本文選題：蚤 + 鼠疫�。� 參考：《石河子大學(xué)》2017年碩士論文

【摘要】：目的:對新疆北疆邊境地區(qū)蚤類(包括部分南疆蚤類)進(jìn)行鑒定和病原檢測,為新疆蚤傳病的發(fā)生做出風(fēng)險評估以及蚤傳病的預(yù)防奠定基礎(chǔ)。除此以外,對溫泉縣鼠疫疫源地所收集的樣本及所有蚤類進(jìn)行鼠疫篩查與分型,為追溯鼠疫的主要宿主、媒介及其生態(tài)地理景觀的生物學(xué)類型提供科學(xué)依據(jù)。方法:1.2015-2016年在新疆北疆邊境地區(qū)(布爾津縣、阿拉山口市、溫泉縣、精河縣、霍城縣、察布查爾錫伯自治縣和新源縣)及和田地區(qū)民豐縣收集哺乳動物體表寄生蚤;通過形態(tài)學(xué)及分子生物學(xué)(18S r DNA,28S r DNA,EF1-a,COII和COI),對所收集的蚤進(jìn)行蚤種鑒定并進(jìn)行基因系統(tǒng)發(fā)育分析。2.選取阿拉山口地區(qū)的長吻角頭蚤、臀突客蚤、葉狀切唇蚤突高亞種、后彎怪蚤及禿病蚤指名亞種組織DNA送至北京諾禾致遠(yuǎn)生物信息科技有限公司,對細(xì)菌16S r DNA基因序列中的V3-V4可變區(qū)進(jìn)行測序,分析其細(xì)菌的群落結(jié)構(gòu)、種群多樣性及豐富度。3.結(jié)合宏細(xì)菌數(shù)據(jù),對所采集蚤類樣本進(jìn)行立克次體(MLST)、沃爾巴克體(wsp和16S r RNA)以及巴通體(glta、ITS和rib C)的篩查,陽性片段進(jìn)行克隆測序分析并構(gòu)建遺傳進(jìn)化樹,進(jìn)行系統(tǒng)發(fā)育分析。4.收集溫泉縣動物血清、組織樣本以及采集點的蚤類樣本,采用間接血凝法對血清樣本進(jìn)行鼠疫血清學(xué)F1抗體監(jiān)測,采用PCR檢測技術(shù)對組織樣本進(jìn)行鼠疫(caf1和pla)篩查并且對陽性樣本進(jìn)行MLVA基因型的分析。結(jié)果:1.2015-2016年共收集蚤類樣本5699匹,隸屬于7科16屬19種,分別為:長吻角頭蚤、人蚤、同型客蚤指名亞種、臀突客蚤、印鼠客蚤、貓櫛首蚤指名亞種、葉狀切唇蚤突高亞種、寬新蚤、修長櫛眼蚤指名亞種、寬臂纖蚤、八櫛蝠蚤、林野細(xì)蚤、真兇中蚤精河亞種、似升額蚤指名亞種、后彎怪蚤、謝氏山蚤、方形黃鼠蚤七河亞種、禿病蚤指名亞種和花蠕形蚤;阿拉山口地區(qū)鼠體外寄生蚤季節(jié)變化顯示,蚤類除1-2月份蚤指數(shù)低,其余10個月蚤指數(shù)較高且呈現(xiàn)一定的波動性。2.阿拉山口地區(qū)5種蚤中共測得726626條Tags被分成5447個OTUs,分析發(fā)現(xiàn)絕大多數(shù)的細(xì)菌分屬于4個細(xì)菌門:放線菌、擬桿菌、厚壁菌和變形菌,豐度呈現(xiàn)明顯差異性。3.立克次體:民豐縣的花蠕形蚤中檢測到暫定的巴貝瑞立克次體(Candidatus Rickettsia barbariae);阿拉山口地區(qū)的臀突客蚤、長吻角頭蚤和后彎怪蚤中檢測到與貝利立克次體(R.bellii)親緣性較高的立克次體,在寬臂纖蚤和禿病蚤指名亞種檢測到更為古老的立克次體。沃爾巴克體:在人蚤、客蚤、貓櫛首蚤指名亞種、似升額蚤、后彎怪蚤、謝氏山蚤、方形黃鼠蚤七河亞種和禿病蚤指名亞種中均檢測到,分子學(xué)分析顯示該內(nèi)共生體與以往在蚤內(nèi)報道存在較大差異性。巴通體:在阿拉山口地區(qū)的5種及溫泉與精河地區(qū)的3種蚤類中檢測到巴通體:1)臀突客蚤和同型客蚤指名亞種中檢測到Bartonella rochalimae;2)在臀突客蚤中檢測到格拉范姆(B.grahamii);3)葉狀切唇蚤突高亞種、后彎怪蚤和禿病蚤指名亞種檢測到伊麗莎白(B.elizabethae);4)來自溫泉縣與精河縣長尾黃鼠體外的似升額蚤和方形黃鼠蚤七河亞種中BZ01與來自溫泉縣旱獺體外的謝氏山蚤中的巴通體BZ02聚為一支且都與B.washoensis聚為一大支。4.2015年溫泉縣收集的血清學(xué)陽性率分別為灰旱獺2.1%(1/47)、牧羊犬6.7%(2/30)、長尾黃鼠0.7%(1/134);分子學(xué)檢測發(fā)現(xiàn)僅在溫泉縣的謝氏山蚤及灰旱獺臟器中檢測到鼠疫核酸陽性,且MLVA類型均為MT86(2-2-2-4-6-8-7-5-2-7-3-3-2-5)。結(jié)論:1.采集的19種蚤,葉狀切唇蚤突高亞種、修長櫛眼蚤指名亞種、寬臂纖蚤、八櫛蝠蚤、真兇中蚤精河亞種、似升額蚤指名亞種、后彎怪蚤、謝氏山蚤、方形黃鼠蚤七河亞種和花蠕形蚤5對基因信息及長吻角頭蚤(28S r DNA,COII,EF1-a,COI)、臀突客蚤(18S r DNA,28S r DNA和EF1-a)和禿病蚤指名亞種(18S r DNA,28S r DNA和COI)均為首次上傳Gen Bank,豐富了數(shù)據(jù)庫的信息并為以后的研究提供便利。2.宏細(xì)菌研究發(fā)現(xiàn)蚤類菌群中存在大量致病菌和條件致病菌屬,提示我們在后續(xù)的研究調(diào)查中,有針對地篩查該地區(qū)的致病性病原,為該地區(qū)風(fēng)險作出評估與預(yù)警。3.在蚤種和地區(qū)上,蚤類的立克次體、沃爾巴克體和巴通體的檢測均為首次發(fā)現(xiàn),并且巴通體均為致病性,提示在這些地區(qū)和蚤種中應(yīng)引起高度重視。4.在阿拉套山和別珍套山又發(fā)現(xiàn)了2個新的鼠疫陽性監(jiān)測點,顯示該區(qū)鼠疫疫情仍在活躍,其范圍有擴(kuò)大的趨勢,應(yīng)高度警惕其向人間的傳播。
