鸭出血性卵巢炎灭活疫苗效力的初步评价

发布时间：2019-01-02 15:35

【摘要】：疫苗免疫是预防传染病的最有效的方法,但鸭出血性卵巢炎(Duck Hemorrhagic Ovaritis,DHO)暂无商品化疫苗产品,现阶段正处于研发阶段。疫苗效力是评价疫苗质量最重要的指标之一。本研究的目的是探讨建立评价本病灭活疫苗效力的方法,以实现对制备的灭活疫苗品质的控制。采用研究室试制的2批次疫苗(A批次、B批次)对北京鸭两次免疫后,人工感染免疫组蛋鸭(共30只)和对照组蛋鸭(共5只),在人工感染后5d采集血液,分离血清,将分离的血清接种于SPF鸡胚,记录SPF鸡胚在24～168h的死亡情况,采用Reed-Muench法计算出不同批次疫苗的半数保护量(PD50)。另取A、B 2批疫苗2次免疫的北京鸭组(共10只)和对照组北京鸭(共5只),对实验鸭感染前和人工感染后2d采血,分离血清,测定血清生化指标；人工感染后7d扑杀,剖检并观察和记录病理变化,同时取材,10%福尔马林固定,制作病理组织切片进行病理组织学观察和免疫组织化学检测。A批次疫苗的PD50为11.84,B批次疫苗的PD50为6.34。血清生化指标中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)含量在人工感染前与人工感染后2d变化显著,免疫组未见明显变化。病理学检查表明：卵巢出血、变形,卵泡膜发生出血和淤血；脾脏肿大明显,表面有花斑状纹理出现,白髓淋巴细胞流失崩解、网状细胞明显增生；肝脏变黄且质地变脆,肝细胞发生脂肪变性并在血管周围有淋巴细胞浸润；脑组织出现噬神经元现象。免疫组化检测结果表明在对照组鸭的心脏、肝脏、脾脏、肾脏、脑、卵巢组织的细胞胞浆内发现阳性颗粒,免疫组在个别组织脏器细胞胞浆内发现阳性颗粒,但免疫组较对照组阳性颗粒明显减少。免疫组之间比较中发现B批次疫苗免疫组较A批次疫苗免疫组出现阳性颗粒明显。通过灭活疫苗半数保护量测定、血清生化指标检测和病理学检查相结合,初步对2批次鸭出血性卵巢炎灭活疫苗完成评价后,证明2批次疫苗有良好的保护效果。
[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.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.32

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