【作者】 陈凯；

【导师】 刁有祥；

【作者基本信息】 山东农业大学 ， 预防兽医学， 2007， 硕士

【摘要】 猪传染性胸膜肺炎(Porcine Pleuropneumonia)是由胸膜肺炎放线杆菌(Actinobacillus Pleuropneumoniae,APP)引起的一种常见的呼吸道传染病,各年龄的猪只均可感染。猪群感染胸膜肺炎放线杆菌后,当遇到气候环境突变,通风不良等应激因素时,猪只抵抗力下降,可诱发本病,主要表现为精神沉郁,体温升高,呼吸困难等临床症状,并出现特征性的出血性肺炎和胸膜炎。本病在世界各养猪国家普遍存在,近年来,流行日趋严重,成为大规模养猪的五大疫病之一。本研究共分为二部分:第一部分:猪传染性胸膜肺炎放线杆菌的分离鉴定本实验对分离自淄博市不同地区的疑似胸膜肺炎放线杆菌进行分离培养、生化鉴定、PCR鉴定、药敏试验、动物实验等研究。研究结果发现该分离菌中有20株细菌为多形态、革兰氏阴性杆菌;生化实验和PCR扩增鉴定结果表明所分离的上述20株菌株为猪传染性胸膜肺炎放线杆菌。对确诊的20株胸膜肺炎放线杆菌进行血清型鉴定,结果发现在淄博地区流行的胸膜肺炎放线杆菌主要为血清5型和血清7型。其中,血清5型13株、血清7型7株,血清5型为相对优势的血清型。第二部分猪传染性胸膜肺炎放线杆菌油乳剂二价灭活疫苗的制备及小鼠免疫保护试验根据淄博地区猪传染性胸膜肺炎放线杆菌的流行情况,制备了血清5型、7型二价灭活疫苗。以该疫苗免疫小白鼠,进行同源攻毒保护试验,结果该疫苗在0.6ml/只的免疫剂量下可对小白鼠产生较强的保护作用,用同血清型菌株对免疫后的小白鼠攻毒,仅表现肺脏轻微出血,无死亡,而对照组未经免疫直接攻毒的小白鼠全部死亡,肺脏严重出血;小白鼠免疫后第5d就可以检测到抗体,并且抗体水平上升较快,到第15d抗体达到最高水平,之后,抗体水平开始下降,但下降幅度不大,可持续1个月左右;用自制的疫苗和市场上购买的疫苗同时对仔猪进行同源攻毒保护实验结果对照组表现明显的胸膜肺炎症状、肺脏严重出血且有两头死亡;市场上购买的疫苗免疫组出现出现明显的厌食、喘气、呼吸困难等急性胸膜肺炎症状;自制疫苗免疫组只有一头出现轻微症状,20h后恢复正常。上述结果表明自制的二价灭活疫苗对胸膜肺炎放线杆菌有坚强的保护作用。更多还原

【Abstract】 Actinobacillus Pleuropneumoniae is the causative agent of Porcine Pleuropneumoniae, a worldwide disease causing tremendous economic loss to the swine industry. It is a common infectious disease which infects the respiratory tract of pigs. The symptoms of infected pigs induced depression, difficult respiratory and pneumonia. With the development of the raising cosmically, Porcine Pleuropneumoniae becomes one of the five primary disease that threaten mordern pig industry.The following two parts were included in the studies:Part I: The isolation of Actinobacillus PleuropneumoniaeThe bacterias isolated from zibo area, were identified by bio-chemistry experiments,PCRand injected the mouse .The results of coloration showed that 20 strains were small bacilus ,the results of bio-chemistry experiments and PCR reveled that these 20 strains were Actinobacillus Pleuropneumoniae. According to morphology observation, culture characteristic, biochemical characteristic, PCR and animal test, the disease breaking out in zi bo aera was Porcine infectious pleuropneumonia. Serotypes of 20 strains were identified,the results showed that 13 strains are serotype 5 and the left are serotype 7, serotype 5 was the dominating serotype. Part II The production of bivalent inactivated vaccine of Actinobacillus Pleuropneaumoniae and the protection test of mouse mmunity.One vaccine out of serotype 7 and serotype 5 was made basing on the epidemic of APP in zi bo aera. And then, the produced vaccine were used as antigens to challenge the mouse with the same serotype bacteria, the results showed that the mouse were well protected immunized with 0.6ml vaccine . The mouse challenged with the same serotype bacteria were not dead at this dose , while the mouse of control group were all dead. Two days after immunization, antibody was examined in the mouse. At the 6th day, the mouse exhibited significantly high antibody against APR Then the antibody slowly began to decline.Different vaccine were used to challenge the pig with the same serotype bacteria, the results revealed that the pig of control group behave clearly symptom,and there were two pigs dead. The pig which immunitied the bought vaccine behave lightly symptom,while the pig whih were injected the vaccine produced by our were well protected. These results revealed that the vaccine produced by ourself were usefull in defending APR更多还原