本文選題：藏豬 + 蛔蟲�。� 參考：《華中農(nóng)業(yè)大學(xué)》2017年博士論文

【摘要】：作為一種以草為主的地方特色品種,藏豬因?qū)Ω吆０蔚貐^(qū)嚴(yán)寒、缺氧等極端氣候具有極好的適應(yīng)能力而成為青藏高原牧民主要的食物來源和經(jīng)濟(jì)來源,其作用不可或缺。由于采取自由放牧的形式,粗放的管理模式,畜群結(jié)構(gòu)不合理以及藏豬品種的退化,再加上薄弱的疾病防控意識,因此近年來藏豬的各種疾病,尤其是寄生蟲疾病的發(fā)病率逐年升高。其中最為常見的寄生蟲是蛔蟲和細(xì)頸囊尾蚴,一旦發(fā)生感染,嚴(yán)重影響藏豬的生長發(fā)育和生產(chǎn)性能,不但給藏豬產(chǎn)業(yè)帶來巨大的經(jīng)濟(jì)損失,同時(shí)也會提高人類感染人畜共患寄生蟲疾病的風(fēng)險(xiǎn)。故本研究在西藏藏豬消化道寄生蟲病流行情況,蛔蟲和細(xì)頸囊尾蚴的分離和鑒定以及線粒體基因組學(xué)等方面展開調(diào)查和研究,結(jié)果如下:1.西藏林芝地區(qū)藏豬消化道寄生蟲流行情況調(diào)查采用寄生蟲學(xué)完全剖檢法、糞便蟲卵檢查法,112頭屠宰藏豬、73份糞便進(jìn)行寄生蟲病流行情況調(diào)查。結(jié)果顯示:共檢測出寄生蟲11種,隸屬于5門、6綱、9目、10科、10屬,其中線蟲5種,絳蟲蚴2種,原蟲2種,吸蟲1種,棘頭蟲1種。優(yōu)勢蟲種為細(xì)頸囊尾蚴(42.9%)、野豬后圓線蟲(38.4%)、棘球蚴(33.0%)、蛔蟲(30.4%)、肝片吸蟲(26.8%)、食道口線蟲(18.8%)、毛首線蟲(15.2%)、球蟲(15.1%)、結(jié)腸小袋纖毛蟲(6.8%)、蛭形巨吻棘頭蟲(5.6%)、六翼泡首線蟲(2.7%)。調(diào)查結(jié)果表明,林芝地區(qū)藏豬胃腸道寄生蟲感染普遍,感染率較高。2.藏豬蛔蟲nad1、cox1和cox2基因擴(kuò)增及序列分析本試驗(yàn)以屠宰藏豬體內(nèi)分離的蛔蟲為研究對象,運(yùn)用PCR方法,以不同引物分別擴(kuò)增nad1、cox1和cox2三個(gè)基因序列,擴(kuò)增后的片段純化后克隆至pGEM?-T Easy載體,轉(zhuǎn)化,并對擴(kuò)增后陽性產(chǎn)物進(jìn)行測序和序列分析,以鑒定藏豬蛔蟲的種類。結(jié)果顯示:藏豬蛔蟲的nad1、cox1和cox2序列均與豬蛔蟲(登錄號:X54253.1,豬蛔蟲)相似性為99%。調(diào)查結(jié)果表明,此次分離的蛔蟲屬于豬蛔蟲。本研究是首次通過藏豬蛔蟲三個(gè)線粒體基因片段nad1、cox1、cox2進(jìn)行分離、鑒定和分子標(biāo)記。3.藏豬細(xì)頸囊尾蚴病流行病學(xué)調(diào)查及cox2基因擴(kuò)增及序列分析本研究以西藏林芝地區(qū)屠宰藏豬體內(nèi)分離出來的細(xì)頸囊尾蚴為研究對象,用ELISA和PCR方法,首次擴(kuò)增藏豬細(xì)頸囊尾蚴的cox2基因序列,并對擴(kuò)增后陽性產(chǎn)物進(jìn)行測序和序列分析,以鑒定藏豬蛔蟲的種類。結(jié)果顯示:藏豬細(xì)頸囊尾蚴的血清抗體陽性率平均為43.93%(2014:42.86%;2015:45.35%),其中公豬和母豬的陽性率分別為43.39%,44.56%,不同年齡段的藏豬細(xì)頸囊尾蚴的陽性率范圍為30.20%~63.79%,結(jié)果顯示:本次分離到細(xì)頸囊尾蚴株與甘肅、青海和四川分離株具有很大的相似性。本研究是首次通過對藏豬細(xì)頸囊尾蚴進(jìn)行流行病學(xué)調(diào)查和cox2基因進(jìn)行分離和鑒定,旨在為西藏地區(qū)藏豬細(xì)頸囊尾蚴的防治提供流行病學(xué)和分子生物學(xué)數(shù)據(jù)。4.藏豬蛔蟲、細(xì)頸囊尾蚴線粒體基因組序列測定與分析本試驗(yàn)對藏豬蛔蟲、細(xì)頸囊尾蚴的線粒體DNA進(jìn)行提取、建庫和測序、功能注釋和生物信息學(xué)分析。結(jié)果表明:豬蛔蟲線粒體基因組大小為14128 bp,編碼基因12個(gè)(cox1-3,nad1-6,nad4L,atp6,cytb)、rRNA 2個(gè)(rrnL、rrnS)、t RNA 35個(gè);細(xì)頸囊尾蚴線粒體基因組大小為13607 bp,編碼基因12個(gè)(cox1-3,nad1-6,nad4L,atp6,cytb)、rRNA2個(gè)(rrnL,rrnS)、t RNA 22個(gè)。豬蛔蟲、細(xì)頸囊尾蚴線粒體基因堿基A、T含量明顯高于C、G含量,分別為71.74%和70.90%。豬蛔蟲、細(xì)頸囊尾蚴線粒體基因密碼子分別為4385(不含終止密碼子304個(gè))和4194個(gè)(不含終止密碼子341個(gè))。兩者密碼子種類均為63個(gè),TTT使用最多,分別為15.51%和10.44%;蛔蟲最少的為CAA(0.05%)、CGC(0.05%),細(xì)頸囊尾蚴最少的為CGC(0.05%)。均翻譯20種氨基酸,豬蛔蟲苯丙氨酸、亮氨酸、纈氨酸比例最高,為17.13%、16.03%、10.86%;細(xì)頸囊尾蚴為亮氨酸、苯丙氨酸、纈氨酸比例最高,為16.48%、11.76%、11.06%。豬蛔蟲氨基酸比列最低的為谷氨酰胺(0.73%)、組氨酸(0.62%);細(xì)頸囊尾蚴氨基酸比列最低的為谷氨酰胺(1.29%)、色氨酸(1.67%)、丙氨酸(1.81%)。進(jìn)化分析表明:藏豬蛔蟲與人蛔蟲(NC-016198.1)同源性高達(dá)99%;與X54253.1同源性高達(dá)99%;與中國湛江分離蟲體(登錄號為HQ704901.1)同源性為98%。該結(jié)果與之前報(bào)道的人蛔蟲與豬蛔蟲高度同源結(jié)果一致,進(jìn)一步證實(shí)了二者可能為同一種蟲體。藏豬細(xì)頸囊尾蚴與參考的甘肅分離株(登錄號:GQ228819.1)同源性均高達(dá)99%。綜上所述,本研究通過對西藏藏豬消化道寄生蟲流行病學(xué)調(diào)查以及2種常見寄生蟲(蛔蟲、細(xì)頸囊尾蚴)進(jìn)行線粒體基因組學(xué)研究,較為全面了解藏豬常見消化道寄生蟲的流行情況,并通過對其進(jìn)行分離鑒定及線粒體基因組測序,這對于了解藏豬寄生蟲的分離鑒定、進(jìn)化分析,種內(nèi)遺傳變異情況,乃至藏豬寄生蟲的防控具有重大的意義。
