本文選題：雙峰駝　切入點：福斯盤尾絲蟲　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2016年博士論文　論文類型：學(xué)位論文

【摘要】：駱駝盤尾絲蟲病是由福斯盤尾絲蟲(Onchocerca fasciata)的成蟲寄生于駱駝的項韌帶、肌腱和皮下組織等身體各部位,形成纖維結(jié)締組織性結(jié)節(jié)病變的一種寄生蟲病。該病不僅影響著駱駝的生長發(fā)育,同時也對駝肉的品質(zhì)和皮革質(zhì)量造成嚴(yán)重危害,因此其給養(yǎng)駝業(yè)帶來不少經(jīng)濟(jì)損失。目前,對于該寄生蟲病的病原以及它的生活史研究甚少,特別是其傳播媒介究竟是什么,國內(nèi)外都不得而知。因此,面臨該寄生蟲病在我國某些地區(qū)駱駝中的高感染現(xiàn)狀,對它的有效防治工作卻是無從下手。本研究結(jié)合內(nèi)蒙古駱駝養(yǎng)殖業(yè)疫病防治的實際需要,主要從以下幾個方面對該寄生蟲病流行及傳播情況,進(jìn)行了相關(guān)研究。首先調(diào)查了內(nèi)蒙古4個地區(qū)(阿拉善盟阿拉善左旗、烏拉察布市四子王旗、巴彥淖爾市烏拉特后旗、錫林郭勒盟蘇尼特右旗)的雙峰駝盤尾絲蟲病的感染情況,先后共檢查了不同性別和不同年齡的雙峰駝(Camelus bactrianus)135峰,發(fā)現(xiàn)福斯盤尾絲蟲感染率為45.9%,最高感染強(qiáng)度為15個/峰,感染率在不同性別中差異不大,但隨著駱駝年齡的增長而上升。對駱駝盤尾絲蟲所造成的組織病理變化進(jìn)行了觀察,結(jié)果顯示蟲體寄生部位多集中在駱駝的項韌帶處,主要的病變表現(xiàn)為形成不同階段的寄生蟲性肉芽腫結(jié)節(jié),蟲體被纖維結(jié)締組織緊密包裹。鏡下觀察,病變初期,囊腔中的蟲體結(jié)構(gòu)完整,周圍組織增生并伴有不同程度的炎性細(xì)胞浸潤；病變中期,囊腔中的壞死蟲體濃縮并被膠原纖維組織吸收和替代；病變后期,囊腔中的蟲體大部分鈣化消失或留有殘片,周圍組織有不同程度的鈣鹽沉積。對收集到的駱駝盤尾絲蟲完整成蟲、微絲蚴和蟲卵,進(jìn)行了形態(tài)學(xué)的詳細(xì)觀察。其成蟲呈白色絲狀,角皮上有螺旋狀脊和橫紋,雄蟲體長為63.00-88.00mm,尾部呈螺旋狀卷曲,有尾乳突；雌蟲較雄蟲長,約為890mm,雌蟲角皮上的螺旋狀脊更為明顯,脊與脊之間隔著四條橫紋,螺旋狀脊和橫紋的寬窄及角皮的厚度隨蟲體部位而變化；微絲蚴細(xì)長,無鞘,長為210～244μm。對駱駝福斯盤尾絲蟲16S rRNA序列進(jìn)行了PCR擴(kuò)增,測得其序列長度為433bp。分析了內(nèi)蒙古福斯盤尾絲蟲病流行地區(qū)雙峰駝生活環(huán)境中水生雙翅目昆蟲的種類組成和分布情況。根據(jù)形態(tài),鑒定出水生雙翅目昆蟲21科42屬,共49種,其中有18種為內(nèi)蒙古新記錄種,有14種為吸血雙翅目昆蟲。據(jù)統(tǒng)計,在水生雙翅目昆蟲中,長角亞目(NEMATOCERA)占比為49.3%；芒角亞目(ARISTOCERA)占比為43.5%；短角亞目(BRACHYCERA)占比為7.2%,在不同的生境和月份,它們的分布不同。對可能的傳播媒介,即14種具有吸血習(xí)性的水生雙翅目昆蟲(新平庫蠓Culicoides xinpingensis、曲囊?guī)祗稢ulicoides puncticollis、淺暗庫蠓Culicoides pallidulus、淡角貝蠓Bezzia albicornis、饒河貝蠓Bezzia raoheensis、棕色裸蠓Atrichopogon fusculus、單囊細(xì)蠓Leptoconops monotheca、騷擾庫蚊Culex pipiens molestus、蒙古白蛉Phlebotomus mongolensis、獵黃虻A(chǔ)tylotus agrestis、脫粉麻虻Haematopota desertorum、莫氏斑虻Chrysops mlokosiewiczi、密斑虻Chrysops suavis和穴虻屬未定種Vermileo sp.),在形態(tài)學(xué)分類鑒定的基礎(chǔ)上,進(jìn)行了線粒體mtDNA的COI基因序列的測定和分析,結(jié)果顯示,5種短角亞目和9種長角亞目的吸血昆蟲的COI序列同源性在80.8%～92.8%和81.2%～97.9%之間,系統(tǒng)進(jìn)化樹不同屬之間差異較顯著,同屬間聚類較好,基本和形態(tài)學(xué)鑒定結(jié)果相同。為了對福斯盤尾絲蟲病可疑傳播媒介進(jìn)行篩選,首先建立了實時熒光PCR方法,并對捕獲的吸血雙翅目昆蟲進(jìn)行了傳播媒介的初步篩選,然后對其進(jìn)行詳細(xì)的生物學(xué)剖檢和分子生物學(xué)驗證。結(jié)果在雌性曲囊?guī)祗?Culicoides puncticollis)腹部發(fā)現(xiàn)了疑似盤尾絲蟲微絲蚴,感染率為7.9%,最高感染強(qiáng)度為5個/只。所獲得的疑似微絲蚴長短不一,尾部微彎曲,體表有橫紋,頭尾清晰。通過普通PCR方法對疑似微絲蚴的rDNA的ITS序列進(jìn)行了擴(kuò)增、測序和比對,結(jié)果表明在曲囊?guī)祗敷w內(nèi)發(fā)現(xiàn)的微絲蚴序列與GenBank中已登錄的福斯盤尾絲蟲ITS序列的同源性為99.6%,證實所檢獲的微絲蚴是福斯盤尾絲蟲微絲蚴,因此基本可以確定曲囊?guī)祗房赡苁俏覈橊劯Ｋ贡P尾絲蟲的傳播媒介。
[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.

【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S858.24

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