Marine Biofilms Research Articles

Bathiapeptides: Polythiazole-Containing Peptides from a Marine Biofilm-Derived Bacillus sp.

Bacteria in marine biofilms are a rich reservoir of natural products. To facilitate novel secondary metabolite discovery, we investigated the metabolic profile of a marine biofilm-derived Bacillus sp. B19-2 by combining bioinformatics and LC-UV-MS analyses. After dereplication and purification of putatively unknown compounds, a new family of compounds 1-8 was uncovered and named bathiapeptides. Structural elucidation using NMR, HRESIMS, ozonolysis, advanced Marfey's analysis, and X-ray diffraction revealed that bathiapeptides are polypeptides that contain a rare polythiazole moiety. These compounds exhibited strong cytotoxicity against Hep G2, HeLa, MCF-7, and MGC-803 cell lines, and the lowest IC50 value was 0.5 μM. An iterative biosynthesis logic in bathiapeptides' biosynthesis was proposed based on the identified chemical structures and putative gene cluster analysis.

Enhancing Biocide Efficacy: Targeting Extracellular DNA for Marine Biofilm Disruption.

Biofilm formation is a global health, safety and economic concern. The extracellular composition of deleterious multispecies biofilms remains uncanvassed, leading to an absence of targeted biofilm mitigation strategies. Besides economic incentives, drive also exists from industry and research to develop and apply environmentally sustainable chemical treatments (biocides); especially in engineered systems associated with the marine environment. Recently, extracellular DNA (eDNA) was implicated as a critical structural polymer in marine biofilms. Additionally, an environmentally sustainable, multi-functional biocide was also introduced to manage corrosion and biofilm formation. To anticipate biofilm tolerance acquisition to chemical treatments and reduce biocide application quantities, the present research investigated eDNA as a target for biofilm dispersal and potential enhancement of biocide function. Results indicate that mature biofilm viability can be reduced by two-fold using reduced concentrations of the biocide alone (1 mM instead of the recommended 10 mM). Importantly, through the incorporation of an eDNA degradation stage, biocide function could be enhanced by a further ~90% (one further log reduction in viability). Biofilm architecture analysis post-treatment revealed that endonuclease targeting of the matrix allowed greater biocide penetration, leading to the observed viability reduction. Biofilm matrix eDNA is a promising target for biofilm dispersal and antimicrobial enhancement in clinical and engineered systems.

Effect of the properties of long afterglow phosphors on the antifouling performance of silicone fouling-release coating

To improve the static antifouling performance of the fouling-release silicone antifouling coating, blue-green (BG), yellow-green (YG), and sky blue (SB) long afterglow phosphors (LAP) with two kinds of luminescence intensities are added into the silicone coating, respectively, and the LAP/polydimethylsiloxane (PDMS) composite antifouling coating with different colors and luminance was prepared. Using a scanning electron microscope (SEM), X-ray diffractometer (XRD), laser particle size analyzer, and fluorescence spectrometer, the crystal structure, particle size, fluorescence properties, and emission wavelength of LAP were studied. Through laser confocal scanning microscope (CLSM), tensile test, contact angle measurement, photometer, marine bacteria, and boat-shaped benthic diatom attachment test, the surface morphology and roughness, tensile properties, wettability, and surface of the coating were studied energy, fluorescence illuminance, and anti-fouling performance. The results showed that the addition of LAP increased the fracture strength, elastic modulus, and surface free energy of the coatings. Each kind of LAP can improve the performance of PDMS coatings to inhibit the adhesion of marine bacterial biofilm and benthic marine algae. The higher the luminous illuminance, the better the antifouling performance of the coating. The SB coating has the most excellent antifouling performance and in particular, the relative adhesion rate (RAR) of fouling organisms of the SB highlight LAP/PDMS composite antifouling coating is only 37.8 %. This research provides new ideas for the development of new fluorescent antifouling coatings and has a certain enlightening significance for improving the static antifouling performance of fouling-release antifouling coatings (FRC).

