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基于饲料的双联全细胞灭活疫苗对奥利亚罗非鱼(尼罗罗非鱼×莫桑比克罗非鱼)链球菌和气单胞菌感染的免疫保护效率。

Immuno-protective efficiency of feed-based whole-cell inactivated bivalent vaccine against Streptococcus and Aeromonas infections in red hybrid tilapia (Oreochromis niloticus × Oreochromis mossambicus).

机构信息

Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.

Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.

出版信息

Fish Shellfish Immunol. 2021 Jun;113:162-175. doi: 10.1016/j.fsi.2021.04.006. Epub 2021 Apr 12.

Abstract

Streptococcosis and motile aeromonad septicemia (MAS) are well-known diseases in tilapia culture, which cause mass mortality with significant economic losses. The development of feed-based bivalent vaccines in controlling these diseases has been initiated, however, the mechanisms of immunities and cross-protection in fish remain unclear. This study was conducted to assess the immuno-protective as well as the cross-protective efficacy of a newly developed feed-based bivalent vaccine against Streptococcus and Aeromonas infections in red hybrid tilapia. A total of five groups of fish were vaccinated orally through two different techniques; bivalent vaccine (inactivated Streptococcus iniae and Aeromonas hydrophila) sprayed on feed pellets (BS group); bivalent vaccine (inactivated S. iniae and A. hydrophila) incorporated in feed (BI group); monovalent inactivated S. iniae and A. hydrophila vaccine separately incorporated into feed as monovalent S. iniae (MS group) and monovalent A. hydrophila (MA group); and control group (without vaccine). The feed-based vaccine was delivered orally at 5% of body weight for five consecutive days. The booster doses were given in the same manner on weeks 2 and 6. Serum and skin mucus samples were collected to assess the IgM responses using indirect ELISA. The first administration of the feed-based vaccine stimulated the IgM levels that lasted until week 3, while the second booster ensured that the IgM levels remained high for a period of 16 weeks in the BI, MS and MA groups. The BI group developed a strong and significantly (P < 0.05) higher systemic and mucosal IgM responses against both S. iniae and A. hydrophila and also cross-protective antigen S. agalactiae and A. veronii compared to the BS and control groups. Quantitative real-time PCR results also showed that the relative expressions of IL-8, INF-γ and IgM in the BI immunized fish spleen, head kidney and hindgut exhibited various significant (P < 0.05) rising trends following both the vaccination and the challenge phase. On weeks 10, all fish were challenged through the intraperitoneal route, where relative percent survivals (RPS) of 82.22 ± 3.85% when challenged with S. iniae, 77.78 ± 3.85% when challenged with A. hydrophila and 77.78 ± 3.85% when co-challenged with both S. iniae and A. hydrophila were observed in the BI group, which were significantly higher (P < 0.05) compared to the other groups. The BI group also showed significantly (P < 0.05) higher partial cross-protections following challenges with S. agalactiae (RPS at 60.00 ± 6.67%) and A. veronii (RPS at 57.78 ± 7.70%). This study demonstrated that immunization with feed-based BI vaccine elicited immune responses that were capable of protecting red hybrid tilapia against streptococcosis and MAS.

摘要

黏球菌病和鳗弧菌败血症(MAS)是罗非鱼养殖中众所周知的疾病,它们会导致大量鱼类死亡,造成重大经济损失。已经开始开发针对这些疾病的基于饲料的双价疫苗,然而,鱼类的免疫和交叉保护机制仍不清楚。本研究旨在评估一种新开发的基于饲料的双价疫苗对红罗非鱼链球菌和气单胞菌感染的免疫保护和交叉保护效果。总共将五组鱼通过两种不同的技术进行口服免疫接种;将双价疫苗(灭活的无乳链球菌和嗜水气单胞菌)喷洒在饲料颗粒上(BS 组);将双价疫苗(灭活的无乳链球菌和嗜水气单胞菌)掺入饲料中(BI 组);单价灭活无乳链球菌和嗜水气单胞菌疫苗分别作为单价无乳链球菌(MS 组)和单价嗜水气单胞菌(MA 组)掺入饲料中;以及对照组(无疫苗)。基于饲料的疫苗以 5%的体重口服给药,连续 5 天。在第 2 周和第 6 周以相同的方式给予加强剂量。收集血清和皮肤黏液样本,使用间接 ELISA 评估 IgM 反应。第一次给予基于饲料的疫苗刺激 IgM 水平,持续到第 3 周,而第二次加强确保 BI、MS 和 MA 组的 IgM 水平在 16 周内保持高水平。BI 组对无乳链球菌和嗜水气单胞菌产生了强烈且显著(P<0.05)更高的系统和黏膜 IgM 反应,并且与 BS 和对照组相比,对链球菌和鳗弧菌交叉保护抗原 S.agalactiae 和 A.veronii 也具有交叉保护作用。实时定量 PCR 结果还表明,BI 免疫鱼的脾脏、头肾和后肠中的 IL-8、INF-γ 和 IgM 的相对表达在接种和攻毒阶段均呈现出各种显著(P<0.05)的上升趋势。在第 10 周,所有鱼都通过腹腔途径进行攻毒,BI 组在攻毒无乳链球菌时的相对存活率(RPS)为 82.22%±3.85%,攻毒嗜水气单胞菌时的 RPS 为 77.78%±3.85%,同时攻毒无乳链球菌和嗜水气单胞菌时的 RPS 为 77.78%±3.85%,均显著高于(P<0.05)其他组。BI 组在攻毒无乳链球菌(RPS 为 60.00%±6.67%)和鳗弧菌(RPS 为 57.78%±7.70%)后也表现出显著(P<0.05)更高的部分交叉保护作用。本研究表明,口服 BI 饲料疫苗免疫可引发免疫反应,使红罗非鱼能够抵抗链球菌病和鳗弧菌败血症。

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