Potential of Polar Lipids Isolated from the Marine Sponge Haliclona (Halichoclona) vansoesti against Melanoma.

Marine sponges represent a good source of natural metabolites for biotechnological applications in the pharmacological, cosmeceutical, and nutraceutical fields. In the present work, we analyzed the biotechnological potential of the alien species Haliclona (Halichoclona) vansoesti de Weerdt, de Kluijver & Gomez, 1999, previously collected in the Mediterranean Sea (Faro Lake, Sicily). The bioactivity and chemical content of this species has never been investigated, and information in the literature on its Caribbean counterpart is scarce. We show that an enriched extract of H. vansoesti induced cell death in human melanoma cells with an IC50 value of 36.36 µg mL-1, by (i) triggering a pro-inflammatory response, (ii) activating extrinsic apoptosis mediated by tumor necrosis factor receptors triggering the mitochondrial apoptosis via the involvement of Bcl-2 proteins and caspase 9, and (iii) inducing a significant reduction in several proteins promoting human angiogenesis. Through orthogonal SPE fractionations, we identified two active sphingoid-based lipid classes, also characterized by nuclear magnetic resonance and mass spectrometry, as the main components of two active fractions. Overall, our findings provide the first evaluation of the anti-cancer potential of polar lipids isolated from the marine sponge H. (Halichoclona) vansoesti, which may lead to new lead compounds with biotechnological applications in the pharmaceutical field.

Molecular Approaches Detect Early Signals of Programmed Cell Death in Hippolyte inermis Leach.

The protandric shrimp Hippolyte inermis is the only known marine invertebrate whose sex determination is strongly influenced by the composition of its food. In H. inermis, a sex reversal is triggered by the ingestion of diatoms of the genus Cocconeis associated with leaves of the seagrass Posidonia oceanica. These diatoms contain compounds that promote programmed cell death (PCD) in H. inermis and also in human cancer cells. Transcriptomic analyses suggested that ferroptosis is the primary trigger of the shrimp's sex reversal, leading to the rapid destruction of the androgen gland (AG) followed by a chain of apoptotic events transforming the testes into ovaries. Here, we propose a molecular approach to detect the effects of compounds stimulating the PCD. An RNA extraction method, suitable for young shrimp post-larvae (five days after metamorphosis; PL5 stage), was established. In addition, six genes involved in apoptosis, four involved in ferroptosis, and seven involved in the AG switch were mined from the transcriptome, and their expression levels were followed using real-time qPCR in PL5 fed on Cocconeis spp., compared to PL5 fed on a basic control feed. Our molecular approach, which detected early signals of sex reversal, represents a powerful instrument for investigating physiological progression and patterns of PCD in marine invertebrates. It exemplifies the physiological changes that may start a few days after the settlement of post-larvae and determine the life destiny of an individual.

The Antiviral Potential of Algal Lectins.

Algae have emerged as fascinating subjects of study due to their vast potential as sources of valuable metabolites with diverse biotechnological applications, including their use as fertilizers, feed, food, and even pharmaceutical precursors. Among the numerous compounds found in algae, lectins have garnered special attention for their unique structures and carbohydrate specificities, distinguishing them from lectins derived from other sources. Here, a comprehensive overview of the latest scientific and technological advancements in the realm of algal lectins with a particular focus on their antiviral properties is provided. These lectins have displayed remarkable effectiveness against a wide range of viruses, thereby holding great promise for various antiviral applications. It is worth noting that several alga species have already been successfully commercialized for their antiviral potential. However, the discovery of a diverse array of lectins with potent antiviral capabilities suggests that the field holds immense untapped potential for further expansion. In conclusion, algae stand as a valuable and versatile resource, and their lectins offer an exciting avenue for developing novel antiviral agents, which may lead to the development of cutting-edge antiviral therapies.

The Red Squat Lobster Pleuroncodes monodon in the Humboldt Current System: From Their Ecology to Commercial Attributes as Marine Bioresource.

