Biofilm-forming Marine Bacteria Research Articles

Structure-Functional Activity of Pyrone Derivatives for Inhibition of Barnacle Settlement and Biofilm Formation.

Naturally occurring 6-pentyl-2H-pyran-2-one and its synthetic analogues greatly inhibit the settlement of Amphibalanus amphitrite cyprids and the growth and biofilm formation of marine bacteria. To optimize the antifouling activities of pyrone derivatives, this study designed pyrone analogues by modifying functional groups, such as the benzyl group, cyclopentane, and halides, substituted on both sides of a pyrone. The antifouling effects of the synthesized pyrone derivatives were subsequently evaluated against five marine biofilm-forming bacteria, Loktanella hongkongensis, Staphylococcus cohnii, S. saprophyticus, Photobacterium angustum, and Alteromonas macleodii, along with barnacle cyprids of Amphibalanus amphitrite. Substituting nonpolar parts-such as the aliphatic, cyclopentyl, or phenyl moieties on C-5 or the furan moieties on C-3-not only increased antibacterial activity and inhibited biofilm formation but also inhibited barnacle cyprid settlement when compared to 6-pentyl-2H-pyran-2-one.

Characterization of Extracellular Polymeric Substances from Biofilm-Forming Marine Bacteria from the Arabian Sea, India

Characterization of Extracellular Polymeric Substances from Biofilm-Forming Marine Bacteria from the Arabian Sea, India

Functional amyloid fibrils of biofilm-forming marine bacterium Pseudomonas aeruginosa PFL-P1 interact spontaneously with pyrene and augment the biodegradation

Bacteria thrive in biofilms embedding in the three-dimensional extracellular polymeric substances (EPS). Functional Amyloid in Pseudomonas (Fap), a protein in EPS, efficiently sequesters polycyclic aromatic hydrocarbons (PAHs). Present study reports the characterization of Fap fibrils from Pseudomonas aeruginosa PFL-P1 and describes the interaction with pyrene to assess the impact on pyrene degradation. Overexpression of fap in E. coli BL21(DE3) cells significantly enhances biofilm formation (p < 0.0001) and amyloid production (p = 0.0002), particularly with pyrene. Defibrillated Fap analysis reveals FapC monomers and increased fibrillation with pyrene. Circular Dichroism (CD), Fourier Transform Infrared Spectroscopy (FTIR), and X-ray Diffraction (XRD) unveil characteristic amyloid peaks and structural changes in Fap fibrils upon pyrene exposure. 3D-EEM analysis identifies a protein-like fluorophore in Fap fibrils, exhibiting pyrene-induced fluorescence quenching. Binding constants range from 5.23 to 7.78 M−1, with ΔG of −5.10 kJ mol−1 at 298 K, indicating spontaneous and exothermic interaction driven by hydrophobic forces. Exogenous Fap fibrils substantially increased the biofilm growth and pyrene degradation by P. aeruginosa PFL-P1 from 46 % to 64 % within 7 days (p = 0.0236). GC–MS identifies diverse metabolites, implying phthalic acid pathway in pyrene degradation. This study deepens insights into structural dynamics of Fap fibrils when exposed to pyrene, offering potential application in environmental bioremediation.

Cell surface hydrophobicity and petroleum hydrocarbon degradation by biofilm-forming marine bacterium Pseudomonas furukawaii PPS-19 under different physicochemical stressors

Biofilm-forming marine bacterium Pseudomonas furukawaii PPS-19 showed strong hydrophobicity under different physicochemical stressors, such as pH and salinity. Strong aggregation of P. furukawaii PPS-19 was observed at hydrophobic interfaces of n-dodecane and crude oil, while uptake of pyrene resulted in blue fluorescence of the bacterium. Changes in biofilm microcolonies were observed under different physicochemical stressors with maximum biofilm thickness of 15.15 µm and 15.77 µm at pH 7% and 1% salinity, respectively. Relative expression analysis of alkB2 gene revealed the maximum expression in n-dodecane (10.5 fold) at pH 7 (1 fold) and 1% salinity (8.3 fold). During the degradation process, a significant drop in surface tension resulted in increased emulsification activity. P. furukawaii PPS-19 showed the respective n-dodecane and pyrene degradation of 94.3% and 81.5% at pH 7% and 94.5% and 83% at 1% salinity. A significant positive correlation was obtained between cell surface hydrophobicity (CSH), biofilm formation, and PHs degradation (P < 0.05) under all the physicochemical stressors, with the highest value at pH 7% and 1% salinity. Analysis of metabolites indicated that mono-terminal oxidation and multiple pathways were followed for n-dodecane and pyrene biodegradation, respectively. Thus, P. furukawaii PPS-19 is an efficient hydrocarbonoclastic bacterium that may be exploited for large-scale oil pollution abatement.

