黄芪多糖对蓝耳弱毒苗免疫调节及抗蓝耳病毒感染的研究

发布时间：2018-04-16 02:10

本文选题：黄芪多糖 + PRRSV　；参考：《安徽农业大学》2015年硕士论文

【摘要】：猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome,PRRS)又名猪蓝耳病,是由猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus.PRRSV)引起猪以繁殖障碍和呼吸系统症状为特征的一种急性、高度传染的病毒性传染病。该病已成为严重威胁我国养猪业发展的重要传染病之一。当前疫苗免疫是防制我国猪群PRRS的重要手段。为了探究黄芪多糖促进PRRSV疫苗免疫效果和抗PRRSV野毒感染能力,本研究开展如下三方面的研究:1.黄芪多糖对PRRSV VR-2332株弱毒苗免疫效果的研究。本研究选取70头后备母猪,根据其血液中PRRSV抗体水平和细胞因子(IL-2和IFN-γ)水平,平均分为两组(试验组A和对照组B)。根据PRRSV抗体S/P值大小,将A组,再分为S/P值1.0以下组(A1组),S/P值1.0-2.0组(A2组),S/P值2.0以上组(A3组),B组分组方法同A。A组连续饲喂黄芪多糖14天,B组不饲喂黄芪多糖。饲喂至第8天,A、B两组免疫PRRSV弱毒疫苗。免疫后第10天和第25天对A、B两组母猪进行前腔静脉采血。利用ELISA方法测定PRRSV抗体和细胞因子(IL-2和IFN-γ)水平。分析A、B两组PRRSV抗体和细胞因子水平的变化。结果表明,(1)A、B组PRRSV疫苗免疫效果无明显差异。但对免疫前S/P1.0的猪群,连续饲喂黄芪多糖14天会对PRRSV疫苗免疫效果有明显的抑制作用;(2)PRRSV疫苗免疫后,A、B组外周血中IL-2和IFN-γ均会上升,但A组IL-2和IFN-γ水平上升的更快,且IFN-γ水平上升持续时间更长。研究结果提示,PRRSV疫苗免疫前后各一周饲喂黄芪多糖,对PRRSV疫苗抗体水平增长没有明显的促进作用,但可以有效促进IL-2和IFN-γ的增长。2.PRRSV病原检测方法选择本探究性试验以稀释不同倍数的PRRSV弱毒活疫苗为样本,分别使用常规RT-PCR、荧光RT-PCR及POCKIT便携式PCR检测系统对稀释不同倍数PRRSV弱毒苗进行PRRSV抗原检测。结果发现:POCKIT便携式PCR检测系统的检测敏感度高,操作简单,使用于研究黄芪多糖对于妊娠母猪抗PRRSV试验PRRSV病原的检测工作。3.黄芪多糖对妊娠母猪抗PRRSV的研究。本研究从未免疫PRRSV疫苗且存在PRRSV带毒的猪群中选取70头妊娠第25天的母猪,根据PRRSV抗体水平和细胞因子(IL-2和IFN-γ)水平及血常规指标,平均分为两组(试验组C和对照组D)。C组连续饲喂黄芪多糖60天,D组不饲喂黄芪多糖。在饲喂黄芪多糖第30天和第60天分别采集C、D组前腔静脉血,检测PRRSV抗体、IL-2、IFN-γ和血常规值。在分娩时采集脐带血和检测PRRSV及统计繁殖指标。结果表明,(1)饲喂黄芪多糖第30天时(即妊娠第55天),C、D两组PRRSV抗体水平无显著差异。饲喂黄芪多糖第60天(即妊娠第85天),C组PRRSV抗体水平显著高于D组;(2)比较脐带血PRRSV检测率,C组为0%(0/35),D组为17%(6/35);(3)相比于妊娠25天,妊娠55天时,C、D两组猪群外周血中IL-2和IFN-γ含量显著性升高,妊娠85天时,C、D两组猪群外周血中IFN-γ含量依然保持高水平。
[Abstract]:Porcine reproductive and respiratory syndrome (PRRSs), also known as porcine blue ear disease, is an acute and highly contagious viral infection characterized by reproductive disorders and respiratory symptoms in pigs caused by porcine reproductive and respiratory syndrome virus.The disease has become a serious threat to the development of pig industry in China one of the important infectious diseases.At present, vaccine immunization is an important method to prevent and control PRRS in swine herd in China.In order to investigate the effect of astragalus polysaccharide on PRRSV vaccine immunity and the ability of resisting PRRSV wild virus infection, this study carried out the following three aspects: 1.Study on the immune effect of Astragalus Polysaccharide on PRRSV VR-2332 attenuated Vaccine.According to the levels of PRRSV antibody and cytokines IL-2 and IFN- 纬 in blood, 70 sows were divided into two groups: experimental group A and control group.According to the S- P value of PRRSV antibody, group A was divided into two groups: group A (< 1.0) and group A1 (P / P = 1.0-2.0), group A _ 2: group A _ 2, group A _ 3 and group A _ 3 were treated with Astragalus polysaccharide for 14 days and group A were not fed with Astragalus polysaccharide for 14 days.Two groups of PRRSV attenuated vaccine were immunized until the 8th day.On the 10th and 25th day after immunization, the blood samples were collected from the anterior vena cava of the two groups of sows.The levels of PRRSV antibody and cytokine IL-2 and IFN- 纬 were measured by ELISA method.The changes of PRRSV antibody and cytokine levels