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This study attempted to evaluate the potential for biofilm formation and the probiotic properties of lactic acid bacteria (LAB) isolated from the Carassius auratus intestine and to determine their antibacterial activity against crucial injurious bacteria of fish. The biological features test was carried out to obtain LAB with well-resistant pH, trypsin, bile and antibiotic challenge, and biofilm formation capacity. The most promising LAB isolates, y11 and y78 were identified as Lactobacillus johnsonii and Weissella confusa using 16S ribosomal RNA gene sequencing. We discovered that oral LAB additives improved immunity in the C. auratus by increasing immune-related expression of immunoglobulin M (IgM), superoxide dismutase (SOD), alkaline phosphatase (AKP), and lysozyme (LYS) in serum and interleukins (IL-10 and IL-1β), interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) in organs. After the introduction of Aeromonas veronii TH0426 for 15 days, the survival rate of C. auratus was determined as follows: the protection rate of group y11 was 45 %, and group y78 was 65 % when challenged with 109 CFU/mL density. This experiment successfully selected two LAB strains as the bioprotective agents and provided theoretical support for further LAB application in aquaculture.

Bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) are a group of antibacterial proteins that play an important role in the host’s innate immune defense against pathogen infection. In this study, two BPI/LBPs, named ToBPI1/LBP (1434 bp in length, 478 amino acids) and ToBPI2/LBP (1422 bp in length, 474 amino acids), were identified from the golden pompano. ToBPI1/LBP and ToBPI2/LBP were significantly expressed in immune-related tissues after being challenged with Streptococcus agalactiae and Vibrio alginolyticus. The two BPI/LBPs showed significant antibacterial activity against Gram-negative Escherichia coli and Gram-positive S. agalactiae and Streptococcus iniae. In contrast, the antibacterial activity against Staphylococcus aureus, Corynebacterium glutamicum, Vibrio parahaemolyticus, V. alginolyticus and Vibrio harveyi was low and decreased with time. The membrane permeability of bacteria treated with recombinant ToBPI1/LBP and ToBPI2/LBP was significantly enhanced. These results suggest that ToBPI1/LBP and ToBPI2/LBP may play important immunological roles in the immune response of the golden pompano to bacteria. This study will provide basic information and new insights into the immune response mechanism of the golden pompano to bacteria and the function of BPI/LBP.

Lactiplantibacillus plantarum could regulate certain physiological functions through the AI-2/LuxS-mediated quorum sensing (QS) system. To explore the regulation mechanism on the growth characteristics and bacteriostatic ability of L. plantarum SS-128, a luxS mutant was constructed by a two-step homologous recombination. Compared with ΔluxS/SS-128, the metabolites of SS-128 had stronger bacteriostatic ability. The combined analysis of transcriptomics and metabolomics data showed that SS-128 exhibited higher pyruvate metabolic efficiency and energy input, followed by higher LDH level and metabolite overflow compared to ΔluxS/SS-128, resulting in stronger bacteriostatic ability. The absence of luxS induces a regulatory pathway that burdens the cysteine cycle by quantitatively drawing off central metabolic intermediaries. To accommodate this mutations, ΔluxS/SS-128 exhibited lower metabolite overflow and abnormal proliferation. These results demonstrate that the growth characteristic and metabolism of L. plantarum SS-128 are mediated by the AI-2/LuxS QS system, which is a positive regulator involved in food safety. It would be helpful to investigate more bio-preservation control potential of L. plantarum, especially when applied in food industrial biotechnology.