Marine dissolved organic matter (DOM) comprises a vast and unexplored molecular space. Most of it resided in the oceans for thousands of years. It is among the most diverse molecular mixtures known, consisting of millions of individual compounds. More than 1 Eg of this material exists on the planet. As such, it comprises a formidable source of natural products promising significant potential for new biotechnological purposes. Great emphasis has been placed on understanding the role of DOM in biogeochemical cycles and climate attenuation, its lifespan, interaction with microorganisms, as well as its molecular composition. Yet, probing DOM bioactivities is in its infancy, largely because it is technically challenging due to the chemical complexity of the material. It is of considerable interest to develop technologies capable to better discern DOM bioactivities. Modern screening technologies are opening new avenues allowing accelerated identification of bioactivities for small molecules from natural products. These methods diminish a priori the need for laborious chemical fractionation. We examine here the application of untargeted metabolomics and multiplexed high-throughput molecular-phenotypic screening techniques that are providing first insights on previously undetectable DOM bioactivities. Key points: • Marine DOM is a vast, unexplored biotechnological resource. • Untargeted bioscreening approaches are emerging for natural product screening. • Perspectives for developing bioscreening platforms for marine DOM are discussed.

The biochemical composition, molecular diversity, and two different bioactivities of Asparagopsis armata and Rugulopteryx okamurae (two alien species with different invasive patterns in the southern Iberian Peninsula) were analyzed through spectrophotometric methods and Fourier transform ion cyclotron mass spectroscopy (FT-ICR-MS). A total of 3042 molecular formulas were identified from the different extracts. The _dH₂O extracts were the most molecularly different. A. armata presented the highest content of nitrogenous compounds (proteins, CHON) and sulphur content, whereas R. okamurae was rich in carbonated compounds (total carbon, lipids, CHO, and CHOP). Antioxidant capacity and phenolic content were higher in R. okamurae than in A. armata. Antimicrobial activity was detected from both species. A. armata showed capacity to inhibit human and fish pathogens (e.g., Staphylococcus aureus or Vibrio anguillarum), whereas R. okamurae only showed inhibition against human bacteria (Staphylococcus aureus and Cutibacterium acnes). In R. okamurae, molecules with a great number of pharmaceutical activities (e.g., anti-inflammatory or antitumoral), antibacterial, biomaterial, and other utilities were found. The main molecules of A. armata had also pharmaceutical applications (e.g., antimalarian, antithrombotic, anti-inflammatory, or antiarthritis). The valorization of these species can help to counteract the environmental effects of the bioinvasions.

Dissolved organic matter (DOM) occupies a huge and uncharted molecular space. Given its properties, DOM can be presented as a promising biotechnological resource. However, research into bioactivities of DOM is still in early stages. In this study, the biotechnological potential of terrestrial and marine DOM, its molecular composition and their relationships are investigated. Samples were screened for their in vitro antibacterial, antifungal, anticancer and antioxidant activities. Antibacterial activity was detected against Staphylococcusaureus in almost all DOM samples, with freshwater DOM showing the lowest IC₅₀ values. Most samples also inhibited Staphylococcusepidermidis, and four DOM extracts showed up to fourfold higher potency than the reference drug. Antifungal activity was limited to only porewater DOM towards human dermatophyte Trichophytonrubrum. No significant in vitro anticancer activity was observed. Low antioxidant potential was exerted. The molecular characterization by FT-ICR MS allowed a broad compositional overview. Three main distinguished groups have been identified by PCoA analyses. Antibacterial activities are related to high aromaticity content and highly-unsaturated molecular formulae (O-poor). Antifungal effect is correlated with highly-unsaturated molecular formulae (O-rich). Antioxidant activity is positively related to the presence of double bonds and polyphenols. This study evidenced for the first time antibacterial and antifungal activity in DOM with potential applications in cosmeceutical, pharmaceutical and aquaculture industry. The lack of cytotoxicity and the almost unlimited presence of this organic material may open new avenues in future marine bioprospecting efforts. Graphical abstract