Microbial Richness of Marine Biofilms Revealed by Sequencing Full-Length 16S rRNA Genes.

Marine biofilms are a collective of microbes that can grow on many different surfaces immersed in marine environments. Estimating the microbial richness and specificity of a marine biofilm community is a challenging task due to the high complexity in comparison with seawater. Here, we compared the resolution of full-length 16S rRNA gene sequencing technique of a PacBio platform for microbe identification in marine biofilms with the results of partial 16S rRNA gene sequencing of traditional Illumina PE250 platform. At the same time, the microbial richness, diversity, and composition of adjacent seawater communities in the same batch of samples were analyzed. Both techniques revealed higher species richness, as reflected by the Chao1 index, in the biofilms than that in the seawater communities. Moreover, compared with Illumina sequencing, PacBio sequencing detected more specific species for biofilms and less specific species for seawater. Members of Vibrio, Arcobacter, Photobacterium, Pseudoalteromonas, and Thalassomonas were significantly enriched in the biofilms, which is consistent with the previous understanding of species adapted to a surface-associated lifestyle and validates the taxonomic analyses in the current study. To conclude, the full-length sequencing of 16S rRNA genes has probably a stronger ability to analyze more complex microbial communities, such as marine biofilms, the species richness of which has probably been under-estimated in previous studies.

Metagenomic analysis of pioneer biofilm-forming marine bacteria with emphasis on Vibrio gigantis adhesion dynamics

Marine biofilms occur frequently and spontaneously in seawater, on almost any submerged solid surface. At the early stages of colonization, it consists of bacteria and evolves into a more complex community. Using 16S rRNA amplicon sequencing and comparative metagenomics, the composition and predicted functional potential of one- to three-day old bacterial communities in surface biofilms were investigated and compared to that of seawater. This confirmed the autochthonous marine bacterium Vibrio gigantis as an early and very abundant biofilm colonizer, also functionally linked to the genes associated with cell motility, surface attachment, and communication via signaling molecules (quorum sensing), all crucial for biofilm formation. The dynamics of adhesion on a solid surface of V. gigantis alone was also monitored in controlled laboratory conditions, using a newly designed and easily implementable protocol. Resulting in a calculated percentage of bacteria-covered surface, a convincing tendency of spontaneous adhering was confirmed. From the multiple results, its quantified and reproducible adhesion dynamics will be used as a basis for future experiments involving surface modifications and coatings, with the goal of preventing adhesion.

Marine biofilms: diversity, interactions and biofouling

Marine biofilms are ubiquitous in the marine environment. These complex microbial communities rapidly respond to environmental changes and encompass hugely diverse microbial structures, functions and metabolisms. Nevertheless, knowledge is limited on the microbial community structures and functions of natural marine biofilms and their influence on global geochemical cycles. Microbial cues, including secondary metabolites and microbial structures, regulate interactions between microorganisms, with their environment and with other benthic organisms, which affects their community succession and metamorphosis. Furthermore, marine biofilms are key mediators of marine biofouling, which greatly affect marine industries. In this Review, we discuss marine biofilm dynamics, including their diversity, abundance and functions. We also highlight knowledge gaps, areas for future research and potential biotechnological applications of marine biofilms.

Insight on the Realizing of Agro-Based Sustainable Low-Cost Fiber Particulate Composite for Thin Film Application in Biofouling Environment

Biofouling is a severe problem in the marine environment, as all hard surfaces, including some macroorganisms, mostly get colonized by microorganisms existing in the maritime. In the past, some substances such as tributyltin and mercury have been used to combat this problem. These substances are still toxic and harmful to aquatic lives and even human beings, leading to these substances being banned globally. It is then highly important to produce anti-biofouling substances from natural products, which are sustainable, economical, environmentally friendly and applied on surfaces conveniently. The inspiration for using these natural substances came from the fact that the microorganisms, which are biofoulers, do not get attached or colonized on marine animals such as whales, dolphins and shellfishes. This review highlights some of the advances in natural and agro-based products that have been used as anti-biofouling. These natural substances are corals, fermented milk, pepper, marine biofilms, seaweeds and shellfishes.