Virtual Screening of a Library of Naturally Occurring Anthraquinones for Potential Anti-Fouling Agents.

Marine biofouling is the undesired accumulation of organic molecules, microorganisms, macroalgae, marine invertebrates, and their by-products on submerged surfaces. It is a serious challenge for marine vessels and the oil, gas, and renewable energy industries, as biofouling can cause economic losses for these industries. Natural products have been an abundant source of therapeutics since the start of civilisation. Their use as novel anti-fouling agents is a promising approach for replacing currently used, harmful anti-fouling agents. Anthraquinones (AQs) have been used for centuries in the food, pharmaceutical, cosmetics, and paint industries. Citreorosein and emodin are typical additives used in the anti-fouling paint industry to help improve the global problem of biofouling. This study is based on our previous study, in which we presented the promising activity of structurally related anthraquinone compounds against biofilm-forming marine bacteria. To help uncover the anti-fouling potential of other AQ-related structures, 2194 compounds from the COCONUT natural products database were analysed. Molecular docking analysis was performed to assess the binding strength of these compounds to the LuxP protein in Vibrio carchariae. The LuxP protein is a vital binding protein responsible for the movements of autoinducers within the quorum sensing system; hence, interrupting the process at an early stage could be an effective strategy. Seventy-six AQ structures were found to be highly docked, and eight of these structures were used in structure-based pharmacophore modelling, resulting in six unique pharmacophore features.

A structure-activity relationship analysis of anthraquinones with antifouling activity against marine biofilm-forming bacteria

Anthraquinones (AQs) are chemical scaffolds that have been used both naturally and synthetically for centuries in the food, pharmaceutical, cosmetic and paint industries. AQs, such as citreorosein and emodin, are common additives in antifouling paints which help prevent the global issue of biofouling. To determine the antifouling potential of a family of structurally related compounds nineteen AQs (1–19), were tested for their microbial growth and biofilm adhesion inhibition activity against three marine biofilm forming bacteria, Vibrio carchariae, Pseudoalteromonas elyakovii and Shewanella putrefaciens. More than three-quarters of the tested AQ compounds exhibited activity against both V. carchariae and P. elyakovii at 10 μg/ml whilst exhibiting low antimicrobial effects. The most active compounds (1, 5, 6, 7, 9, 10, 14, 15, 18, 19) were tested for their minimum inhibitory concentrations (MICs) which highlighted that all the tested compounds were highly effective at inhibiting the biofilm growth of P. elyakovii at a very low concentration of 0.001 μg/ml. The variability in MIC for inhibiting the biofilm growth of V. carchariae was assessed by analysing the structure-activity relationships (SARs) between the AQ compounds, and the key structural features leading to improved biofilm growth inhibition activity are reported. Molecular docking analysis was also performed to assess whether interruption of quorum sensing in V. carchariae could be a possible mode of action for the antifouling activity observed.

Variable pH and subsequent change in pCO2 modulates the biofilm formation, synthesis of extracellular polymeric substances, and survivability of a marine bacterium Bacillus stercoris GST-03

Biofilm-forming bacteria adhere to the substrates and engage in the nutrient cycling process. However, environmental conditions may interrupt the biofilm formation ability, which ultimately may affect various biogeochemical cycles. The present study reports the effect of varying pH and subsequent change in pCO2 on the survivability, biofilm formation, and synthesis of extracellular polymeric substances (EPS) of a biofilm-forming marine bacterium Bacillus stercoris GST-03 isolated from the Bhitarkanika mangrove ecosystem, Odisha, India. Understanding the pH-dependent alteration in EPS constituents, and associated functional groups of a marine bacterium will provide better insight into the adaptability of the bacteria in future ocean acidification scenarios. The strain was found to tolerate and form biofilm up to pH 4, with the maximum biofilm formation at pH 6. EPS yield and the synthesis of the key components of the EPS, including carbohydrate, protein, and lipid, were found maximum at pH 6. Changes in biofilm formation patterns and various topological parameters at varying pH/pCO2 conditions were observed. A cellular chaining pattern was observed at pH 4, and maximum biofilm formation was obtained at pH 6 with biomass of 5.28582 ± 0.5372 μm3/μm2 and thickness of 9.982 ± 1.5288 μm. Structural characterization of EPS showed changes in various functional groups of constituent macromolecules with varying pH. The amorphous nature of the EPS and the changes in linkages and associated functional groups (-R2CHOR, –CH3, and –CH2) with pH variation was confirmed. EPS showed a two-step degradation with a maximum weight loss of 59.147% and thermal stability up to 480 °C at pH 6. The present work efficiently demonstrates the role of EPS in providing structural and functional stability to the biofilm in varying pH conditions. The findings will provide a better understanding of the adaptability of marine bacteria in the future effect of ocean acidification.

