The Identification of Peptide Inhibitors of the Coronavirus 3CL Protease from a Fucus ceranoides L. Hydroalcoholic Extract Using a Ligand-Fishing Strategy.

Brown seaweeds of the Fucus genus represent a rich source of natural antiviral products. In this study, a Fucus ceranoides hydroalcoholic extract (FCHE) was found to inhibit 74.2 ± 1.3% of the proteolytic activity of the free SARS-CoV-2 3CL protease (3CLpro), an enzyme that plays a pivotal role in polyprotein processing during coronavirus replication and has been identified as a relevant drug discovery target for SARS- and MERS-CoVs infections. To purify and identify 3CLpro ligands with potential inhibitory activity using a one-step approach, we immobilized the enzyme onto magnetic microbeads (3CLpro-MPs), checked that the enzymatic activity was maintained after grafting, and used this bait for a ligand-fishing strategy followed by a high-resolution mass spectrometry analysis of the fished-out molecules. Proof of concept for the ligand-fishing capacity of the 3CLpro-MPs was demonstrated by doping the FCHE extract with the substrate peptide TSAVLQ-pNA, resulting in the preferential capture of this high-affinity peptide within the macroalgal complex matrix. Ligand fishing in the FCHE alone led to the purification and identification via high-resolution mass spectrometry (HRMS) of seven hepta-, octa-, and decapeptides in an eluate mix that significantly inhibited the free 3CLpro more than the starting FCHE (82.7 ± 2.2% inhibition). Molecular docking simulations of the interaction between each of the seven peptides and the 3CLpro demonstrated a high affinity for the enzyme's proteolytic active site surpassing that of the most affine peptide ligand identified so far (a co-crystallographic peptide). Testing of the corresponding synthetic peptides demonstrated that four out of seven significantly inhibited the free 3CLpro (from 46.9 ± 6.4 to 76.8 ± 3.6% inhibition at 10 µM). This study is the first report identifying peptides from Fucus ceranoides with high inhibitory activity against the SARS-CoV-2 3CLprotease which bind with high affinity to the protease's active site. It also confirms the effectiveness of the ligand-fishing strategy for the single-step purification of enzyme inhibitors from complex seaweed matrices.

Cultivable epiphytic bacteria of the Chlorophyta Ulva sp.: diversity, antibacterial, and biofilm-modulating activities.

AIMS: Macroalgae harbor a rich epiphytic microbiota that plays a crucial role in algal morphogenesis and defense mechanisms. This study aims to isolate epiphytic cultivable microbiota from Ulva sp. surfaces. Various culture media were employed to evaluate a wide range of cultivable microbiota. Our objective was to assess the antibacterial and biofilm-modulating activities of supernatants from isolated bacteria. METHODS AND RESULTS: Sixty-nine bacterial isolates from Ulva sp. were identified based on 16S rRNA gene sequencing. Their antibacterial activity and biofilm modulation potential were screened against three target marine bacteria: 45%, mostly affiliated with Gammaproteobacteria and mainly grown on diluted R2A medium (R2Ad), showed strong antibacterial activity, while 18% had a significant impact on biofilm modulation. Molecular network analysis was carried out on four bioactive bacterial supernatants, revealing new molecules potentially responsible for their activities. CONCLUSION: R2Ad offered the greatest diversity and proportion of active isolates. The molecular network approach holds promise for both identifying bacterial isolates based on their molecular production and characterizing antibacterial and biofilm-modulating activities.

The Use of FTIR Spectroscopy as a Tool for the Seasonal Variation Analysis and for the Quality Control of Polysaccharides from Seaweeds.

