产气荚膜梭菌NetB毒素基因重组植物乳酸杆菌疫苗免疫保护性研究  

Study on Immune Protection of Lactobacillus plantarum Recombinant Vaccine with NetB Toxin Gene of Clostridium perfringens

作　　者：张艳[1,2] 周庆民[1] 冯万宇[1] 徐馨[1] 黄健[1] 王靓靓 李兰兰[2] 王莹莹[2] 郑晓星[2] 李广兴[2] ZHANG Yan ZHOU Qingmin FENG Wanyu XU Xin HUANG Jian WANG Jingjing LI Lanlan WANG Yingying ZHENG Xiaoxing LI Guangxing(Heilongjiang Institute of Veterinary Science,Qiqihar,Heilongjiang 161006 College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang 150030 Animal Husbandry and Veterinary Bureau of Daqing City,Daqing,Helongjiang 163000)

机构地区：[1]黑龙江省兽医科学研究所,黑龙江齐齐哈尔161006 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030 [3]大庆市畜牧兽医局,黑龙江大庆163000

出　　处：《中国家禽》2017年第14期15-20,共6页China Poultry

摘　　要：为了评估表达产气荚膜梭菌NetB毒素的重组植物乳酸杆菌口服活菌疫苗对鸡的免疫保护效果,分别进行了重组活菌疫苗的免疫和产气荚膜梭菌攻菌试验。免疫组雏鸡分为3组,于7~28日龄分别连续免疫p SIP409空载体植物乳酸杆菌、p SIP409-NetB重组植物乳酸杆菌和灭菌PBS;攻菌组雏鸡免疫方法同上,于43~47日龄连续口服产气荚膜梭菌(设置PBS免疫对照组和不攻菌只口服等量PBS对照组)。通过采用ELISA检测抗体动态变化,观察雏鸡体重变化和免疫器官发育情况以评价疫苗的免疫效果。结果显示,NetB组特异性抗体Ig G和s Ig A水平随日龄增长呈现上升趋势,且与未免疫对照组存在显著或极显著差异(P<0.05或P<0.01);NetB攻菌组各项指标在短暂下降后很快升高,且明显高于未免疫对照组雏鸡(P<0.01)。NetB组周增重14日龄后均高于其他组,且35日龄开始与其他组差异极显著(P<0.01);攻菌组体重变化总趋势是NetB攻菌组最高,其次是空载体攻菌组(P<0.05),PBS攻菌组最低(P<0.01)。免疫组在免疫结束时胸腺和脾脏指数呈明显上升趋势,NetB组的这两项指数分别在56日龄和49日龄达到最高,法氏囊指数在28日龄下降,随后稳步上升;虽然攻菌后各免疫器官指数上升趋势变缓,但NetB攻菌组的各项指数在各时间点均与其他组差异极显著(P<0.01)。表明重组口服活菌疫苗对产气荚膜梭菌攻击雏鸡起到很好的保护作用,所构建的重组疫苗安全有效,结果为重组植物乳酸杆菌表达系统应用于口服疫苗提供了理论依据。To evaluate the protective effect of recombinant Lactobacillus plantarum oral vaccine with NetB toxin of Clostridium perfringens in chickens, the recombinant live vaccine immunization and Clostridium perfringens challenge test were carried out respectively. The immune treatment was divided into 3 groups at 7 to 28 days of age to continuously inoculate pSIP409 empty vector Lactobacillus, pSIP409-NetB recombinant Ix^ctobacillus and sterile PBS, respectively： Immune-challenge treatment was grouped as above, which was continuously oral inoculated C. perfringens at 43 to 47 days of age. ELISA was used to detect antibody change, and chick weight changes and immune organs development were used to evaluate the overall immunity of the vaccine. The results showed that NetB-specific. [gG and sIgA level of recombinant vaccine group were increased with age, and there were significant or very significant differences between the immune group and the control groups （P〈0.05 or P〈0.01）. The weekly weight gain of NetB group from 14 days of age was higher than that of other groups, and there were significant differences between NetB group and other groups from 35 days of age （P〈0.01）; The general trend of weight change in challenging groups was that NetB-challenging group was the highest, empty-vector-challenging group was higher （P〈0.05）, and PBS-challenging group was the lowesl （P〈0.01）. The thymus index and spleen index of immnue treatment were obviously increased at the end of immunization, and which of NetB group reached the peak at 56 and 49 days of age, respectively, and bursa index was declined at 28 days of age then increased steadily; After bacteria challenging, although the upturn trend of immune organ indexes were slowed down, which of NetB-challenging group were significantly different with other groups at all time （P〈0.01）. The results showed that the recombinant live bacteria oral vaccine could provide good protection for C. perfringens chagellging chicks. It＇s indicated that the re

关 键 词：产气荚膜梭菌 NetB毒素 植物乳酸杆菌 免疫保护 

分 类 号：S855.1[农业科学—临床兽医学]