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基于饲料的重组疫苗的研制及其对奥利亚罗非鱼无乳链球菌细胞表面锚定家族蛋白的功效

Development and efficacy of feed-based recombinant vaccine encoding the cell wall surface anchor family protein of Streptococcus agalactiae against streptococcosis in Oreochromis sp.

机构信息

Department of Veterinary Pathology & Microbiology, Faculty of Veterinary Medicine, University Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.

Department of Veterinary Pathology & Microbiology, Faculty of Veterinary Medicine, University Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.

出版信息

Fish Shellfish Immunol. 2014 Mar;37(1):193-200. doi: 10.1016/j.fsi.2014.01.011. Epub 2014 Jan 30.

DOI:10.1016/j.fsi.2014.01.011
PMID:24486904
Abstract

This study was carried out to determine the antibody responses and protective capacity of an inactivated recombinant vaccine expressing the cell wall surface anchor family protein of Streptococcus agalactiae following oral vaccination against streptococcosis in tilapia. Tilapia were vaccinated orally with 10(6) CFU/mL of the recombinant vaccine incorporated in feed (feed-based recombinant vaccine) (vaccinated group or Group 1), 10(6) CFU/mL of pET-32 Ek/LIC vector without cell wall surface anchor family protein (control group or Group 2), 10(6) CFU/mL of formalin-killed cells of S. agalactiae vaccine incorporated in feed was also prepared (feed-based vaccine) (vaccinated group or Group 3), and unvaccinated control group or Group 4 (fed with commercial pellets). During the course of study, serum, mucus and gut lavage fluid were collected to evaluate the antibody levels via enzyme-linked immunosorbent assay (ELISA). The results showed that tilapia immunized with the feed-based recombinant vaccine developed a strong and significantly (P < 0.05) higher IgM antibody response in serum, mucus and gut lavage fluid samples compared to groups 2, 3 and 4. Following heat intervenes and intraperitoneal challenge, the rate of survivors (RPS) was 70% for the vaccinated group, and 0% for the rest of the groups. Therefore, the study revealed that the feed-based recombinant vaccine significantly provides high protection against high dose challenge in heat stress environment and enhances the production of the mucosal and humoral immunity.

摘要

本研究旨在确定经口服接种表达无乳链球菌细胞壁表面锚定家族蛋白的灭活重组疫苗后,牙鲆对链球菌病的抗体反应和保护能力。牙鲆经口投喂 10(6)CFU/mL 的重组疫苗(饲料型重组疫苗)(接种组或第 1 组)、10(6)CFU/mL 的不含细胞壁表面锚定家族蛋白的 pET-32 Ek/LIC 载体(对照组或第 2 组)、10(6)CFU/mL 的含甲醛灭活的无乳链球菌疫苗的饲料(饲料型疫苗)(接种组或第 3 组),以及未接种的对照组或第 4 组(投喂商业颗粒饲料)进行免疫。在研究过程中,采集血清、黏液和肠灌洗液,通过酶联免疫吸附试验(ELISA)评估抗体水平。结果表明,与第 2、3 和 4 组相比,投喂饲料型重组疫苗的牙鲆在血清、黏液和肠灌洗液样本中产生了强烈且显著(P<0.05)更高的 IgM 抗体反应。经过热干预和腹腔攻毒后,接种组的存活率(RPS)为 70%,而其余组均为 0%。因此,本研究表明,饲料型重组疫苗在热应激环境下对高剂量攻毒具有显著的高保护力,并增强了黏膜和体液免疫的产生。

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