山莨菪碱提高嗜水气单胞菌灭活疫苗浸泡免疫鲫的效果

Anisodamine increases efficacy of Aeromonas hydrophila inactivated vaccine in gibel carp Carassius auratus gibelio by immersion immunization

为研究山莨菪碱作为佐剂在细菌灭活疫苗浸泡免疫中的作用,将山莨菪碱和嗜水气单胞菌全菌灭活疫苗联合浸泡免疫异育银鲫,首次浸泡免疫7 d后加强免疫1次。通过实时荧光定量PCR检测第2、4、7、11、14和21天脾脏中Ig M、IL-1β、C3、C-凝集素以及溶菌酶的mRNA表达量,并在第21天进行同源菌株活菌攻击实验。结果显示,山莨菪碱组第4天Ig M和IL-1β的表达量达到最高值,分别为28.3和332.7;而无佐剂疫苗组第11天Ig M和IL-1β达到最高值,分别为56.1和791.8。山莨菪碱组的补体C3、C-凝集素以及溶菌酶的表达量都显著高于无佐剂组和对照组,且表达持续时间长。活菌攻击实验表明山莨菪碱组的相对免疫保护率可达80.0%,显著高于无佐剂组的56.0%。结果表明,山莨菪碱与嗜水气单胞菌灭活疫苗共同浸泡免疫银鲫,可以增强脾脏中Ig M、IL-1β、C3、C-凝集素以及溶菌酶基因表达,提高银鲫相对免疫保护率。

In order to study the role of anisodamine as an adjuvant in the immersion administration together with inactivated w hole cell vaccines,gibel carps w ere administrated by immersion with inactivated Aeromonas hydrophila vaccine（ inactivated Ah vaccine） with anisodamine on days 1 and 7. Spleens were sampled on days 2,4,7,11,14 and 21 after the second immersion immunization,and the expression levels of IgM,IL-1β,C3,C-lectin and lysozyme mRNAs in spleen were detected through the real-time fluorescence quantitative PCR（ RT-PCR）. The gibel carps from the control and immunization groups w ere challenged intraperitoneally w ith A. hydrophila YYK090901 at the dose of LD50 concentration on day 21 and then mortality was determined and relative percent survival（ RPS） was calculated. The results show ed that the expression levels of Ig M and IL-1β mRNAs of anisodamine-adjuvant vaccine group reached the peak on day4,i. e. 28. 3 and 332. 7 respectively,but the adjuvant-free vaccine group reached the peak on day 11. The expression levels of C3,C-lectin and lysozyme mRNAs of the anisodamine-adjuvant vaccine group were significantly higher than those of the adjuvant-free group and the control,and those of the anisodamineadjuvant group had longer expression duration than the others. The efficacy of inactivated Ah vaccine could be significantly improved by anisodamine through immersion administration,and RPS rate was 80. 0%. The results show ed that anisodamine immersion administrated gibel carps together w ith inactivated whole cell vaccines can significantly promote the expresstion of IgM,IL-1β,C3,C-lectin and lysozyme mRNAs in spleen,and protect gibel carps better against A. hydrophila infection.