Background. The effluents of the mono-specific aquaculture contain high concentrations of dissolved nutrients and organic matter, which affect negatively water quality of the recipient aquatic ecosystems. A key feature of water quality is its transparency. Chromophoric dissolved organic matter (CDOM) determines most of the light transmission in the ultraviolet and blue bands in the aquatic ecosystems. A sustainable alternative to mono-specific aquaculture is the integrated multitrophic aquaculture that includes species trophically complementary named “extractive” species. Sea cucumbers are recognized as efficient extractive species, with a high potential to improve water quality, due to the consumption of particulate organic matter (POM). However, the effects of sea cucumbers on CDOM are still unknown. Methods. During one year, we biweekly monitored CDOM in two aquaculture tanks with different trophic structure. One of the tanks (-holothurian tank) only contained the primary species, Anemonia sulcata, whereas the other tank (+ holothurian tank) also contained individuals of Holothuria tubulosa and H. forskali. We routinely performed CDOM absorption spectra from 200 nm to 750 nm and determined quantitative (absorption coefficients at 325 nm) and qualitative (spectral slopes and molar absorption coefficients at 325 nm) optical parameters in the inlet waters, in the tanks, and in their corresponding effluents. Results. Absorption coefficients at 325 nm (a₃₂₅) and spectral slopes from 275 to 295 nm (S_275-295) were significantly lower in the effluents of the +holothurian tank (average: 0.33 and 16 μm^-1, respectively) than in the effluents of the −holothurian tank (average: 0.69 m^-1 and 34 μm^-1, respectively), being the former similar to those found in the inlet waters (average: 0.32 m^-1 and 22 μm^-1, respectively). This reduction in CDOM absorption appears to be mediated by the POM consumption by the holothurians. The reduction of POM concentration in the +holothurian tank may weaken the process of POM disaggregation into dissolved organic matter, which ultimately might have generated CDOM in the –holothurian tank. Discussion. Extractive species such as holothurians improve water transparency through POM consumption, likely because reduces POM disaggregation into CDOM. We suggest that CDOM monitoring in aquaculture facilities, using automatic probes or even remote sensing, could be a useful tool to trace the effectiveness of extractive species at large scales of time and space.

Background. The effluents of the mono-specific aquaculture contain high concentrations of dissolved nutrients and organic matter, which affect negatively water quality of the recipient aquatic ecosystems. A key feature of water quality is its transparency. Chromophoric dissolved organic matter (CDOM) determines most of the light transmission in the ultraviolet and blue bands in the aquatic ecosystems. A sustainable alternative to mono-specific aquaculture is the integrated multitrophic aquaculture that includes species trophically complementary named “extractive” species. Sea cucumbers are recognized as efficient extractive species, with a high potential to improve water quality, due to the consumption of particulate organic matter (POM). However, the effects of sea cucumbers on CDOM are still unknown. Methods. During one year, we biweekly monitored CDOM in two aquaculture tanks with different trophic structure. One of the tanks (-holothurian tank) only contained the primary species, Anemonia sulcata, whereas the other tank (+ holothurian tank) also contained individuals of Holothuria tubulosa and H. forskali. We routinely performed CDOM absorption spectra from 200 nm to 750 nm and determined quantitative (absorption coefficients at 325 nm) and qualitative (spectral slopes and molar absorption coefficients at 325 nm) optical parameters in the inlet waters, in the tanks, and in their corresponding effluents. Results. Absorption coefficients at 325 nm (a₃₂₅) and spectral slopes from 275 to 295 nm (S_275-295) were significantly lower in the effluents of the +holothurian tank (average: 0.33 and 16 μm^-1, respectively) than in the effluents of the −holothurian tank (average: 0.69 m^-1 and 34 μm^-1, respectively), being the former similar to those found in the inlet waters (average: 0.32 m^-1 and 22 μm^-1, respectively). This reduction in CDOM absorption appears to be mediated by the POM consumption by the holothurians. The reduction of POM concentration in the +holothurian tank may weaken the process of POM disaggregation into dissolved organic matter, which ultimately might have generated CDOM in the –holothurian tank. Discussion. Extractive species such as holothurians improve water transparency through POM consumption, likely because reduces POM disaggregation into CDOM. We suggest that CDOM monitoring in aquaculture facilities, using automatic probes or even remote sensing, could be a useful tool to trace the effectiveness of extractive species at large scales of time and space.