Lipophilic toxins occurrence in non-traditional invertebrate vectors from North Atlantic Waters (Azores, Madeira, and Morocco): Update on geographical tendencies and new challenges for monitoring routines.

In the last decades, due to monitoring programs and strict legislation poisoning incidents occurrence provoked by ingestion of naturally contaminated marine organisms has decreased. However, climate change and anthropogenic interference contributed to the expansion and establishment of toxic alien species to more temperate ecosystems. In this work, the coasts of Madeira, São Miguel islands and the northwestern Moroccan coast were surveyed for four groups of lipophilic toxins (yessotoxins, azaspiracids, pectenotoxins, and spirolides), searching for new vectors and geographical tendencies. Twenty-four species benthic organisms were screened using UHPLC-MS/MS technique. We report 19 new vectors for these toxins, six of them with commercial interest (P. aspera, P. ordinaria, C. lampas, P. pollicipes, H. tuberculata and P. lividus). Regarding toxin uptake a south-north gradient was detected. This study contributes to the update of monitoring routines and legislation policies, comprising a wider range of vectors, to better serve consumers and ecosystems preservation.

Restructuring of the 'Macaronesia' biogeographic unit: A marine multi-taxon biogeographical approach.

The Azores, Madeira, Selvagens, Canary Islands and Cabo Verde are commonly united under the term "Macaronesia". This study investigates the coherency and validity of Macaronesia as a biogeographic unit using six marine groups with very different dispersal abilities: coastal fishes, echinoderms, gastropod molluscs, brachyuran decapod crustaceans, polychaete annelids, and macroalgae. We found no support for the current concept of Macaronesia as a coherent marine biogeographic unit. All marine groups studied suggest the exclusion of Cabo Verde from the remaining Macaronesian archipelagos and thus, Cabo Verde should be given the status of a biogeographic subprovince within the West African Transition province. We propose to redefine the Lusitanian biogeographical province, in which we include four ecoregions: the South European Atlantic Shelf, the Saharan Upwelling, the Azores, and a new ecoregion herein named Webbnesia, which comprises the archipelagos of Madeira, Selvagens and the Canary Islands.

Tetrodotoxins Occurrence in Non-Traditional Vectors of the North Atlantic Waters (Portuguese Maritime Territory, and Morocco Coast).

Tetrodotoxin (TTX) is a potent alkaloid typically from tropical ecosystems, but in the last decade its presence has been more pronounced in the temperate waters of the Atlantic. In its last scientific opinion, the European Food Safety Authority (EFSA) stressed the need for data regarding TTX prevalence in European waters. To address EFSA's concerns, benthic organisms such as mollusks, crustaceans, echinoderms and fish with different feeding habits were collected along the Portuguese continental coast, islands (São Miguel, Azores, and Madeira) and the northwestern Moroccan coast. A total of 165 samples were analyzed by ultra high performance liquid chromatography high resolution mass spectrometry (UHPLC-HRMS) and ultra high performance chromatography mass spectrometry (UHPLC-MS/MS). Geographical tendencies were detected as follows, by descending order: S. Miguel Island (Azores), Moroccan coast, Madeira Island and Portuguese continental coast. The toxin amounts detected were significant, above the Dutch limit value established in 2017, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, unraveling new vectors, in order to better assess human health risk. This work represents a general overview of new TTX bearers (7) most of them in gastropods (Patella depressa, Nucella lapillus, Onchidella celtica and Aplysia depilans), followed by echinoderms (Echinus esculentus and Ophidiaster ophidianus) and puffer fish Sphoeroides marmoratus.

Paralytic Shellfish Toxins Occurrence in Non-Traditional Invertebrate Vectors from North Atlantic Waters (Azores, Madeira, and Morocco).

Paralytic shellfish toxins (PSTs) are potent alkaloids of microalgal and cyanobacterial origin, with worldwide distribution. Over the last 20 years, the number of poisoning incidents has declined as a result of the implementation of legislation and monitoring programs based on bivalves. In the summer of 2012 and 2013, we collected a total of 98 samples from 23 different species belonging to benthic and subtidal organisms, such as echinoderms, crustaceans, bivalves, and gastropods. The sampling locations were Madeira, São Miguel Island (Azores archipelago), and the northwestern coast of Morocco. The samples were analyzed using post-column oxidation liquid chromatography with a fluorescence detection method. Our main goal was to detect new vectors for these biotoxins. After reporting a total of 59 positive results for PSTs with 14 new vectors identified, we verified that some of the amounts exceeded the limit value established in the EU. These results suggest that routine monitoring of saxitoxin and its analogs should be extended to more potential vectors other than bivalves, including other edible organisms, for a better protection of public health.

