【作者】 刘安石；

【导师】 刘进辉； 张长弓；

【作者基本信息】 湖南农业大学 ， 预防兽医， 2014， 硕士

【摘要】 目的：首先,为了探讨草分枝杆菌对猪繁殖与呼吸综合征活疫苗的免疫应答效果；其次,为了探讨灭活的草分枝杆菌溶液(水剂)作为免疫佐剂对猪繁殖与呼吸综合征活疫苗免疫的影响。方法：试验选取30日龄健康的断奶仔猪,20头,随机分成4组,1个对照组和3个试验组,每组5头。对照组正常饲养并进行猪繁殖与呼吸综合征活疫苗免疫,试验组Ⅰ、试验组Ⅱ和试验组Ⅲ在猪繁殖与呼吸综合征活疫苗免疫的前后7天于饲料中分别添加0.1%、0.2%和0.5%的灭活草分枝杆菌粉,于免疫前1天和免疫后的第14天、21天、28天和35天测其抗体水平,并于疫苗注射后观察猪只的临床表现。免疫佐剂试验时,选取60头30日龄的健康断奶仔猪,分成对照组和试验组,每组各30头。对照组使用繁殖与呼吸综合征活疫苗自带稀释液稀释后于仔猪耳后颈部外侧肌肉注射,免疫剂量为2ml/头份；试验组使用0.2%的灭活草分枝杆菌溶液稀释疫苗后于仔猪耳后颈部外侧肌肉注射,免疫剂量为2ml/头份。两组猪只于免疫的前1天和免疫后的第15天、30天、45天、60天和90天,对所有试验仔猪进行前腔静脉采血2ml,置于离心机中以1500r/min离心5min,取血清,冷藏保存备用。将分离的血清用猪繁殖与呼吸综合征ELISA抗体检测试剂盒检测抗体的OD630nm值。试验结果表明：三个试验组猪只的抗体OD630-值均极显著的高于对照组(P<0.01),试验组Ⅱ和试验组Ⅲ抗体效价高于试验组Ⅰ,差异显著(P<0.05)。本研究结果表明在猪饲料中添加0.2%以上的草分枝杆菌,可有效的提高猪繁殖与呼吸综合征的疫苗免疫效果但不能减轻猪只的疫苗免疫应激反应。免疫佐剂试验结果表明：对照组猪只在繁殖与呼吸综合征疫苗免疫后的第15天,产生有效保护抗体的仔猪比率为50%左右,免疫后的30天,产生有效保护抗体的仔猪比率为86.67%。试验组第15天的抗体OD630nm值都在上升,第30天保护率达到了76.67%,但第45天和第60天的抗体保护率都达到了100%(30/30),第90天时实验组的抗体保护率还有90%(27/30),而对照组仅为43.3%(13/30)。结论：试验结果表明,使用灭活草分枝杆菌溶液可有效的提高猪繁殖与呼吸综合征疫苗的抗体效价,且高抗体水平维持时间较长,与对照组相比差异极显著(P<0.01),在生产上具有实际意义。更多还原

【Abstract】 Purpose:Firstly, the experiment aims at exploring Mycobacterium phlei’s effect of the immune response for PRRS vaccine. Secondly, the study explores the inactivated Mycobacterium phlei (agent)’s effects on PRRS live vaccine as immune adjuvant.Method:In the experiment,20 30-day healthy weaning piglets are selected and randomly divided into four groups, a control group and three experimental groups. There are five piglets in each group. The control group is provided daily feed and PRRS live vaccined. The test Group Ⅰ, test Group Ⅱ and test group Ⅲ are provided of the feed which respectively added 0.1%,0.2% and 0.5% of inactivated Mycobacterium phlei powder seven days before and after the PRRS live vaccined. Observing the piglets’ clinical manifestations after vaccination, the antibody levels are tested when one day before immunization and 14 days,21st days,35 after immunization. In the part two, there are 60 30-day healthy weaning piglets selected and divided them into a control group and a test group. Each group includs 30 piglets. The control group uses PRRS vaccine with dilutions,2ml each piglet, to inject the piglets’ posterior neck muscles behind their ears. The test group uses 0.2% of inactivated Mycobacterium phlei solution to dilute the vaccine and injected the piglets’ posterolateral neck muscles behind their ears,2ml each piglet. Then carrying out before vena cava blood of all the test piglets one day before immunization,15th days,30th days,45th and 60th days after immunization, putting the blood in a centrifuge and centrifuged five minutes at the speed of 1500r/min. Finally, taking the serum and refrigerated for use. At last, the PRRS ELISA antibody test kit is used to detect the OD630nm value of the separated serum’s antibody.Result:The experiment shows that antibody OD630nm value of the piglets of three test groups are significantly higher than control group (P<0.01), the antibody titers of test group Ⅱ and test group Ⅲ were higher than test group Ⅰ and the difference is significant (P<0.05). This study shows that adding more than 0.2% mycobacterium phlei in the feed could effectively improve the immune effect of PRRS vaccine but can not reduce the vaccinate stress response of piglets. The results shows that about 50% of the piglets in the control group produced effective protection antibody at the 15th day after PRRS vaccines and immunization. At the 30th day after immunization, 86.67% of the piglets produce effective protection antibody. The OD630nm of the piglets in the test group are all on the rise at the 15th day and the protection force reached 76.67 at the 30th day. Moreover, it reaches even 100%(30/30)at the 45th day and the 60th day. It is 90% at the 90th day, while the control group is only 43.3% (13/30).Conclusion:The experiment results show that using inactivated Mycobacterium phlei solution can effectively improve the antibody titers of PRRS vaccine, and the high antibody level could maintain a longer time. It was extremely significant different that compared with the control group (P<0.01). The Mycobacterium phlei make sense.更多还原