Anti-Osteoclastogenic and Antibacterial Effects of Chlorinated Polyketides from the Beibu Gulf Coral-Derived Fungus Aspergillus unguis GXIMD 02505.

One new depsidone derivative, aspergillusidone H (3), along with seven known biosynthetically related chlorinated polyketides, were obtained from the Beibu Gulf coral-derived fungus Aspergillus unguis GXIMD 02505. Their structures were determined by comprehensive physicochemical and spectroscopic data interpretation. Notably, the X-ray crystal structure of 2 and the previously unknown absolute configuration of 8, assigned by ECD calculations, are described here for the first time. Compounds 1–5, 7 and 8 exhibited inhibition of lipopolysaccharide (LPS)-induced NF-κB in RAW 264.7 macrophages at 20 μM. In addition, the two potent inhibitors (2 and 7) dose-dependently suppressed RANKL-induced osteoclast differentiation without any evidence of cytotoxicity in bone marrow macrophages cells (BMMs). This is the first report of osteoclastogenesis inhibitory activity for the metabolites of these kinds. Besides, compounds 1, 2, 4, and 6–8 showed inhibitory activity against marine biofilm-forming bacteria, methicillin-resistant Staphylococcus aureus, Microbulbifer variabilis, Marinobacterium jannaschii, and Vibrio pelagius, with their MIC values ranging from 2 to 64 μg/mL. These findings provide a basis for further development of chlorinated polyketides as potential inhibitors of osteoclast differentiation and/or for use as anti-fouling agents.

The effect of a biofilm-forming bacterium Tenacibaculum mesophilum D-6 on the passive film of stainless steel in the marine environment

The microbiologically influenced corrosion of 304 stainless steel in the presence of a marine biofilm-forming bacterium Tenacibaculum mesophilum D-6 was systematically investigated by means of electrochemical techniques and surface analyses to reveal the effect of the selective attachment and adsorption of the biofilms on the passivity breakdown of the stainless steel. It was found that the T. mesophilum D-6 was electroactive and could oxidize low valent cations and metal, facilitating the local dissolution of the passive film and the substrate in the film defects, nearly doubling the surface roughness. The biofilms of T. mesophilum D-6 with mucopolysaccharide secreta and chloride ions tended to preferentially adsorb at the defects of the passive film on the steel, yielding non-homogeneous microbial aggregates and local Cl− enrichment there. The adsorption of the bacteria and chloride ions reduced the thickness of passive film by 23.9%, and generate more active sites for pitting corrosion on the passive film and more semiconducting carrier acceptors in the film. The maximum current density of the 304 SS in the presence of T. mesophilum D-6 was over one order of magnitude higher than that in the sterile medium, and the largest pit was deepened 3 times.

Structural and mechanical characterization of biofilm-associated bacterial polymer in the emulsification of petroleum hydrocarbon.

The online version contains supplementary material available at 10.1007/s13205-021-02795-8.

Isolation and identification of early marine biofilm-forming bacteria on commercial paint surface

Isolation and identification of early marine biofilm-forming bacteria on commercial paint surface

Flavonoid Glycosides with a Triazole Moiety for Marine Antifouling Applications: Synthesis and Biological Activity Evaluation.

