the acute toxicity test was carried out with two kinds of plant extracts, allicin and matrine. The effective drugs to kill intermediate host snails were screened under laboratory conditions. The results are as follows: 1. The cercariae and cercariae were collected from the vertebrae and the cysticercaria was collected in the water of the cultured turnip. The three species of cercariae were identified by morphology as paragonimiasis hepatica. The genomic DNA of the three species were further extracted, the 18s r RNA gene fragment was cloned and sequenced, and the collected cercariae were identified by molecular taxonomy. The length of the fragment was 1805bp.The sequence alignment analysis with DNAMAN showed that the similarity with the sequence of Fasciola gigantica)18S r RNA gene in Gen Bank was 92.03 and 99.12% with that of Fasciola hepatica. According to the phylogenetic tree map, we can further determine that the collected metacercaria is a metacercaria of clonorchiasis liver. The traditional morphological identification method could not classify and identify the middle species of paragonimiasis and paragonimiasis. In this experiment, 18s r RNA gene cloning and sequence analysis were used to detect the target fragment size of 1834bp. The sequence was compared with the 18s r RNA gene sequence of the trematode published in Gen Bank by DNAMAN software. The similarity was 92.92%, and the similarity of 18s r RNA gene was 99.78 with that of paragonimus hepatica. The adult samples were determined to be hepatic paragonimiasis by homologous phylogenetic tree map. It was found that there are three main species in the investigated area, Radix cucnnorica, Radix swinhoeii and Radix lagotis. The results showed that there was no parasitism of paragonimiasis in oval radish snail, while that of Rayleidae in radish snail was lower than that in radish snail, and the infection rate was lower than that in radish snail. The parasitic larvae and cercariae were found in the body of turnip snails in Qinghai province, and the infection rate reached 41.33. The cysticercaria was collected successfully in the vessel of cultivating turnip snail, but no cysticercaria was collected in the culture process of radish snail and elliptical snail. The results showed that the main intermediate host snail of the Tibetan sheep liver clonorchiasis in Qinghai was the turnip snail. 4. Two plant extracts, allicin and matrine, were used to study the effect of imbibition on molluscicidal effect in laboratory. The results showed that the mortality rate of allicin was 70 under the condition of 48 h and concentration of 8mg/L, and the concentration of matrine was 24mg/L for 96 h. The mortality rate is 100. The LC50 for 48h of the two extracts was 6.02mg/L and 45.55 mg / L, respectively. The results showed that garlic extract was more effective than Sophora flavescens extract in killing vermicelli, that is to say, the resistance ability of garlic extract was weaker than that of Sophora flavescens extract.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S852.7

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