Unveiling the Cultivation of Nostoc sp. under Controlled Laboratory Conditions.

Cyanobacteria, photoautotrophic Gram-negative bacteria, play a crucial role in aquatic and terrestrial environments, contributing significantly to fundamental ecological processes and displaying potential for various biotechnological applications. It is, therefore, critical to identify viable strains for aquaculture and establish accurate culture parameters to ensure an extensive biomass supply for biotechnology purposes. This study aims to establish optimal laboratory batch culture conditions for Nostoc 136, sourced from Alga2O, Coimbra, Portugal. Preliminary investigations were conducted to identify the optimal culture parameters and to perform biomass analysis, including protein and pigment content. The highest growth was achieved with an initial inoculum concentration of 1 g.L-1, using modified BG11 supplemented with nitrogen, resulting in a Specific Growth Rate (SGR) of 0.232 ± 0.017 µ.day-1. When exposed to white, red, and blue LED light, the most favourable growth occurred under a combination of white and red LED light exhibiting an SGR of 0.142 ± 0.020 µ.day-1. The protein content was determined to be 10.80 ± 2.09%. Regarding the pigments, phycocyanin reached a concentration of 200.29 ± 30.07 µg.mL-1, phycoerythrin 148.29 ± 26.74 µg.mL-1, and allophycocyanin 10.69 ± 6.07 µg.mL-1. This study underscores the influence of light and nutrient supplementation on the growth of the Nostoc biomass.

Valorization of the Red Seaweed Gracilaria gracilis Through a Biorefinery Approach.

The interest in seaweeds as an abundant feedstock to obtain valuable and multitarget bioactive ingredients is continuously growing. In this work, we explore the potential of Gracilaria gracilis, an edible red seaweed cultivated worldwide for its commercial interest as a source of agar and other ingredients for cosmetic, pharmacological, food, and feed applications. G. gracilis growth conditions were optimized through vegetative propagation and sporulation while manipulating the physicochemical conditions to achieve a large biomass stock. Green extraction methodologies with ethanol and water were performed over the seaweed biomass. The bioactive potential of extracts was assessed through a set of in vitro assays concerning their cytotoxicity, antioxidant, and antimicrobial properties. Additionally, dried seaweed biomass was incorporated into pasta formulations to increase food's nutritional value. Pigments extracted from G. gracilis have also been incorporated into yogurt as a natural colorant, and their stability was evaluated. Both products were submitted to the appreciation of a semi-trained sensorial panel aiming to achieve the best final formulation before reaching the market. Results support the versatility of G. gracilis whether it is applied as a whole biomass, extracts and/or pigments. Through implementing several optimized protocols, this work allows the development of products with the potential to profit the food, cosmetic, and aquaculture markets, promoting environmental sustainability and a blue circular economy. Moreover, and in line with a biorefinery approach, the residual seaweed biomass will be used as biostimulant for plant growth or converted to carbon materials to be used in water purification of the in-house aquaculture systems of MARE-Polytechnic of Leiria, Portugal.

Gelidiales Are Not Just Agar-Revealing the Antimicrobial Potential of Gelidium corneum for Skin Disorders.

In recent decades, seaweeds have proven to be an excellent source of bioactive molecules. Presently, the seaweed Gelidium corneum is harvested in a small area of the Portuguese coast exclusively for agar extraction. The aim of this work was to fully disclosure Gelidium corneum as a sustainable source of antimicrobial ingredients for new dermatological formulations, highlighting its potential to be explored in a circular economy context. For this purpose, after a green sequential extraction, these seaweed fractions (F1-F5) were chemically characterized (1H NMR) and evaluated for their antimicrobial potential against Staphylococcus aureus, Staphylococcus epidermidis and Cutibacterium acnes. The most active fractions were also evaluated for their effects on membrane potential, membrane integrity and DNA damage. Fractions F2 and F3 displayed the best results, with IC50 values of 16.1 (7.27-23.02) µg/mL and 51.04 (43.36-59.74) µg/mL against C. acnes, respectively, and 53.29 (48.75-57.91) µg/mL and 102.80 (87.15-122.30) µg/mL against S. epidermidis, respectively. The antimicrobial effects of both fractions seem to be related to membrane hyperpolarization and DNA damage. This dual mechanism of action may provide therapeutic advantages for the treatment of skin dysbiosis-related diseases.

Seasonal Nutritional Profile of Gelidium corneum (Rhodophyta, Gelidiaceae) from the Center of Portugal.

