本文選題：蜱媒傳染病 + 伯氏疏螺旋體��； 參考：《中國人民解放軍軍事醫(yī)學科學院》2017年碩士論文

【摘要】：一.研究背景蜱媒傳染病(Tick-borne infectious diseases)是一類由媒介蜱傳播、感染所引發(fā)的自然疫源性疾病。蜱攜帶和傳播的病原體種類數量繁多(包括病毒、細菌、立克次體、螺旋體等)、宿主動物分布廣泛、流行環(huán)節(jié)復雜交錯,且易受到自然環(huán)境和社會經濟條件的影響,幾種蜱傳疾病往往共存于同一疫源地,一種媒介蜱也可同時攜帶兩種或兩種以上的病原體,這極大的增加了宿主動物及人類復合感染的機率,極易造成蜱傳疾病的流行和暴發(fā)。自1982年以來,我國已發(fā)現33種新發(fā)蜱媒傳染病病原體[1-7],包括3種無形體(Anaplasma)、3種埃里克體(Ehrlichia)、8種斑點熱群立克次體(spotted fever group rickettsiae,SFGR)、6種伯氏疏螺旋體(Borrelia burgdorferi sensu lato)、11種巴貝西蟲(Babesia)及發(fā)熱伴血小板減少綜合征病毒(severe fever with thrombocytopenia syndrome virus,SFTSV),其中15種新發(fā)蜱傳病原體可以導致人類感染(新發(fā)蜱媒傳染病第一時間被確認的地點和時間,附錄1[82])。萊姆病(Lyme disease,LD)是20世紀70年代發(fā)現的由伯氏疏螺旋體(Borrelia burgdorferi sensu lato,Bb)引起的一種蜱媒傳染病。現報告已有70多個國家和地區(qū)存在本病或自然疫源地,在美國萊姆病又被稱作“第二艾滋病”,世界衛(wèi)生組織已將其列為需要重點進行研究和防治的新發(fā)傳染病。我國自1986年發(fā)現萊姆病自然疫源地以來,通過病原學檢測已證實有19個省、市和自治區(qū)存在萊姆病的自然疫源地,9種蜱可攜帶和傳播伯氏疏螺旋體,8種野鼠、狗、牛、羊和野兔體內存在自然感染。近年來,我國萊姆病患者每年都有上萬余例報道,其危害性和嚴重性日益受到人們關注。斑點熱是由立克次體屬(Rickettsia)的斑點熱群立克次體(spotted fever group Rickettsiae,SFGR)引起的一類以急性發(fā)熱和全身皮疹為主要特征的疾病總稱。SFGR是立克次體目中最大最復雜的一個群種,是一種全球性分布、嚴格血管內皮細胞寄生的小桿菌,革蘭氏染色陰性,到目前為止能引起人類感染的SFGR至少19種。我國已在10多個省、市、自治區(qū)證實有SFGR感染,并且發(fā)病地區(qū)及范圍不斷擴大和變遷。近年來,新種SFGR及其所引致的斑點熱在我國和世界各地不斷發(fā)現和報道。內蒙大興安嶺林區(qū)自然資源豐富,生態(tài)系統(tǒng)物種多樣,并以其獨特的地理優(yōu)勢和自然景觀適宜蜱類棲息,是我國蜱媒傳染病主要疫區(qū)之一[8],隨著林區(qū)天然林保護工程實施和產業(yè)結構調整,林下資源開發(fā)逐漸增多,森林采伐量逐年減少,帶動當地經濟不斷發(fā)展的同時,林區(qū)旅游業(yè)也迅速升溫。隨著進入林區(qū)人群的增加,人們被蜱蟲叮咬、感染的機率就不斷增多,發(fā)病率也越來越來高。因此,蜱媒傳染病對在內蒙古林區(qū)從事經濟開發(fā)、野外生產活動、旅游、野營、軍事訓練以及國防建設的人群威脅極大。二.研究目的通過本研究進一步了解我國內蒙古大興安嶺林區(qū)蜱中疏螺旋體和SFGR感染情況,補充和完善媒介蜱攜帶病原體的多樣性和復雜性,為更好的防治蜱媒傳染病提供可以借鑒的新思路。三.研究方法本研究選取內蒙古畢拉河林區(qū)(北緯N49°25′48.52″,東經E123°32′15.82″)、庫都爾林區(qū)(北緯N50°01′35.64″,東經E121°36′58.73″)和奇乾林區(qū)(北緯N52°11′45.30″,東經E120°46′21.42″)為調查點,廣泛采集全溝硬蜱(Ixodes persulcatus),森林革蜱(Dermacentor silvarum)和嗜群血蜱(Haemaphysalis conicinna)等多種蜱種,采用聚合酶鏈式反應(PCR)方法對疏螺旋體和SFGR感染及其基因分型進行研究。四.研究結果本研究我們在畢拉河、庫都爾和奇乾三個林區(qū)共采集蜱標本792只,其中全溝硬蜱723只(91.29%)、森林革蜱48只(6.06%),嗜群血蜱21只(2.65%)。庫都爾林區(qū)和奇乾林區(qū)均以全溝硬蜱為優(yōu)勢蜱種(c2=202.405,p=0.000),提示全溝硬蜱是該林區(qū)蜱媒傳染病的主要傳播媒介之一。PCR檢測發(fā)現,蜱標本中疏螺旋體陽性攜帶率為35.35%,全溝硬蜱、森林革蜱、嗜群血蜱中攜帶疏螺旋體感染分別為276只(38.17%)、2只(4.17%)、2只(9.52%);基因分型結果顯示有5種基因型別:B.garinii型,陽性蜱212只(陽性率26.77%);B.afzelii型,陽性蜱53只(陽性率6.69%);B.miyamotoi型,陽性蜱13只(陽性率1.64%);B.spielmanii型,陽性蜱1只(陽性率0.12%);Borrelia sp.NM517(B.sp.NM517)型,陽性蜱1只(陽性率為0.12%)。其中B.miyamotoi、B.spielmanii和B.sp.NM517三種基因型均為我國首次發(fā)現。經統(tǒng)計學分析表明B.garinii為我國大興安嶺林區(qū)萊姆病伯氏疏螺旋體優(yōu)勢基因型(c2=19.433,p=0.013)。另外,結果發(fā)現蜱標本中SFGR陽性攜帶率為40.4%,全溝硬蜱、森林革蜱、嗜群血蜱中SFGR感染分別為283只(39.14%)、28只(58.33%)、9只(42.86%),基因分型結果顯示有3種基因型:新塔拉塞維奇立克次體(Candidatus Rickettsia tarasevichiae,CRT),陽性蜱284只(陽性率35.86%);勞氏立克次體(Rickettsia raoultii,R.raoultii),陽性蜱33只(陽性率4.17%);黑龍江立克次體(Rickettsiahelongjiangii,R.helongjiangii),陽性蜱3只(陽性率0.38%)。經統(tǒng)計學檢驗,內蒙古大興安嶺三個林區(qū)SFGR不同基因型陽性感染差異存在統(tǒng)計學意義(c2=82.474,p=0.000),庫都爾林區(qū)和奇乾林區(qū)均以CRT為SFGR優(yōu)勢基因型,畢拉河林區(qū)以R.raoultii為優(yōu)勢基因型。經統(tǒng)計學分析顯示,三個蜱種不同基因型SFGR的感染差異存在統(tǒng)計學意義(c2=251.838,p=0.000),全溝硬蜱和嗜群血蜱中CRT基因型為蜱攜SFGR優(yōu)勢基因型,森林革蜱中R.raoultii為蜱攜SFGR優(yōu)勢基因型。研究也發(fā)現媒介蜱存在疏螺旋體和SFGR的復合感染,復合感染率為14.52%(115只)。五.