Complete genome sequence of Halomonas boliviensis strain kknpp38, a chlorine-resistant bacterium isolated from the early-stage marine biofilm.

H. boliviensis strain kknpp38 is a dense exopolysaccharide (EPS) producing bacterium, isolated from the early-stage (72-h-old) of marine biofilm. Laboratory experiments demonstrated that this isolate forms a potent biofilm on various artificial substrata viz. polystyrene, stainless steel as well as titanium and possesses high tolerance to chlorine disinfection. To determine the genes and biosynthetic pathways involved in the EPS production, whole-genome sequencing was performed using high-throughput Illumina tag sequencing. The high-quality reads were first de novo assembled using Unicycler genome assembler (version 0.4.9b) and then annotated using Prokka (version 1.13). The complete genome comes from one circular chromosome containing 4.96 Mbp DNA with G+C content of 55%, and encompasses genes encoding 4476 proteins, 2 rRNAs, and 57 tRNAs. Intriguingly, genomic analysis revealed the existence of genes involved in ATP-binding cassette (ABC) transporter-dependent EPS biosynthesis pathways (ugd, ugd2, galU). In addition, we identified genes involved in ectoine (ectA, ectB, ectC, ectD) and polyhydroxyalkanoates (PHAs; fabA, fabB, fabD, fabF, fabH, fabV, fabZ, phaC, phaD, phaG, phaR, phaZ1) production, which are known to involve in bacterial adaptation in saline environment. The outcomes of this study expand scientific understanding on the genes and pathways involved in EPS biosynthesis by marine bacteria.

Bio-approach ZnO/Ag nano-flowers: enhanced photocatalytic and photoexcited anti-microbial activities towards pathogenic bacteria

Zinc oxide (ZnO) and silver (Ag) nanoparticles (NPs) are well proven as one of the most common anti-microbial agents used for many environmental applications. However, in the present study, Sargassum muticum (SM) -mediated nanoparticles (NPs) and its their nanocomposites (NCs) were proved to be photocatalytic and photoexcited anti-microbial agents. Drug-resistant superbugs (DRS) were isolated, cultured, and 16s rDNA identified as B. filamentosus; B. flexus; P. stutzeri; and A. baumannii. The anti-microbial and photoexcited anti-microbial studied by using various microbes such as pathogenic bacteria (PB),drug-resistant superbugs (DRS), and marine biofilm bacteria (MBB), respectively. The anti-microbial activity showed 80 μg/mL of flower-like SMZnO/Ag–NCs (S5) displayed a maximum growth inhibition of 21 mm against PB and a minimum of 11 mm against DRS. The photoexcited anti-microbial killing efficacy was observed in Gram-positive PB strains showed a maximum activity of 16.7 [ × 102] and a minimum in DRS 14.6 [ × 103] CFUs mL−1 and Gram-negative strains 9.4 [ × 102] and minimum in DRS 10.1 [ × 103] CFUs mL−1 display a reduced density within 20 min interval. The results indicate, that green NCs with UV-light phototherapy to generating reactive oxygen species (ROS) that cause bacterial cell death. In addition, the UV-light phototherapy combined with nano-flowers S5 attained an effective degradation of harmful toxic methylene blue (MB) dyes up to 98.6% upon 45 min of irradiation. Photostability/reusability of prepared nano-flowers S5 provided up to 6 consecutive cycles of usage towards MB decolorization for the aquatic environment as well as anti-microbial effects.

Impact of phosphate concentration on the metabolome of biofilms of the marine bacterium Pseudoalteromonas lipolytica.

