Data_Sheet_1_Multi-Strain Tropical Bacillus spp. as a Potential Probiotic Biocontrol Agent for Large-Scale Enhancement of Mariculture Water Quality.docx

Aquaculture is suffering from long-term water eutrophication in intensive models, whereas the knowledge of multi-strain/specie for improving water quality is extremely limited. Herein, we aimed to develop multi-strain tropical Bacillus spp. as a potential probiotic biocontrol agent for large-scale enhancement of mariculture water quality. Given the practical application, the optimum multi-strain tropical Bacillus spp. (B. flexus QG-3, B. flexus NS-4, and B. licheniformis XCG-6 with the proportion 5: 5: 4) as a probiotic biocontrol agent was screened and obtained, which effectively improved water quality by removing chemical oxygen demand (COD), ammonia-nitrogen, and nitrate and significantly inhibited Vibrio spp. even at relatively low bacterial concentrations (104 CFU/ml) in artificial feed wastewater and large-scale shrimp aquaculture ponds. More importantly, we found that the initial proportion of these three Bacillus sp. strains of multi-strain tropical Bacillus spp. markedly affected the final purification effects, whereas the initial concentration of that only influenced the purification rates at the early stage (0–48 h) instead of final purification effects. We reason that this multi-strain tropical Bacillus spp. as a good probiotic biocontrol agent could perform multiple actions, such as COD-degrading, nitrifying, denitrifying, and antagonistic actions, for large-scale enhancement of tropical aquaculture water. Additionally, the multi-strain tropical Bacillus spp. was safe for shrimp and could be stored for at least 240 days in spore form at room temperature. This multi-strain probiotic biocontrol agent may facilitate its adoption for further marine recirculating aquaculture system development and large-scale commercial application.