Barley as a production platform for oral vaccines in sustainable fish aquaculture

Vaccination is the most effective measure to prevent disease outbreaks in fish aquaculture, with oral vaccine administration emerging as the most practical approach. However, oral vaccines face a notable limitation due to insufficient stimulation of the complex gut-associated lymphoid tissue caused by factors such as vaccine degradation, poor absorption, and recognition by the immune cells. An innovative solution to these limitations lies in the plant-based production of recombinant vaccines. Plant cells enable the production and targeted storage of recombinant vaccines in specific cell organelles which ensure superior protection from degradation and contain natural compounds acting as adjuvants. Our study explores the potential of barley (Hordeum vulgare), a globally significant cereal crop, for producing orally administered subunit vaccines against viral infections affecting economically important fish species in the Salmonidae and Cyprinidae families. Through Agrobacterium-mediated transformation of immature barley embryos, we have generated homozygous T₂ generation of transgenic barley expressing recombinant antigens of spring viremia of carp virus and infectious salmon anaemia virus. The expression of these plant-based recombinant vaccines was confirmed by immunodetection, which was supported by fluorescence observation, specifically in the seed endosperm. The antigenicity of transgenic plant material containing recombinant antigens was evaluated using an intubation model of common carp (Cyprinus carpio), revealing a substantial upregulation of the immunoglobulin transcripts in both systemic and mucosal tissues over a period of 28 days following a single dose of transgenic antigens. Collectively, these results underscore the potential of barley-based recombinant vaccines for disease prevention in fish aquaculture.