[Abstract]:As a local characteristic variety with grass mainly, the Tibetan Pig has become the main food source and economic source for the herdsmen of the Qinghai Tibet Plateau because of its excellent adaptability to the extreme cold and anoxic extreme climate in high altitude areas, and its function is indispensable. In recent years, the incidence of various diseases in Tibetan pigs, especially parasitic diseases, is increasing year by year. The most common parasites are Ascaris Ascaris and cysticercus cellulosae, which seriously affect the growth and production performance of Tibetan pigs, not only to the Tibetan Pig Industry, but also to the pig industry. Huge economic losses will also increase the risk of human zoonosis parasitic diseases. Therefore, this study investigates and studies the epidemic of parasitic diseases in the digestive tract of the Tibetan pigs in Tibet, the isolation and identification of Ascaris and cercariae and the mitochondrial genomics. The results are as follows: 1. the digestive tract of the Tibetan pigs in Linzhi, Tibet Parasitology epidemiological survey using parasitology complete examination, fecal egg examination, 112 slaughtered hogs and 73 faeces for parasitic diseases. The results showed that 11 species of parasites were detected in 5 doors, 6 classes, 9 orders, 10 families and 2 species, 2 species of tapeworms, 2 species of protozoa, 1 species of acanthosis and 1 species of echinohead. Insect species are cysticercus cellulosae (42.9%), hydatidis (38.4%), hydatid (33%), Ascaris (30.4%), Fasciola hepatica (26.8%), esophagus nematode (18.8%), nematode (15.2%), coccidia (15.1%), small colonic ciliates (6.8%), leech giant spinous echinohead (5.6%), and six winged nematode (2.7%). The results of the investigation showed that the gastrointestinal parasitism of Tibetan pigs in Linzhi area was parasitized. The infection rate is common, the infection rate is high.2. hid NaD1, cox1 and COX2 gene amplification and sequence analysis. This experiment takes the Ascaris Ascaris isolated in the slaughtered pig as the research object. The PCR method is used to amplify three gene sequences of NaD1, cox1 and COX2 with different primers. After the amplification, the fragment is cloned to pGEM? -T Easy carrier. The positive products were sequenced and sequenced to identify the species of Ascaris Ascaris. The results showed that the NaD1, cox1 and COX2 sequences of Ascaris Ascaris were all similar to that of Ascaris Ascaris (X54253.1, Ascaris Ascaris). The results showed that the Ascaris belonged to Ascaris Ascaris. This study was the first three mitochondrial bases of Ascaris Ascaris. NaD1, cox1, COX2 were isolated, identified and labeled by molecular marker.3., the epidemiological investigation of cysticercosis in porcine cysticercosis and the amplification and sequence analysis of COX2 gene were studied. The COX2 gene of the cysticercus cercariae was amplified by ELISA and PCR method for the study of the cysticercus isolated from the slaughtered and Tibetan pigs in Linzhi, Tibet. Sequence analysis and sequence analysis of positive products were carried out to identify the species of Ascaris Ascaris. The