中华鳖对温和气单胞菌体液免疫应答规律的研究
STUDIES ON HUMORAL IMMUNE RESPONSE OF TRIONYX SINENSIS TO AEROMONAS SOBRIA INACTIVED VACCINE
-
摘要: 从健康中华鳖血清中提取抗体并用Sephadex及DEAE-52进行了纯化,再免疫新西兰白兔制备兔抗鳖抗体的抗血清,纯化后以辣根过氧化物酶(HRP)标记酶标复合物,方阵滴定法确定酶标复合物的最适稀释度为1/3200.用温和气单胞菌TL9702株制成铝胶佐剂灭活苗免疫健康中华鳖,经一次免疫和二次免疫后,分别在不同时间采集血清,用ELISA法测定血清抗体变化规律,结果表明中华鳖对该菌苗产生良好的体液免疫应答,免疫后20d抗体水平达到高峰,在测定期间内二次免疫组的抗体水平高于一次免疫组,表明中华鳖存在着二次免疫应答,但与哺乳类相比,中华鳖的二次应答则相对较弱.免疫后90d对一次免疫组的鳖进行了攻毒试验,10d后免疫鳖100%存活,而非免疫对照组鳖则100%死亡,表明温和气单胞菌TL9702株灭活苗免疫效果良好.Abstract: An enzyme linked-immunosorbent assay(ELISA)for detection of Aeromonas sobria(As)antibody in Chinese soft-shelled turtle(Trionyx sinensis,Ts)was developed. Chinese soft-shelled turtle antibody was purified by sephadex and DEAE-52.Then inoculated into the New Zealand white rabbit with Freund complete adjuvant.Rabbit anti-turtle IgG was also purified and coagulated with Horseradish peroxidase(HRP).It was determined by square titration method that a dilution of 1:3200 of this enzyme complex was optimal in terms of test sensitivity.Immune serum was collected at different time after the turtles were immunized with Aeromonas sobria(As)-TL970424 inactived vaccine once or twice.ELISA methods were applied to detecting the change of antibody in serum.The result indicated that Chinese soft-shelled turtle could mount good humoral immune response as well as the second immune response to As inactived vaccine.However this second immune response is weak when compared to that of mammals.Challenge test was carried out 90 days after immunization.No mortality was recorded with turtles injected once whereas there was 100%mortality from group of non-immunized turtles.This result indicated that As inactived vaccine was effective in protecting immunized turtles from attack of this bacteria within 3 monthes post immunization.
-
Keywords:
- Humoral immune /
- Trionyx sinensis /
- Aeromonas sobria
-
-
[1] Yu L, Technique of Immunology [M]. HangZhou: Zhejiang University Press, 1992. [于涟.免疫学实验技术.杭州:浙江大学出版社,1992][2] Wang S R. Modern Animal Immunology [M]. Jielin: Jielin Science and Technology Press. 1996.[王世若.现代动物免疫学.长春:吉林科学技术出版社,1996][3] Xu Y W. Detecting Technique of Immunology(second edition)[M]. Beijing: Science Press. 1997.[徐宜为.免疫检测技术[M]. 北京:科学出版社,1997][4] Yang X L, Zhou J G, Chai W Q, et al. Studien on the memory immune response of soft-shelled turtle to T3 bacterin [J]. J. of Aquaculture, 2000, 24(2): 156-160. [杨先乐,周剑光,蔡完其等.中华鳖对T3菌苗的回忆应答[J].水产学报,2002,24(2):156-160][5] Grey H M.Phylogeny of the immune response.Studies on some physical,chemical and serological characteristics of antibody production in Turtles[J].J Immunol,1963,91:819-825[6] Chartrand S L.The evolution of the immune response-The immunoglobulins of the Turtles.Molecular requirements for biologic activity of the 5.7S immunoglobulin[J].J Immunol,1971,107:1-11[7] Maung H T.Immunity in the tortoise,Testudo ibera[J].J Pathol Bacteriol,1963,85:51-66[8] Jian J C, Wu T T, Yang H et al. Prielimary studies of humoral immune response of soft-shelled turtle[J]. J. of Fishery Science of China, 1998,5(4): 6-10. [简纪常,吴婷婷,杨弘,等.中华鳖体液免疫应答的初步研究. 中国水产科学,1998,5(4):6-10][9] Yang Xianle, J G Zhou, X H Ei, et al. Studies on effect of weight, age,nutrion of sotf-shelled turtle to immune response [J]. J. of Aquaculture, 1999, 24(4): 375-380. [杨先乐,周剑光,艾晓辉等.鳖的体重、年龄及营养状况对中华鳖免疫应答的影响. 水产学报,1999,24(4)375-380][10] Sun Qihuan, Sun Peifang, Jin Lihua et al. On the Pathogen Bacteria of the Hemolytic Ascitesosis of Allogynogenetic Crucian Carp[J]. J. of Aquaculture,1991, 15(2): 130-139. [孙其焕,孙佩芳,金丽华,等.异育银鲫溶血性腹水病病原的研究. 水产学报,1991,15(2):130-139] Yu L, Technique of Immunology [M]. HangZhou: Zhejiang University Press, 1992. [于涟.免疫学实验技术.杭州:浙江大学出版社,1992][2] Wang S R. Modern Animal Immunology [M]. Jielin: Jielin Science and Technology Press. 1996.[王世若.现代动物免疫学.长春:吉林科学技术出版社,1996][3] Xu Y W. Detecting Technique of Immunology(second edition)[M]. Beijing: Science Press. 1997.[徐宜为.免疫检测技术[M]. 北京:科学出版社,1997][4] Yang X L, Zhou J G, Chai W Q, et al. Studien on the memory immune response of soft-shelled turtle to T3 bacterin [J]. J. of Aquaculture, 2000, 24(2): 156-160. [杨先乐,周剑光,蔡完其等.中华鳖对T3菌苗的回忆应答[J].水产学报,2002,24(2):156-160][5] Grey H M.Phylogeny of the immune response.Studies on some physical,chemical and serological characteristics of antibody production in Turtles[J].J Immunol,1963,91:819-825[6] Chartrand S L.The evolution of the immune response-The immunoglobulins of the Turtles.Molecular requirements for biologic activity of the 5.7S immunoglobulin[J].J Immunol,1971,107:1-11[7] Maung H T.Immunity in the tortoise,Testudo ibera[J].J Pathol Bacteriol,1963,85:51-66[8] Jian J C, Wu T T, Yang H et al. Prielimary studies of humoral immune response of soft-shelled turtle[J]. J. of Fishery Science of China, 1998,5(4): 6-10. [简纪常,吴婷婷,杨弘,等.中华鳖体液免疫应答的初步研究. 中国水产科学,1998,5(4):6-10][9] Yang Xianle, J G Zhou, X H Ei, et al. Studies on effect of weight, age,nutrion of sotf-shelled turtle to immune response [J]. J. of Aquaculture, 1999, 24(4): 375-380. [杨先乐,周剑光,艾晓辉等.鳖的体重、年龄及营养状况对中华鳖免疫应答的影响. 水产学报,1999,24(4)375-380][10] Sun Qihuan, Sun Peifang, Jin Lihua et al. On the Pathogen Bacteria of the Hemolytic Ascitesosis of Allogynogenetic Crucian Carp[J]. J. of Aquaculture,1991, 15(2): 130-139. [孙其焕,孙佩芳,金丽华,等.异育银鲫溶血性腹水病病原的研究. 水产学报,1991,15(2):130-139]
计量
- 文章访问数: 883
- HTML全文浏览量: 0
- PDF下载量: 467
下载: