骆驼盘尾丝虫病及其传播媒介的研究

发布时间：2018-03-04 10:02

本文选题：双峰驼　切入点：福斯盘尾丝虫　出处：《内蒙古农业大学》2016年博士论文　论文类型：学位论文

【摘要】：骆驼盘尾丝虫病是由福斯盘尾丝虫(Onchocerca fasciata)的成虫寄生于骆驼的项韧带、肌腱和皮下组织等身体各部位,形成纤维结缔组织性结节病变的一种寄生虫病。该病不仅影响着骆驼的生长发育,同时也对驼肉的品质和皮革质量造成严重危害,因此其给养驼业带来不少经济损失。目前,对于该寄生虫病的病原以及它的生活史研究甚少,特别是其传播媒介究竟是什么,国内外都不得而知。因此,面临该寄生虫病在我国某些地区骆驼中的高感染现状,对它的有效防治工作却是无从下手。本研究结合内蒙古骆驼养殖业疫病防治的实际需要,主要从以下几个方面对该寄生虫病流行及传播情况,进行了相关研究。首先调查了内蒙古4个地区(阿拉善盟阿拉善左旗、乌拉察布市四子王旗、巴彦淖尔市乌拉特后旗、锡林郭勒盟苏尼特右旗)的双峰驼盘尾丝虫病的感染情况,先后共检查了不同性别和不同年龄的双峰驼(Camelus bactrianus)135峰,发现福斯盘尾丝虫感染率为45.9%,最高感染强度为15个/峰,感染率在不同性别中差异不大,但随着骆驼年龄的增长而上升。对骆驼盘尾丝虫所造成的组织病理变化进行了观察,结果显示虫体寄生部位多集中在骆驼的项韧带处,主要的病变表现为形成不同阶段的寄生虫性肉芽肿结节,虫体被纤维结缔组织紧密包裹。镜下观察,病变初期,囊腔中的虫体结构完整,周围组织增生并伴有不同程度的炎性细胞浸润；病变中期,囊腔中的坏死虫体浓缩并被胶原纤维组织吸收和替代；病变后期,囊腔中的虫体大部分钙化消失或留有残片,周围组织有不同程度的钙盐沉积。对收集到的骆驼盘尾丝虫完整成虫、微丝蚴和虫卵,进行了形态学的详细观察。其成虫呈白色丝状,角皮上有螺旋状脊和横纹,雄虫体长为63.00-88.00mm,尾部呈螺旋状卷曲,有尾乳突；雌虫较雄虫长,约为890mm,雌虫角皮上的螺旋状脊更为明显,脊与脊之间隔着四条横纹,螺旋状脊和横纹的宽窄及角皮的厚度随虫体部位而变化；微丝蚴细长,无鞘,长为210～244μm。对骆驼福斯盘尾丝虫16S rRNA序列进行了PCR扩增,测得其序列长度为433bp。分析了内蒙古福斯盘尾丝虫病流行地区双峰驼生活环境中水生双翅目昆虫的种类组成和分布情况。根据形态,鉴定出水生双翅目昆虫21科42属,共49种,其中有18种为内蒙古新记录种,有14种为吸血双翅目昆虫。据统计,在水生双翅目昆虫中,长角亚目(NEMATOCERA)占比为49.3%；芒角亚目(ARISTOCERA)占比为43.5%；短角亚目(BRACHYCERA)占比为7.2%,在不同的生境和月份,它们的分布不同。对可能的传播媒介,即14种具有吸血习性的水生双翅目昆虫(新平库蠓Culicoides xinpingensis、曲囊库蠓Culicoides puncticollis、浅暗库蠓Culicoides pallidulus、淡角贝蠓Bezzia albicornis、饶河贝蠓Bezzia raoheensis、棕色裸蠓Atrichopogon fusculus、单囊细蠓Leptoconops monotheca、骚扰库蚊Culex pipiens molestus、蒙古白蛉Phlebotomus mongolensis、猎黄虻Atylotus agrestis、脱粉麻虻Haematopota desertorum、莫氏斑虻Chrysops mlokosiewiczi、密斑虻Chrysops suavis和穴虻属未定种Vermileo sp.),在形态学分类鉴定的基础上,进行了线粒体mtDNA的COI基因序列的测定和分析,结果显示,5种短角亚目和9种长角亚目的吸血昆虫的COI序列同源性在80.8%～92.8%和81.2%～97.9%之间,系统进化树不同属之间差异较显著,同属间聚类较好,基本和形态学鉴定结果相同。为了对福斯盘尾丝虫病可疑传播媒介进行筛选,首先建立了实时荧光PCR方法,并对捕获的吸血双翅目昆虫进行了传播媒介的初步筛选,然后对其进行详细的生物学剖检和分子生物学验证。结果在雌性曲囊库蠓(Culicoides puncticollis)腹部发现了疑似盘尾丝虫微丝蚴,感染率为7.9%,最高感染强度为5个/只。所获得的疑似微丝蚴长短不一,尾部微弯曲,体表有横纹,头尾清晰。通过普通PCR方法对疑似微丝蚴的rDNA的ITS序列进行了扩增、测序和比对,结果表明在曲囊库蠓体内发现的微丝蚴序列与GenBank中已登录的福斯盘尾丝虫ITS序列的同源性为99.6%,证实所检获的微丝蚴是福斯盘尾丝虫微丝蚴,因此基本可以确定曲囊库蠓可能是我国骆驼福斯盘尾丝虫的传播媒介。
[Abstract]:The camel is composed of Fuchs onchocerciasis Onchocerca volvulus (Onchocerca fasciata) adult parasites in the camel's ligament, tendon and subcutaneous tissue of various parts of the body, a parasitic disease of the connective tissue nodules. This disease not only affects the camel's growth and development, but also on the quality of camel meat and leather quality caused serious harm, so it supplies the camel industry bring a lot of economic loss. At present, the parasitic disease pathogen and life history studies it is very little, especially what is the media, both at home and abroad can make nothing of it. Therefore, facing the high infection status of the parasites in the camel in some areas of our country, to it is impossible to start the effective prevention and control work. Based on the actual needs of Inner Mongolia camel breeding disease prevention and control, mainly from the following aspects of the epidemic and spread of parasites, Relevant research has been carried out. The first survey of the Inner Mongolia 4 area (Alashanzuoqi of Alashan City, Siziwangqi Bayannaoer City ural Chahar, wulatehouqi, Xilinguole Meng banner infection) in onchocerciasis, were examined in different gender and different age of Bactrian camel (Camelus bactrianus) 135 peaks, found Fuchs Onchocerca infection rate 45.9%, the highest intensity of infection is 15 / peak, the infection rate in different gender differences is not big, but increased with age. The camel pathological changes caused by camel Onchocerca volvulus were observed, results showed that the parasites were located in the nuchal ligament at the camel, the main lesions for the formation of parasitic granuloma nodules in different stages, the body tightly wrapped by fibrous connective tissue. Microscopically, early lesions, cystic cavity in the body structure Complete surrounding tissue hyperplasia