研究結論本研究結合現場調查、實驗室檢測,綜合應用描述性和分析性流行病學等方法,對內蒙古大興安嶺林區(qū)媒介蜱中攜帶病原體的感染情況進行了深入調查,調查發(fā)現全溝硬蜱為當地優(yōu)勢蜱種。蜱中可單獨或混合存在疏螺旋體(B.garinii型、B.afzelii型、B.miyamotoi、B.spielmanii、B.sp.NM517)和SFGR(CRT型、R.raoultii型、R.heilongjiangii型)感染。本研究首次在我國發(fā)現了疏螺旋體B.miyamotoi、B.spielmanii和B.sp.NM517三個基因型,其中B.miyamotoi屬于疏螺旋體屬中的回歸熱螺旋體,主要寄生在硬蜱中,常與B.afzelii、B.garinii等基因型在蜱中混合感染。B.spielmanii屬于疏螺旋體屬中的伯氏疏螺旋體,是新型人類致病性病原體。B.sp.NM517型序列同B.Theileri strain KAT 16s ribosomal RNA的序列相似度為98.77%,是一種新的基因型,但還未對其進行分離培養(yǎng),這有待于進一步深入研究。本研究結果不僅為我國內蒙古大興安嶺林區(qū)媒介蜱攜帶病原體的多樣性起到了補充和完善的作用,同時也可為更好的防治蜱媒傳染病提供可以借鑒的新思路。
[Abstract]:Tick-borne infectious diseases is a natural epidemic disease caused by the transmission and infection of ticks. The number of pathogens carried and transmitted by ticks is numerous (including viruses, bacteria, Rickettsia, spirals, etc.), the host is widely distributed, the epidemic is complicated and interlaced, and is vulnerable to natural rings. Several tick borne diseases often coexist in the same source area, and a medium tick can also carry two or more than two pathogens at the same time, which greatly increases the probability of complex infection between host animals and human beings, which is very easy to cause the epidemic and outbreak of tick borne diseases. Since 1982, China has found 33 new types of new diseases. The tick borne infectious disease pathogen [1-7], including 3 kinds of intangibles (Anaplasma), 3 kinds of Eric body (Ehrlichia), 8 kinds of spotted fever group rickettsiae, SFGR, 6 kinds of Borrelia (Borrelia burgdorferi sensu lato), 11 kinds of BABEI and fever accompanied thrombocytopenia syndrome virus Th thrombocytopenia syndrome virus, SFTSV), of which 15 new tick borne pathogens can cause human infection (the location and time identified for the first time of the new tick infectious disease, appendix 1[82]). Lyme disease (Lyme disease, LD) was found in 1970s by a species of Borrelia burgdorferi (Borrelia burgdorferi sensu). Tick borne infectious diseases have been reported in more than 70 countries and regions. In the United States, Lyme disease is also known as "second AIDS". The WHO has listed it as a new infectious disease which needs to be studied and controlled. Since the discovery of the natural epidemic of Lyme disease in 1986, it has been tested by pathogeny. There are 19 provinces, cities and autonomous regions have natural foci of Lyme disease, 9 ticks can carry and spread Borrelia burgdorferi, 8 kinds of wild mice, dogs, cattle, sheep and rabbits have natural infection. In recent years, there are more than 10000 cases of Lyme disease patients in China each year, and their harmfulness and severity are increasingly concerned. Rickettsia (spotted fever group Rickettsiae, SFGR), a group of diseases characterized by acute fever and systemic rash, is the largest and most complex group in the rickettsia, a global distribution, microbacilli with strict vascular endothelial cells, and gram dye. Color negative, at least 19 species of SFGR can cause human infection so far. China has been in more than 10 provinces, municipalities and autonomous regions confirmed SFGR infection, and the area and