Department of Biotechnology, Faculty of Applied Sciences, UCSI University, Kuala Lumpur, Malaysia.
Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
Fish Shellfish Immunol. 2024 Jun;149:109572. doi: 10.1016/j.fsi.2024.109572. Epub 2024 Apr 16.
Streptococcosis outbreaks caused by Streptococcus agalactiae infection in tilapia aquaculture have been consistently reported and associated with high mortality and morbidity leading to significant economic losses. Existing vaccine candidates against Streptococcus spp. are designed for intraperitoneal injections that are not practical and labor-intensive which have prompted farmers to protect aquatic animals with antibiotics, thus encouraging the emergence of multidrug resistant bacteria. In this study, a live recombinant L. lactis vaccine expressing a 1403 bp surface immunogenic protein (SIP) and a 1100 bp truncated SIP (tSIP) gene was developed and evaluated against S. agalactiae infection in tilapia. Both SIP and tSIP sequences were cloned and transformed into L. lactis. The recombinant L.lactis vaccine was orally administered to juvenile tilapia for a month. Detection of SIP-specific serum IgM in vaccinated groups compared to control groups indicated that recombinant proteins expressed from L. lactis could elicit immunogenic reactions in tilapia. Fish immunized with the tSIP vaccine also showed the highest level of protection compared to other test groups, and the mortality rate was significantly reduced compared to both control groups. The relative percentage of survival (RPS) against S. agalactiae for both SIP and tSIP-vaccinated groups was 50 % and 89 %, respectively, at 14 days post-challenge. Significant up-regulation of IgM, IL-1β, IL-10, TNF-α and IFN-γ were observed at day 34 between the vaccinated and control groups. These results indicated that the recombinant lactococcal tSIP vaccine can elicit both cell-mediated and humoral responses and is recommended as a potential oral vaccine against S. agalactiae infection. Future work will include further in vivo challenge assessments of this vaccine candidate fused with adjuvants to boost immunogenicity levels in tilapia.
无乳链球菌感染罗非鱼养殖中的链球菌病爆发一直有报道,并与高死亡率和发病率相关,导致重大的经济损失。现有的链球菌候选疫苗是为腹腔内注射设计的,这种方法不实际且劳动强度大,这促使农民用抗生素来保护水生动物,从而鼓励了多药耐药菌的出现。在这项研究中,开发并评估了一种表达 1403bp 表面免疫蛋白(SIP)和 1100bp 截断 SIP(tSIP)基因的活重组乳球菌疫苗,以对抗罗非鱼的无乳链球菌感染。SIP 和 tSIP 序列均被克隆并转化到乳球菌中。重组乳球菌疫苗口服施用于罗非鱼幼鱼一个月。与对照组相比,接种组中 SIP 特异性血清 IgM 的检测表明,重组蛋白可以在罗非鱼中引发免疫反应。与其他测试组相比,用 tSIP 疫苗免疫的鱼也显示出最高的保护水平,死亡率与对照组相比显著降低。SIP 和 tSIP 疫苗接种组对无乳链球菌的相对存活率(RPS)分别为 50%和 89%,攻毒后 14 天。在接种组和对照组之间,第 34 天观察到 IgM、IL-1β、IL-10、TNF-α 和 IFN-γ 的显著上调。这些结果表明,重组乳球菌 tSIP 疫苗可以引发细胞介导和体液反应,可作为无乳链球菌感染的潜在口服疫苗。未来的工作将包括进一步评估这种疫苗候选物与佐剂融合后的体内攻毒评估,以提高罗非鱼的免疫原性水平。