本文關(guān)鍵詞：貴州矮馬主要傳染性疾病的血清學(xué)和病原學(xué)研究　出處：《貴州大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 貴州矮馬 西南馬 馬泰勒蟲(chóng) 馬駑巴貝斯蟲(chóng) 馬傳染性貧血病毒 馬動(dòng)脈炎病毒 馬皰疹病毒I

【摘要】：貴州矮馬是目前全球僅存的少數(shù)天然矮馬品種之一,是在自然生態(tài)環(huán)境和人工選擇的雙重影響下形成的珍貴遺傳資源,主要分布在黔西南一帶的偏遠(yuǎn)山區(qū),其中以安順市紫云縣火花鄉(xiāng)的矮馬最為著名。2013年間,紫云縣的馬匹出現(xiàn)大量死亡現(xiàn)象,尤其是以懂桑村養(yǎng)殖的貴州矮馬死亡率最高,發(fā)病率70.1%,死亡率60%,死亡病因不明。針對(duì)病馬臨床癥狀的分析,懷疑可能是馬泰勒蟲(chóng)(Theileria equi)、馬駑巴貝斯蟲(chóng)(Babesia caballi)、馬傳染性貧血病毒(Equine Infectious Anemia virus,EIAV)、馬動(dòng)脈炎病毒(Equine arteritis virus,EAV)或者馬皰疹病毒I型(Equine herpesvirus type-1,EHV-1)所引發(fā)的傳染性疾病。本研究采用血液涂片鏡檢、酶聯(lián)免疫吸附實(shí)驗(yàn)(Enzyme-linked immunosorbent assay,ELISA)和聚合酶鏈?zhǔn)椒磻?yīng)(Polymerase Chain Reaction,PCR)的方法對(duì)可疑的病原進(jìn)行血清學(xué)和分子病原學(xué)研究,以期找到貴州矮馬大量死亡的真正病因。采用競(jìng)爭(zhēng)性酶聯(lián)免疫吸附試驗(yàn)(cELISA)和PCR的方法對(duì)馬泰勒蟲(chóng)病和馬駑巴貝斯蟲(chóng)病的血清學(xué)和分子病原學(xué)進(jìn)行研究,結(jié)果表明,cELISA方法檢測(cè)貴州矮馬、西南馬和伊犁馬血漿樣品中馬泰勒蟲(chóng)抗體陽(yáng)性率分別為23.86%、11.43%、31.71%,而只有伊犁馬樣品中檢測(cè)到馬駑巴貝斯蟲(chóng)抗體,陽(yáng)性率為85.37%。三個(gè)馬群中均檢測(cè)到馬泰勒蟲(chóng)和馬駑巴貝斯蟲(chóng)的18s rRNA基因,貴州矮馬、西南馬和伊犁馬血液樣品中馬泰勒蟲(chóng)18s rRNA基因的檢出率分別為78.41%、62.86%、68.29%,而馬駑巴貝斯蟲(chóng)的18s rRNA基因檢出率分別為11.36%、11.43%、90.24%。同時(shí),對(duì)新鮮血液樣品制作血液涂片,經(jīng)顯微鏡觀察,發(fā)現(xiàn)馬泰勒蟲(chóng)和馬駑巴貝斯蟲(chóng)蟲(chóng)體的存在,其大小及形態(tài)特征與文獻(xiàn)中的描述相符。PCR和cELISA兩種方法的檢測(cè)結(jié)果存在一定的差異,PCR的陽(yáng)性率普遍高于cELISA的陽(yáng)性率,這可能是由于PCR方法檢測(cè)靈敏度較高,另外,可能是由于采樣時(shí)部分馬匹正處于感染的早期階段,還沒(méi)有抗體的產(chǎn)生。有些樣品的cELISA檢測(cè)結(jié)果為陽(yáng)性,但PCR方法檢測(cè)卻為陰性,這可能是由于馬匹體內(nèi)的馬泰勒蟲(chóng)蟲(chóng)體已被自身的吞噬系統(tǒng)消除,但其抗體還會(huì)在體內(nèi)存在一段時(shí)間。有部分樣品cELISA檢測(cè)結(jié)果為陰性,但PCR方法檢測(cè)卻為陽(yáng)性,可能是因?yàn)榧闹鲃偙桓腥?或者是在抗體消失后又出現(xiàn)了新的感染。以貴州矮馬、西南馬和伊犁馬的血清為材料,采用ELISA的方法對(duì)EIAV、EAV和EHV-1進(jìn)行血清學(xué)研究,3個(gè)馬群中均未檢測(cè)到EIAV抗體陽(yáng)性樣品;EAV的陽(yáng)性率為48.08%、22.86%、10.81%;EHV-1的陽(yáng)性率為99.38%、97.14%、100%。對(duì)EHV-1的DNA進(jìn)行PCR擴(kuò)增,貴州矮馬、西南馬和伊犁馬EHV-1的檢出率分別為94.23%、80%、83.78%。由于馬傳貧與馬泰勒蟲(chóng)病的臨床癥狀比較相似,所以EIAV抗體陰性的結(jié)果為馬泰勒蟲(chóng)病的確診提供了理論依據(jù)。EHV-1的陽(yáng)性率很高,但是其所導(dǎo)致的疾病死亡率并不高,可能一種繼發(fā)性感染。貴州矮馬EAV血清學(xué)陽(yáng)性率較西南馬和伊犁馬要高,而且馬病毒性動(dòng)脈炎的死亡率較高,表明EAV可能是導(dǎo)致貴州矮馬大量死亡的病原之一。研究結(jié)果表明,馬泰勒蟲(chóng)病和馬病毒性動(dòng)脈炎可能是導(dǎo)致貴州矮馬大量死亡的主要原因。對(duì)尚存活的患病馬匹根據(jù)其臨床癥狀施以伊維菌素、血蟲(chóng)迪以及附紅二合一藥物進(jìn)行及時(shí)的治療,并同時(shí)施用抗生素藥物對(duì)繼發(fā)性感染進(jìn)行治療,直至馬匹痊愈,最終有效的降低了當(dāng)?shù)伛R群的死亡率。綜上,本研究應(yīng)用顯微鏡、ELISA和PCR的方法對(duì)貴州矮馬主要傳染性疾病的血清學(xué)和病原學(xué)的研究,不僅減少了當(dāng)?shù)剞r(nóng)戶的經(jīng)濟(jì)損失,避免了貴州矮馬這一珍貴品種即將滅絕噩夢(mèng)的發(fā)生,更是為當(dāng)?shù)伛R屬動(dòng)物疾病的診斷、治療和預(yù)防提供了可靠的理論依據(jù)。
[Abstract]:Guizhou pony pony is one of the few natural varieties currently the world's only, is the precious genetic resources formed in the double influence of natural ecological environment and artificial selection in the remote mountainous areas are mainly distributed in the southwest area, Ziyun County of Anshun City, which sparks township is the most famous pony. 2013 years, Ziyun County horses appear a large number of death phenomenon, especially to understand the Guizhou pony village of mulberry breeding the highest mortality rate, the incidence rate was 70.1%. The mortality rate was 60%, the death of unknown etiology. According to the analysis of the clinical symptoms of the disease of horses, is suspected of theileriaequi (Theileria equi), BABEI, Ma nu (Babesia, caballi) from equine infectious anemia virus (Equine Infectious Anemia virus, EIAV), equine arteritis virus (Equine arteritis virus, EAV) or equine herpes virus type I (Equine herpesvirus type-1, EHV-1) caused by infectious diseases. This study used blood smear microscopy, enzyme-linked immunosorbent assay (Enzyme-linked immunosorbent, assay, ELISA) and polymerase chain reaction (Polymerase Chain Reaction, PCR) method