This study focused on gathering available information on Pleuroncodes monodon, a widely distributed crustacean in the Humboldt Current System. Off the Chilean coast, this species presents benthic habits and constitutes the main resource of the industrial crustacean fishery; many studies have been carried out on its life cycle during the last century. In contrast, off the coast of Peru, this species exhibits mainly pelagic habits, with latent information gaps on aspects of its life history and no commercial fishery activities, such as catching, taking or harvesting from the marine environment. P. monodon is an ecologically important species, as a source of energy for its predators, which include invertebrates, birds, marine mammals and fish of commercial interest. Thus, P. monodon seems to play a key role in this ecosystem, mainly as an intermediate link between top predators and the first links in the food chain. In addition, this species presents various adaptation strategies to the changing oceanographic parameters of the areas it inhabits, even tolerating hypoxic environments and great depths in order to avoid being predated. Likewise, from an economic viewpoint, it has a high commercial value as a marine bioresource with great potential in the pharmaceutical and food industries. Considering this, more studies must be carried out to corroborate the biological, ecological, and fishing importance of this species in order to generate efficient management measures and ensure a sustainable fishery.

Marine Resources Offer New Compounds and Strategies for the Treatment of Skin and Soft Tissue Infections.

Bioprospecting of the marine environment for drug development has gained much attention in recent years owing to its massive chemical and biological diversity. Drugs for the treatment of skin and soft tissue infections have become part of the search, mainly with respect to enlarging the number of available antibiotics, with a special focus on multidrug-resistant Gram-positive bacteria, being the major causative agents in this field. Marine resources offer novel natural products with distinct biological activities of pharmaceutical importance, having the chance to provide new chemical scaffolds and new modes of action. New studies advance the field by proposing new strategies derived from an ecosystemic understanding for preventive activities against biofilms and new compounds suitable as disinfectants, which sustain the natural flora of the skin. Still, the development of new compounds is often stuck at the discovery level, as marine biotechnology also needs to overcome technological bottlenecks in drug development. This review summarizes its potential and shows these bottlenecks and new approaches.

Identification of Marine Biotechnology Value Chains with High Potential in the Northern Mediterranean Region.

Marine (blue) biotechnology is an emerging field enabling the valorization of new products and processes with massive potential for innovation and economic growth. In the Mediterranean region, this innovation potential is not exploited as well as in other European regions due to a lack of a clear identification of the different value chains and the high fragmentation of business innovation initiatives. As a result, several opportunities to create an innovative society are being missed. To address this problem, eight Northern Mediterranean countries (Croatia, France, Greece, Italy, Montenegro, Portugal, Slovenia and Spain) established five national blue biotechnology hubs to identify and address the bottlenecks that prevent the development of marine biotechnology in the region. Following a three-step approach (1. Analysis: setting the scene; 2. Transfer: identification of promising value chains; 3. Capitalization: community creation), we identified the three value chains that are most promising for the Northern Mediterranean region: algae production for added-value compounds, integrated multi-trophic aquaculture (IMTA) and valorization aquaculture/fisheries/processing by-products, unavoidable/unwanted catches and discards. The potential for the development and the technical and non-technical skills that are necessary to advance in this exciting field were identified through several stakeholder events which provided valuable insight and feedback that should be addressed for marine biotechnology in the Northern Mediterranean region to reach its full potential.

Unravelling the Lipids Content and the Fatty Acid Profiles of Eight Recently Described Halophytophthora Species and H. avicennae from the South Coast of Portugal.

