【摘要】：禽白血病病毒(Avian Leukosis Virus,ALV)自首次被報道以來已在世界各地爆發(fā)和流行,因其高致病性、高致死率、高腫瘤并發(fā)率給各國養(yǎng)禽業(yè)造成了巨大的損失。ALV主要通過垂直傳播方式呈指數(shù)級擴大,種蛋和精液在病毒垂直傳播過程中起著決定性的作用。在很長的時期內(nèi),我們主要通過檢測母雞的血漿以及種蛋中是否有ALV感染來實施凈化,而常常忽略了對公雞的檢測,然而對于凈化中公雞檢測究竟以血漿病毒分離為準還是以精液分離為準也無明確結(jié)論。另外,ALV經(jīng)垂直傳播感染后,會引起感染雞體重、產(chǎn)蛋率等各種生產(chǎn)機能下降,也會導(dǎo)致很高的致死率,給雞群造成很大的危害,對于一些高陽性率的地方品系雞群還存在過度淘汰影響育種進度的現(xiàn)實問題,我們希望通過多種途徑能夠降低高陽性率雞群的ALV帶毒率或至少減少由于垂直傳播帶毒對雛雞的致病作用,為此本研究通過雞胚卵黃囊接種模擬雛雞感染ALV,然后給感染雛雞添加AZT等逆轉(zhuǎn)錄酶抑制劑類藥物觀察了此類藥物對降低ALV垂直傳播危害的作用。1.不同種雞群禽白血病凈化中的種公雞檢測本研究對我國兩個正在實施禽白血病凈化的種雞場的公雞分別采取血漿病毒分離和精液病毒分離兩種方式觀察和比較了兩種檢測途徑對評價種公雞ALV感染狀態(tài)時的對應(yīng)性,并對部分陽性毒株做了序列測定和分析。結(jié)果顯示,對正在實施凈化的某蛋雞場檢測公雞400份,其中精液樣品病毒分離陽性為11份,血漿樣品病毒分離陽性為13份,精液和血漿病毒分離同時為陽性的有8份。對正在實施凈化的某野山雞場檢測公雞1800份,其中精液樣品病毒分離陽性為45份,血漿樣品病毒分離陽性為188份,精液和血漿病毒分離同時為陽性的僅有16份。上述數(shù)據(jù)說明僅僅通過單純的血漿病毒分離或者單純的精液病毒分離均不能有效的檢出所有的ALV陽性公雞,從凈化效率角度考慮應(yīng)該同時對公雞的血漿和精液做全面的病毒分離來確定陽性個體進而淘汰,這將有助于在最短的時間內(nèi)淘汰陽性公雞。對部分毒株gp85序列的測定和同源性分析結(jié)果顯示,所分離到的20個毒株之間同源性高達99.0%~100%,這些樣品與A亞群參考株之間的同源性最高,為88.6%~99.2%,顯著高于與B、E、F、J、K亞群的同源性,與B、E、F、J、K等不同亞群同源性分別為83.2%~83.7%、85.4%~89.6%、83.0%~83.9%、50.0%~51.1%和84.8%~85.9%。進化樹比對分析也發(fā)現(xiàn),分離到的所有毒株均與A亞群毒株在一個分支上。2.逆轉(zhuǎn)錄酶抑制劑類藥物對垂直傳播感染ALV的SPF雞的保護作用本研究通過對SPF雞胚卵黃囊接種NX0101毒株模擬雞胚經(jīng)垂直傳播感染ALV-J,待雞胚孵育出殼后,將雞胚分為兩組,只對其中一組在出殼后的連續(xù)一周內(nèi)肌肉注射5mg的Zidovudine(AZT)和2mg的Lamivudine(LAM),另一組在出殼后的連續(xù)一周內(nèi)肌肉注射等劑量的生理鹽水,與卵黃囊接種空白DMEM并且出殼后連續(xù)一周內(nèi)肌肉注射等劑量生理鹽水的完全空白對照組對比,觀察逆轉(zhuǎn)錄酶抑制劑類藥物對SPF雞的保護作用以及對降低垂直傳播危害的作用。結(jié)果顯示,未使用藥物的病毒感染組其體重、死亡率以及免疫NDV和AIV-H9所產(chǎn)生的抗體水平均顯著低于使用藥物的病毒感染組,更顯著低于未接毒的生理鹽水對照組,但是藥物的使用未能阻止感染組ALV病毒血癥的形成,說明藥物無法替代針對禽白血病的凈化工作。上述相關(guān)數(shù)據(jù)不僅進一步展示了ALV-J垂直傳播對雞群的危害,也顯示了AZT和LAM等逆轉(zhuǎn)錄酶抑制劑類藥物的使用有助于降低ALV-J垂直傳播感染對雞群的危害。
[Abstract]:Avian Leukosis Virus (ALV) has been outbursts and prevalent all over the world since it was first reported. Because of its high pathogenicity, high mortality, and high incidence of cancer, it has caused huge losses to poultry industry in various countries..ALV is expanded exponentially through vertical transmission, and eggs and semen play a role in the vertical transmission of the virus. In a very long period of time, we have done the purification mainly by detecting the plasma of hens and whether there is ALV infection in the eggs, and we often ignore the detection of the cocks. However, there is no definite conclusion whether the detection of the cock is based on the separation of the plasma virus or the semen separation. In addition, the ALV is vertical. After transmission of infection, it can cause the infection of chicken weight, the production function of laying eggs and other production functions, which will cause a high mortality rate and cause great harm to the chicken group. For some local chicken groups with high positive rates, there are some practical problems affecting the breeding progress. We hope to reduce the high positive rate of chicken through a variety of ways. The virulence rate of ALV in the group or at least reduces the pathogenic effect of the vertical transmission band poison on the chicks. Therefore, the chicken embryo yolk sac was inoculated with the simulated chicks infected with ALV, and then the infected chicks were added AZT and other reverse transcriptase inhibitors to observe the effect of this kind of drug on reducing the vertical transmission