Marine biofilms are the most widely distributed mode of life on Earth and drive biogeochemical cycling processes of most elements. Phosphorus (P) is essential for many biological processes such as energy transfer mechanisms, biological information storage and membrane integrity. Our aim was to analyze the effect of a gradient of ecologically relevant phosphate concentrations on the biofilm-forming capacity and the metabolome of the marine bacterium Pseudoalteromonas lipolytica TC8. In addition to the evaluation of the effect of different phosphate concentration on the biomass, structure and gross biochemical composition of biofilms of P. lipolytica TC8, untargeted metabolomics based on liquid chromatography-mass spectrometry (LC-MS) analysis was used to determine the main metabolites impacted by P-limiting conditions. Annotation of the most discriminating and statistically robust metabolites was performed through the concomitant use of molecular networking and MS/MS fragmentation pattern interpretation. At the lowest phosphate concentration, biomass, carbohydrate content and three-dimensional structures of biofilms tended to decrease. Furthermore, untargeted metabolomics allowed for the discrimination of the biofilm samples obtained at the five phosphate concentrations and the highlighting of a panel of metabolites mainly implied in such a discrimination. A large part of the metabolites of the resulting dataset were then putatively annotated. Ornithine lipids were found in increasing quantity when the phosphate concentration decreased, while the opposite trend was observed for oxidized phosphatidylethanolamines (PEs). This study demonstrated the suitability of LC-MS-based untargeted metabolomics for evaluating the effect of culture conditions on marine bacterial biofilms. More precisely, these results supported the high plasticity of the membrane of P. lipolytica TC8, while the role of the oxidized PEs remains to be clarified.

Surface Characteristics Together With Environmental Conditions Shape Marine Biofilm Dynamics in Coastal NW Mediterranean Locations

Microbial colonization of artificial substrates in coastal areas, which concerns hull ships, sensors as well as plastic debris, is of huge significance to attain a rational environmental management. Some surface and environmental drivers of biofilm development have previously been described but their relative impact on the formation of biofilms remains unknown while crucial. Especially, there is no evidence of the relative importance of physical surface properties (wettability, roughness, smoothness) compared to seawater characteristics in driving biofilm abundance and diversity. In addition, few studies have considered the temporal evolution of this complex form of colonization, which often prevent to globally understand the process. Using experimental facilities in two Mediterrranean locations, a multidisciplinary approach including surface characterizations as well as seawaterquality analyses, flow cytometry and 16S rDNA metabarcoding, allowed for the identification of the main drivers of colonization for two antifouling (AF) coatings. One AF coating released copper (SPC1) while the other limit colonization thanks to physical properties, namely a low surface energy, roughness and smoothness (FRC1). Results were obtained over 75 days and compared to a control surface (PVC). Biofilm development was observed on all surfaces, with increasing density from AF coatings to PVC. Pionneer bacteria were dissimilar within all three surface types, however, communities observed on FRC1 converged toward PVC ones overtime, whereas SPC1 communities remained highly specific. A remarkably low and unique diversity was found on SPC1 during the experiment as Alteromonas accounted for more than 90% of the community colonizing this substrate until 12 days, and remained one of the co-dominant taxa of mature biofilms. Moreover, clear differences were found between geographical locations. Low nutrients and higher hydrodymanics in Banyuls bay resulted in less dense biofilms overall compared to Toulon, but also in a the slower dynamic of biofilm formation. This is illustrated by the persistence of pioneer Alteromonas but also Hyphomonadacae after 75 days on SPC1. We concluded that, even if local environmental conditions influenced the composition of biofilm communities, particular physical features may control the biofilm density but not the diversity, while copper releasing coating controlled both. In addition, it is evident from these results that sequential biofilm dynamics should carefully be considered as initial processes of formation differed from the long-term ones.