[Abstract]:Objective: to identify and detect the fleas (including some of the southern Xinjiang fleas) in the northern border area of Xinjiang, and to lay the foundation for the risk assessment and prevention of flea disease in Xinjiang. Besides, the plague screening and typing of all the fleas in hot spring county plague foci were screened and classified to trace the main plague. To provide the scientific basis for the host, the media and the biological types of the ecological landscape. Methods: in 1.2015-2016, the parasitic fleas of mammalian body were collected in the northern frontier region of Xinjiang (Buerjin County, Ara Shankou City, hot spring county, Jinghe County, Huo Cheng County, Ca Bbu Charles Sieber autonomous County and Xinyuan county). And Molecular Biology (18S R DNA, 28S R DNA, EF1-a, COII and COI), the fleas were identified and analyzed by gene phylogenetic analysis..2. selected fleas, fleas, fleas, leaflike fleas, subspecies of fleas, and subspecies of bald fleas, DNA sent to the biologic information department of NOA, Beijing. Technology limited, sequencing the V3-V4 variable region in the sequence of the bacterial 16S R DNA gene, analyzed the community structure of the bacteria, the population diversity and the richness.3. combined with the macrobacterial data, and screened the flea samples for the rickettsia (MLST), the Wolbachia (WSP and 16S R RNA) and the Bartonella (GLTA, ITS, and excluded). The phylogenetic analysis and phylogenetic tree were constructed, and phylogenetic analysis.4. collected animal serum, tissue samples and flea samples in hot spring county. The serological F1 antibody was monitored by indirect hemagglutination, and the PCR detection technique was used to screen the plague (Caf1 and PLA) and to the sun. Analysis of the MLVA genotype of sex samples. Results: 5699 samples of fleas were collected in 1.2015-2016, belonging to 19 species of 16 genera and 19 species of fleas, the fleas, fleas of the same type, the fleas of the gluteus, the fleas of the cat, the subspecies of the fleas, the wide new fleas, the name subspecies of the long Chlamys, and the wide brachial fleas. Eight Daphnia fleas, wild fleas, Jinghe subspecies of fleas, similar to subspecies of fleas, fleas, fleas, squirrel fleas, seven subspecies of squirrel fleas, bald fleas name subspecies and flower permiform fleas; the seasonal changes of fleas in the ARA mountain mouth area mice in vitro parasitical fleas show that the fleas index is low in 1-2 months, and the other 10 months flea index is higher and presents a certain degree. 5 species of fleas of.2. in aramashi region were divided into 5447 OTUs. The analysis found that most of the bacteria were divided into 4 bacterial Gates: actinomycetes, bacteriobacteria, pamacomycetes and deformable bacteria, which showed significant difference in abundance of.3. Rickettsia. In Minfeng County, the temporary Barber Ray Ricketts Ti (Candi) was detected in the Daphnia flea of Minfeng County. Datus Rickettsia barbariae); the Rickettsia with higher affinity with the Bailey rickettsia (R.bellii) was detected in the fleas of the buttocks and fleas of the ARA Yamaguchi, the fleas of the rickettsia (R.bellii), and the older Rickettsia in the subspecies of the wide brachial fleas