scope of the disease has been expanding and changing. In recent years, new species of SFGR and its spotted fever have been found and reported in China and all over the world. Natural capital in the Greater Khingan Range forest area of Inner Mongolia. Rich in source and diversity of ecosystem species, and with its unique geographical advantage and natural landscape suitable for tick species, it is one of the main epidemic areas of tick borne diseases in China, [8]. With the implementation of the forest protection project and the adjustment of industrial structure, the development of the forest resources is gradually increasing, the forest harvesting is decreasing year by year, and the local economy is developing continuously. At the same time, the tourism industry in the forest area has also increased rapidly. As people enter the forest area, people are bitten by ticks, and the incidence of infection is increasing and the incidence is increasing. Therefore, tick borne infectious diseases are a great threat to people engaged in economic development, field production, tourism, camping, military training and national defense construction in the Inner Mongolia forest area. Two. The purpose of this study is to further understand the infection of sparsely spirals and SFGR in Greater Khingan Range forest ticks in Inner Mongolia, to supplement and improve the diversity and complexity of vector borne pathogens, and to provide new ideas for better prevention and control of tick borne infectious diseases. Three. The research method selected the Inner Mongolia Biira forest The area (north latitude N49 25 '48.52 ", east longitude E123 32' 15.82"), Qulu forest region (north latitude N50 / 01 '35.64 ", East Jing E121 36' 58.73") and Qi Qian forest region (north latitude N52 11 '45.30 "), E120 degree 46' 21.42") as investigation points, widely collect whole ditch Ixodes (Ixodes persulcatus), chlorpyrifos (Dermacentor silvarum) and eosinophilia (H) Aemaphysalis conicinna) and other ticks, using polymerase chain reaction (PCR) method to study the infection and genotyping of Treponema and SFGR. Four. We collected 792 tick specimens in three forest areas of the Biira River, koudar and Qi Qian, of which 723 (91.29%), 48 (6.06%), three (6.06%). 21 (2.65%) of the blood ticks (c2=202.405, p=0.000) in the coudine forest area and Qi Qian forest area, suggesting that the whole trench tick is one of the main vectors of the tick borne infectious disease in the forest area. The positive rate of the spiral body in the ticks is 35.35%, the whole furrow tick, the leeticos, and the eosinophilic tick carry the sparse helix. The body infection was 276 (38.17%), 2 (4.17%) and 2 (9.52%). The genotyping results showed that there were 5 genotypes: B.garinii type, 212 positive ticks (positive rate 26.77%); B.afzelii, positive ticks 53 (positive rate 6.69%); B.miyamotoi type, positive ticks 13 (positive rate 1.64%); B.spielmanii, positive ticks (positive rates); Borrelia sp.NM51 7 (B.sp.NM517) and 1 positive ticks (positive rate 0.12%). Among them, three genotypes of B.miyamotoi, B.spielmanii