Marine Vibrio spp. protect carbon steel against corrosion through secreting extracellular polymeric substances

The protection of marine materials against corrosion using marine bacterial biofilms is a promising strategy. However, little is known about the mechanisms of this attractive corrosion prevention method. In this work, the corrosion behaviors of X80 carbon steel (CS) in the presence of three different marine Vibrio species were studied. The results demonstrated that all the three Vibrio spp. displayed significant corrosion protection with a weight loss reduction of up to 68%. Moreover, their corrosion prevention performance was tightly related to their abilities to form biofilms, which was in the order of Vibrio sp. EF187016 > Vibrio alginolyticus > Vibrio parahaemolyticus. To further investigate the corrosion prevention mechanism caused by marine biofilms, the extracellular polymeric substances (EPS) of Vibrio sp. EF187016 was extracted and added to 3.5 wt% NaCl for abiotic corrosion testing. The results suggested that the EPS inhibited corrosion, which means EPS can play a significant role in corrosion protection by biofilm.

Profiling Signal Transduction in Global Marine Biofilms

Microbes use signal transduction systems in the processes of swarming motility, antibiotic resistance, virulence, conjugal plasmid transfer, and biofilm formation. However, the signal transduction systems in natural marine biofilms have hardly been profiled. Here we analyzed signal transduction genes in 101 marine biofilm and 91 seawater microbial metagenomes. The abundance of almost all signal transduction-related genes in biofilm microbial communities was significantly higher than that in seawater microbial communities, regardless of substrate types, locations, and durations for biofilm development. In addition, the dominant source microbes of signal transduction genes in marine biofilms were different from those in seawater samples. Co-occurrence network analysis on signal communication between microbes in marine biofilms and seawater microbial communities revealed potential inter-phyla interactions between microorganisms from marine biofilms and seawater. Moreover, phylogenetic tree construction and protein identity comparison displayed that proteins related to signal transductions from Red Sea biofilms were highly similar to those from Red Sea seawater microbial communities, revealing a possible biological basis of interspecies interactions between surface-associated and free-living microbial communities in a local marine environment. Our study revealed the special profile and enrichment of signal transduction systems in marine biofilms and suggested that marine biofilms participate in intercellular interactions of the local ecosystem where they were seeded.

Impacts of UV-C Irradiation on Marine Biofilm Community Succession.

Marine biofilms are diverse microbial communities and important ecological habitats forming on surfaces submerged in the ocean. Biofilm communities resist environmental disturbance, making them a nuisance to some human activities ("biofouling"). Antifouling solutions rarely address the underlying stability or compositional responses of these biofilms. Using bulk measurements and molecular analyses, we examined temporal and UV-C antifouling-based shifts in marine biofilms in the coastal western North Atlantic Ocean during early fall. Over a 24-day period, bacterial communities shifted from early dominance of Gammaproteobacteria to increased proportions of Alphaproteobacteria, Bacteroidia, and Acidimicrobiia. In a network analysis based on temporal covariance, Rhodobacteraceae (Alphaproteobacteria) nodes were abundant and densely connected with generally positive correlations. In the eukaryotic community, persistent algal, protistan, and invertebrate groups were observed, although consistent temporal succession was not detected. Biofilm UV-C treatment at 13 and 20 days resulted in losses of chlorophyll a and transparent exopolymer particles, indicating biomass disruption. Bacterial community shifts suggested that UV-C treatment decreased the biofilm maturation rate and was associated with proportional shifts among diverse bacterial taxa. UV-C treatment was also associated with increased proportions of protists potentially involved in detritivory and parasitism. Older biofilm communities had increased resistance to UV-C, suggesting that early biofilms are more susceptible to UV-C-based antifouling. The results suggest that UV-C irradiation is potentially an effective antifouling method in marine environments in terms of biomass removal and in slowing maturation. However, as they mature, biofilm communities may accumulate microbial members that are tolerant or resilient under UV treatment. IMPORTANCE Marine biofilms regulate processes ranging from organic matter and pollutant turnover to eukaryotic settlement and growth. Biofilm growth and eukaryotic settlement interfering with human activities via growth on ship hulls, aquaculture operations, or other marine infrastructure are called "biofouling." There is a need to develop sustainable antifouling techniques by minimizing impacts to surrounding biota. We use the biofouling-antifouling framework to test hypotheses about marine biofilm succession and stability in response to disturbance, using a novel UV-C light-emitting diode (LED) device. We demonstrate strong bacterial biofilm successional patterns and detect taxa potentially contributing to stability under UV-C stress. Despite UV-C-associated biomass losses and varying UV susceptibility of microbial taxa, the overall bacterial community composition remained relatively stable, suggesting decoupling of disruption in biomass and community composition following UV-C irradiation. We also report microbial covariance patterns over 24 days of biofilm growth, pointing to areas for study of microbial interactions and targeted antifouling.