In this study, mycelia of eight recently described species of Halophytophthora and H. avicennae collected in Southern Portugal were analysed for lipids and fatty acids (FA) content to evaluate their possible use as alternative sources of FAs and understand how each species FAs profile relates to their phylogenetic position. All species had a low lipid percentage (0.06% in H. avicennae to 0.28% in H. frigida). Subclade 6b species contained more lipids. All species produced monounsaturated (MUFA), polyunsaturated (PUFA) and saturated (SFA) FAs, the latter being most abundant in all species. H. avicennae had the highest FA variety and was the only producer of γ-linolenic acid, while H. brevisporangia produced the lowest number of FAs. The best producer of arachidonic acid (ARA) and eicosapentaenoic acid (EPA) was H. thermoambigua with 3.89% and 9.09% of total FAs, respectively. In all species, palmitic acid (SFA) was most abundant and among the MUFAs produced oleic acid had the highest relative percentage. Principal component analysis (PCA) showed partial segregation of species by phylogenetic clade and subclade based on their FA profile. H. avicennae (Clade 4) differed from all other Clade 6 species due to the production of γ-linolenic and lauric acids. Our results disclosed interesting FA profiles in the tested species, adequate for energy (biodiesel), pharmaceutical and food industries (bioactive FAs). Despite the low amounts of lipids produced, this can be boosted by manipulating culture growth conditions. The observed interspecific variations in FA production provide preliminary insights into an evolutionary background of its production.

Seaweed Extracts to Control Postharvest Phytopathogenic Fungi in Rocha Pear.

Fungal infections cause losses amounting to between 20 and 25% of the fruit industry's total outcome, with an escalating impact on agriculture in the last decades. As seaweeds have long demonstrated relevant antimicrobial properties against a wide variety of microorganisms, extracts from Asparagopsis armata, Codium sp., Fucus vesiculosus, and Sargassum muticum were used to find sustainable, ecofriendly, and safe solutions against Rocha pear postharvest fungal infections. Alternaria alternata, Botrytis cinerea, Fusarium oxysporum, and Penicillium expansum mycelial growth and spore germination inhibition activities were tested in vitro with five different extracts of each seaweed (n-hexane, ethyl acetate, aqueous, ethanolic, and hydroethanolic). An in vivo assay was then performed using the aqueous extracts against B. cinerea and F. oxysporum in Rocha pear. The n-hexane, ethyl acetate, and ethanolic extracts from A. armata showed the best in vitro inhibitory activity against B. cinerea, F. oxysporum, and P. expansum, and promising in vivo results against B. cinerea using S. muticum aqueous extract were also found. The present work highlights the contribution of seaweeds to tackle agricultural problems, namely postharvest phytopathogenic fungal diseases, contributing to a greener and more sustainable bioeconomy from the sea to the farm.

Microbiome enrichment from contaminated marine sediments unveils novel bacterial strains for petroleum hydrocarbon and heavy metal bioremediation.

Petroleum hydrocarbons and heavy metals are some of the most widespread contaminants affecting marine ecosystems, urgently needing effective and sustainable remediation solutions. Microbial-based bioremediation is gaining increasing interest as an effective, economically and environmentally sustainable strategy. Here, we hypothesized that the heavily polluted coastal area facing the Sarno River mouth, which discharges >3 tons of polycyclic aromatic hydrocarbons (PAHs) and ∼15 tons of heavy metals (HMs) into the sea annually, hosts unique microbiomes including marine bacteria useful for PAHs and HMs bioremediation. We thus enriched the microbiome of marine sediments, contextually selecting for HM-resistant bacteria. The enriched mixed bacterial culture was subjected to whole-DNA sequencing, metagenome-assembled-genomes (MAGs) annotation, and further sub-culturing to obtain the major bacterial species as pure strains. We obtained two novel isolates corresponding to the two most abundant MAGs (Alcanivorax xenomutans strain-SRM1 and Halomonas alkaliantarctica strain-SRM2), and tested their ability to degrade PAHs and remove HMs. Both strains exhibited high PAHs degradation (60-100%) and HMs removal (21-100%) yield, and we described in detail >60 genes in their MAGs to unveil the possible genetic basis for such abilities. Most promising yields (∼100%) were obtained towards naphthalene, pyrene and lead. We propose these novel bacterial strains and related genetic repertoire to be further exploited for effective bioremediation of marine environments contaminated with both PAHs and HMs.

In situ devices can culture the microbial dark matter of corals.

