發(fā)布時間：2019-01-02 15:35

【摘要】：疫苗免疫是預防傳染病的最有效的方法,但鴨出血性卵巢炎(Duck Hemorrhagic Ovaritis,DHO)暫無商品化疫苗產(chǎn)品,現(xiàn)階段正處于研發(fā)階段。疫苗效力是評價疫苗質(zhì)量最重要的指標之一。本研究的目的是探討建立評價本病滅活疫苗效力的方法,以實現(xiàn)對制備的滅活疫苗品質(zhì)的控制。采用研究室試制的2批次疫苗(A批次、B批次)對北京鴨兩次免疫后,人工感染免疫組蛋鴨(共30只)和對照組蛋鴨(共5只),在人工感染后5d采集血液,分離血清,將分離的血清接種于SPF雞胚,記錄SPF雞胚在24～168h的死亡情況,采用Reed-Muench法計算出不同批次疫苗的半數(shù)保護量(PD50)。另取A、B 2批疫苗2次免疫的北京鴨組(共10只)和對照組北京鴨(共5只),對實驗鴨感染前和人工感染后2d采血,分離血清,測定血清生化指標；人工感染后7d撲殺,剖檢并觀察和記錄病理變化,同時取材,10%福爾馬林固定,制作病理組織切片進行病理組織學觀察和免疫組織化學檢測。A批次疫苗的PD50為11.84,B批次疫苗的PD50為6.34。血清生化指標中谷丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST)、總膽紅素(TBIL)含量在人工感染前與人工感染后2d變化顯著,免疫組未見明顯變化。病理學檢查表明：卵巢出血、變形,卵泡膜發(fā)生出血和淤血；脾臟腫大明顯,表面有花斑狀紋理出現(xiàn),白髓淋巴細胞流失崩解、網(wǎng)狀細胞明顯增生；肝臟變黃且質(zhì)地變脆,肝細胞發(fā)生脂肪變性并在血管周圍有淋巴細胞浸潤；腦組織出現(xiàn)噬神經(jīng)元現(xiàn)象。免疫組化檢測結(jié)果表明在對照組鴨的心臟、肝臟、脾臟、腎臟、腦、卵巢組織的細胞胞漿內(nèi)發(fā)現(xiàn)陽性顆粒,免疫組在個別組織臟器細胞胞漿內(nèi)發(fā)現(xiàn)陽性顆粒,但免疫組較對照組陽性顆粒明顯減少。免疫組之間比較中發(fā)現(xiàn)B批次疫苗免疫組較A批次疫苗免疫組出現(xiàn)陽性顆粒明顯。通過滅活疫苗半數(shù)保護量測定、血清生化指標檢測和病理學檢查相結(jié)合,初步對2批次鴨出血性卵巢炎滅活疫苗完成評價后,證明2批次疫苗有良好的保護效果。
[Abstract]:Vaccine immunization is the most effective method to prevent infectious diseases, but there is no commercial vaccine product for duck hemorrhagic ovary disease (Duck Hemorrhagic Ovaritis,DHO). Vaccine effectiveness is one of the most important indicators to evaluate vaccine quality. The aim of this study was to establish a method to evaluate the efficacy of inactivated vaccine for the purpose of controlling the quality of the inactivated vaccine. Two batches of vaccine (A lot, B batch) were used to immunize Beijing ducks twice. The blood samples were collected and the serum was isolated from the infected ducks (30 ducks) and the control ducks (5 ducks) on the 5th day after artificial infection. The isolated serum was inoculated into SPF chicken embryo and the death rate of SPF chicken embryo was recorded at 24 ~ 168h. The half protection dose (PD50) of different batches of vaccine was calculated by Reed-Muench method. In addition, 10 Beijing ducks (10 ducks) and 5 Beijing ducks (control group) were immunized twice with AMB-2 vaccine. Blood samples were collected before infection and 2 days after artificial infection. Serum samples were isolated and serum biochemical indexes were measured. After 7 days of artificial infection, culling, dissection, observation and recording of pathological changes, taking of materials, fixation of 10% formalin, histopathological observation and immunohistochemical examination were made. The PD50 of A batch vaccine was 11.84. The PD50 of batch B vaccine was 6.34. The content of total bilirubin (TBIL) of serum alanine aminotransferase (ALT),) and aspartate aminotransferase (AST),) was significantly changed before and 2 days after artificial infection, but there was no significant change in immunization group. Pathological examination showed: ovarian hemorrhage, deformation, follicular membrane bleeding and congestion, splenomegaly, appearance of blotchy surface, loss and disintegration of white pulp lymphocytes, proliferation of reticular cells; The liver became yellow and brittle, the liver cells had fatty degeneration and lymphocytic infiltration around the blood vessels, and the neurons in the brain tissue were phagocytomized. The results of immunohistochemistry showed that positive granules were found in the cytoplasm of the heart, liver, spleen, kidney, brain and ovary of the control group, and in the cytoplasm of individual organ cells in the immunized group. But the positive granules in the immunized group were significantly lower than those in the control group. It was found that the positive granules in group B were significantly higher than those in group A. After the evaluation of 2 batches of duck hemorrhagic ovaritis inactivated vaccine was preliminarily evaluated, it was proved that the two batches of inactivated duck ovaritis vaccine had a good protective effect through the determination of half protective dose of inactivated vaccine, the detection of serum biochemical index and pathological examination.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.32