of ALV in.1.. In this study, we observed and compared the two methods of plasma virus isolation and separation of semen virus from two roosters who were carrying out avian leukaemia in our country, and compared two detection approaches to the evaluation of the ALV infection status of the cock, and the sequencing and classification of some positive strains. The results showed that 400 cocks were detected in a chicken farm which was being purified, of which 11 were isolated from the semen sample virus, 13 in the plasma sample virus isolation, and 8 in the semen and plasma virus isolation. 1800 roosters were detected in a wild mountain chicken field being purify, of which the semen sample virus was isolated. The positive was 45, the plasma sample virus isolation was 188, the semen and the plasma virus were isolated and positive only 16. The above data showed that all ALV positive cocks could not be detected only by pure plasma virus isolation or pure semen virus isolation, and the cocks should be considered at the same time from the angle of purification efficiency. The plasma and semen were separated from the virus to determine the positive individuals and then eliminated, which would help to eliminate the positive cocks in the shortest time. The determination of the gp85 sequence and the homology analysis showed that the homology of the 20 isolated strains was up to 99.0%~ 100%, and these samples were the same as the A subgroup reference strain. The homology of the subgroups of B, E, F, J, K is the highest, and the homology of the subgroups of B, E, F, J, K, respectively, are 83.2%~83.7%. The protective effect of drugs on SPF chicken with vertical transmission infection ALV was studied by inoculating the NX0101 strain of the yolk sac of the SPF chicken embryo to simulate the vertical transmission of ALV-J in chicken embryo, and the chicken embryo was divided into two groups after the chicken embryo was incubated and incubated, and the Zidovudine (AZT) and 2mg Lamivudine (LAM) of a group of 5mg were injected into the shell for a week. The other group was intramuscularly injected with normal saline within one week after the outer shell, compared with the complete blank control group of the yolk sac inoculated with blank DMEM and the intramuscular injection of normal saline within a week after the outer shell. The protective effect of the reverse transcriptase inhibitor on the SPF chicken and the reduction of the vertical transmission damage were observed. The results showed that the body weight, mortality, and the level of antibody produced by immune NDV and AIV-H9 were significantly lower than those in the virus infected group, which were significantly lower than those in the uninfected saline control group, but the use of the drug failed to prevent the formation of ALV virus in the infection group, indicating that the drug was not possible. Instead of purifying the avian leukaemia, these data not only further demonstrate the harm of vertical transmission of ALV-J to the chickens, but also show that the use of reverse transcriptase inhibitors such as AZT and LAM helps to reduce the harm of vertical transmission of ALV-J to chickens.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S858.31

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