Fishery College of Jimei University/Engineering Research Center of Modern Eel Industrial Technology of the Ministry of Education, PRC, Jimei University, Yindou Road, Fujian, Xiamen 361021, China.
Fish Shellfish Immunol. 2013 Aug;35(2):213-20. doi: 10.1016/j.fsi.2013.04.027. Epub 2013 May 2.
The specific and non-specific immune parameters and protection of American eels (Anguilla rostrata) were evaluated after immunized eels with a bivalent expressed out membrane protein (OMP) of porin Ⅱ of Aeromonas hydrophila and ompS2 of Edwardsiella tarda. One hundred eighty eels were distributed into 3 equal groups and intraperitoneal (i.p) injection with phosphate-buffered saline (PBS group), formalin-killed-whole-cell (FKC) of A. hydrophila and E. tarda (FKC group) or the bivalent OMP (OMP group). The lymphocytes and red blood cells collected on 14, 21 and 42 days post-vaccination were used to evaluate the stimulation index (SI) and the sera collected on 14, 21, 28 and 42 days were used to assize the titers of specific antibody as well as lysozyme activity. Lysozyme activities in skin mucus, suspension of liver and kidney were also recorded on 14, 21 and 28 days. On 28 d post-vaccination, eels from all three groups were challenged by i.p injection of live A. hydrophila or E. tarda. The results show that, compared with the PBS group, proliferation of lymphocytes in OMP group was significantly (P < 0.05) enhanced on 21 days, and the serum titers of anti-A. hydrophila and anti- E. tarda antibody in eels of FKC and OMP group were significant increased (P < 0.05 or P < 0.01) on 14, 21 and 28 days. Activity of the lysozyme in serum, skin mucus, liver and kidney were significant changed (P < 0.05 or P < 0.01) between the three groups. Relative Percent Survival (RPS) after challenged with A. hydrophila on 28 days post immunization in two vaccinated groups vs. PBS group were 50%, and the RPS challenge E. tarda in FKC and OMP vs. PBS group were 50% and 37.5% respectively. These results suggest that American eels immunized with the bivalent OMP would positively affect specific as well as non-specific immune parameters and protect against infection by the two pathogens in freshwater farming.
本研究评估了经二价表达外膜蛋白(OMP)的铜绿假单胞菌 Porin Ⅱ和迟缓爱德华氏菌 ompS2免疫后的美洲鳗(Anguilla rostrata)的特异性和非特异性免疫参数和保护作用。将 180 条美洲鳗随机分为 3 组,分别经腹腔(i.p.)注射磷酸盐缓冲盐水(PBS 组)、福尔马林杀死的铜绿假单胞菌和迟缓爱德华氏菌全细胞(FKC 组)或二价 OMP(OMP 组)。在免疫后第 14、21 和 42 天采集淋巴细胞和红细胞,评估刺激指数(SI);在免疫后第 14、21、28 和 42 天采集血清,检测特异性抗体滴度和溶菌酶活性。在免疫后第 14、21 和 28 天还记录了皮肤黏液、肝和肾悬浮液中的溶菌酶活性。在免疫后第 28 天,所有三组鳗鲡均通过腹腔注射活的铜绿假单胞菌或迟缓爱德华氏菌进行攻毒。结果表明,与 PBS 组相比,OMP 组的淋巴细胞增殖在第 21 天显著增强(P < 0.05),FKC 和 OMP 组鳗鲡的血清抗铜绿假单胞菌和抗迟缓爱德华氏菌抗体滴度在第 14、21 和 28 天显著升高(P < 0.05 或 P < 0.01)。三组间血清、皮肤黏液、肝和肾中的溶菌酶活性均发生显著变化(P < 0.05 或 P < 0.01)。免疫后第 28 天攻毒铜绿假单胞菌后,两个免疫组相对于 PBS 组的相对存活率(RPS)分别为 50%,攻毒迟缓爱德华氏菌后 FKC 和 OMP 组相对于 PBS 组的 RPS 分别为 50%和 37.5%。这些结果表明,用二价 OMP 免疫的美洲鳗可积极影响特异性和非特异性免疫参数,并防止在淡水养殖中感染这两种病原体。