Macroalgae are a potentially novel source of nutrition and biologically active molecules. Proliferative species such as Eucheuma denticulatum, Solieria chordalis (red algae) and Sargassum muticum (brown alga) constitute a huge biomass that can be exploited. In this study, we focus on the extraction of polysaccharides from these three macroalgae species and the characterization of cell wall polysaccharides such as carrageenans, fucoidans and alginates by Fourier Transform Infrared spectroscopy with Attenuated Reflectance Module (FTIR-ATR). The comparison of purified extracts with commercial solutions of fucoidans, alginates or carrageenans shows a strong similarity between the spectra. It demonstrates that the methods of extraction that have been used are also suitable purifying technics. Moreover, it validates infrared spectroscopy as a quick, simple and non-destructive method for the accurate analysis of polysaccharides. The FTIR technique applied to samples collected at different periods of the year allowed us to highlight differences in the composition of fucoidans, alginates and carrageenans. Different classes corresponding to the season can be distinguished by statistical multidimensionnal analysis (Principal Component Analysis) showing that the structure of algal polysaccharides, related to bioactivity, depends on the period of harvest. FTIR results showed that S. chordalis and E. denticulatum possess a dominant type of carrageenan called iota-carrageenan. This type of carrageenan is in the majority when the alga is at maturity in its development cycle. During its growth phase, iota-carrageenan precursors can be detected by FTIR spectra, enabling a better control of the extraction and an application of these compounds in various economic sectors. When the alga E. denticulatum is in its juvenile stage, we found traces of kappa-carrageenan and nu-carrageenan polysaccharides in some extracts.

Primary structural features, physicochemical and biological properties of two water-soluble polysaccharides extracted from the brown Tunisian seaweed Halopteris scoparia.

Marine algae are the most abundant resource in the marine environment and are still a promising source of bioactive compounds including hydrocolloids. This study contributes to the evaluation of the biological and biotechnological potentials of two water soluble polysaccharides, namely alginates (AHS) and fucoidan (FHS), extracted and purified from Halopteris scoparia, an abundant Tunisian brown macroalgae collected in Tunisia (Tabarka region). The total sugars, neutral monosaccharides, uronic acids, proteins, polyphenols, and sulfate groups contents were quantified for both fractions, as well as their functional groups and primary structural features by Fourier transform infrared spectroscopy, ionic and/or gas chromatography and nuclear magnetic resonance analyses. AHS and FHS showed significant anti-inflammatory (IC50 ≈ 1 mg/mL), anticoagulant (e.g., 27-61.7 for the activated partial thromboplastin time), antihyperglycemic (0.1-40 µg/mL) and anti-trypsin (IC50 ≈ 0.3-0.4 mg/mL) effects. FHS and a hydrolyzed fraction showed a very promising potential against herpes viruses (HSV-1) (IC50 < 28 µg/mL). Besides, AHS and two hydrolyzed fractions were able to stimulate the natural defenses of tomato seedlings, assessing their elicitor capacity, by increasing the activity of phenylalanine ammonia-lyase (66-422 %) but also significantly changing the polyphenol content in the leaves (121-243 %) and roots (30-104 %) of tomato plants.

Anti-inflammatory potential of ulvan.

Green seaweeds are a widespread group of marine macroalgae that could be regarded as biorenewable source of valuable compounds, in particular sulfated polysaccharides like ulvans with interesting biological properties. Among them, anti-inflammatory activity represents an interesting target, since ulvans could potentially avoid side effects of conventional therapies. However, a great variability in ulvan content, composition, structure and properties occurs depending on seaweed specie and growth and processing conditions. All these aspects should be carefully considered in order to have reproducible and well characterized products. This review presents some concise ideas on ulvan composition and general concepts on inflammation mechanisms. Then, the main focus is on the importance of adequate selection of extraction, depolymerization and purification technologies followed by an updated survey on anti-inflammatory properties of ulvans through modulation of different signaling pathways. The potential application in a number of diseases, with special emphasis on inflammaging, gut microbiota dysbiosis, wound repair, and metabolic diseases is also discussed. This multidisciplinary overview tries to present the potential of ulvans considering not only mechanistic, but also processing and applications aspects, trusting that it can aid in the development and application of this widely available and renewable resource as an efficient and versatile anti-inflammatory agent.

Anti-SARS-CoV-2 Activity of Polysaccharides Extracted from Halymenia floresii and Solieria chordalis (Rhodophyta).

