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Some aspects of traditional aquaculture have negative impacts on the aquatic environment, leading to pollution and disease outbreaks in farmed organisms. Biofloc technology (BFT) is a closed aquaculture system that utilizes specific microbial communities to remove ammonia emitted from aquaculture organisms or adds carbon to the aquaculture system to improve water quality. BFT has benefits, such as increasing production and improving water quality, and reducing disease spread and pollution, without the need for water exchange. However, there are disadvantages, such as rapid changes in water quality due to accumulation of dissolved nutrients and total suspended soils (TSS) and the requirement for expensive aeration equipment to maintain dissolved oxygen. BFT can be enhanced in value and efficiency by combining it with other aquaculture technologies, such as aquaponics and vertical aquaculture to overcome the disadvantages. The integration of biofloc with technologies from the fourth industrial revolution holds potential for further development, while aquaponics and vertical farming can eliminate geographical limitations and accelerate the urbanization of aquaculture. The integration of aquaponics and vertical aquaculture with BFT has potential for development, accelerating the urbanization of aquaculture and removing geographic limitations.

The South Korean aquaculture industry has incurred considerable production losses due to various infectious diseases. Artificially synthesized polyinosinic–polycytidylic acid (poly I:C), structurally similar to double-stranded RNA (dsRNA) and cytidine–phosphate–guanosine oligodeoxynucleotides, can enhance immune responses and protect against diseases. Here, we investigated dsRNA molecules with different cytidine–phosphate–guanosine (CpG) motifs (dsRNA-CpGMix) as fused agents to treat Miamiensis avidus and viral hemorrhagic septicemia virus (VHSV) infection in olive flounders. We further investigated the efficacy of specific sequence motifs in dsRNA in modulating immunostimulatory effects. Fish treated with poly I:C or dsRNA-CpGMix exhibited higher survival rates than the control group. Olive flounder leukocytes stimulated with poly I:C or dsRNA-CpGMix showed increased scuticocidal activity in the presence of inactivated immune sera. dsRNA with CpG motif sequences induced higher resistance against M. avidus and VHSV infections than dsRNA without CpG motif sequences, and the dsRNA-CpGMix group showed upregulated ISG15 or Mx compared to the dsRNA-GFP group. Thus, dsRNA containing CpG motifs can be used as effective immunostimulants to enhance resistance against viral and parasitic diseases in olive flounder. The specific sequences of the CpG motifs in dsRNA may be important for enhancing immune responses and resistance against M. avidus and VHSV infections in olive flounders.