Coral reef biofilm bacterial diversity and successional trajectories are structured by reef benthic organisms and shift under chronic nutrient enrichment

Work on marine biofilms has primarily focused on host-associated habitats for their roles in larval recruitment and disease dynamics; little is known about the factors regulating the composition of reef environmental biofilms. To contrast the roles of succession, benthic communities and nutrients in structuring marine biofilms, we surveyed bacteria communities in biofilms through a six-week succession in aquaria containing macroalgae, coral, or reef sand factorially crossed with three levels of continuous nutrient enrichment. Our findings demonstrate how biofilm successional trajectories diverge from temporal dynamics of the bacterioplankton and how biofilms are structured by the surrounding benthic organisms and nutrient enrichment. We identify a suite of biofilm-associated bacteria linked with the orthogonal influences of corals, algae and nutrients and distinct from the overlying water. Our results provide a comprehensive characterization of marine biofilm successional dynamics and contextualize the impact of widespread changes in reef community composition and nutrient pollution on biofilm community structure.

Colonization characteristics of pioneer surface-associated eukaryotes during natural biofilm formation on PDMS-based composites via 18S rRNA gene sequencing methods

Marine biofilms formed on submerged substrates are important sources of biofouling. Despite extensive studies dedicated to the surface-biofilm bacteria, few studies focused exclusively on the colonization characteristics of pioneer surface-associated eukaryotes (PSE) in biofilms on composite surfaces. Given this, the colonization dynamics of the biofilm-forming PSE on different carbon nanotubes (CNTs)-filled polydimethylsiloxane (PDMS) composites were investigated via 18S rRNA gene sequencing. Diverse PSE with differential taxonomic compositions, structures and diversity characteristics were colonized on different CNTs/PDMS composites, and their colonization was greatly influenced after CNTs incorporation in PDMS. Bacillariophyta, Annelida and Arthropoda were the core phyla, taking 44.17% of the total; Aphelochaeta, Navicula and Pseudovorticella were the dominant genera, taking 38% of the total. These dominant taxa showed different sensitivities to the CNT incorporation. Some dominant PSE involved multiple biosynthesis and degradation pathways (e.g. Nucleoside and Nucleotide Biosynthesis, Amino Acid Biosynthesis and Fatty Acid and Lipid Degradation), and two key functional modules (Module 50 and Module 88) related to the initial colonization of Navicula were excavated via the correlation network analysis. Our study revealed the potential mechanisms on how different CNTs/PDMS composites influenced the colonization dynamics of PSE and provided fundamental guidance for future development of the anti-biofouling materials.

Arginine Induced Streptococcus gordonii Biofilm Detachment Using a Novel Rotating-Disc Rheometry Method.

Oral diseases are one of the most common pathologies affecting human health. These diseases are typically associated with dental plaque-biofilms, through either build-up of the biofilm or dysbiosis of the microbial community. Arginine can disrupt dental plaque-biofilms, and maintain plaque homeostasis, making it an ideal therapeutic to combat the development of oral disease. Despite our understanding of the actions of arginine towards dental plaque-biofilms, it is still unclear how or if arginine effects the mechanical integrity of the dental plaque-biofilm. Here we adapted a rotating-disc rheometry assay, a method used to quantify marine biofilm fouling, to study how arginine treatment of Streptococcus gordonii biofilms influences biofilm detachment from surfaces. We demonstrate that the assay is highly sensitive at quantifying the presence of biofilm and the detachment or rearrangement of the biofilm structure as a function of shear stress. We demonstrate that arginine treatment leads to earlier detachment of the biofilm, indicating that arginine treatment weakens the biofilm, making it more susceptible to removal by shear stresses. Finally, we demonstrate that the biofilm disrupting affect is specific to arginine, and not a general property of amino acids, as S. gordonii biofilms treated with either glycine or lysine had mechanical properties similar to untreated biofilms. Our results add to the understanding that arginine targets biofilms by multifaceted mechanisms, both metabolic and physical, further promoting the potential of arginine as an active compound in dentifrices to maintain oral health.