Even after hundreds of clinical trials, the search for new antivirals to treat COVID-19 is still relevant. Carrageenans are seaweed sulfated polysaccharides displaying antiviral activity against a wide range of respiratory viruses. The objective of this work was to study the antiviral properties of Halymenia floresii and Solieria chordalis carrageenans against SARS-CoV-2. Six polysaccharide fractions obtained from H. floresii and S. chordalis by Enzyme-Assisted Extraction (EAE) or Hot Water Extraction (HWE) were tested. The effect of carrageenan on viral replication was assessed during infection of human airway epithelial cells with a clinical strain of SARS-CoV-2. The addition of carrageenans at different times of the infection helped to determine their mechanism of antiviral action. The four polysaccharide fractions isolated from H. floresii displayed antiviral properties while the S. chordalis fractions did not. EAE-purified fractions caused a stronger reduction in viral RNA concentration. Their antiviral action is likely related to an inhibition of the virus attachment to the cell surface. This study confirms that carrageenans could be used as first-line treatment in the respiratory mucosa to inhibit the infection and transmission of SARS-CoV-2. Low production costs, low cytotoxicity, and a broad spectrum of antiviral properties constitute the main strengths of these natural molecules.

Sulfated Polysaccharides from Seaweed Strandings as Renewable Source for Potential Antivirals against Herpes simplex Virus 1.

Herpes simplex virus 1 (HSV-1) remains a prominent health concern widespread all over the world. The increasing genital infections by HSV-1 that might facilitate acquisition and transmission of HIV-1, the cumulative evidence that HSV-1 promotes neurodegenerative disorders, and the emergence of drug resistance signify the need for new antiviral agents. In this study, the in vitro anti-herpetic activity of sulfated polysaccharides (SPs) extracted by enzyme or hot water from seaweeds collected in France and Mexico from stranding events, were evaluated. The anti-herpetic activity evaluation of the semi-refined-polysaccharides (sr-SPs) and different ion exchange purified fractions showed a wide range of antiviral activity. Among them, the sr-SPs from the Rhodophyta Halymenia floresii showed stronger activity EC50 0.68 µg/mL with SI 1470, without cytotoxicity. Further, the antiviral activity of the sr-SPs evaluated at different treatment schemes showed a high EC50 of 0.38 µg/mL during the viral adsorption assays when the polysaccharide and the virus were added simultaneously, whilst the protection on Vero cell during the post-infection assay was effective up to 1 h. The chemical composition, FTIR and 1H NMR spectroscopic, and molecular weights of the sr-SPs from H. floresii were determined and discussed based on the anti-herpetic activity. The potential utilization of seaweed stranding as a source of antiviral compounds is addressed.

Effects of Ulva sp. Extracts on the Growth, Biofilm Production, and Virulence of Skin Bacteria Microbiota: Staphylococcus aureus, Staphylococcus epidermidis, and Cutibacterium acnes Strains.

Ulva sp. is known to be a source of bioactive compounds such as ulvans, but to date, their biological activity on skin commensal and/or opportunistic pathogen bacteria has not been reported. In this study, the effects of poly- and oligosaccharide fractions produced by enzyme-assisted extraction and depolymerization were investigated, for the first time in vitro, on cutaneous bacteria: Staphylococcus aureus, Staphylococcus epidermidis, and Cutibacterium acnes. At 1000 µg/mL, poly- and oligosaccharide fractions did not affect the growth of the bacteria regarding their generation time. Polysaccharide Ulva sp. fractions at 1000 µg/mL did not alter the bacterial biofilm formation, while oligosaccharide fractions modified S. epidermidis and C. acnes biofilm structures. None of the fractions at 1000 µg/mL significantly modified the cytotoxic potential of S. epidermidis and S. aureus towards keratinocytes. However, poly- and oligosaccharide fractions at 1000 µg/mL induced a decrease in the inflammatory potential of both acneic and non-acneic C. acnes strains on keratinocytes of up to 39.8%; the strongest and most significant effect occurred when the bacteria were grown in the presence of polysaccharide fractions. Our research shows that poly- and oligosaccharide Ulva sp. fractions present notable biological activities on cutaneous bacteria, especially towards C. acnes acneic and non-acneic strains, which supports their potential use for dermo-cosmetic applications.

Poly- and Oligosaccharide Ulva sp. Fractions from Enzyme-Assisted Extraction Modulate the Metabolism of Extracellular Matrix in Human Skin Fibroblasts: Potential in Anti-Aging Dermo-Cosmetic Applications.

