重组产气荚膜梭菌plc基因植物乳酸杆菌的免疫效果评价  

Study on Immune Protection of Recombinant Lactobacillus plantarum Expressing α-toxin plc Gene of Clostridium perfringens

作　　者：张艳[1,2] 周庆民[1] 冯万宇[1] 徐馨[1] 黄健[1] 王新[1] 李兰兰[2] 王莹莹[2] 郑晓星 宁秀云 李广兴[2] ZHANG Yan;ZHOU Qing-min;FENG Wan-yu;XU Xin;HUANG Jian;WANG Xin;LI Lan-lan;WANG Ying-ying;ZHENG Xiao-xing;NING Xiu-yun;LI Guang-xing(Heilongjiang Veterinary Science Research Institute,Qiqihar 161006,China;College of Veterinary Medicine,NortheastAgricultural University,Harbin 150030,China;Animal epidemic prevention station of Nehe,Nehe 161300,China)

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]黑龙江省讷河市动物防疫站,黑龙江讷河161300

出　　处：《中国兽医杂志》2018年第8期3-8,129,130,共8页Chinese Journal of Veterinary Medicine

摘　　要：为了评估表达产气荚膜梭菌α毒素plc基因的重组植物乳酸杆菌口服活菌疫苗的免疫保护效果,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 d分别连续免疫p SIP409空载体植物乳酸杆菌、pSIP409-plc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 d连续口服产气荚膜梭菌(设置PBS免疫组不攻菌只口服等量PBS对照)。通过病理组织学检查疫苗对雏鸡组织器官的损害作用; ELISA检测抗体动态变化、攻菌后细菌分离计数、剖检肠道病变评分、雏鸡体重变化和免疫器官发育情况来整体评价疫苗的免疫水平。结果显示,与对照组相比重组疫苗的免疫没有对雏鸡组织器官产生损害作用,plc特异性抗体IgG和s IgA水平检测显示,抗体水平随日龄增长呈现上升趋势,且与未免疫组有显著和极显著差异(P <0.05,P <0. 01);攻菌后免疫组雏鸡各项指标在短暂下降后很快升高,且明显高于未免疫组雏鸡。攻菌组各项试验也证明,我们的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。To assess the expression of recombinant α-toxin plc gene of C. Perfringens live bacteria Lactobacillus plantarum oralvaccine immune protection, they were immune attack and C. perfringens bacteria test of viable recombinant vaccines. Immunohisto-chemical chickens divided into three groups at 7 - 28 d continuous immune pSIP409 empty vector Lactobacillus respectively, pSIP409- plc recombinant Lactobacillus and sterile PBS; Immune attack bacteria group as above, at 43 - 47 d continuous oral C. perfrin-gens. ELISA detection antibody dynamic change, after the attack of bacteria isolated bacteria count, necropsy intestinal lesionscores, chick weight changes and immune organs development situation to evaluate the overall immunity of vaccines; vaccine by his-topathological examination damaging effects on the tissues and organs of the chicks. The results showed that compared with the controlgroup did not produce recombinant vaccine immune damaging effects on the tissues and organs of chicks, plc specific IgG and sIgA leveltests showed antibody levels increase with age on the rise, and there is a significant and very non-immunized group significant differ-ences （P 〈 0. 05, P 〈 0. 01）; after the attack bacteria immunohistochemical chicks indexes after a brief decline rapidly increased, andwas significantly higher than the group immunized chickens. Challenge also proved that we live bacteria recombinant oral vaccine a-gainst C. perfringens attack chicks play a very good protection. Description recombinant bacterium Lactobacillus constructed safe and ef-fective, provides a theoretical basis for the recombinant expression systems as Lactobacillus oral vaccine applications.

关 键 词：产气荚膜梭菌 plc毒素 植物乳酸杆菌 免疫保护 

分 类 号：S8[农业科学—畜牧兽医]