Gelidium corneum is a well-known agarophyte, harvested worldwide for its high agar quality. However, the species also exhibits an interesting nutritional profile, but with seasonal variations. Therefore, to evaluate the nutritional value of G. corneum, ash, crude protein, total lipids, and carbohydrates were analyzed at different times of the year. The heavy metals mercury, arsenic, lead, cadmium, and tin, as well as iodine were also measured. Finally, the seasonal antioxidant capacity of G. corneum extracts was evaluated. Our results indicate that the biomass is rich in protein (up to 16.25 ± 0.33%) and carbohydrates (up to 39.5 ± 3.29%), and low in lipids (up to 2.75 ± 0.28%), and especially in the summer, the AI, TI indexes, n-6/n-3 and h/H ratios (0.93, 0.6, 0.88 and 1.08, respectively) are very interesting. None of the contaminants exceeded the legally established limits, and the iodine values were adequate for a healthy diet. Finally, the antioxidant capacity is fair, with the DPPH ≤ 10.89 ± 1.46%, and ABTS ≤ 13.90 ± 1.54% inhibition, FRAP ≤ 0.91 ± 0.22 AAE.g-1, and TPC ≤ 6.82 ± 0.26 GAE.g-1. The results show that G. corneum is an attractive resource, with potential use as food or as a food supplement.

Seasonal Changes in the Nutritional Composition of Agarophyton vermiculophyllum (Rhodophyta, Gracilariales) from the Center of Portugal.

Seaweeds exhibit high nutritional value due to a balanced concentration of proteins, vitamins and minerals, a high concentration of low digestibility polysaccharides, and reduced levels of lipids, many of which are n-3 and n-6 fatty acids. The species Agarophyton vermiculophyllum is no exception and, as such, a comprehensive study of the chemical and nutritional profile of this red seaweed was carried out for 1 year. Seasonal variations in moisture, ash, protein and amino acids content, crude fibers, ascorbic acid, agar, lipids, and the corresponding fatty acid profile, were analyzed. We found low levels of fatty acids and a high protein content, but also noticed interesting seasonal change patterns in these compounds. The present study gives insights on the environmental conditions that can lead to changes in the nutritional composition of this species, aiming, therefore, to bring new conclusions about the manipulation of environmental conditions that allow for maximizing the nutritional value of this seaweed.

Recent Efforts to Recover Armeria berlengensis, an Endemic Species from Berlengas Archipelago, Portugal.

Berlengas archipelago is a UNESCO world heritage site and the only location where Armeria berlengensis is found. This species faces various threats, namely, human disturbance, the presence of Carpobrotus edulis, yellow-legged gull, common-rabbit, and black-rat populations. Thus, exclusion areas were installed, which blocked the access of most Gulls, aiming to promote the recovery of A. berlengensis. Additionally, rabbits and rats were removed from the island. After six years of surveys, there has been an increase in the number of individuals of A. berlengensis in the exclusion areas, and a clear shift in the size structure of the A. berlengensis population. Significant changes in the height and diameter of the individuals were also noted. These findings indicate that the population of A. berlengensis is changing and becoming a healthier population. Principal component analysis results show a straightforward dissimilarity between the areas with A. berlengensis and those without the species and allowed the clustering of two groups: the rupicolous species and the nitrophilous species. A. berlengensis produces few seeds (seed set 3.4%), which raises concern regarding the long-term survival of the species. Thus, further conservation efforts must be implemented, such as the control of invasive species, gulls, and ruderals, to allow for the recovery of A. berlengensis.

Unravelling the Dermatological Potential of the Brown Seaweed Carpomitra costata.

The ever-increasing interest in keeping a young appearance and healthy skin has leveraged the skincare industry. This, coupled together with the increased concern regarding the safety of synthetic products, has boosted the demand for new and safer natural ingredients. Accordingly, the aim of this study was to evaluate the dermatological potential of the brown seaweed Carpomitra costata. The antioxidant, anti-enzymatic, antimicrobial, photoprotective and anti-inflammatory properties of five C. costata fractions (F1-F5) were evaluated. The ethyl acetate fraction (F3) demonstrated the most promising results, with the best ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (EC50 of 140.1 µg/mL) and the capacity to reduce reactive oxygen species (ROS) production promoted by UVA and UVB radiation in 3T3 cells, revealing its antioxidant and photoprotective potential. This fraction also exhibited the highest anti-enzymatic capacity, inhibiting the activities of collagenase, elastase and tyrosinase (IC50 of 7.2, 4.8 and 85.9 µg/mL, respectively). Moreover, F3 showed anti-inflammatory potential, reducing TNF-α and IL-6 release induced by LPS treatment in RAW 264.7 cells. These bioactivities may be related to the presence of phenolic compounds, such as phlorotannins, as demonstrated by NMR analysis. The results highlight the potential of C. costata as a source of bioactive ingredients for further dermatological applications.