Ulva sp. is known to be a source of bioactive compounds such as ulvans, but their biological activity on human dermal fibroblast extracellular matrix (ECM) is poorly reported. In this work, the regulation of ECM has been investigated for the first time at both proteomic and transcriptomic levels in normal human skin dermal fibroblasts, after 48 h of incubation with poly- and oligosaccharide fractions from Ulva sp. obtained after enzyme-assisted extraction and depolymerization. Cell proliferation enhancement (up to +68%) without exhibiting any cytotoxic effect on fibroblasts was demonstrated at 50 and 1000 µg/mL by both fractions. At the proteomic level, polysaccharide fractions at 1000 µg/mL enhanced the most the synthesis of glycosaminoglycans (GAGs, up to +57%), total collagen, especially types I (up to +217%) and III, as well as the synthesis and activity of MMP-1 (Matrix Metalloproteinase-1, up to +309%). In contrast, oligosaccharide fractions had no effect on GAGs synthesis but exhibited similarities for collagens and MMP-1 regulation. At the transcriptomic level, the decrease of COL1A1 and COL1A2 expression, and increase of COL3A1 and MMP-1 expression, confirmed the modulation of ECM metabolism by both fractions. Our research emphasizes that poly- and oligosaccharide Ulva sp. fractions exhibit interesting biological activities and supports their potential use in the area of skin renewal for anti-aging dermo-cosmetic applications.

An Analysis of the Nutritional and Health Values of Caulerpa racemosa (Forsskål) and Ulva fasciata (Delile)-Two Chlorophyta Collected from the Philippines.

Polysaccharides, lipids and amino acid profiles were investigated to understand the nutritional value of Caulerpa racemosa and Ulva fasciata from the Philippines. The results revealed that both species contain high amounts of proteins (8.8-19.9% for C. racemosa and 8.0-11.1% for U. fasciata). The portions of the total amino acids that were essential amino acids (EAAs) (45.28 ± 0.12% for C. racemosa and 42.17 ± 0.12% for U. fasciata) out were comparable to FAO/WHO requirements. Leucine, valine, isoleucine, and lysine are the dominant EAAs in C. racemosa, while leucine, valine, lysine, and phenylalanine are those in U. fasciata. The fatty acid profiles are dominated by monounsaturated fatty acids and polyunsaturated fatty acids in C. racemosa (56.2%), while saturated fatty acids (72.1%) are dominant in U. fasciata. High C18/C20 polyunsaturated fatty acid ratios were recorded in both species. Mineral contents for both seaweeds were within levels considered safe for functional foods. Total pigment content of C. racemosa (140.84 mg/g dw) was almost 20 times higher than that of U. fasciata (7.54 mg/g dw). Hot water extract (HWE) from C. racemosa showed in vitro antiherpetic activity without cytotoxicity. Nutritional characteristics confirmed that C. racemosa could be potentially used as a nutritious and functional food items for human consumption.

Bioactivity of compounds secreted by symbiont bacteria of Nudibranchs from Indonesia.

The aims of this work are to isolate bacterial symbionts from nudibranchs and subsequently to determine anti-Methicillin resistant Staphylococcus aureus (MRSA), cytotoxicity and anti-Herpes simplex virus type 1 (HSV-1) activities of bio compounds. A total of 15 species of nudibranchs were collected from Karimunjawa and five species from Bali, respectively. A total of 245 bacteria isolates were obtained. The anti-MRSA activity screening activity indicated two active bacteria. Ethyl acetate extracts from supernatants, indicating extracelullar compounds, showed an inhibition zone against MRSA at concentrations of 500-1,000 µg/ml. DNA sequence analysis showed that the strain KJB-07 from Phyllidia coelestis was closely related to Pseudoalteromonas rubra, whereas the strain NP31-01 isolated from Phyllidia varicosa was closely related to Virgibacillus salarius. The extract of Pseudoalteromonas rubra was cytotoxic to Vero cells at a concentration of 75 µg/ml. The extract of V. salarius presented no cytotoxicity at concentrations of 5-1,000 µg/ml. No anti HSV-1 was observed for both isolated bacteria. This is the first study describing research on anti-MRSA, cytotoxicity and anti HSV-1 activity of bacterial symbionts from the viscera of nudibranch. Compounds produced by Pseudoalteromonas rubra and V. salarius, had potential anti-MRSA activity. However, only extracts from Pseudoalteromonas rubra showed cytotoxic effects on Vero cells. Three compounds were identified by LC/MS after purification from culture supernatant.

Production of Active Poly- and Oligosaccharidic Fractions from Ulva sp. by Combining Enzyme-Assisted Extraction (EAE) and Depolymerization.