of serological and molecular pathogenic suspicious pathogen research, in order to find the real cause of death of a large number of Guizhou pony. By competitive enzyme-linked immunosorbent assay (cELISA) for Martel and BABEI, jade horse worm disease worm disease serology and molecular etiology research methods and results of PCR showed that the cELISA method for the detection of Guizhou pony, southwest Martel horse and Yili horse in plasma samples of antibody positive rate were 23.86%, 11.43%, 31.71% but, only Yili horse was detected in the samples of BABEI, Ma Nu antibody positive rate was 85.37%. Three horses were detected in 18S rRNA gene of theileriaequi horse and jade BABEI worm, Guizhou pony, southwest Mataile horse and Yili horse blood samples from 18S rRNA gene were detected in 78.41%, 62.86%, 68.29%, and 18S rRNA gene, insect jade horse BABEI detection rate was 11.36%, respectively. 11.43%, 90.24%. At the same time, the fresh blood samples making blood smears, through a microscope observation, found of theileriaequi and jade horse BABEI worm body exists, its size and morphology and the literature description. There are some differences between the results of PCR and cELISA of the two methods, the positive rate of PCR is higher than cELISA, this may be because the PCR method has higher detection sensitivity, in addition, may be due to sampling when some horses are in the early stages of infection, no antibodies. CELISA test results of some samples were positive, but PCR detection is negative, it may be because Martel purpose of horses in vivo has been their phagocytic system eliminated, but the antibody will continue to exist for some time in the body. Some samples were negative for cELISA test, but PCR method was positive, probably because the host was just infected, or after the antibody disappeared, there was a new infection. In Guizhou, Yili and southwest horse pony horse serum as the material, uses the method of ELISA serological research on EIAV, EAV and EHV-1, the 3 horses were not detected in EIAV antibody positive samples; the positive rate of EAV was 48.08%, 22.86%, 10.81%; the positive rate of EHV-1 was 99.38%, 97.14%, 100%. The EHV-1 DNA was amplified by PCR, the Guizhou pony, southwest horse and Yili horse EHV-1 detection rates were 94.23%, 80%, 83.78%. As the clinical symptoms of horse transmission and Martel's disease are similar, the results of EIAV antibody negative provide a theoretical basis for the diagnosis of Martel's disease. The positive rate of EHV-1 is high, but the mortality of the disease is not high, and it may be a secondary infection. The positive rate of serum EAV is southwest of Guizhou Pony Ma and Yili horse must be high, and equine viral arteritis with high mortality, suggest that EAV may be one of the causes of death of a large number of pathogenic Guizhou pony. The results show that the Martel worm disease and equine viral arteritis was likely to cause the death of a large number of Guizhou pony. For the remaining sick horses according to their clinical symptoms with ivermectin, blood worms and drug combo with red Di timely treatment and application of antibiotics for the treatment of secondary infection, until the horse recovered, eventually reduce the death rate of local horses. In summary, this study used microscope and ELISA and PCR on the main Guizhou pony infectious disease serology and etiology, not only reduces the farmers' economic loss, avoid the Guizhou pony this precious varieties will be extinct nightmare, is when the diagnosis, equine animal diseases provide theory a reliable basis for the prevention and treatment of.
【學(xué)位授予單位】：貴州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S858.21

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