本文選題：肝片形吸蟲 + 大片形吸蟲　； 參考：《吉林農(nóng)業(yè)大學(xué)》2017年博士論文

【摘要】：片形吸蟲病是由片形吸蟲(肝片形吸蟲、大片形吸蟲)感染引起的危害十分嚴(yán)重的人畜共患寄生蟲病,給畜牧業(yè)帶來重大經(jīng)濟(jì)損失。肝片吸蟲病主要發(fā)生在溫帶地區(qū),而大片形吸蟲病主要發(fā)生在熱帶和亞熱帶地區(qū)。據(jù)統(tǒng)計,全世界240多萬人感染,一億八千萬人存在感染風(fēng)險。在我國,有大量關(guān)于肝片吸蟲感染的病例報道,但是關(guān)于牦牛肝片吸蟲感染的信息非常有限。此外,大片形吸蟲是片形吸蟲病的重要病原之一,但對大片形吸蟲轉(zhuǎn)錄組、基因組及小RNA組等數(shù)據(jù)比較缺乏,影響了對該病致病機(jī)制的深入研究。因此,有必要對我國牦牛肝片形吸蟲的感染情況進(jìn)行流行病學(xué)調(diào)查并對不同發(fā)育期大片形吸蟲轉(zhuǎn)錄組和小RNA組進(jìn)行系統(tǒng)的研究。本研究應(yīng)用ELISA方法對我國甘肅省瑪曲縣、碌曲縣和天�？h牦牛肝片形吸蟲的感染情況進(jìn)行了調(diào)查。通過RNA-Seq高通量測序和生物信息學(xué)分析等不同的研究手段,對不同發(fā)育期大片形吸蟲轉(zhuǎn)錄組及小RNA組進(jìn)行了系統(tǒng)研究,鑒定出大片形吸蟲不同發(fā)育期的轉(zhuǎn)錄組和小RNA組特異表達(dá)和差異表達(dá)的基因,并預(yù)測了其功能。結(jié)果顯示:(1)本研究中的牦牛肝片形吸蟲血清學(xué)總陽性率為28.7%(454/1,584),其中白牦牛陽性率為29.2%(284/974),黑牦牛陽性率為27.9%(170/610)。牦牛的年齡及季節(jié)與片形吸蟲感染相關(guān),是重要的風(fēng)險因素。(2)通過RNA-Seq高通量測序和生物信息學(xué)分析等不同手段,對不同發(fā)育期大片形吸蟲轉(zhuǎn)錄組進(jìn)行了系統(tǒng)的研究,鑒定出不同發(fā)育期大片形吸蟲unigenes共319261個,平均長度1038個核苷酸,N50值為1552�？偣�153573個unigenes注釋到Nr(122405,38.34%),Nt(35756,11.91%),KOG(48889,15.31%),KO(32384,10.14%),Swiss-Prot(66543,20.84%),GO(86793,27.18%)和Pfam(86211,27%)數(shù)據(jù)庫。鑒定出若干大片形吸蟲不同發(fā)育期差異表達(dá)基因,其中毛蚴vs蟲卵組29183個,雷蚴vs毛蚴組27341個,尾蚴vs雷蚴組323075個,囊蚴vs尾蚴組16388個,42天童蟲vs囊蚴組30601個,70天童蟲vs42天童蟲組42個,成蟲vs70天童蟲組3847個。通過GO和KEGG富集分析發(fā)現(xiàn)兩條排卵相關(guān)的信號通路(cGMP-PKG信號通路and TGF-β信號通路)。(3)通過RNA-Seq高通量測序和生物信息學(xué)分析等不同手段,對不同發(fā)育期大片形吸蟲小RNA組進(jìn)行了系統(tǒng)的研究,鑒定出160個大片形吸蟲不同發(fā)育期差異表達(dá)miRNAs,毛蚴vs蟲卵組69個,雷蚴vs毛蚴組59個,尾蚴vs雷蚴組95個,囊蚴vs尾蚴組44個,42天童蟲vs囊蚴組95個,70天童蟲vs42天童蟲組35個,成蟲vs70天童蟲組39個。其中,24個已知miRNAs和126個新miRNAs。通過GO和KEGG富集分析鑒別出兩個(sja-miR-1和sja-miR-7-5p)在成蟲性成熟過程中起到至關(guān)重要的miRNAs。(4)通過對轉(zhuǎn)錄組和miRNA組數(shù)據(jù)進(jìn)行關(guān)聯(lián)分析發(fā)現(xiàn),差異表達(dá)miRNA靶向的差異表達(dá)基因總共有毛蚴vs蟲卵組8868個,雷蚴vs毛蚴組7792個,尾蚴vs雷蚴組11646個,囊蚴vs尾蚴組3025個,42天童蟲vs囊蚴組11760個,70天童蟲vs42天童蟲組14個,成蟲vs70天童蟲組1097個。本研究首次對我國甘肅地區(qū)牦牛肝片形吸蟲感染情況進(jìn)行了血清學(xué)調(diào)查。同時,應(yīng)用RNA-Seq技術(shù)、生物信息學(xué)技術(shù)等首次研究了大片形吸蟲不同發(fā)育期轉(zhuǎn)錄組和小RNA組的變化,同時對mRNA和miRNA組數(shù)據(jù)進(jìn)行了關(guān)聯(lián)分析。研究結(jié)果為深入了解我國牦牛片形吸蟲感染情況、闡明大片形吸蟲感染性變化的分子基礎(chǔ)及致病機(jī)制提供了重要基礎(chǔ)數(shù)據(jù)。
[Abstract]:Schistosomiasis is a serious zoonotic parasitic disease caused by Fasciola Fasciola (Fasciola hepatica) infection. It brings great economic loss to animal husbandry. The main occurrence of Fasciola hepatica in the temperate region, and large Fasciola in the tropics and subtropics. According to statistics, about 2400000 people all over the world Infection, one hundred and eighty million people have a risk of infection. In our country, there are a large number of cases of Fasciola hepatica infection, but the information about the infection of the yak liver flake is very limited. In addition, large Fasciola is one of the important pathogens of Fasciola Fasciola, but the data of large Fasciola, genome and small RNA group are relatively lack, Therefore, it is necessary to make an epidemiological investigation on the infection of the yak liver Fasciola hepatica in China and systematically study the different development stages of large tractable Fasciola and small RNA groups. This study applies the ELISA method to yak liver slices in Maqu, Gansu, and Tianzhu County of China. The infection of clonorimiasis was investigated. Through RNA-Seq