Valuable compounds in macroalgae extracts.

Bioactive compounds present in ethanolic extracts from 18 macroalgae of the Portuguese coast were analysed by gas chromatography-mass spectrometry (GC-MS), leading to the characterization of 14 compounds: proline, phloroglucinol, mannitol, 8 fatty acids and 3 sterols. A dose-dependent response against enzymes with biological significance (α-glucosidase, acetylcholinesterase and butyrylcholinesterase) and free radicals (DPPH, nitric oxide, superoxide and hydroxyl) was found, Phaeophyta being the most promising group. A PCA analysis was performed and allowed the establishment of a correlation between the algae chemical composition and the biological activity. Cystoseira tamariscifolia (Hudson) Papenfuss, Cystoseira nodicaulis (Withering) M. Roberts, Cystoseira usneoides (Linnaeus) M. Roberts and Fucus spiralis Linnaeus are among the most active species, which is in accordance with their higher contents in phloroglucinol, mannitol, oleic, arachidonic and eicosapentaenoic acids, and fucosterol. The results point to the potential interest of the use of Phaeophyta species as food additives, due to their potent antiradical activities, and especially highlights the importance of F. spiralis in the food chain of Mediterranean countries. Moreover, the incorporation of the extracts of these species in food products, nutraceutical and pharmaceutical preparations for human health should also be instigated, since they can suppress hyperglycemia and inhibit cholinesterases.

Phlorotannin extracts from fucales characterized by HPLC-DAD-ESI-MSn: approaches to hyaluronidase inhibitory capacity and antioxidant properties.

Purified phlorotannin extracts from four brown seaweeds (Cystoseira nodicaulis (Withering) M. Roberts, Cystoseira tamariscifolia (Hudson) Papenfuss, Cystoseira usneoides (Linnaeus) M. Roberts and Fucus spiralis Linnaeus), were characterized by HPLC-DAD-ESI-MSn. Fucophloroethol, fucodiphloroethol, fucotriphloroethol, 7-phloroeckol, phlorofucofuroeckol and bieckol/dieckol were identified. The antioxidant activity and the hyaluronidase (HAase) inhibitory capacity exhibited by the extracts were also assessed. A correlation between the extracts activity and their chemical composition was established. F. spiralis, the species presenting higher molecular weight phlorotannins, generally displayed the strongest lipid peroxidation inhibitory activity (IC50 = 2.32 mg/mL dry weight) and the strongest HAase inhibitory capacity (IC50 = 0.73 mg/mL dry weight). As for superoxide radical scavenging, C. nodicaulis was the most efficient species (IC50 = 0.93 mg/mL dry weight), followed by F. spiralis (IC50 = 1.30 mg/mL dry weight). These results show that purified phlorotannin extracts have potent capabilities for preventing and slowing down the skin aging process, which is mainly associated with free radical damage and with the reduction of hyaluronic acid concentration, characteristic of the process.

A gas chromatography-mass spectrometry multi-target method for the simultaneous analysis of three classes of metabolites in marine organisms.

In this work a fast and simple multi-target gas chromatography-mass spectrometry (GC-MS) method for the simultaneous detection and absolute quantification of amino acids, fatty acids, sterols and lupanes in marine organisms is proposed. The methodology was applied to the characterization of the echinoderm Marthasterias glacialis Linnaeus spiny sea star extracts. The main factors influencing the extraction of target compounds were evaluated by using different extraction procedures, solvent systems and temperature conditions and a comparison with a reference technique was performed. The most suitable procedure, capable of successfully extract the three classes of target compounds, was ethanol as solvent at 40°C under magnetic stirring. Good analytical parameters were obtained since calibrations curves for the 40 compounds under analysis (15 amino acids, 16 fatty acids, 6 sterols and 3 lupanes) showed regression coefficients (r(2)) ranging from 0.9844 to 0.9978, with low RSD (from 0.00 to 9.45%), and detection limits varying from 0.03 to 15.40 µg/L. The RSD values for intra- and interday variations studies were also good (RSD<13.5%, for both) and recoveries were higher than 92%. Variation in samples from different harvests and origins and their chemical composition during the year is reported. The fact that no previous treatment of samples is required can make this a useful technique for metabolite profiling in marine organisms, among others, both in biomedical and nutritional studies. Moreover, due to the fast and robust character of the proposed method it seems to be suitable for the implementation as routine analysis.