Data on fractionation and depolymerization of the matrix ulvan polysaccharides, and studies on the biological activities on skin cells, are very scarce. In this work, crude ulvans were produced by using EAE (enzyme-assisted extraction) and compared to maceration (an established procedure). After different fractionation procedures-ethanolic precipitation, dialysis, or ammonium sulfate precipitation-the biochemical composition showed that EAE led to an increased content in ulvans. Coupling EAE to sulfate ammonium precipitation led to protein enrichment. Oligosaccharides were obtained by using radical depolymerization by H2O2 and ion-exchange resin depolymerization. Sulfate groups were partially cleaved during these chemical treatments. The potential bioactivity of the fractions was assessed using a lipoxygenase inhibition assay for anti-inflammatory activity and a WST-1 assay for human dermal fibroblast viability and proliferation. All ulvans extracts, poly- and oligosaccharidic fractions from EAE, expanded the fibroblast proliferation rate up to 62%. Our research emphasizes the potential use of poly- and oligosaccharidic fractions of Ulva sp. for further development in cosmetic applications.

Environmentally Friendly Valorization of Solieria filiformis (Gigartinales, Rhodophyta) from IMTA Using a Biorefinery Concept.

Marine macroalgae (seaweed) are an excellent source of novel bioactive metabolites. The biorefinery concept applied to seaweed facilitates the extraction of many chemical constituents from the same biomass ensuring that the resource is used fully, generating few residues through a succession of extraction steps. In the present study, the biomass of the carragenophyte Solieria filiformis (Rhodophyta, Gigartinales) cultured in an integrated multi-trophic aquaculture (IMTA) system was evaluated to obtain valuable products by a biorefinery approach. Enzymatic-assisted extraction (EAE) and microwave-assisted extraction (MAE) were the eco-friendly technologies used to ensure an environmentally friendly valorization of the biomass. Three valuable products were successfully recovered: a water-soluble extract rich in proteins and sulfated polysaccharides suitable as a food supplement; a lipid fraction rich in polyunsaturated fatty acids (PUFAs) with potential to be used in the nutraceutical industry; and a pure ι-carrageenan with a powerful antiviral activity against Herpes simplex virus (EC50 = 6.3 µg mL-1) comparable to the commercial antiviral acyclovir (EC50 = 3.2⁻5.4 µg mL-1).

Seasonal biomass and alginate stock assessment of three abundant genera of brown macroalgae using multispectral high resolution satellite remote sensing: A case study at Ekas Bay (Lombok, Indonesia).

The potential of Indonesian bays as alginate producers was assessed by determining the stock of wild brown algae and exploring their biomass as alginophytes at the scale of entire bay, using a combination of field observations, remote sensing high resolution data and GIS tools. Ekas Bay in Lombok Island presented a stock of brown macroalgae which varied with season and species: for Padina the biomass reached 97.85±12.63 and 79.54±2.53tons in May/June and November respectively; for Sargassaceae species, it reached 669.70±109.64 and 147.70±77.97tons in May/June and November respectively. The best alginate yields occurred during the May/June period: Padina could produce 9.10±0.06tons DW of alginates. Interestingly, Sargassum/Turbinaria together allow 207.61±0.42tons DW of alginates. This study suggests that wild Sargassaceae represent an interesting stock in terms of biomass, alginate yield and M/G ratio.

Phenolics, fatty acids composition and biological activities of various extracts and fractions of Malaysian Aaptos aaptos

Objective: To investigate phenolics, fatty acids composition and biological activities of various extracts and fractions of Malaysian Aaptos aaptos. Methods: Fatty acid methyl ester was analyzed by gas chromatography-flame ionization detector. Antioxidant activity was determined using 2,2-diphenyl-picrylhydrazyl radical scavenging assay and total phenolics content by Folin-Ciocalteu procedure. Vero cells viability was evaluated using methyl thiazole tetrazolium and the inactivation of herpes simplex virus type 1 by neutral red uptake assay. p-Hydroxybenzamide isolated by column chromatography was characterized by utilizing nuclear magnetic resonance spectroscopy and electron impact mass spectrometry. Results:The chloroform, ethyl acetate and methanol extracts of Aaptos aaptos produced higher portions of straight-chain saturated fatty acid, while hexane extract mainly consisted of unsaturated fatty acid. The five majors of fatty acid methyl ester were identified as behenic acid, cis-10-heptadecenoic acid and cis-10-pentadecenoic acids, palmitic acid and tricosanoic acid. In addition, among all organic extracts, chloroform extract inactivated herpes simplex virus type 1 while exhibited weak cytotoxic activity against normal Vero cells and also exhibited strong cytotoxic activity on HL-60, MCF-7, K562, CEM-SS and WEHI-3B cells. A phenolic compound, p-hydroxybenzamide was also isolated from the sponge. Conclusions: Aaptos aaptos could be a source to derive the potential antiviral and anticancer agents. However, further studies are needed to determine the mechanism involved in the process.