with different degree of inflammatory cell infiltration lesions; mid concentrated necrosis cysts in the cavity and is absorbed and replaced the collagen fibers; at the early stage of the disease, cystic cavity in the body most have disappeared or calcified debris, calcium salt deposition in different degree of surrounding tissue. The collected camel Onchocerca volvulus microfilariae complete adults and eggs were examined in detail. The adult morphology is white filamentous, spiral ridges and horizontal angle of the skin, the body length is 63.00-88.00mm, the tail of a spiral coiled tail papillae; females than males, approximately 890mm, spiral the female angle ridge skin is more obvious, and the ridge ridge between four stripes, width of spiral ridges and transverse and angular skin thickness varies with body parts; microfilaria sheath, long slender, 210~244 M. of camel Fuchs Onchocerca 16S The rRNA sequence was amplified by PCR, measured the length of sequence analysis of species composition of living environment in Inner Mongolia Fuchs onchocerciasis endemic areas in Bactrian camel aquatic dipteral insects and the distribution of 433bp.. According to morphology, identification of aquatic dipteral insects of 21 families and 42 genera, 49 species, 18 of which are new records in Inner Mongolia there are 14 kinds, for the blood sucking Diptera insects. According to statistics, in the aquatic dipteral insects, nemocera (NEMATOCERA) accounted for 49.3%; Mong Kok suborder (ARISTOCERA) accounted for 43.5%; Brachycera (BRACHYCERA) accounted for 7.2%, in different habitats and months, their distribution in different. The media, which has 14 kinds of blood sucking habits of the aquatic dipteral insects (Xinping Culicoides Culicoides xinpingensis, Qu sac Culicoides Culicoides puncticollis dark Culicoides Culicoides pallidulus light DENTALIUM midge Bezzia albicornis, Raohe Bezzia Bezzia raoheensis, brown atrichopogon Atrichopogon fusculus single capsule leptoconops Leptoconops monotheca Culex pipiens molestus, Culex pipiens molestus, Mongolia Phlebotomus Phlebotomus mongolensis, Atylotus agrestis hunt atylotus, shedding HAEMATOPOTA Haematopota desertorum Chrysops mlokosiewiczi, Morse deerfly, deerfly suavis and Chrysops dense point hybomitra Vermileo sp.), in the uncertain species based on the morphological identification, the mitochondrial mtDNA COI gene sequence determination and analysis, results showed that 5 kinds of Brachycera and 9 kinds of NEMATOCERA bloodsucking insects COI sequence homology between 80.8% ~ 92.8% and 81.2% ~ 97.9%, the phylogenetic tree among different genera were significantly different between clustering, belong to well, the basic and the morphological identification of the same. In order to Fuchs onchocercosis suspicious media were screened, first established the real time PCR method, and the Preliminary screening of blood sucking Diptera Insect capture of the media, and then the biological dissection and molecular biology. The results with results in the female song Culicoides (Culicoides puncticollis) were found suspected abdominal Onchocerca volvulus microfilariae, the infection rate was 7.9%, the highest intensity of infection is 5 / only suspected. Microfilaria lengths obtained by tail bending, covered with stripes, head and tail clear. Through PCR method of ITS sequence of suspected microfilaria rDNA were amplified, sequenced and compared, results show that logged in the song in the bursa of Culicoides microfilariae and GenBank sequence in Fuchs Onchocerca ITS sequence homology was 99.6%, confirmed the seizure of microfilariae of Onchocerca volvulus microfilariae is Fuchs, so basically you can determine the song capsule Culicoides may be China's camel Fuchs Onchocerca media.

【学位授予单位】：内蒙古农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S858.24

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