and B.sp.NM517 were found for the first time in China. The statistical analysis showed that B.garinii was the dominant genotype of Borrelia burgdorferi (c2= 19.433, p=0.013) in Greater Khingan Range forest region of China. In addition, it was found that SFGR positive was found in the ticks. The rate of SFGR was 40.4%, 283 (39.14%), 28 (58.33%) and 9 (42.86%) infected ticks of ticks, 28 (58.33%) and 9 (42.86%). The results of genotyping showed that new Tara Savage Ricketts Ti (Candidatus Rickettsia tarasevichiae, CRT), positive ticks 284 (positive rate 35.86%); Rickettsia Lloyd (Rickettsia Raoult). II, R.raoultii), 33 positive ticks (positive rate 4.17%); Heilongjiang rickettsia (Rickettsiahelongjiangii, R.helongjiangii), 3 positive ticks (positive rate 0.38%). By statistical test, the difference of positive infection of different genotypes of SFGR in three forest areas in Greater Khingan Range of Inner Mongolia was statistically significant (c2=82.474, p=0.000), and in the region of Hulu forest and Qi Qian forest area CRT was the dominant genotype of SFGR, and R.raoultii was the dominant genotype in the Biya river forest region. The statistical analysis showed that the difference of infection between three ticks of different genotypes was statistically significant (c2=251.838, p=0.000). The CRT genotypes of ticks and eosinophilic ticks were ticks carrying SFGR dominant genotypes, and R.raoultii was tick carrying SF. GR dominant genotypes. The study also found that the compound infection of Treponema and SFGR was found in the medium ticks, and the rate of compound infection was 14.52% (115). Five. Conclusion this study combined with field investigation, laboratory testing, comprehensive application of descriptive and analytical epidemiology to carry out the infection of pathogens in Greater Khingan Range forest area of Inner Mongolia. The survey found that the whole ditch was a local dominant tick. The ticks could be isolated or mixed in the ticks (B.garinii, B.afzelii, B.miyamotoi, B.spielmanii, B.sp.NM517) and SFGR (CRT, R.raoultii, R.heilongjiangii). This study was first found in our country for B.miyamotoi, B.spiel. Three genotypes of manii and B.sp.NM517, of which B.miyamotoi belongs to the regression heat spirals in the genus heliospira, mainly parasitic in ticks, often mixed with B.afzelii, B.garinii and other genotypes in ticks and.B.spielmanii belongs to burgospira Borrelia, a new human pathogenic pathogen.B.sp.NM517 sequence and B.Th. The sequence similarity of eileri strain KAT 16S ribosomal RNA is 98.77%, which is a new genotype, but it has not been isolated and cultured. It needs to be further studied. The results of this study not only complement and improve the diversity of the pathogen of the media ticks in the Greater Khingan Range forest area of Inner Mongolia, but also can be used as a result. Better prevention and treatment of tick borne infectious diseases can provide new ideas for reference.
【學位授予單位】：中國人民解放軍軍事醫(yī)學科學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R181.3;R53

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