新疆北疆边境地区蚤种的鉴定与病原检测

发布时间：2018-05-20 07:47

本文选题：蚤 + 鼠疫　；参考：《石河子大学》2017年硕士论文

【摘要】：目的:对新疆北疆边境地区蚤类(包括部分南疆蚤类)进行鉴定和病原检测,为新疆蚤传病的发生做出风险评估以及蚤传病的预防奠定基础。除此以外,对温泉县鼠疫疫源地所收集的样本及所有蚤类进行鼠疫筛查与分型,为追溯鼠疫的主要宿主、媒介及其生态地理景观的生物学类型提供科学依据。方法:1.2015-2016年在新疆北疆边境地区(布尔津县、阿拉山口市、温泉县、精河县、霍城县、察布查尔锡伯自治县和新源县)及和田地区民丰县收集哺乳动物体表寄生蚤;通过形态学及分子生物学(18S r DNA,28S r DNA,EF1-a,COII和COI),对所收集的蚤进行蚤种鉴定并进行基因系统发育分析。2.选取阿拉山口地区的长吻角头蚤、臀突客蚤、叶状切唇蚤突高亚种、后弯怪蚤及秃病蚤指名亚种组织DNA送至北京诺禾致远生物信息科技有限公司,对细菌16S r DNA基因序列中的V3-V4可变区进行测序,分析其细菌的群落结构、种群多样性及丰富度。3.结合宏细菌数据,对所采集蚤类样本进行立克次体(MLST)、沃尔巴克体(wsp和16S r RNA)以及巴通体(glta、ITS和rib C)的筛查,阳性片段进行克隆测序分析并构建遗传进化树,进行系统发育分析。4.收集温泉县动物血清、组织样本以及采集点的蚤类样本,采用间接血凝法对血清样本进行鼠疫血清学F1抗体监测,采用PCR检测技术对组织样本进行鼠疫(caf1和pla)筛查并且对阳性样本进行MLVA基因型的分析。结果:1.2015-2016年共收集蚤类样本5699匹,隶属于7科16属19种,分别为:长吻角头蚤、人蚤、同型客蚤指名亚种、臀突客蚤、印鼠客蚤、猫栉首蚤指名亚种、叶状切唇蚤突高亚种、宽新蚤、修长栉眼蚤指名亚种、宽臂纤蚤、八栉蝠蚤、林野细蚤、真凶中蚤精河亚种、似升额蚤指名亚种、后弯怪蚤、谢氏山蚤、方形黄鼠蚤七河亚种、秃病蚤指名亚种和花蠕形蚤;阿拉山口地区鼠体外寄生蚤季节变化显示,蚤类除1-2月份蚤指数低,其余10个月蚤指数较高且呈现一定的波动性。2.阿拉山口地区5种蚤中共测得726626条Tags被分成5447个OTUs,分析发现绝大多数的细菌分属于4个细菌门:放线菌、拟杆菌、厚壁菌和变形菌,丰度呈现明显差异性。3.立克次体:民丰县的花蠕形蚤中检测到暂定的巴贝瑞立克次体(Candidatus Rickettsia barbariae);阿拉山口地区的臀突客蚤、长吻角头蚤和后弯怪蚤中检测到与贝利立克次体(R.bellii)亲缘性较高的立克次体,在宽臂纤蚤和秃病蚤指名亚种检测到更为古老的立克次体。沃尔巴克体:在人蚤、客蚤、猫栉首蚤指名亚种、似升额蚤、后弯怪蚤、谢氏山蚤、方形黄鼠蚤七河亚种和秃病蚤指名亚种中均检测到,分子学分析显示该内共生体与以往在蚤内报道存在较大差异性。巴通体:在阿拉山口地区的5种及温泉与精河地区的3种蚤类中检测到巴通体:1)臀突客蚤和同型客蚤指名亚种中检测到Bartonella rochalimae;2)在臀突客蚤中检测到格拉范姆(B.grahamii);3)叶状切唇蚤突高亚种、后弯怪蚤和秃病蚤指名亚种检测到伊丽莎白(B.elizabethae);4)来自温泉县与精河县长尾黄鼠体外的似升额蚤和方形黄鼠蚤七河亚种中BZ01与来自温泉县旱獭体外的谢氏山蚤中的巴通体BZ02聚为一支且都与B.washoensis聚为一大支。4.2015年温泉县收集的血清学阳性率分别为灰旱獭2.1%(1/47)、牧羊犬6.7%(2/30)、长尾黄鼠0.7%(1/134);分子学检测发现仅在温泉县的谢氏山蚤及灰旱獭脏器中检测到鼠疫核酸阳性,且MLVA类型均为MT86(2-2-2-4-6-8-7-5-2-7-3-3-2-5)。结论:1.采集的19种蚤,叶状切唇蚤突高亚种、修长栉眼蚤指名亚种、宽臂纤蚤、八栉蝠蚤、真凶中蚤精河亚种、似升额蚤指名亚种、后弯怪蚤、谢氏山蚤、方形黄鼠蚤七河亚种和花蠕形蚤5对基因信息及长吻角头蚤(28S r DNA,COII,EF1-a,COI)、臀突客蚤(18S r DNA,28S r DNA和EF1-a)和秃病蚤指名亚种(18S r DNA,28S r DNA和COI)均为首次上传Gen Bank,丰富了数据库的信息并为以后的研究提供便利。2.宏细菌研究发现蚤类菌群中存在大量致病菌和条件致病菌属,提示我们在后续的研究调查中,有针对地筛查该地区的致病性病原,为该地区风险作出评估与预警。3.在蚤种和地区上,蚤类的立克次体、沃尔巴克体和巴通体的检测均为首次发现,并且巴通体均为致病性,提示在这些地区和蚤种中应引起高度重视。4.在阿拉套山和别珍套山又发现了2个新的鼠疫阳性监测点,显示该区鼠疫疫情仍在活跃,其范围有扩大的趋势,应高度警惕其向人间的传播。
[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..
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R384.3

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