Marine microplastic pollution is a growing problem for ecotoxicology that needs to be resolved. In particular, microplastics may be carriers of “dangerous hitchhikers,” pathogenic microorganisms, i.e., Vibrio. Microplastics are colonized by bacteria, fungi, viruses, archaea, algae and protozoans, resulting in the biofilm referred to as the “plastisphere.” The microbial community composition of the plastisphere differs significantly from those of surrounding environments. Early dominant pioneer communities of the plastisphere belong to primary producers, including diatoms, cyanobacteria, green algae and bacterial members of the Gammaproteobacteria and Alphaproteobacteria. With time, the plastisphere mature, and the diversity of microbial communities increases quickly to include more abundant Bacteroidetes and Alphaproteobacteria than natural biofilms. Factors driving the plastisphere composition include environmental conditions and polymers, with the former having a much larger influence on the microbial community composition than polymers. Microorganisms of the plastisphere may play key roles in degradation of plastic in the oceans. Up to now, many bacterial species, especially Bacillus and Pseudomonas as well as some polyethylene degrading biocatalysts, have been shown to be capable of degrading microplastics. However, more relevant enzymes and metabolisms need to be identified. Here, we elucidate the potential roles of quorum sensing on the plastic research for the first time. Quorum sensing may well become a new research area to understand the plastisphere and promote microplastics degradation in the ocean.

Vibrios are among the most diverse and ecologically important marine bacteria, which have evolved many characteristics and lifestyles to occupy various niches. The relationship between genome features and environmental adaptation strategies is an essential part for understanding the ecological functions of vibrios in the marine system. The advent of complete genome sequencing technology has provided an important method of examining the genetic characteristics of vibrios on the genomic level. Two Vibrio genomes were sequenced and found to occupy many unique orthologues families which absent from the previously genes pool of the complete genomes of vibrios. Comparative genomics analysis found vibrios encompass a steady core-genome and tremendous pan-genome with substantial gene gain and horizontal gene transfer events in the evolutionary history. Evolutionary analysis based on the core-genome tree suggested that V. fischeri emerged ~ 385 million years ago, along with the occurrence of cephalopods and the flourish of fish. The relatively large genomes, the high number of 16S rRNA gene copies, and the presence of R-M systems and CRISPR system help vibrios live in various marine environments. Chitin-degrading related genes are carried in nearly all the Vibrio genomes. The number of chitinase genes in vibrios has been extremely expanded compared to which in the most recent ancestor of the genus. The chitinase A genes were estimated to have evolved along with the genus, and have undergone significant purifying selective force to conserve the ancestral state. Vibrios have experienced extremely genome expansion events during their evolutionary history, allowing them to develop various functions to spread globally. Despite their close phylogenetic relationships, vibrios were found to have a tremendous pan-genome with a steady core-genome, which indicates the highly plastic genome of the genus. Additionally, the existence of various chitin-degrading related genes and the expansion of chitinase A in the genus demonstrate the importance of the chitin utilization for vibrios. Defensive systems in the Vibrio genomes may protect them from the invasion of external DNA. These genomic features investigated here provide a better knowledge of how the evolutionary process has forged Vibrio genomes to occupy various niches.

Many bacteria, such as Proteobacteria, Cyanobacteria and Bacteroidetes, use N-acylhomoserine lactones (AHLs) as quorum-sensing (QS) signal molecules for communication. Enzymatic degradation of AHLs, such as AHL acylase and AHL lactonase, can degrade AHLs (quorum quenching, QQ) to attenuate or disarm the virulence of pathogens. QQ is confirmed to be common in marine bacterial communities. Many genes encoding AHL acylases are found in marine bacteria and metagenomic collections, but only a few of these have been characterized in detail. We have reported that the marine bacterium Pseudoalteromonas flavipulchra JG1 can degrade AHLs. In the present study, a novel AHL acylase PfmA, which can degrade AHLs with acyl chains longer than 10 carbons, was identified from strain JG1. Ultra-performance liquid chromatography (UPLC) and electrospray ionization mass spectrometry (ESI-MS) analysis demonstrated that PfmA functions as an AHL acylase, which hydrolysed the amide bond of AHL. The purified PfmA of P. flavipulchra JG1 showed optimum activity at 30 °C and pH 7.0. PfmA belongs to the N-terminal nucleophile (Ntn) hydrolase superfamily and showed homology to a member of penicillin amidases, but PfmA can degrade ampicillin but not penicillin G. The residue Ser256 in PfmA is the active site according to site-directed mutagenesis. Furthermore, PfmA reduced AHL accumulation and the production of virulence factors in Vibrio anguillarum VIB72 and Pseudomonas aeruginosa PAO1, and attenuated the virulence of P. aeruginosa to increase Artemia survival, which suggested that PfmA can be considered as a therapeutic agent to control AHL-mediated pathogenicity.