results showed that the positive rate of serum antibody was 43.93% (2014:42.86%; 2015:45.35%), and the positive rate of boar and sow was 43.39%, 44.56%, and the positive rate of Cysticercus cellulosae in different age groups. The range was 30.20%~63.79%. The results showed that the isolated strains of Cysticercus cellulosae were very similar to Gansu, Qinghai and Sichuan isolates. This study was the first time to isolate and identify the COX2 gene by epidemiological investigation of the cysticercus cercariae and to provide the epidemic prevention and control of Cysticercus cercariae in Tibet. Disease and molecular biology data.4. hid Ascaris Ascaris, mitochondrial genome sequencing of Cysticercus cercariae and analysis of mitochondrial DNA from Ascaris Ascaris and cysticercus cercariae were extracted, constructed and sequenced, functional annotation and bioinformatics analysis. The results showed that the size of the mitochondrial genome of Ascaris Ascaris was 14128 BP and 12 (co X1-3, nad1-6, nad4L, Atp6, cytb), rRNA 2 (rrnL, rrnS) and t RNA 35. The size of the mitochondrial genome of Cysticercus cercariae is 13607 BP, and 12 encoded genes. The codon of the mitochondrial gene of Ascaris Ascaris and cercariae is 4385 (304 without terminating codon) and 4194 (without terminating codon 341). Both codon is 63, TTT is used most, 15.51% and 10.44% respectively. The Ascaris is CAA (0.05%), CGC (0.05%), and the cercariae at least CGC (0.05%). Both translates 20 ammonia. The ratio of phenylalanine, leucine and valine in Ascaris Ascaris is the highest, 17.13%, 16.03%, 10.86%. The proportion of Cysticercus is leucine, phenylalanine and valine is the highest, 16.48%, 11.76%. The lowest amino acid of Ascaris 11.06%. is glutamine (0.73%), histidine (0.62%), and the lowest amino acid in Cysticercus cercariae is glutamine. (1.29%), tryptophan (1.67%) and alanine (1.81%). The phylogenetic analysis showed that the homology of Ascaris Ascaris (NC-016198.1) was up to 99%, and the homology of X54253.1 was 99%, and the homology of the Chinese Zhanjiang isolate (login number HQ704901.1) was 98%., which was consistent with the high homologous results of Ascaris Ascaris and Ascaris Ascaris reported before. The two species may be the same type of insect. The homology of the porcine cysticercus and the reference Gansu isolate (login number: GQ228819.1) are all up to 99%.. The study on the mitochondrial genomics of the parasites of the digestive tract of the Tibetan pigs in Tibet and the 2 common parasites (Ascaris Ascaris, cysticercus cercariae) are studied in this study. It is of great significance for understanding the isolation, identification, evolution analysis, intraspecific genetic variation and even the prevention and control of parasites in Tibetan pigs, through the isolation and identification of the common digestive tract parasites in Tibetan pigs and the sequencing of the mitochondrial genome.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S858.28

