The efficacy of new oral vaccine feeds against Salmonid novirhabdovirus in rainbow trout

Bartolomeo Gorgoglione , Juan-Ting Liu , Jie Li , Vikram N. Vakharia
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Abstract

Salmonid novirhabdovirus (IHNV) causes infectious haematopoietic necrosis (IHN) in salmonid species. Despite an injectable plasmid-based DNA vaccine of the glycoprotein (G) gene is effective, there are no oral vaccines for mass vaccination of rainbow trout (Oncorhynchus mykiss) fry. Recombinant baculoviruses were generated, used in cabbage looper (Trichoplusia ni) insect larvae to produce IHNV G and IHNV G-C5a proteins. Western blotting and chemiluminescence assays confirmed the expression of recombinant proteins, which were added to the fish feeding and top-coated with unflavored gelatin binder. Commercial rainbow trout were fed with experimental diets containing either IHNV G or IHNV G-C5a proteins for 2 weeks, and boosted 4 weeks after. Four weeks post-booster, fish were challenged with IHNV by immersion. Survival upon the infection challenge was evaluated. Spleen were sampled at 7 and 14 days post infection (dpi). Non-vaccinated and IHNV G fed trout reached a mortality of 91.7 and 97.6%, and 70.9 and 88.4%, respectively at 8 and 15 dpi. The IHNV G-C5a fed group exhibited a reduced mortality of 51.2% at 8 dpi, reaching 81.7% at 15 dpi, suggesting some level of antiviral protection. The individual viral load was measured by RT-qPCR detection of IHNV N gene, showing no significant difference across experimental groups. The transcription modulation of selected immune response markers was evaluated across experimental groups, including Type I IFN-a, Mx-1, CD4, and IgM. Further study is needed to assess how new oral vaccines may become effective to mitigate IHNV pathogenesis in juvenile trout by modulating the host immune response to protect towards IHNV exposure.

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虹鳟新冠病毒口服疫苗饲料的效果
沙门氏菌新杆状病毒(IHNV)在沙门氏菌属物种中引起感染性造血坏死(IHN)。尽管糖蛋白(G)基因的可注射的基于质粒的DNA疫苗是有效的,但还没有用于大规模接种虹鳟鱼苗的口服疫苗。构建了重组杆状病毒,并将其用于甘蓝环虫(Trichoplusia ni)幼虫中产生IHNV G和IHNV G-C5a蛋白。蛋白质印迹和化学发光分析证实了重组蛋白的表达,这些蛋白被添加到鱼类饲料中,并用未经处理的明胶粘合剂进行顶部涂布。商业虹鳟用含有IHNV G或IHNV G-C5a蛋白的实验日粮喂养2周,并在4周后增强。加强剂注射后四周,通过浸泡的方式对鱼类进行IHNV攻击。评估感染激发后的存活率。在感染后7天和14天对脾脏进行取样。未接种疫苗和IHNV G喂养的鳟鱼在8和15dpi时的死亡率分别为91.7%和97.6%,70.9%和88.4%。IHNV G-C5a喂养组在8 dpi时的死亡率降低了51.2%,在15 dpi时达到81.7%,这表明有一定程度的抗病毒保护作用。通过IHNV N基因的RT-qPCR检测来测量个体病毒载量,显示实验组之间没有显著差异。在包括I型IFN-a、Mx-1、CD4和IgM在内的实验组中评估所选免疫反应标志物的转录调节。需要进一步的研究来评估新的口服疫苗如何通过调节宿主免疫反应来保护对IHNV暴露的保护,从而有效减轻幼年鳟鱼的IHNV发病机制。
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2.60
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12 weeks
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