and the bald fleas. The name of the subspecies of the fleas, fleas, and cat fleas was identified. It was found that the fleas, the fleas, the fleas, the fleas, the seven subspecies of the squirrel fleas, and the bald fleas were all detected. The subspecies analysis showed that the intersymbionts were significantly different from those in the flea. Bartonella was detected in the 5 species of the ARA mountain mouth and 3 of the fleas in the Jinghe region: 1) gluteus. Bartonella rochalimae; 2) detected in the subspecies of fleas and the same species of fleas; 2) detected in the fleas of gluteus gluteus (B.grahamii); 3) the subspecies of the leaf shaped labial fleas, and the subspecies of the strange fleas and the bald fleas were detected by the subspecies of Elizabeth (B.elizabethae); 4) came from the in vitro fleas and squirrel squirrels of wequan county and Jinghe. The BZ01 in the seven River subspecies of flea and the Barton body BZ02 from the flea flea from the outside of the hot spring county were gathered into one branch and all were gathered with B.washoensis to be a large branch of.4.2015 year hot spring county. The serological positive rates were 2.1% (1/47), 6.7% (2/30), and 0.7% (1/134) of the long tailed rat, respectively. In the viscera of mountain fleas and Marmota Marmota, the positive of Yersinia pestis were detected and MLVA type was MT86 (2-2-2-4-6-8-7-5-2-7-3-3-2-5). Conclusion: 1. species of fleas, 19 species of fleas, leaflike fleas, fleas, fleas, eight fleas, the subspecies of fleas, the fleas, the fleas, the fleas of Mount Xie, square The 5 pairs of genetic information and the 28S R DNA, COII, EF1-a, COI) of the seven subspecies of the rat flea and the fleas (COII, EF1-a, COI), and the first subspecies of the gluteus flea (18S R DNA, 28S R DNA) and the bald fleas, enrich the information of the data base and facilitate the future research to facilitate the research of macrobacteria. There are a large number of pathogenic and conditional pathogenic bacteria in the flea microflora. It is suggested that in the follow-up investigation, we have screened the pathogenic pathogenic pathogens in this area, and the risk assessment and early warning for the region,.3. in flea species and areas, flea Rickettsia, the walbaker body and Bartonella, and the first detection of the disease, and the bartlike body for the first time. All of them are pathogenicity, which suggests that in these areas and flea species, 2 new plague positive monitoring points have been found in Alan mountain and farewell Zhen Shan mountain. It shows that the epidemic situation of the plague is still active in this area, and the scope of the epidemic is expanding, and it should be highly vigilant for the spread of the.4..
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R384.3

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