Marine resources such as marine invertebrates and mangrove plants favor the production of secondary metabolites that exhibit antifouling properties. These natural-derived compounds are considered environmentally friendly compared to synthetic compounds with similar activity and technological applications. The current study was conducted to determine the antifouling properties of Diadema setosum (DS) and Sonneratia lanceolata (SL) crude extracts and their incorporated paints, in addition to the identification of the metabolites involved. Both crude extracts were tested against Pseudomonas aeruginosa via a crystal violet assay, while the incorporated paints with 5% (SL5% and DS5%) and 10% (SL10% and DS10%) weight per volume (w/v) were tested in an aquarium and submerged in the seawater at Kemaman and Pulau Redang (Malaysia) for field testing. The identification of the bioactive compounds from the crude extracts was carried out using Liquid Chromatography-Mass Spectrometry (LC-MS). The results of the crystal violet assay showed that both of the crude extracts reduced the biofilm formed by Pseudomonas aeruginosa. The marine bacteria growths contained in natural seawater were inhibited the most by SL5%, followed by DS5%, DS10%, and SL10% in the aquarium testing. Based on the photographic observation, all of the paints incorporated with the crude extracts successfully reduced the settlement of fouling organisms compared to the blank paint, as lesser macroalgae were found growing on the SL5%, DS5%, and DS10%. The LC-MS results showed 3-Methyloxiranyl phosphonic acid; (2RS,3SR)-form from the SL crude extract, while the 8-Decene-1,3,5-triol, 3-Hydroxyundecanoic acid, and 1-O-(6-Deoxy-6-sulfoglucopyranosyl)glycerol; α-D-form, 3-Hexadecanoyl from the DS crude extract were involved in the antifouling properties. In conclusion, both crude extracts have the potential to be developed as antifouling agents.

Biofouling is defined as the excessive colonization process of epibiotic organisms, ranging from microfoulers to macrofoulers, on any submerged surface in water. Previous research has attempted to explore the antifouling activity of bacterial isolates due to the biofouling problems occurring worldwide. One solution is to inhibit the early stage of fouling using secondary metabolites produced by marine bacteria. This study aims to determine the antifouling activities of the marine microorganism P. aeruginosa and to characterize the bacteria isolated as a potential anti-biofouling agent. The bacterial isolate was cultured and isolated on a media culture. The bacteria culture extract was extracted using ethyl acetate and concentrated prior to the bioassay method. It was screened for antibacterial activities against Gram-positive and Gram-negative bacteria, such as Bacillus cereus, Streptococcus uberis, Pseudomonas sp., and Vibrio parahaemolyticus, using the disk diffusion technique. The extract was investigated to verify its bioactivity in the prevention of biofilm formation following the crystal violet assay and aquarium test. The results indicated the inhibition of activity through biofilm formation, with the highest percentage at 83% of biofilm inhibition at a concentration of 0.1563 mg/mL. The bacterial isolate at a concentration of 5% showed the highest reduction in bacteria colonies in the aquarium test (161.8 × 10³ CFU/mL compared to 722.5 × 10³ CFU/mL for the blank sample). The bacterial isolate was characterized through phenotypic and genotypic tests for species identification. It was identified as a Gram-stain-negative, aerobic, and long-rod-shaped bacteria, designated as RLimb. Based on the 16S rDNA gene sequencing analysis, RLimb was identified as Pseudomonas aeruginosa (accession number: OP522351), exhibiting a similarity of 100% to the described neighbor P. aeruginosa strain DSM 50071. These results indicated that these isolated bacteria can potentially be used as a substitute for toxic antifoulants to prevent the formation of microfoulers.

Marine natural products especially seaweeds have gained much attention to combat biofouling. Ulva lactuca was determined for its antifouling activity and characterized the isolated non-polar metabolite involved. The methanolic crude extract (MCE) of U. lactuca was screened using crystal violet assay against biofilm-forming bacteria Pseudomonas aeruginosa and was further tested on laboratory and field tests. Then, it was fractionated and isolated using Liquid-Liquid Fractionation (LLE) and Column Chromatography (CC). The isolated compound was characterized using Liquid Chromatography-Mass Spectrometry (LC-MS), Nuclear Magnetic Resonance (NMR), and Fourier Transform-Infrared Spectroscopy (FTIR). The current study showed that the growth of biofilm produced by P. aeruginosa was inhibited by MCE at concentrations of 0.0156 mg/mL. The laboratory test indicated UL5% demonstrated a higher bacterial reduction of bacterial colonies with 1.903 × 10⁶ CFU/mL better than blank paint. According to the field test, crude panels of UL5% were successful in reducing the settlement of fouling organisms due to less macrofouler growth compared to blank paint. The isolated compound A4 was identified as hexadecanoic acid (C₁₆H₃₂O₂) through NMR with a molecular mass of 256 g/mol detected using LC-MS. The characterization through FTIR obtained functional groups consisting of CH₃, CH₂, C=O, and OH. Therefore, U. lactuca produced hexadecanoic acid as one of the promising compounds from the seaweed group as an eco-friendly antifouling agent.