摘要
为了探究添加蜂胶佐剂、弗氏佐剂和铝胶佐剂后对大鲵（Andrias davidianus）嗜水气单胞菌（Aeromonas hy-drophila）灭活疫苗的免疫效果，取已制备好的大鲵嗜水气单胞菌灭活疫苗，分别加入3种佐剂，充分混匀后制备成含菌量为1×108 CFU ／mL 的佐剂疫苗，通过腹腔注射免疫健康大鲵（平均体长40 cm、平均体质量85 g），每尾1 mL，并设不加佐剂组和对照组；分别于免疫后第1、4、7、14、21、28、35天尾静脉采血，利用凝集反应检测血清抗体效价，于免疫后第35天进行攻毒感染试验。结果表明，与对照组相比，蜂胶佐剂组抗体效价于免疫后第1～21天呈快速上升趋势并在第21天达到峰值，为1∶469．33，弗氏佐剂组于免疫后一直呈上升趋势，在第35天达到最大值，为1∶448．00，随后下降；铝胶佐剂组抗体效价于免疫后第1～28天呈上升趋势并在第28天达到最大值，为1∶362．67，不加佐剂组于免疫后第21天达到峰值，为1∶384．00，随后呈下降趋势，而对照组基本稳定在1∶4．00。攻毒感染试验表明，蜂胶佐剂组、弗氏佐剂组、铝胶佐剂组和不加佐剂组的死亡率为10％、20％、25％和35％，相对免疫保护率分别为88．9％、77．8％、72．2％和61．1％，对照组大鲵死亡率为90％；3种佐剂疫苗均有较好的免疫保护效果，其中以蜂胶佐剂疫苗效果最佳。
Andrias davidianus （the Chinese giant salamander）is a unique and rare amphibian with important value for biodiversity,scientific research and medicine and is also considered a delicacy.However,in recent years,the occurrence of an infectious disease has seriously hampered the development of large-scale cultivation of A.davidia-nus,resulting in economic loss to farmers.Aeromonas hydrophila is the pathogenic bacteria responsible for Virulent ascitesosic disease in A.davidianus,a serious disease that can result in the death of the salamander on a large scale.The extensive use of antibacterial agents could lead to a drug resistance in Aeromonas hydrophila,so it is im-portant to develop a safe and effective vaccine to prevent the disease.In this study,we explored the adjuvant effect of propolis,Freund and aluminum on the immune response produced in the giant salamander by inactivated Aero-monas hydrophila vaccine.Formalin inactivated Aeromonas hydrophila （1 ×108 CFU /mL）was prepared and mixed with propolis,Freund and aluminum.In addition a no adjuvant group and control group were prepared.Two hun-dred and fifty healthy giant salamanders of （40 ±5 cm,85 ±6 g）were randomly assigned to five treatment groups for inoculation by intraperitoneal injection,with the control group vaccinated with an equal volume of DPBS.Pe-ripheral blood samples for each group were collected from the giant salamanders on day 1,4,7,14,21,28 and 35 post-immunization for a serum agglutinating antibody assay.The challenge test with Aeromonas hydrophila was per-formed on d 35 post-immunization and 20 giant salamanders in each group were selected for the test.After two weeks,death and prevalence rates were calculated.Results show that the serum agglutinating antibody titers of the giant salamanders in the propolis adjuvant group followed an upward trend from day 1 -21 and peaked at 1 ∶469.33 on d 21 post-immunization.The antibody titers of the giant salamanders in the Freund adjuvant group followed an upward trend during the whole test and reached the maximum value of 1 ∶448.00 on d 35 post-immuni-zation,then followed a downward trend.The aluminum adjuvant group followed an upward trend from day 1 -28 and reached the maximum value of 1 ∶362.67 on d 28 post-immunization.The no adjuvant group peaked at 1 ∶384.00 on d 21 post-immunization,then decreased continuously and antibody titers of the giant salamanders in the control group was at a constant value of 1 ∶4.00.The challenge test with Aeromonas hydrophila shows that the mortality of the propolis adjuvant group,Freund adjuvant group,aluminum adjuvant group and the no adjuvant group were 10%,20%,25% and 35%,respectively,and the relative percent survival were 88.9%,77.8%, 72.2% and 61.1%,respectively.The mortality of the control group was 90%.Thus,our study demonstrates that the immune protection of the bacterial inactivated vaccine can be improved by using immunoadjuvants,especially by propolis.
出处
《水生态学杂志》
CSCD
北大核心
2015年第5期69-73,共5页
Journal of Hydroecology
基金
农业部公益性行业科研专项经费项目(201203086)
关键词
大鲵
嗜水气单胞菌
灭活疫苗
免疫应答
免疫佐剂
Andrias davidianus
Aeromonas hydrophila
inactivated vaccine
immune response
immunoadjuvants