Total phenolic content and biological activities of enzymatic extracts from Sargassum muticum (Yendo) Fensholt.

Seaweeds are potentially excellent sources of bioactive metabolites that could represent useful leads in the development of new functional ingredients in pharmaceutical and cosmetic industries. In the last decade, new marine bioprocess technologies have allowed the isolation of substances with biological properties. The brown alga Sargassum muticum (Yendo) Fensholt (Ochrophyta) was enzymatically hydrolyzed to prepare water-soluble extracts by using six different commercially available carbohydrate-degrading enzymes and two proteases. Evaluation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) antioxidant, tyrosinase, elastase, and biofilm inhibition, antibacterial and antiviral activities as well as evaluation of cytotoxicity were realized for each extract. Total phenolic content was measured for extract characterization, and solid-phase extraction was useful to purify the enzymatic extract. Soluble total phenolic content of S. muticum Viscozyme extract was highest with 6.4% of dry weight. Enzymatic Celluclast and Viscozyme extracts had the lowest value of DPPH IC50 indicating a strong antiradical activity, 0.6 mg mL-1, in comparison with other enzymes. The ferric reducing antioxidant power ranged between 48.7 µM Fe2+ Eq, digested with Viscozyme, and 60.8 µM Fe2+ Eq, digested with Amyloglucosidase. Tyrosinase inhibition activity of S. muticum Neutrase extract was 41.3% higher compared to other enzymes. Elastase inhibition activity of S. muticum Shearzyme extract had highest activity (32.8%). All enzymatic extracts showed no cytotoxic effect towards the kidney Vero cells. Meanwhile, only S. muticum Neutrase and Alcalase extracts exhibited potential antiviral activity. In addition, S. muticum Viscozyme and Shearzyme extracts showed promising activity in suppressing the biofilm formation against Pseudomonas aeruginosa and Escherichia coli, respectively. Purification of S. muticum Viscozyme extracts by solid-phase extraction managed to concentrate the phenolic content and improve the bioactivity. These results indicate the promising potential of enzyme-assisted followed by solid-phase extraction in recovering phenolic content and in improving its bioactivity.

Lipid Composition, Fatty Acids and Sterols in the Seaweeds Ulva armoricana, and Solieria chordalis from Brittany (France): An Analysis from Nutritional, Chemotaxonomic, and Antiproliferative Activity Perspectives.

Lipids from the proliferative macroalgae Ulva armoricana (Chlorophyta) and Solieria chordalis (Rhodophyta) from Brittany, France, were investigated. The total content of lipids was 2.6% and 3.0% dry weight for U. armoricana and S. chordalis, respectively. The main fractions of S. chordalis were neutral lipids (37%) and glycolipids (38%), whereas U. armoricana contained mostly neutral lipids (55%). Polyunsaturated fatty acids (PUFA) represented 29% and 15% of the total lipids in U. armoricana and S. chordalis, respectively. In both studied algae, the phospholipids were composed of PUFA for 18%. In addition, PUFA were shown to represent 9% and 4.5% of glycolipids in U. armoricana and S. chordalis, respectively. The essential PUFA were 16:4n-3, 18:4n-3, 18:2n-3, 18:2n-6, and 22:6n-3 in U. armoricana, and 20:4n-6 and 20:5n-3 in S. chordalis. It is important to notice that six 2-hydroxy-, three 3-hydroxy-, and two monounsaturated hydroxy fatty acids were also identified and may provide a chemotaxonomic basis for algae. These seaweeds contained interesting compounds such as squalene, α-tocopherol, cholest-4-en-3-one and phytosterols. The antiproliferative effect was evaluated in vitro on human non-small-cell bronchopulmonary carcinoma line (NSCLC-N6) with an IC50 of 23 µg/mL for monogalactosyldiacylglycerols isolated from S. chordalis and 24 µg/mL for digalactosyldiacylglycerols from U. armoricana. These results confirm the potentialities of valorization of these two species in the fields of health, nutrition and chemotaxonomy.

