本文關(guān)鍵詞： 駱駝 斯氏副柔線蟲(chóng) 角蠅 發(fā)育過(guò)程 18S rDNA　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：駱駝斯氏副柔線蟲(chóng)病(Parabronemosis)是山斯氏副柔線蟲(chóng)(Parabronema skrjabini)寄生于駱駝?wù)嫖敢鸬囊环N嚴(yán)重危害養(yǎng)駝業(yè)的寄生性線蟲(chóng)病,該病的傳播媒介為西方角蠅(Haematobia irritans)和截脈角蠅(Haematobia titillans)。為了弄清斯氏副柔線蟲(chóng)在角蠅體內(nèi)的發(fā)育過(guò)程,本研究從駱駝生活環(huán)境的駝糞和牛糞中,大量收集疑似角蠅幼蟲(chóng),依據(jù)角蠅幼蟲(chóng)齡期劃分標(biāo)準(zhǔn),并通過(guò)分子生物學(xué)方法鑒定其為角蠅各齡期幼蟲(chóng)；繼而大量剖解角蠅各齡期幼蟲(chóng),在其體內(nèi)尋找疑似斯氏副柔線蟲(chóng)幼蟲(chóng),利用分子生物學(xué)方法對(duì)找到的疑似斯氏副柔線蟲(chóng)幼蟲(chóng)進(jìn)行鑒定,統(tǒng)計(jì)角蠅各齡期幼蟲(chóng)體內(nèi)斯氏副柔線蟲(chóng)幼蟲(chóng)的感染率并觀察其形態(tài)特征。與此同時(shí),對(duì)內(nèi)蒙古地區(qū)西方角蠅和截脈角蠅18S rDNA基因全序列進(jìn)行克隆和測(cè)定,比較兩種角蠅18S rDNA基因序列及其與GenBank中已登錄的9種雙翅目昆蟲(chóng)18S rDNA基因序列的同源性,并利用它們的全序列和最保守同源區(qū)構(gòu)建系統(tǒng)進(jìn)化樹(shù),以確定兩種角蠅在雙翅目昆蟲(chóng)中的位置,并為雙翅目不同科屬昆蟲(chóng)之間的差異性及其分子系統(tǒng)進(jìn)化特點(diǎn)的相關(guān)研究提供依據(jù)。結(jié)果表明：①?gòu)鸟橊勆瞽h(huán)境的駝糞和牛糞中收集的角蠅幼蟲(chóng)被證實(shí)是角蠅各齡期幼蟲(chóng),通過(guò)分子生物學(xué)方法鑒定角蠅幼蟲(chóng)時(shí),改良酚/氯仿抽提法是提取角蠅各齡期幼蟲(chóng)DNA的最佳方法。②角蠅感染斯氏副柔線蟲(chóng)蟲(chóng)卵始于其幼蟲(chóng)階段,其Ⅰ、Ⅱ、Ⅲ期幼蟲(chóng)均可感染,且平均感染率分別為4.48%、5.66%和4.78%。角蠅各齡期幼蟲(chóng)體內(nèi)的斯氏副柔線蟲(chóng)幼蟲(chóng)在光學(xué)顯微鏡下的形態(tài)結(jié)構(gòu)相似,蟲(chóng)體頭端鈍圓,口針明顯并位于其頂端；尾部尖細(xì)向腹面彎曲,肛門開(kāi)口于蟲(chóng)體近末端；其內(nèi)部結(jié)構(gòu)比較簡(jiǎn)單,可以看到食道,且能分出食道肌質(zhì)部和腺質(zhì)部,腸與食道后端相連。③西方角蠅和截脈角蠅的18S rDNA基因全序列長(zhǎng)度均為1984bp,二者的同源性為96.4%,存在73個(gè)識(shí)別位點(diǎn),其中截脈角蠅的18S rDNA基因序列系國(guó)內(nèi)外首次報(bào)道。11種雙翅目昆蟲(chóng)18S rDNA基因序列具有三段保守程度較高的同源區(qū),分別相當(dāng)于西方角蠅18S rDNA基因序列的320bp～693bp、848bp～1181bp、1569bp～1849bp,其中第一段為最保守同源區(qū)；利用其構(gòu)建的系統(tǒng)發(fā)育樹(shù)對(duì)11種雙翅目昆蟲(chóng)進(jìn)行分類,比18SrDNA基因全序列構(gòu)建的系統(tǒng)發(fā)育樹(shù)更符合傳統(tǒng)形態(tài)學(xué)分類結(jié)果。本研究首次在角蠅各齡期幼蟲(chóng)中分離鑒定出斯氏副柔線蟲(chóng)幼蟲(chóng),并對(duì)其形態(tài)特征進(jìn)行了描述,這不僅填補(bǔ)了國(guó)內(nèi)外對(duì)上述研究的空白,還為弄清斯氏副柔線蟲(chóng)在傳播媒介角蠅體內(nèi)的發(fā)育過(guò)程奠定了基礎(chǔ)。
[Abstract]:Parabronema skrjabinii of Camel is Parabronema skrjabinii. Parasitic nematode parasitic disease caused by camel eurogastric disease that seriously endangers camel farming. The transmission vectors of the disease are Haematobia irritanss and Haematobia titillanss. In order to understand the developmental process of nematode skeldahl in hornfly. In this study, a large number of suspected hornfly larvae were collected from camel dung and cow dung in the living environment of camels. Then, a large number of larvae of different ages of hornfly were dissected, and the suspected larvae of Pararinus skrjabini were searched in its body, and the suspected larvae were identified by molecular biological method. The infection rate and morphological characteristics of nematode skeldahl larvae in different larvae of hornfly were analyzed. The 18s rDNA gene was cloned and sequenced from western keratflies and C. truncus in Inner Mongolia. To compare the 18s rDNA gene sequence of two species of hornfly and its homology with the 18s rDNA gene sequence of 9 species of Diptera insects registered in GenBank. The phylogenetic tree was constructed by using their whole sequence and most conserved homologous region to determine the position of two species of hornflies in Diptera insects. It also provides a basis for the study of the differences among different families and genera of Diptera and the characteristics of molecular phylogeny. The results show that:. 1 the larvae collected from camel dung and cow dung were proved to be larvae of different ages. The modified phenol / chloroform extraction method is the best method for extracting DNA from the larvae of keratflies by molecular biological method. Stage 鈪，

本文編號(hào)：1479706