in two groups were analyzed.The results showed that there was no significant difference in the immune effect of PRRSV vaccine in group A and B.However, the immune effect of Astragalus Polysaccharide on PRRSV vaccine was significantly inhibited by Astragalus Polysaccharide for 14 days before immunization. The levels of IL-2 and IFN- 纬 in peripheral blood of group A were higher than those of group A, but the levels of IL-2 and IFN- 纬 in group A increased more rapidly.The level of IFN- 纬 increased for a longer time.The results showed that Astragalus polysaccharide was fed with Astragalus membranaceus polysaccharide for one week before and after immunization with PRRSv vaccine, and the antibody level of PRRSV vaccine did not increase significantly.However, it can effectively promote the growth of IL-2 and IFN- 纬. 2. PRRSv pathogen detection method selected this inquiry test to dilute different times of PRRSV attenuated live vaccine as a sample,Routine RT-PCR, fluorescent RT-PCR and POCKIT portable PCR system were used to detect the PRRSV antigen of diluted different times PRRSV attenuated vaccine.The results showed that the portable PCR detection system was highly sensitive and easy to operate. It was used to study the detection of Astragalus polysaccharides against PRRSV in pregnant sows.Study on the effect of Astragalus Polysaccharide on PRRSV in pregnant sows.In this study, 70 sows on the 25th day of gestation were selected from a group of pigs that had never been immunized with PRRSV vaccine and were infected with PRRSV virus. According to the level of PRRSV antibody, the levels of cytokines IL-2 and IFN- 纬, and the blood routine indexes,The rats were divided into two groups: experimental group C and control group DX. C were fed with Astragalus polysaccharide for 60 days and group D was not fed with Astragalus polysaccharide.The blood samples of anterior vena cava of group C were collected on the 30th and 60th day after feeding with Astragalus membranaceus polysaccharides. The serum levels of IL-2mIFN- 纬 and blood routine were detected.Umbilical cord blood was collected during delivery, PRRSV was detected and reproductive index was counted.The results showed that there was no significant difference in the level of PRRSV antibody between the two groups on the 30th day (that is, the 55th day of pregnancy) fed with Astragalus polysaccharides.The level of PRRSV antibody in group C was significantly higher than that in group D on day 60 (that is, 85 days after pregnancy). The PRRSV detection rate in group C was higher than that in group C, and the rate of detection of PRRSV in group C was higher than that in group D (group D was 17.6 / 35) compared with that in group D on the 25th day of gestation, compared with that of group D, the level of PRRSV antibody in group C was significantly higher than that in group D on day 85 of pregnancy.The levels of IL-2 and IFN- 纬 in peripheral blood of two groups were significantly increased at day 55, and the levels of IFN- 纬 in peripheral blood of two groups were still high at day 85 of gestation.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.28

【相似文献】