Gram-negative bacteria utilize N-acylhomoserine lactones (AHLs) as quorum sensing (QS) signaling molecules for intercellular communication. Cell-to-cell communication depends on cell population density, and AHL-dependent QS is related to the production of multiple genes including virulence factors. Quorum quenching (QQ), signal inactivation by enzymatic degradation, is a potential strategy for attenuating QS regulated bacterial infections. Both Gram-positive and -negative bacteria have QQ enzymes that can degrade AHLs. In our previous study, strain Ruegeria mobilis YJ3, isolated from healthy shrimp, showed strong AHLs degradative activity. In the current study, an AHL lactonase (designated RmmL) was cloned and characterized from Ruegeria mobilis YJ3. Amino acid sequence analysis showed that RmmL has a conserved “HXHXDH” motif and clusters together with lactonase AidC that belongs to the metallo-β-lactamase superfamily. Recombinant RmmL could degrade either short- or long-chain AHLs in vitro. High-performance liquid chromatography analysis indicated that RmmL works as an AHL lactonase catalyzing AHL ring-opening by hydrolyzing lactones. Furthermore, RmmL can reduce the production of pyocyanin by Pseudomonas aeruginosa PAO1, while for the violacein and the extracellular protease activities by Chromobacterium violaceum CV026 and Vibrio anguillarum VIB72, no significant reduction was observed. This study suggests that RmmL might be used as a therapeutic agent in aquaculture.

Chitinase, as one of the most important extracellular enzymes in the marine environment, has great ecological and applied values. In this study, two chitinases (Chi1557 and Chi4668) with 97.33% amino acid sequences identity were individually found in Vibrio rotiferianus and Vibrio harveyi. They both were encoding by 561 amino acids, but differed in 15 amino acids and showed different enzymatic properties. The optimal temperature and pH ranges were 45–50 °C and pH 5.0–7.0 for Chi1557, while ~50 °C and pH 3.0–6.0 for Chi4668. K⁺, Mg²⁺, and EDTA increased the enzymatic activity of Chi4668 significantly, yet these factors were inhibitory to Chi1557. Moreover, Chi1557 degraded colloidal chitin to produce (GlcNAc)₂ and minor GlcNAc, whereas Chi4668 produce (GlcNAc)₂ with minor (GlcNAc)₃ and (GlcNAc)₄. The Kcat/Km of Chi4668 was ~4.7 times higher than that of Chi1557, indicating that Chi4668 had stronger catalytic activity than Chi1557. Furthermore, site-directed mutagenesis was performed on Chi1557 focusing on seven conserved amino acid residues of family GH18 chitinases. Chi1557 was almost completely inactive after Glu154, Gln219, Tyr221, or Trp312 was individually mutated, retained ~50% activity after Tyr37 was mutated, and increased two times activity after Asp152 was mutated, indicating that these six amino acids were key sites for Chi1557.

Site F is the most vigorous cold seep known on the continental slope of the northern South China Sea. Up to now, the microbial community structures in sediments of Site F based on the high-throughput sequencing of the 16S rRNA genes have been studied extensively. However, few studies investigated the microbial community structures at fine vertical scales of Site F and control stations outside Site F. In this study, a comprehensive investigation of microbial communities in sediments of Site F along the depths varying from 0 to 24 cm below sea floor (cmbsf) of four sampling sites—SRS (Southern Reduced Sediment), NRS (Northern Reduced Sediment), Control 1 (close to Site F), and Control 2 (far from Site F)—was carried out. The high relative abundances of anaerobic methanotrophic archaea (ANME),Desulfobacterota[sulfate-reducing bacteria (SRB)], andCampylobacteria[sulfur-oxidizing bacteria (SOB)] in SRS and NRS indicated that these two sites were newborn cold seep sites compared with non-seep sites, Control 1, and Control 2. A positive correlation between ANME-1b, ANME-2, and SEEP-SRB and an enrichment ofSulfurovumandMethlomonadaceaewere found in the surface sediments of both SRS and NRS, indicating that the processes of anaerobic oxidation of methane (AOM), sulfur oxidation, and sulfate reduction might occur in seep sites. SRS was enriched with ANME-1b and SEEP-SRB2 with a proposed sulfate-methane transition zone (SMTZ) approximately located at 8 cmbsf. The high abundance of ANME in SRS may due to the high concentration of methane. NRS was enriched with ANME-2,Desulfatiglans,Sulfurovum, andMethanosarcinaceaewith a proposed SMTZ at about 10 cmbsf. According to the analyses of microbial community structure and environmental factors, NRS could be described as a notable cold seep reduced sediment site with low sulfate and high H₂S that nourished abundant SEEP-SRB1, ANME-2,Methanosarcinales, andSulfurovum, which showed similar distribution pattern. Our study expands the current knowledge on the differences of microbial communities in cold seep sites and non-seep sites and sheds light on the horizontal and vertical heterogeneity of sediment microbial community in Site F.