Mohamad Aslah, Zamri-Saad Mohd, Amal Mohammad Noor Azmai, Al-Saari Nurhidayu, Monir Md Shirajum, Chin Yong Kit, Md Yasin Ina-Salwany
Aquatic Animal Health and Therapeutics Laboratory, Institute of Bioscience, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia.
Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia.
Vaccines (Basel). 2021 Apr 10;9(4):368. doi: 10.3390/vaccines9040368.
Multiple infections of several bacterial species are often observed under natural farm conditions. The infections would cause a much more significant loss compared to a single infectious agent. Vaccination is an essential strategy to prevent diseases in aquaculture, and oral vaccination has been proposed as a promising technique since it requires no handling of the fish and is easy to perform. This research attempts to develop and evaluate a potential feed-based polyvalent vaccine that can be used to treat multiple infections by spp., , and , simultaneously. The oral polyvalent vaccine was prepared by mixing formalin-killed vaccine of , and strains with commercial feed pellet, and palm oil as an adjuvant was added to improve their antigenicity. Thereafter, a vaccinated feed pellet was tested for feed quality analysis in terms of feed stability in water, proximate nutrient analysis, and palatability, safety, and growth performance using Asian seabass, as a fish host model. For immune response analysis, a total of 300 Asian seabass juveniles (15.8 ± 2.6 g) were divided into two groups in triplicate. Fish of group 1 were not vaccinated, while group 2 was vaccinated with the feed-based polyvalent vaccine. Vaccinations were carried out on days 0 and 14 with oral administration of the feed containing the bacterin at 5% body weight. Samples of serum for antibody and lysozyme study and the spleen and gut for gene expression analysis were collected at 7-day intervals for 6 weeks. Its efficacy in protecting fish was evaluated in aquarium challenge. Following vaccination by the polyvalent feed-based vaccine, IgM antibody levels showed a significant ( < 0.05) increase in serum against , , and and reached the peak at week 3, 5, and 6, respectively. The high-stimulated antibody in the serum remained significantly higher than the control ( < 0.05) at the end of the 6 weeks vaccination trial. Not only that, but the serum lysozyme level was also increased significantly at week 4 ( < 0.05) as compared to the control treatment. The immune-related gene, dendritic cells, C3, Chemokine ligand 4 (CCL4), and major histocompatibility complex class I (MHC I) showed significantly higher expression ( < 0.05) after the fish were vaccinated with the oral vaccine. In the aquarium challenge, the vaccine provided a relative percentage survival of 75 ± 7.1%, 80 ± 0.0%, and 80 ± 0.0% after challenge with , , and , respectively. Combining our results demonstrate that the feed-based polyvalent vaccine could elicit significant innate and adaptive immunological responses, and this offers an opportunity for a comprehensive immunization against vibriosis, streptococcosis, and motile aeromonad septicemia in Asian seabass, Nevertheless, this newly developed feed-based polyvalent vaccination can be a promising technique for effective and large-scale fish immunization in the aquaculture industry shortly.
在自然养殖条件下,经常会观察到几种细菌的多重感染。与单一感染源相比,这些感染会造成更严重的损失。疫苗接种是水产养殖中预防疾病的重要策略,口服疫苗因其无需处理鱼且易于实施,已被视为一种有前景的技术。本研究试图开发并评估一种潜在的基于饲料的多价疫苗,该疫苗可用于同时治疗由[具体菌种1]、[具体菌种2]和[具体菌种3]引起的多重感染。口服多价疫苗是通过将[具体菌种1]、[具体菌种2]和[具体菌种3]菌株的福尔马林灭活疫苗与商业饲料颗粒混合制备而成,并添加棕榈油作为佐剂以提高其抗原性。此后,使用尖吻鲈作为鱼类宿主模型,对接种疫苗的饲料颗粒进行了饲料质量分析,包括在水中的稳定性、近似养分分析、适口性、安全性和生长性能测试。为了进行免疫反应分析,总共300尾尖吻鲈幼鱼(15.8±2.6克)被分成两组,每组一式三份。第1组的鱼未接种疫苗,而第2组则接种了基于饲料的多价疫苗。在第0天和第14天进行口服接种,投喂含菌苗的饲料,投喂量为鱼体重的5%。每隔7天采集血清样本用于抗体和溶菌酶研究,采集脾脏和肠道样本用于基因表达分析,持续6周。通过水族箱攻毒试验评估其对鱼的保护效果。接种基于饲料的多价疫苗后,血清中针对[具体菌种1]、[具体菌种2]和[具体菌种3]的IgM抗体水平显著升高(P<0.05),分别在第3周、第5周和第6周达到峰值。在6周的疫苗接种试验结束时,血清中高刺激产生的抗体仍显著高于对照组(P<0.05)。不仅如此,与对照处理相比,血清溶菌酶水平在第4周也显著升高(P<0.05)。在鱼口服疫苗后,免疫相关基因、树突状细胞、补体C3、趋化因子配体4(CCL4)和主要组织相容性复合体I类(MHC I)的表达显著升高(P<0.05)。在水族箱攻毒试验中,接种疫苗后,在分别用[具体菌种1]、[具体菌种2]和[具体菌种3]攻毒后,疫苗提供的相对存活率分别为75±7.1%、80±0.0%和80±0.0%。综合我们的结果表明,基于饲料的多价疫苗可引发显著的先天性和适应性免疫反应,这为亚洲海鲈的弧菌病、链球菌病和运动性气单胞菌败血症的综合免疫提供了机会。尽管如此,这种新开发的基于饲料的多价疫苗接种在不久的将来可能成为水产养殖业中有效且大规模鱼类免疫的一项有前景的技术。