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 羅潤波;貢嘎;王剛;索朗斯珠;;西藏林芝市藏豬豬流感H1、H3亞型流行病學(xué)調(diào)查初報(bào)[J];甘肅畜牧獸醫(yī);2016年19期

2 羅厚強(qiáng);汪小強(qiáng);張輝;蘭彥芳;邱剛;李家奎;強(qiáng)巴央宗;;西藏林芝地區(qū)藏豬消化道寄生蟲流行情況調(diào)查[J];中國獸醫(yī)學(xué)報(bào);2016年09期

3 鄒勇;吳飛;郭雅旭;耿曉蕊;張文慧;王玉霞;李希來;林青;;西北部分地區(qū)豬蛔蟲rDNAITS基因序列測定及遺傳變異分析[J];動(dòng)物醫(yī)學(xué)進(jìn)展;2016年06期

4 馬興斌;吳金措姆;拉巴次旦;曾江勇;;墨竹工卡和工布江達(dá)兩縣的川藏公路沿線藏豬流感病毒的血清學(xué)調(diào)查[J];養(yǎng)豬;2016年03期

5 張輝;羅厚強(qiáng);李坤;李家奎;強(qiáng)巴央宗;;林芝部分地區(qū)藏豬豬瘟的血清學(xué)調(diào)查[J];畜牧與獸醫(yī);2016年06期

6 曹旺平;;豬囊尾蚴病的致病作用及其防治措施[J];畜牧獸醫(yī)雜志;2016年02期

7 曹玉娟;付聰;田徑軍;曾東柱;顏運(yùn)和;;豬蛔蟲病的診斷與防治[J];湖北畜牧獸醫(yī);2016年01期

8 唐建華;陳曉英;拉巴次旦;吳金措姆;馬興斌;;拉林公路沿線藏豬副豬嗜血桿菌血清抗體流行病學(xué)調(diào)查[J];養(yǎng)豬;2015年05期

9 趙寧;;淺談藏香豬傳染病流行的原因及控制措施[J];山東畜牧獸醫(yī);2015年08期

10 徐雪萍;張?jiān)品?;藏香豬養(yǎng)殖技術(shù)及現(xiàn)狀[J];新疆農(nóng)墾科技;2015年05期

相關(guān)博士學(xué)位論文 前3條

1 高云航;豬瘟超前免疫效果綜合評價(jià)[D];吉林農(nóng)業(yè)大學(xué);2007年

2 應(yīng)三成;藏豬幾個(gè)重要遺傳特性基因的研究[D];四川大學(xué);2007年

3 李明偉;弓首蛔蟲分子鑒定和線粒體基因組的研究[D];華南農(nóng)業(yè)大學(xué);2006年

相關(guān)碩士學(xué)位論文 前4條

1 鄒勇;西北部分地區(qū)豬蛔蟲形態(tài)學(xué)觀察及其種群遺傳變異分析[D];西北農(nóng)林科技大學(xué);2016年

2 時(shí)新春;西藏林芝地區(qū)藏豬群6種傳染病的血清流行病學(xué)調(diào)查[D];西北農(nóng)林科技大學(xué);2013年

3 肖文萍;藏豬腸道微生物多樣性的研究[D];西北農(nóng)林科技大學(xué);2012年

4 張穎;東方巴貝斯蟲EST序列分析及其線粒體基因的測定[D];華中農(nóng)業(yè)大學(xué);2009年

，

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