Over the last decades, antifouling coatings containing biocidal compounds as active ingredients were used to prevent biofouling, and eco-friendly alternatives are needed. Previous research from our group showed that polymethoxylated chalcones and glycosylated flavones obtained by synthesis displayed antifouling activity with low toxicity. In this work, ten new polymethoxylated flavones and chalcones were synthesized for the first time, including eight with a triazole moiety. Eight known flavones and chalcones were also synthesized and tested in order to construct a quantitative structure-activity relationship (QSAR) model for these compounds. Three different antifouling profiles were found: three compounds (1b, 11a and 11b) exhibited anti-settlement activity against a macrofouling species (Mytilus galloprovincialis), two compounds (6a and 6b) exhibited inhibitory activity against the biofilm-forming marine bacteria Roseobacter litoralis and one compound (7b) exhibited activity against both mussel larvae and microalgae Navicula sp. Hydrogen bonding acceptor ability of the molecule was the most significant descriptor contributing positively to the mussel larvae anti-settlement activity and, in fact, the triazolyl glycosylated chalcone 7b was the most potent compound against this species. The most promising compounds were not toxic to Artemia salina, highlighting the importance of pursuing the development of new synthetic antifouling agents as an ecofriendly and sustainable alternative for the marine industry.

Naphthalene catabolism by biofilm forming marine bacterium Pseudomonas aeruginosa N6P6 and the role of quorum sensing in regulation of dioxygenase gene.

This study aims to establish the role of quorum sensing (QS) system on the regulation of naphthalene ring cleaving gene ndo (encoding naphthalene dioxygenase) in biofilm forming marine bacterium Pseudomonas aeruginosa N6P6 for naphthalene degradation. Total cell count of P. aeruginosa N6P6 during biofilm mode of growth was slightly higher (7·3×108 CFU per ml) than its planktonic mid-exponential phase culture (4·7×108 CFU per ml). Naphthalene degradation in 20h by biofilm (48-h old) and planktonic culture was 99·4±0·002% and 77±3·25%, respectively. Pseudomonas aeruginosa N6P6 was able to degrade 64·3± 4·7% naphthalene in sterile soil microcosm in 24h. The bacterium showed the presence of 136bp ndo gene which was upregulated in a dose-dependent manner in presence of naphthalene. QS inhibitor (QSI) tannic acid downregulated the expression of ndo gene, naphthalene 1, 2-dioxygenase (N12O) enzyme activity and naphthalene degradation (by biofilm culture). P. aeruginosa N6P6 shows chemotaxis towards naphthalene and adapts well in terrestrial environment for naphthalene degradation. This study provides the information that the QS plays crucial role in biofilm formation in P. aeruginosa N6P6 and QS regulatory genes subsequently control the ndo gene for enzymatic degradation of naphthalene.

Whole genome characterization and phenanthrene catabolic pathway of a biofilm forming marine bacterium Pseudomonas aeruginosa PFL-P1

Pseudomonas aeruginosa is a small rod shaped Gram-negative bacterium of Gammaproteobacteria class known for its metabolic versatility. P. aeruginosa PFL-P1 was isolated from Polycyclic Aromatic Hydrocarbons (PAHs) contaminated site of Paradip Port, Odisha Coast, India. The strain showed excellent biofilm formation and could retain its ability to form biofilm grown with different PAHs in monoculture as well as co-cultures. To explore mechanistic insights of PAHs metabolism, the whole genome of the strain was sequenced. Next generation sequencing unfolded a genome size of 6,333,060 bp encoding 5857 CDSs. Gene ontology distribution assigned to a total of 2862 genes, wherein 2235 genes were allocated to biological process, 1549 genes to cellular component and 2339 genes to molecular function. A total of 318 horizontally transferred genes were identified when the genome was compared with the reference genomes of P. aeruginosa PAO1 and P. aeruginosa DSM 50071. Further comparison of P. aeruginosa PFL-P1 genome with P. putida containing TOL plasmids revealed similarities in the meta cleavage pathway employed for degradation of aromatic compounds like xylene and toluene. Gene annotation and pathway analysis unveiled 145 genes involved in xenobiotic biodegradation and metabolism. The biofilm cultures of P. aeruginosa PFL-P1 could degrade ~74% phenanthrene within 120 h while degradation increased up to ~76% in co-culture condition. GC-MS analysis indicated presence of diverse metabolites indicating the involvement of multiple pathways for one of the PAHs (phenanthrene) degradation. The strain also possesses the genetic machinery to utilize diverse toxic aromatic compounds such as naphthalene, benzoate, aminobenzoate, fluorobenzoate, toluene, xylene, styrene, atrazine, caprolactam etc. Common catabolic gene clusters such as benABCD, xylXYZ and catAB were observed within the genome of P. aeruginosa PFL-P1 which play key roles in the degradation of various toxic aromatic compounds.