Enzyme-assisted extraction of bioactive material from Chondrus crispus and Codium fragile and its effect on herpes simplex virus (HSV-1).

Codium fragile and Chondrus crispus are, respectively, green and red seaweeds which are abundant along the North Atlantic coasts. We investigated the chemical composition and antiviral activity of enzymatic extracts of C. fragile (CF) and C. crispus (CC). On a dry weight basis, CF consisted of 11% protein, 31% neutral sugars, 0.8% sulfate, 0.6% uronic acids, and 49% ash, while CC contained 27% protein, 28% neutral sugars, 17% sulfate, 1.8% uronic acids, and 25% ash. Enzyme-assisted hydrolysis improved the extraction efficiency of bioactive materials. Commercial proteases and carbohydrases significantly improved (p ≤ 0.001) biomass yield (40%-70% dry matter) as compared to aqueous extraction (20%-25% dry matter). Moreover, enzymatic hydrolysis enhanced the recovery of protein, neutral sugars, uronic acids, and sulfates. The enzymatic hydrolysates exhibited significant activity against Herpes simplex virus (HSV-1) with EC50 of 77.6-126.8 µg/mL for CC and 36.5-41.3 µg/mL for CF, at a multiplicity of infection (MOI) of 0.001 ID50/cells without cytotoxity (1-200 µg/mL). The extracts obtained from proteases (P1) and carbohydrases (C3) were also effective at higher virus MOI of 0.01 ID50/cells without cytotoxity. Taken together, these results indicate the potential application of enzymatic hydrolysates of C. fragile and C. crispus in functional food and antiviral drug discovery.

Ostreid herpesvirus type 1 replication and host response in adult Pacific oysters, Crassostrea gigas.

Since 2008, massive mortality outbreaks associated with OsHV-1 detection have been reported in Crassostrea gigas spat and juveniles in several countries. Nevertheless, adult oysters do not demonstrate mortality in the field related to OsHV-1 detection and were thus assumed to be more resistant to viral infection. Determining how virus and adult oyster interact is a major goal in understanding why mortality events are not reported among adult Pacific oysters. Dual transcriptomics of virus-host interactions were explored by real-time PCR in adult oysters after a virus injection. Thirty-nine viral genes and five host genes including MyD88, IFI44, IkB2, IAP and Gly were measured at 0.5, 10, 26, 72 and 144 hours post infection (hpi). No viral RNA among the 39 genes was detected at 144 hpi suggesting the adult oysters are able to inhibit viral replication. Moreover, the IAP gene (oyster gene) shows significant up-regulation in infected adults compared to control adults. This result suggests that over-expression of IAP could be a reaction to OsHV-1 infection, which may induce the apoptotic process. Apoptosis could be a main mechanism involved in disease resistance in adults. Antiviral activity of haemolymph against herpes simplex virus (HSV-1) was not significantly different between infected adults versus control.

Marennine, promising blue pigments from a widespread Haslea diatom species complex.

In diatoms, the main photosynthetic pigments are chlorophylls a and c, fucoxanthin, diadinoxanthin and diatoxanthin. The marine pennate diatom Haslea ostrearia has long been known for producing, in addition to these generic pigments, a water-soluble blue pigment, marennine. This pigment, responsible for the greening of oysters in western France, presents different biological activities: allelopathic, antioxidant, antibacterial, antiviral, and growth-inhibiting. A method to extract and purify marennine has been developed, but its chemical structure could hitherto not be resolved. For decades, H. ostrearia was the only organism known to produce marennine, and can be found worldwide. Our knowledge about H. ostrearia-like diatom biodiversity has recently been extended with the discovery of several new species of blue diatoms, the recently described H. karadagensis, H. silbo sp. inedit. and H. provincialis sp. inedit. These blue diatoms produce different marennine-like pigments, which belong to the same chemical family and present similar biological activities. Aside from being a potential source of natural blue pigments, H. ostrearia-like diatoms thus present a commercial potential for aquaculture, cosmetics, food and health industries.