Most microorganisms found in environmental samples have never been cultured and can often only be explored through molecular or microscopic approaches. Here, we adapt the use of in situ diffusion-based devices to culture "yet-to-be-cultured" microorganisms associated with coral mucus and compare this with a traditional culturing method. The culturability of microorganisms associated with mucus of the coral Pocillopora damicornis increased by 420% and 570% with diffusion growth chambers and microwell chip devices, respectively, compared with the traditional method tested. The obtained cultures represent up to 64.4% of the total diversity of amplicon sequence variants (ASVs) found in the mucus of the coral P. damicornis. In addition, some previously uncultured microorganisms, such as members of the family Nitrosopumilaceae and halophilic/halotolerant bacteria were cultured. Our results validate alternative microbial culturing strategies to culture coral-associated microorganisms, while significantly increasing the culturability of previous microbial dark matter.

Marine Bioprospecting, Biocatalysis and Process Development.

Oceans possess tremendous diversity in microbial life. The enzymatic machinery that marine bacteria present is the result of extensive evolution to assist cell survival under the harsh and continuously changing conditions found in the marine environment. Several bacterial cells and enzymes are already used at an industrial scale, but novel biocatalysts are still needed for sustainable industrial applications, with benefits for both public health and the environment. Metagenomic techniques have enabled the discovery of novel biocatalysts, biosynthetic pathways, and microbial identification without their cultivation. However, a key stage for application of novel biocatalysts is the need for rapid evaluation of the feasibility of the bioprocess. Cultivation of not-yet-cultured bacteria is challenging and requires new methodologies to enable growth of the bacteria present in collected environmental samples, but, once a bacterium is isolated, its enzyme activities are easily measured. High-throughput screening techniques have also been used successfully, and innovative in vitro screening platforms to rapidly identify relevant enzymatic activities continue to improve. Small-scale approaches and process integration could improve the study and development of new bioprocesses to produce commercially interesting products. In this work, the latest studies related to (i) the growth of marine bacteria under laboratorial conditions, (ii) screening techniques for bioprospecting, and (iii) bioprocess development using microreactors and miniaturized systems are reviewed and discussed.

Cultivating marine bacteria under laboratory conditions: Overcoming the "unculturable" dogma.

Underexplored seawater environments may contain biological resources with potential for new biotechnological applications. Metagenomic techniques revolutionized the study of bacterial communities but culture dependent methods will still be important to help the biodiscovery of new products and enzymes from marine bacteria. In this context, we promoted the growth of bacteria from a marine rock pond by culture dependent techniques and compared the results with culture independent methods. The total number of bacteria and diversity were studied in different agar plate media during 6 weeks. Agar plate counting was of the same order of magnitude of direct microscopy counts. The highest efficiency of cultivation was 45% attained in marine agar medium. Molecular analysis revealed 10 different phyla of which only four were isolated by the culture dependent method. On the other hand, four taxonomic orders were detected by cultivation but not by the molecular technique. These include bacteria from the phyla Bacillota and Actinomycetota. Our study shows that it is possible to grow more than the traditionally considered 1% of bacteria from a seawater sample using standard agar plate techniques and laboratorial conditions. The results also demonstrate the importance of culture methods to grow bacteria not detected by molecular approaches for future biotechnological applications.

Functional proteins through green refining of seafood side streams.

Scarcity of nutritive protein is a major global problem, the severity of which is bound to increase with the rising population. The situation demands finding additional sources of proteins that can be both safe as well as acceptable to the consumer. Food waste, particularly from seafood is a plausible feedstock of proteins in this respect. Fishing operations result in appreciable amounts of bycatch having poor food value. In addition, commercial processing results in 50 to 60% of seafood as discards, which consist of shell, head, fileting frames, bones, viscera, fin, skin, roe, and others. Furthermore, voluminous amounts of protein-rich effluents are released during commercial seafood processing. While meat from the bycatch can be raw material for proteinous edible products, proteins from the process discards and effluents can be recovered through biorefining employing upcoming, environmental-friendly, low-cost green processes. Microbial or enzyme treatments release proteins bound to the seafood matrices. Physico-chemical processes such as ultrasound, pulse electric field, high hydrostatic pressure, green solvent extractions and others are available to recover proteins from the by-products. Cultivation of photosynthetic microalgae in nutrient media consisting of seafood side streams generates algal cell mass, a rich source of functional proteins. A zero-waste marine bio-refinery approach can help almost total recovery of proteins and other ingredients from the seafood side streams. The recovered proteins can have high nutritive value and valuable applications as nutraceuticals and food additives.