high throughput sequencing and bioinformatics analysis, a systematic study was carried out on different developmental stages of tracts and small RNA groups in different developmental stages. The results showed that: (1) the total positive rate of yak liver Fasciola hepatica was 28.7% (454/1584), the positive rate of white yak was 29.2% (284/974), and the positive rate of black yak was 27.9% (170/610). The age and season of yak were related to the infection of lamellar fluke. (2) high throughput sequencing by RNA-Seq and high throughput sequencing. Bioinformatics analysis and other means of different developmental stages of large tractable Fasciola transcriptional groups were systematically studied. There were 319261 unigenes, with an average length of 1038 nucleotides, and a total of 153573 unigenes annotations to Nr (122405,38.34%), Nt (35756,11.91%), KOG (48889,15.31%), KO (3238). 4,10.14%), Swiss-Prot (66543,20.84%), GO (86793,27.18%) and Pfam (86211,27%) database, identified a number of differentially expressed genes in different developmental stages of large tracts, including 29183 eggs of cercariae eggs, 27341 of cercariae in VS, 323075 in the cercariae, 16388 in the cercariae, 30601 in 42 of the cercariae, and 70 in 70 days. 2 days of 2 days, 42 adults and 3847 adult worms. Two ovulation related signaling pathways (cGMP-PKG signaling pathway and TGF- beta signaling pathway) were found through GO and KEGG enrichment analysis. (3) through RNA-Seq high throughput sequencing and bioinformatics analysis, a systematic study was carried out on small RNA groups of large tracts in different development stages. There were 160 different developmental stages of miRNAs, 69 eggs of cercariae, 69 eggs, 59 of cercariae vs, 95 in cercariae, 95 in the cercariae, 44 in the cercariae, 44 in the cercariae of the cercariae, 95 in the 42 cysts of the cercariae, 35 in the vs42 Tiantong group of 70, and 39 for the adult Vs70 Tiantong group, of which 24 known miRNAs and 126 new miRNAs. passed GO and 59. KEGG enrichment analysis identified two (sja-miR-1 and sja-miR-7-5p) plays an important role in adult sexual maturation (4) (4) by correlation analysis of the data of the transcriptional and miRNA groups. The differential expression genes of the differentially expressed miRNA targets were 8868 of the eggs of the larvae of the cercariae, 7792 in the cercariae, and 1164 in the cercariae. 6 cercariae vs cercariae group 3025, 42 Tiantong vs cyst group 11760, 70 Tiantong vs42 Tiantong worm group 14, adult Vs70 Tiantong worm group 1097. This study for the first time in Gansu area of China yak liver Fasciola infection situation conducted a serological investigation. At the same time, the application of RNA-Seq technology, bioinformatics technology for the first time study of large bugs The changes in the different development stage and the small RNA group, and the correlation analysis of the data of mRNA and miRNA group. The results provide an important basic data for understanding the infection of the yak shaped fluke in our country, and clarifying the molecular basis and pathogenesis of the infectious changes of large tracts.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S858.23

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