甘肃省牦牛肝片形吸虫病血清学流行病学调查及不同发育期大片形吸虫转录组和MicroRNA组研究

发布时间：2018-06-17 03:40

本文选题：肝片形吸虫 + 大片形吸虫　；参考：《吉林农业大学》2017年博士论文

【摘要】：片形吸虫病是由片形吸虫(肝片形吸虫、大片形吸虫)感染引起的危害十分严重的人畜共患寄生虫病,给畜牧业带来重大经济损失。肝片吸虫病主要发生在温带地区,而大片形吸虫病主要发生在热带和亚热带地区。据统计,全世界240多万人感染,一亿八千万人存在感染风险。在我国,有大量关于肝片吸虫感染的病例报道,但是关于牦牛肝片吸虫感染的信息非常有限。此外,大片形吸虫是片形吸虫病的重要病原之一,但对大片形吸虫转录组、基因组及小RNA组等数据比较缺乏,影响了对该病致病机制的深入研究。因此,有必要对我国牦牛肝片形吸虫的感染情况进行流行病学调查并对不同发育期大片形吸虫转录组和小RNA组进行系统的研究。本研究应用ELISA方法对我国甘肃省玛曲县、碌曲县和天祝县牦牛肝片形吸虫的感染情况进行了调查。通过RNA-Seq高通量测序和生物信息学分析等不同的研究手段,对不同发育期大片形吸虫转录组及小RNA组进行了系统研究,鉴定出大片形吸虫不同发育期的转录组和小RNA组特异表达和差异表达的基因,并预测了其功能。结果显示:(1)本研究中的牦牛肝片形吸虫血清学总阳性率为28.7%(454/1,584),其中白牦牛阳性率为29.2%(284/974),黑牦牛阳性率为27.9%(170/610)。牦牛的年龄及季节与片形吸虫感染相关,是重要的风险因素。(2)通过RNA-Seq高通量测序和生物信息学分析等不同手段,对不同发育期大片形吸虫转录组进行了系统的研究,鉴定出不同发育期大片形吸虫unigenes共319261个,平均长度1038个核苷酸,N50值为1552。总共153573个unigenes注释到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%)数据库。鉴定出若干大片形吸虫不同发育期差异表达基因,其中毛蚴vs虫卵组29183个,雷蚴vs毛蚴组27341个,尾蚴vs雷蚴组323075个,囊蚴vs尾蚴组16388个,42天童虫vs囊蚴组30601个,70天童虫vs42天童虫组42个,成虫vs70天童虫组3847个。通过GO和KEGG富集分析发现两条排卵相关的信号通路(cGMP-PKG信号通路and TGF-β信号通路)。(3)通过RNA-Seq高通量测序和生物信息学分析等不同手段,对不同发育期大片形吸虫小RNA组进行了系统的研究,鉴定出160个大片形吸虫不同发育期差异表达miRNAs,毛蚴vs虫卵组69个,雷蚴vs毛蚴组59个,尾蚴vs雷蚴组95个,囊蚴vs尾蚴组44个,42天童虫vs囊蚴组95个,70天童虫vs42天童虫组35个,成虫vs70天童虫组39个。其中,24个已知miRNAs和126个新miRNAs。通过GO和KEGG富集分析鉴别出两个(sja-miR-1和sja-miR-7-5p)在成虫性成熟过程中起到至关重要的miRNAs。(4)通过对转录组和miRNA组数据进行关联分析发现,差异表达miRNA靶向的差异表达基因总共有毛蚴vs虫卵组8868个,雷蚴vs毛蚴组7792个,尾蚴vs雷蚴组11646个,囊蚴vs尾蚴组3025个,42天童虫vs囊蚴组11760个,70天童虫vs42天童虫组14个,成虫vs70天童虫组1097个。本研究首次对我国甘肃地区牦牛肝片形吸虫感染情况进行了血清学调查。同时,应用RNA-Seq技术、生物信息学技术等首次研究了大片形吸虫不同发育期转录组和小RNA组的变化,同时对mRNA和miRNA组数据进行了关联分析。研究结果为深入了解我国牦牛片形吸虫感染情况、阐明大片形吸虫感染性变化的分子基础及致病机制提供了重要基础数据。
[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.
【学位授予单位】：吉林农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S858.23

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