Marine Compounds for Melanoma Treatment and Prevention.

Melanoma is considered a multifactorial disease etiologically divided into melanomas related to sun exposure and those that are not, but also based on their mutational signatures, anatomic site, and epidemiology. The incidence of melanoma skin cancer has been increasing over the past decades with 132,000 cases occurring globally each year. Marine organisms have been shown to be an excellent source of natural compounds with possible bioactivities for human health applications. In this review, we report marine compounds from micro- and macro-organisms with activities in vitro and in vivo against melanoma, including the compound Marizomib, isolated from a marine bacterium, currently in phase III clinical trials for melanoma. When available, we also report active concentrations, cellular targets and mechanisms of action of the mentioned molecules. In addition, compounds used for UV protection and melanoma prevention from marine sources are discussed. This paper gives an overview of promising marine molecules which can be studied more deeply before clinical trials in the near future.

A Comparative Analysis of Mucus Immunomodulatory Properties from Seven Marine Gastropods from the Mediterranean Sea.

The treatment of inflammatory and immune-related diseases due to dysfunctioning of the immune system necessitates modulation of the immune response through immunomodulatory compounds. Marine environments are considered as a new frontier for health benefit product implementations. Marine biodiversity is still a low explored resource, despite it is expected to represent an important platform for chemical bioactive compounds. Within the phylum Mollusca, gastropods are known to synthetize mucus, the latter presenting relevant bioactive properties, e.g., related to immunomodulant molecules able to activate the innate and acquired immune system. This study proposes a bioprospecting of the immunomodulant activity of mucus isolated from seven common gastropod species from the Gulf of Naples (Mediterranean Sea). Results showed that not all mucus displayed a significant cytotoxic activity on the two human cancer cell lines A549 and A2058. On the other hand, the mucus from Bolinus brandaris was strongly bioactive and was therefore thoroughly investigated at cellular, molecular, and protein levels on the human monocytes U937 line. It can conclusively induce monocyte differentiation in vitro and significantly stimulate natural immunity response.

Responsible Research and Innovation Framework, the Nagoya Protocol and Other European Blue Biotechnology Strategies and Regulations: Gaps Analysis and Recommendations for Increased Knowledge in the Marine Biotechnology Community.

As the quest for marine-derived compounds with pharmacological and biotechnological potential upsurges, the importance of following regulations and applying Responsible Research and Innovation (RRI) also increases. This article aims at: (1) presenting an overview of regulations and policies at the international and EU level, while demonstrating a variability in their implementation; (2) highlighting the importance of RRI in biodiscovery; and (3) identifying gaps and providing recommendations on how to improve the market acceptability and compliance of novel Blue Biotechnology compounds. This article is the result of the work of the Working Group 4 "Legal aspects, IPR and Ethics" of the COST Action CA18238 Ocean4Biotech, a network of more than 130 Marine Biotechnology scientists and practitioners from 37 countries. Three qualitative surveys ("Understanding of the Responsible Research and Innovation concept", "Application of the Nagoya Protocol in Your Research", and "Brief Survey about the experiences regarding the Nagoya Protocol") indicate awareness and application gaps of RRI, the Nagoya Protocol, and the current status of EU policies relating to Blue Biotechnology. The article categorises the identified gaps into five main categories (awareness, understanding, education, implementation, and enforcement of the Nagoya Protocol) and provides recommendations for mitigating them at the European, national, and organisational level.

Comparative transcriptomic analysis of in situ and onboard fixed deep-sea limpets reveals sample preparation-related differences.

