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以温和气单胞菌灭活菌苗为免疫原,平均体重(100±25)g的健康中华倒刺鲃为实验对象,免疫组腹腔注射0.2mL浓度为1.0×108CFU/mL的免疫原,对照组注射等量灭菌生理盐水,分别在单次注射0、1、2、4、7、14、21、28、35d后随机从两组各取6尾实验鱼,尾静脉采血,测定外周血的血细胞数量、白细胞分类计数、吞噬活性、抗体效价和蛋白质含量等免疫指标的变化,第35天活菌攻毒。结果表明:温和气单胞菌灭活菌苗(F-AS)可诱导中华倒刺鲃红细胞和白细胞数量增加,并引起各种白细胞分类百分比变化,提高吞噬活性和抗体效价,血清中总蛋白及球蛋白含量增加,红细胞也具有免疫功能,灭活茵苗的相对免疫保护力达65.21%。免疫早期(第1周)主要是红细胞、单核细胞和中性粒细胞数量明显增加,吞噬细胞的吞噬活性迅速提高,吞噬百分比和吞噬指数第4天达峰值;随后则是淋巴细胞大量增殖,第21天淋巴细胞、抗体效价及球蛋白达峰值。可见灭活菌苗通过促进中华倒刺鲃血细胞增殖、提高吞噬细胞的吞噬活性、产生特异性抗体等方式提高免疫保护力;免疫早期非特异性细胞免疫起重要作用,之后特异性免疫起主要作用。
In the experiment, 0.2 ml immunogen was injected intraperitoneally to the immunized group with the concentration of 1.0 × 108CFU / mL, and the control group was immunized with A. aeruginosa bacterin with the average body weight of (100 ± 25) g. The rats were injected with the same amount of sterilized normal saline. Blood samples were obtained from 6 experimental fish and tail vein at 0, 1, 2, 4, 7, 14, 21, 28 and 35 d after a single injection respectively. The number of blood cells, white blood cell count, phagocytic activity, antibody titer and protein content and other immune indicators of change, the first 35 days live bacteria challenge. The results showed that: A. aeruginosa inactivated vaccine (F-AS) could induce the increase of the number of red blood cells and white blood cells of Spinibarbus sinensis, and caused a variety of leucocyte classification percentages, increased phagocytic activity and antibody titer, total protein in serum And globulin increased, red blood cells also have immune function, inactivation of vaccine relative immunity protection of 65.21%. The number of erythrocytes, monocytes and neutrophils increased significantly in the early stage of immunization (week 1), the phagocytic activity of phagocytes rapidly increased, the percentage of phagocytosis and phagocytic index peaked on the 4th day, and then the lymphocytes proliferated significantly, Day 21 Lymphocytes, antibody titer and globulin peak. It can be seen that the inactivated bacterin plays an important role in promoting immune protection by promoting phagocyte’s phagocytic activity, producing specific antibodies and the like by promoting the proliferation of blood cells of Spinibarbus sinensis, and the specific immunity plays an important role after immunization.