Investigating the microbial-influenced corrosion of UNS S32750 stainless-steel base alloy and weld seams by biofilm-forming marine bacterium Macrococcus equipercicus

This study investigates the microbial-influenced corrosion of UNS S32750 super-duplex stainless-steel joints fabricated using different welding methods. Herein, the samples were introduced into a medium inoculated with Macrococcus equipercicus isolated from a marine environment. Confocal laser scanning microscopy and atomic force microscopy were used to characterise the topography and formation of pits in the corroded samples, respectively. Potentiodynamic polarisation studies were conducted on both the base alloy and weld seams exposed for 30 and 60 days in the experimental system inoculated with M. equipercicus and un-inoculated system. Results indicate that the thickness of the biofilm formed due to this bacterium increased and became heterogeneous with an increase in the exposure time, thereby resulting in micro-pits. Bacterial colonisation was observed in all the coupons after exposure to the inoculated medium. Although micro-pits were observed in all the coupons, the base metal and flux-cored arc weld seams showed highest sensitivity to bacterial attack.

Antifouling Napyradiomycins from Marine-Derived Actinomycetes Streptomyces aculeolatus.

The undesired attachment of micro and macroorganisms on water-immersed surfaces, known as marine biofouling, results in severe prevention and maintenance costs (billions €/year) for aquaculture, shipping and other industries that rely on coastal and off-shore infrastructures. To date, there are no sustainable, cost-effective and environmentally safe solutions to address this challenging phenomenon. Therefore, we investigated the antifouling activity of napyradiomycin derivatives that were isolated from actinomycetes from ocean sediments collected off the Madeira Archipelago. Our results revealed that napyradiomycins inhibited ≥80% of the marine biofilm-forming bacteria assayed, as well as the settlement of Mytilus galloprovincialis larvae (EC50 < 5 µg/ml and LC50/EC50 >15), without viability impairment. In silico prediction of toxicity end points are of the same order of magnitude of standard approved drugs and biocides. Altogether, napyradiomycins disclosed bioactivity against marine micro and macrofouling organisms, and non-toxic effects towards the studied species, displaying potential to be used in the development of antifouling products.

Overcoming environmental problems of biocides: Synthetic bile acid derivatives as a sustainable alternative

Marine biofouling represents a global economic and ecological challenge. Some marine organisms produce bioactive metabolites, such as steroids, that inhibit the settlement and growth of fouling organisms. The aim of this work was to explore bile acids as a new scaffold with antifouling (AF) activity by using chemical synthesis to produce a series of bile acid derivatives with optimized AF performance and understand their structure-activity relationships. Seven bile acid derivatives were successfully synthesized in moderate to high yields, and their structures were elucidated through spectroscopic methods. Their AF activities were tested against both macro- and microfouling communities. The most potent bile acid against the settlement of Mytilus galloprovincialis larvae was the methyl ester derivative of cholic acid (10), which showed an EC50 of 3.7 μM and an LC50/EC50 > 50 (LC50 > 200 μM) in AF effectiveness vs toxicity studies. Two derivatives of deoxycholic acid (5 and 7) potently inhibited the growth of biofilm-forming marine bacteria with EC50 values < 10 μM, and five bile acids (1, 5, and 7–9) potently inhibited the growth of diatoms, showing EC50 values between 3 and 10 μM. Promising AF profiles were achieved with some of the synthesized bile acids by combining antimacrofouling and antimicrofouling activities. Initial studies on the incorporation of one of these promising bile acid derivatives in polymeric coatings, such as a marine paint, demonstrated the ability of these compounds to generate coatings with antimacrofouling activity.

Expression of metallothionein encoding gene bmtA in biofilm-forming marine bacterium Pseudomonas aeruginosa N6P6 and understanding its involvement in Pb(II) resistance and bioremediation.

