【摘要】：疫苗免疫是預(yù)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(wú)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(píng)價(jià)疫苗質(zhì)量最重要的指標(biāo)之一。本研究的目的是探討建立評(píng)價(jià)本病滅活疫苗效力的方法,以實(shí)現(xiàn)對(duì)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(duì)北京鴨兩次免疫后,人工感染免疫組蛋鴨(共30只)和對(duì)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(jì)算出不同批次疫苗的半數(shù)保護(hù)量(PD50)。另取A、B 2批疫苗2次免疫的北京鴨組(共10只)和對(duì)照組北京鴨(共5只),對(duì)實(shí)驗(yàn)鴨感染前和人工感染后2d采血,分離血清,測(cè)定血清生化指標(biāo)；人工感染后7d撲殺,剖檢并觀察和記錄病理變化,同時(shí)取材,10%福爾馬林固定,制作病理組織切片進(jìn)行病理組織學(xué)觀察和免疫組織化學(xué)檢測(cè)。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(biāo)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(jiàn)明顯變化。病理學(xué)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(xì)胞流失崩解、網(wǎng)狀細(xì)胞明顯增生；肝臟變黃且質(zhì)地變脆,肝細(xì)胞發(fā)生脂肪變性并在血管周圍有淋巴細(xì)胞浸潤(rùn)；腦組織出現(xiàn)噬神經(jīng)元現(xiàn)象。免疫組化檢測(cè)結(jié)果表明在對(duì)照組鴨的心臟、肝臟、脾臟、腎臟、腦、卵巢組織的細(xì)胞胞漿內(nèi)發(fā)現(xiàn)陽(yáng)性顆粒,免疫組在個(gè)別組織臟器細(xì)胞胞漿內(nèi)發(fā)現(xiàn)陽(yáng)性顆粒,但免疫組較對(duì)照組陽(yáng)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(yáng)性顆粒明顯。通過(guò)滅活疫苗半數(shù)保護(hù)量測(cè)定、血清生化指標(biāo)檢測(cè)和病理學(xué)檢查相結(jié)合,初步對(duì)2批次鴨出血性卵巢炎滅活疫苗完成評(píng)價(jià)后,證明2批次疫苗有良好的保護(hù)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(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S858.32

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