Can phlorotannins purified extracts constitute a novel pharmacological alternative for microbial infections with associated inflammatory conditions?

Bacterial and fungal infections and the emerging multidrug resistance are driving interest in fighting these microorganisms with natural products, which have generally been considered complementary to pharmacological therapies. Phlorotannins are polyphenols restricted to brown seaweeds, recognized for their biological capacity. This study represents the first research on the antibacterial, antifungal, anti-inflammatory and antioxidant activity of phlorotannins purified extracts, which were obtained from ten dominant brown seaweeds of the occidental Portuguese coast.Phlorotannins content was determined by the specific dimethoxybenzaldehyde (DMBA) method and a yield between 75 and 969 mg/Kg phloroglucinol units (dry matter) was obtained. Fucus spiralis ranked first, followed by three Cystoseira species. The anti-inflammatory potential of the purified extracts was assessed via inhibitory effect on nitric oxide (NO) production by lipopolysaccharide-stimulated RAW 264.7 macrophage cells, Cystoseira tamariscifolia being the one showing promising activity for the treatment of inflammation. NO scavenging ability was also addressed in cell free systems, F. spiralis being the species with highest capacity. The antimicrobial potential of the extracts was checked against five Gram-positive and four Gram-negative bacteria and three fungi strains, that commonly colonize skin and mucosa and are responsible for food contamination. The different extracts were more effective against Gram-positive bacteria, Staphylococcus epidermidis being the most susceptible species. Concerning antifungal activity, Trichophyton rubrum was the most sensitive species.Although the molecular mechanisms underlying these properties remain poorly understood, the results obtained turn phlorotannins purified extracts a novel and potent pharmacological alternative for the treatment of a wide range of microbial infections, which usually also present an inflammatory component. In addition to the biological properties demonstrated herein, phlorotannins extracts may also be preferred, in order to avoid side effects and allergic reactions commonly associated with synthetic drugs.

STEROL PROFILES IN 18 MACROALGAE OF THE PORTUGUESE COAST(1).

The sterol profiles of dominant macroalgae occurring in the western Portuguese coast were evaluated. An analytical procedure, involving alkaline hydrolysis and extraction followed by separation by reversed-phase HPLC-diode array detection (HPLC-DAD), was optimized for the study of their sterols composition. The validated methodology is short in analysis time (as the compounds are determined in <20 min), sensitive, reproducible, and accurate. It was then successfully applied to the determination of campesterol, cholesterol, desmosterol, ergosterol, fucosterol, stigmasterol, and ß-sitosterol in 18 species (three Chlorophyta, five Rhodophyta, and 10 Phaeophyta). The profiles obtained for the several macroalgal species were considerably different. C29 sterols were predominant in Phaeophyta and Chlorophyta (71%-95% of total sterol content), while in Rhodophyta cholesterol content is significantly higher (34%-87%). Among the studied species, Asparagopsis armata Harv. contained the lowest sterol amount (555 mg · kg(-1) dry weight), and Cystoseira tamariscifolia (Huds.) Papenf. the highest one (6,502 mg · kg(-1) dry weight). Data obtained may be helpful in identifying suitable marine sources of sterols, with potential applications in the food and pharmaceutical industries.

HPLC-PAD-atmospheric pressure chemical ionization-MS metabolite profiling of cytotoxic carotenoids from the echinoderm Marthasterias glacialis (spiny sea-star).

An HPLC-PAD-atmospheric pressure chemical ionization-MS metabolite profiling analysis was conducted on the marine echinoderm Marthasterias glacialis (spiny sea-star). Bio-guided purification of the methanolic extract led to the isolation of several carotenoids, namely zeaxanthin, astaxanthin and lutein. These compounds were characterized using both UV-Vis characteristics and MS spectra interpretation. No previous works addressed the MS analysis of carotenoids present in this organism. The purified carotenoid fraction displayed a strong cell proliferation inhibition against rat basophilic leukemia RBL-2H3 (IC(25)=268 microg/mL) cancer cell line. Against healthy V79 (rat lung fibroblasts (IC(25)=411 microg/mL)) cell line, however, toxicity was lower, as it is desired for anti-cancer molecules. This study suggests that M. glacialis may constitute a good source of bioactive compounds that can be used as lead compounds for the pharmaceutical industry.