【參考文獻】

相關(guān)期刊論文 前10條

1 周曉波;李銀;楊凌;朱明;卞紅春;;鴨坦布蘇病毒病發(fā)病模型的建立[J];山東畜牧獸醫(yī);2015年02期

2 謝佳;韓春華;林健;段會娟;楊志遠;趙際成;潘潔;劉月煥;;鴨出血性卵巢炎病毒血癥研究[J];中國農(nóng)業(yè)科學;2014年23期

3 吳曉剛;高旭元;余磊;李雪松;閆麗萍;滕巧泱;李國新;李澤君;;鴨坦布蘇病毒病活疫苗(FX2010-180P株)毒種保存期的研究[J];中國動物傳染病學報;2014年03期

4 陸新浩;劉友生;白露;陳秋英;劉鴻;廖敏;周繼勇;任祖伊;;體外抗鴨坦布蘇病毒的藥物篩選[J];畜牧與獸醫(yī);2013年10期

5 靳宇田;黃顯明;許秀梅;尹秀鳳;徐延偉;朱鵬;張小飛;;鴨坦布蘇病毒實時熒光定量RT-PCR檢測方法的建立及應(yīng)用[J];中國動物傳染病學報;2013年04期

6 萬春和;朱海俠;施少華;黃瑜;程龍飛;傅光華;陳紅梅;;鴨坦布蘇病毒SYBR GreenⅠ實時熒光定量檢測方法的建立[J];中國獸醫(yī)學報;2013年07期

7 李振華;李小康;郭香玲;張春杰;劉鳳軍;王臣;;鴨坦布蘇病毒滅活油乳苗的制備及免疫效力測定[J];中國預防獸醫(yī)學報;2013年05期

8 劉浩飛;黃顯明;張小飛;楊倩;;鴨黃病毒病的臨床病理學觀察及病原學檢測[J];畜牧與獸醫(yī);2013年02期

9 張金強;王勝;張青嬋;玄甲峰;王蕾;胡北俠;許明清;李霞;張秀美;;種鴨黃病毒病疫苗免疫效果觀察[J];中國家禽;2013年02期

10 李國平;;鴨坦布蘇病毒RT-PCR診斷方法的建立與應(yīng)用[J];中國畜牧獸醫(yī);2013年01期

相關(guān)會議論文 前1條

1 黃迪海;劉霞;張再輝;張華;徐秀榮;張從敬;秦卓明;;“百毒寧”對蛋鴨黃病毒病的療效研究[A];中國畜牧獸醫(yī)學會禽病學分會第十六次學術(shù)研討會論文集[C];2012年

相關(guān)碩士學位論文 前2條

1 孫曉艷;不同日齡肉鴨對坦布蘇病毒的易感性研究[D];山東農(nóng)業(yè)大學;2014年

2 顏丕熙;鴨坦布蘇病毒分離鑒定及其生物學特性的研究[D];中國農(nóng)業(yè)科學院;2012年

，

本文編號：2398680