Zwitterionic Peptides Reduce Accumulation of Marine and Freshwater Biofilm Formers.

Zwitterionic peptides are facile low-fouling compounds for environmental applications as they are biocompatible and fully biodegradable as their degradation products are just amino acids. Here, a set of histidine (H) and glutamic acid (E), as well as lysine (K) and glutamic acid (E) based peptide sequences with zwitterionic properties were synthesized. Both oligopeptides (KE)4K and (HE)4H were synthesized in d and l configurations to test their ability to resist the nonspecific adsorption of the proteins lysozyme and fibrinogen. The coatings were additionally tested against the attachment of the marine organisms Navicula perminuta and Cobetia marina as well as the freshwater bacterium Pseudomonas fluorescens on the developed coatings. While the peptides containing lysine performed better in protein resistance assays and against freshwater bacteria, the sequences containing histidine were generally more resistant against marine organisms. The contribution of amino acid-intrinsic properties such as side chain pKa values and hydrophobicity, as well as external parameters such as pH and salinity of fresh water and seawater on the resistance of the coatings is discussed. In this way, a detailed picture emerges as to which zwitterionic sequences show advantages in future generations of biocompatible, sustainable, and nontoxic fouling release coatings.

Temporal dynamic of biofilms enhances the settlement of the central-Mediterranean reef-builder Dendropoma cristatum (Biondi, 1859)

Research on marine invertebrate settlement provides baseline knowledge for restoration technique implementation, especially for biogenic engineers with limited dispersion ability. Previously, we determined that the maturity of a biofilm strongly enhances the settlement of the vermetid reef-builder Dendropoma cristatum. To elucidate settlement-related biofilm features, here we analyse the structure and composition of marine biofilms over time, through microscopic observations, eukaryotic and prokaryotic fingerprinting analyses and 16S rDNA Illumina sequencing.The vermetid settlement temporal increase matched with the higher biofilm coverage on the substratum and the reduction of the eukaryotic abundance and diversity. The prokaryotic assemblage become, over time, more similar to that found on the reef-associated biofilm.Vermetids may detect these differences and selectively settle on those biofilms which show an advantageous structure and composition.These outcomes may support the production of ideal substrates for vermetid colonization and their further translocation to repopulate degraded reefs.

Marine Biofilms with Significant Corrosion Inhibition Performance by Secreting Extracellular Polymeric Substances.

The development of environmentally friendly and sustainable corrosion protection technologies is a longstanding yet difficult problem, especially for the marine environment. The utilization of living biofilms isolated from local environments is an effective strategy for infrastructure protection. In this study, three aerobic marine bacteria, Tenacibaculum mesophilum D-6, Tenacibaculum litoreum W-4, and Bacillus sp. Y-6, with strong biofilm-forming abilities were isolated and evaluated for the corrosion protection of X80 carbon steel. The corrosion inhibitory effect of the bacteria was found to be closely related to their biofilm-forming abilities. This conclusion was corroborated by biofilm characterization, electrochemical tests, weight loss analysis, and corrosion product analysis. Moreover, secreted extracellular polymeric substances were identified to play significant roles in corrosion inhibition. Herein, we proposed a novel, eco-friendly, and cost-effective method for corrosion protection of carbon steels in the marine environment, providing guiding principles for identifying corrosion inhibitory bacteria from the local marine environment.