Precise gene expression reflects the molecular response of deep-sea organisms to their harsh living environments. However, changes in environmental factors during lifting samples from the deep sea to a research vessel can also affect gene expression. By using the transcriptomic approach, we compared the gene expression profiles of the onboard fixed with the in situ fixed samples of the deep-sea limpet Bathyacmaea lactea. Our results revealed that the concomitant stress during conventional deep-sea sampling without RNA in situ fixation greatly influenced the gene expression. Various biological activities, such as cell and tissue structure, lysosomal activity, fluid balance, and unsaturated fatty acid metabolism, were perturbed, suggesting that the sampling stress has exerted systemic impacts on the life of the limpets. These findings clearly illustrate that deep-sea samples without RNA in situ fixation can easily lead to biased results in gene expression analysis, which requires to be appropriately addressed in future studies.

Drug Delivery Systems for Cancer Treatment: A Review of Marine-derived Polysaccharides.

Cancer is a disease characterized by uncontrolled cell proliferation and the spread of cells to other tissues and remains one of the worldwide problems waiting to be solved. There are various treatment strategies for cancer, such as chemotherapy, surgery, radiotherapy, and immunotherapy, although it varies according to its type and stage. Many chemotherapeutic agents have limited clinical use due to lack of efficacy, off-target toxicity, metabolic instability, or poor pharmacokinetics. One possible solution to this high rate of clinical failure is to design drug delivery systems that deliver drugs in a controlled and specific manner and are not toxic to normal cells. Marine systems contain biodiversity, including components and materials that can be used in biomedical applications and therapy. Biomaterials such as chitin, chitosan, alginate, carrageenan, fucoidan, hyaluronan, agarose, and ulvan obtained from marine organisms have found use in DDSs today. These polysaccharides are biocompatible, non-toxic, biodegradable, and cost-effective, making them ideal raw materials for increasingly complex DDSs with a potentially regulated release. In this review, the contributions of polysaccharides from the marine environment to the development of anticancer drugs in DDSs will be discussed.

Accessing Nystatin through Mariculture.

Understanding our oceans and their marine ecosystems has enabled the development of sustainable systems for mariculture. While the bulk of studies to date have focused on the production of food, its remarkable expanse has inspired the translation of other markets towards aquatic environments. This manuscript outlines an approach to pharmaceutical mariculture, by demonstrating a benchmark for future prototyping. Here, design, field evaluation and natural product chemistry are united to successfully produce nystatin at sea. This study begins by evaluating new designs for culture flasks, illustrating a next step towards developing self-contained bioreactors for culturing in marine environments. Through pilot studies, an underwater system was developed to cost effectively produce cultures that yielded 200 mg of nystatin per deployment. Overall, this study demonstrates the potential for the practical culturing of microbes in a marine environment and provides an important next step for the fledgling field of molecular mariculture.

Potential of Asparagopsis armata as a Biopesticide for Weed Control under an Invasive Seaweed Circular-Economy Framework.

Marine macroalgae have been increasingly targeted as a source of bioactive compounds to be used in several areas, such as biopesticides. When harvesting invasive species, such as Asparagopsis armata, for this purpose, there is a two-folded opportunity: acquiring these biomolecules from a low-cost resource and controlling its spreading and impacts. The secondary metabolites in this seaweed's exudate have been shown to significantly impact the physiology of species in the ecosystems where it invades, indicating a possible biocidal potential. Considering this in the present work, an A. armata exudate cocktail was applied in the model weed Thellungiella halophila to evaluate its physiological impact and mode of action, addressing its potential use as a natural biocide. A. armata greatly affected the test plants' physiology, namely, their photochemical energy transduction pathway (impairing light-harvesting and chemical energy production throughout the chloroplast electron transport chain), carotenoid metabolism and oxidative stress. These mechanisms of action are similar to the ones triggered when using the common chemical pesticides, highlighting the potential of the A. armata exudate cocktail as an eco-friendly biopesticide.