The genetic basis and biochemical aspects of heavy metal endurance abilities have been precisely studied in planktonic bacteria; however, in nature, bacteria mostly grows as surface-attached communities called biofilms. A hallmark trait of biofilm is increased resistance to heavy metals compared with the resistance of planktonic bacteria. A proposed mechanism that contributes to this increased resistance is the enhanced expression of metal-resistant genes. bmtA gene coding for metallothionein protein is one such metal-resistant gene found in many bacterial spp. In the present study, lead (Pb) remediation potential of a biofilm-forming marine bacterium Pseudomonas aeruginosa N6P6 was explored. Biofilm-forming marine bacterium P. aeruginosa N6P6 possess bmtA gene and shows resistance towards many heavy metals, i.e., Pb, Cd, Hg, Cr, and Zn. The expression of metallothionein encoding gene bmtA is significantly high in 48-h-old biofilm culture (11. 4 fold) followed by 24-h-old biofilm culture of P. aeruginosa N6P6 (4.7 fold) (P < 0.05). However, in the case of planktonically grown culture of P. aeruginosa N6P6, the highest expression of bmtA gene was observed in 24-h-old culture. The expression of bmtA also increased significantly with increase in Pb concentration up to 800 ppm. CSLM analysis indicated significant reduction in the raw integrated density of biofilm-associated lipids and polysaccharides (PS) of P. aeruginosa N6P6 biofilm grown in Pb (sub-lethal concentration)-amended medium (P < 0.05), whereas no significant reduction was observed in the raw integrated density of EPS-associated protein. The role of bmtA gene as Pb(II)-resistant determinant was characterized by overexpressing the bmtA gene derived from P. aeruginosa N6P6 in Escherichia coli BL21(DE3). ESI-MS and SDS-PAGE analyses validated the presence of 11.5-kDa MT protein isolated from Pb(II)-induced recombinant E. coli BL21(DE3) harboring bmtA gene.

Metabolomic and proteomic changes induced by growth inhibitory concentrations of copper in the biofilm-forming marine bacterium Pseudoalteromonas lipolytica.

Copper is an essential element for living cells but this metal is present in some marine environments at such high concentrations that it can be toxic for numerous organisms. In polluted areas, marine organisms may develop specific adaptive responses to prevent cell damage. To investigate the influence of copper on the metabolism of a single organism, a dual approach combining metabolomics and proteomics was undertaken on the biofilm-forming bacterial strain Pseudoalteromonas lipolytica TC8. In order to highlight differential adaptation according to the phenotype, the response of P. lipolytica TC8 to copper stress was studied in planktonic and biofilm culture modes under growth inhibitory copper concentrations. As expected, copper exposure led to the induction of defense and detoxification mechanisms. Specific metabolite and protein profiles were thus observed in each condition (planktonic vs. biofilm and control vs. copper-treated cultures). Copper exposure seems to induce drastic changes in the lipid composition of the bacterial cell membrane and to modulate the abundance of proteins functionally known to be involved in copper cell homeostasis in both planktonic and biofilm culture modes. Much more proteins differentially expressed after copper treatment were observed in biofilms than in planktonic cells, which could indicate a more heterogeneous response of biofilm cells to this metallic stress.

Exploring the chemodiversity of tropical microalgae for the discovery of natural antifouling compounds

Marine microalgae and cyanobacteria have largely been studied for their biotechnological potential and proved their ability to produce a wide array of bioactive molecules. We investigated the antifouling potential of unexplored benthic tropical microalgae using anti-adhesion and toxicity bioassays against two major micro- and ma crobiofoulers, namely bacteria and barnacles. Fifty strains belonging to six phyla [Cyanobacteria, Miozoa (Dinoflagellata), Bacillariophyta, Cryptophyta, Rhodophyta and Haptophyta] were isolated from southwestern Islands of the Indian Ocean. They were chosen in order to represent as much as possible the huge biodiversity of such a rich tropical ecosystem. The associated chemodiversity was highlighted by both NMR- and LC-MS-based metabolomics. The screening of 84 algal fractions revealed that the anti-adhesion activity was concentrated in methanolic ones (i.e. 93% of all active fractions). Our results confirmed that microalgae constitute a promising source of natural antimicrofoulants as 17 out of the 30 active fractions showed high or very high capacity to inhibit the adhesion of three biofilm-forming marine bacteria. Dinoflagellate-derived fractions were the most active, both in terms of number and intensity. However, dinoflagellates were also more toxic and may not be suitable as a source of environmentally friendly antifouling compounds, in contrast to diatoms, e.g. Navicula mollis. The latter and two dinoflagellates of the genus Amphidinium also had interesting anti-settlement activities while being moderately toxic to barnacle larvae. Our approach, combining the bioprospecting of a large number of tropical microalgae for their anti-settlement potential and metabolomics analyses, constituted a first step towards the discovery of alternative ecofriendly antifoulants.