Coastal pollution, global warming, ocean acidification, and other reasons lead to the imbalance of the coral reef ecosystem, resulting in the increasingly serious problem of coral degradation. Coral bleaching is often accompanied by structural abnormalities of coral symbiotic microbiota, among which Vibrio is highly concerned. In this study, Vibrio fortis S10-1 (MCCC 1H00104), isolated from sea cucumber, was used for the bacterial infection on coral Seriatopora guttatus and Pocillopora damicornis. The infection of S10-1 led to coral bleaching and a significant reduction of photosynthetic function in coral holobiont, and the pathogenicity of V. fortis was regulated by quorum sensing. Meanwhile, Vibrio infection also caused a shift of coral symbiotic microbial community, with significantly increased abundant Proteobacteria and Actinobacteria and significantly reduced abundant Firmicutes; on genus level, the abundance of Bacillus decreased significantly and the abundance of Rhodococcus, Ralstonia, and Burkholderia–Caballeronia–Paraburkholderia increased significantly; S10-1 infection also significantly impacted the water quality in the micro-ecosystem. In contrast, S10-1 infection showed less effect on the microbial community of the live stone, which reflected that the microbes in the epiphytic environment of the live stone might have a stronger ability of self-regulation; the algal symbionts mainly consisted of Cladocopium sp. and showed no significant effect by the Vibrio infection. This study verified that V. fortis is the primary pathogenic bacterium causing coral bleaching, revealed changes in the microbial community caused by its infection, provided strong evidence for the “bacterial bleaching” hypothesis, and provided an experimental experience for the exploration of the interaction mechanism among microbial communities, especially coral-associated Vibrio in the coral ecosystem, and potential probiotic strategy or QS regulation on further coral disease control.

Biofilm in dental unit water lines may pose a health risk to patients and dental practitioners. An AdiC-like quorum quenching enzyme, YtnP, was cloned from a deep-sea probiotic Bacillus velezensis, and heterologously expressed in E. coli to examine the application on the improvement of hygiene problems caused by biofilm infection of Pseudomonas aeruginosa in dental units. Pseudomonas bacteria were isolated from dental chair units and used to grow static biofilms in the laboratory. A water filter system was designed to test the antifouling activity of YtnP in Laboratory, to simulate the biofilm contamination on water filter in dental unit water lines. The results demonstrated that the enzyme of YtnP was able to degrade the N-acyl homoserine lactones, significantly inhibited the EPS generation, biofilm formation, and virulence factors production (pyocyanin and rhamnolipid) of P. aeruginosa, and was efficient on the antifouling against P. aeruginosa. The findings in this study indicated the possibility of YtnP as novel disinfectant reagent for hygiene treatment in dental units.

Aquatic pathogens such as Vibrio parahaemolyticus cause a bacterial infection that reduces the economic benefits of aquaculture and affects the food quality and safety of human beings. Quorum quenching (QQ) is considered a novel strategy of microbial antagonism that inhibits pathogens and reduces the abuse of antibiotics. This study investigates a QQ bacterial strain, Bacillus velezensis DH82 from the deep sea Yap trench, in vitro to examine the effects of DH82 and its functional products against V. parahaemolyticus, focusing on the Quorum sensing (QS) regulation and the inhibition of pathogenicity and bacterial growth. The study also conducted in vivo investigation in the aquaculture of Litopenaeus vannamei challenged with V. parahaemolyticus by immersion and injection challenge. The results of the QS regulator transcription level demonstrated the multiple QQ enzymes in DH82 regulated the pathogenicity but could not fully control the biofilm formation; the effective antibacterial activity of extracellular peptides on microbial antagonism verified the inhibition on bacterial growth of V. parahaemolyticus. The in vivo experiment in aquaria demonstrated the effective enrichment of DH82 and inhibition of Vibrio in both the aquatic system and the shrimp intestine. The dietary DH82 relieved the negative effect of Vibrio on the activity of enzyme acid phosphatase (ACP), alkaline phosphatase (AKP), superoxide dismutase (SOD) under challenge of Vibrio pathogens, and was not harmful to host according to lysozyme (LZM) activity. DH82 also ameliorated the damage to the intestine and muscles induced by V. parahaemolyticus infection according to tissue imaging. Though DH82 did present some dose-dependent adverse effects to the host, the findings revealed the effective QQ and antibacterial activity of DH82 on emerging biocontrol against V. parahaemolyticus, therefore indicating the potential application of DH82 as a biological control reagent in the sustainable and green production of aquaculture.

Pseudomonas aeruginosa in water lines may cause bacteria pollution indrinking fountains that could affect the quality of potable water, thus posing a risk to public health. A clean and efficient strategy is required for drinking water treatment for food safety. In this study, an AiiA-homologous lactonase was cloned from a deep-sea probiotics Bacillus velezensis (DH82 strain), and was heterologously expressed so that the capacity of the enzyme on the N-acyl-L-homoserine lactone (AHL)-degrading, effect of bacterial proliferation, biofilm formation and toxic factors release, and membrane pollution from P. aeruginosa could each be investigated to analyze the effect of the enzyme on water treatment. The enzyme effectively degraded the signal molecules of P. aeruginosa (C6-HSL and C12-HSL), inhibited early proliferation and biofilm formation, significantly reduced toxic products (pyocyanin and rhamnolipid), and inhibited bacterial fouling on the filter membrane, which prevented the secondary contamination of P. aeruginosa in drinking water. The findings demonstrated that the quorum quenching enzyme from probiotics could prevent bacteria pollution and improve potable water quality, and that the enzyme treatment could be used as a probable strategy for drinking water treatment.

Quorum quenching (QQ) is of interest for potential application as a sustainable strategy for bacterial disease control via communication interruption. The QQ enzyme can be used as a good alternative antagonist to combat antibiotic abuse and bacterial resistance. Here, genomic DNA sequencing was performed on N-acyl homoserine lactonase from the deep-sea strain Bacillus velezensis DH82 with Cluster of Orthologous Groups of proteins (COGs) annotation. The homologous sequences with β-lactamase domain-containing protein were predicted to be potential QQ enzymes and were cloned and expressed to study their quorum quenching properties by comparing them with the reported enzyme AiiA_3DHB. The experimental results of enzyme activity analysis and steady-state kinetics, as well as enzyme structure and substrate docking simulations and predictions, all consistently demonstrated that YtnP_DH82 presented superior enzyme structural stability and higher degradation efficiency of N-acyl homoserine lactones than AiiA_DH82 under the effects of pH, and temperature, and performed better on short -chain and 3-O-substituted AHSLs. The findings revealed the structural and biochemical characterization of YtnP_DH82 from the deep sea, which provide the capacity for further application in sustainable aquaculture as an alternative to antibiotics.

Gaussia luciferase (GLuc) is a secreted protein with significant potential for use as a reporter of gene expression in bacterial pathogenicity studies. To date there are relatively few examples of its use in bacteriology. In this study we show that GLuc can be functionally expressed in the human pathogen Staphylococcus aureus and furthermore show that it can be used as a biosensor for the agr quorum sensing (QS) system which employs autoinducing peptides to control virulence. GLuc was linked to the P3 promoter of the S. aureusagr operon. Biosensor strains were validated by evaluation of chemical agent-mediated activation and inhibition of agr. Use of GLuc enabled quantitative assessment of agr activity. This demonstrates the utility of Gaussia luciferase for in vitro monitoring of agr activation and inhibition.