Seasonal Variability of the Biochemical Composition and Antioxidant Properties of Fucus spiralis at Two Azorean Islands.

This study evaluates, for the first time, the seasonal (winter and summer) and geographical (São Miguel⁻SMG and Santa Maria⁻SMA Islands) variability of Fucus spiralis (Fs) biochemical composition (dry weight basis) and antioxidant properties. Protein and carbohydrates presented higher values in Fs-SMGwinter, lipids, total dietary fiber, and energy value in Fs-SMAsummer, and ash and soluble dietary fiber/insoluble dietary fiber ratio in Fs-SMAwinter. The fatty acid (FA) profiles showed a lower SFA in Fs-SMGsummer, whereas MUFA and PUFA presented higher values in Fs-SMGsummer and Fs-SMGwinter, respectively. Excellent dietary ratios of n6/n3 PUFA and hypocholesterolemic/hypercholesterolemic FA were found, with lower values in Fs-SMAwinter and higher in Fs-SMGsummer, respectively. The highest total phenolics was found in Fs-SMAsummer acetone:water extract and total flavonoids showed the higher value in Fs-SMGwinter methanol extract. The best free radical-scavenging activity was observed in the Fs-SMAwinter methanol (EC50 = 0.045 mg/mL) and acetone:water (EC50 = 0.059 mg/mL) extracts. The ferric-reducing antioxidant power showed the best results in Fs-SMAwinter methanol extract (EC50 = 0.016 mg/mL) and Fs-SMAsummer acetone:water extract (EC50 = 0.017 mg/mL). The best ferrous ion-chelating activity was found in Fs-SMGwinter acetone:water extract. Overall, results revealed that F. spiralis nutritional and functional bioactivity values have geographical and seasonal variations and that its regular consumption may add benefits to human health.

Angiotensin I-Converting Enzyme (ACE) Inhibitory Activity, Antioxidant Properties, Phenolic Content and Amino Acid Profiles of Fucus spiralis L. Protein Hydrolysate Fractions.

Food protein-derived hydrolysates with multi-bioactivities such as antihypertensive and antioxidant properties have recently received special attention since both activities can play significant roles in preventing cardiovascular diseases. This study reports, for the first time, the angiotensin I-converting enzyme (ACE)-inhibition and antioxidant properties of ultrafiltrate fractions (UF) with different molecular weight ranges (<1, 1-3 and ≥3 kDa) obtained from Fucus spiralis protein hydrolysate (FSPH) digested with cellulase-bromelain. The amino acids profile, recovery yield, protein, peptide and total phenolic contents of these FSPH-UF, and the in vitro digestibility of F. spiralis crude protein were also investigated. FSPH-UF ≥3 kDa presented remarkably higher ACE-inhibition, yield, peptide and polyphenolic (phlorotannins) contents. Antioxidant analysis showed that FSPH-UF <1 kDa and ≥3 kDa exhibited significantly higher scavenging of 2,2-diphenyl-1-picrylhydrazyl radical and ferrous ion-chelating (FIC) activity. FSPH-UF ≥3 kDa had also notably higher ferric reducing antioxidant power (FRAP). Strong correlations were observed between ACE-inhibition and antioxidant activities (FIC and FRAP). The results suggest that ACE-inhibition and antioxidant properties of FSPH-UF may be due to the bioactive peptides and polyphenols released during the enzymatic hydrolysis. In conclusion, this study shows the potential use of defined size FSPH-UF for the prevention/treatment of hypertension and/or oxidative stress-related diseases.

Nutritional and Functional Bioactivity Value of Selected Azorean Macroalgae: Ulva compressa, Ulva rigida, Gelidium microdon, and Pterocladiella capillacea.

This study presents information on the biochemical composition (dry weight basis), nutritional aspects, and angiotensin I-converting enzyme (ACE) inhibitory properties of selected macroalgae (Ulva compressa, Ulva rigida, Gelidium microdon, and Pterocladiella capillacea) from Azores. Moisture content was very high (83.2% to 90.0% of fresh weight). Total dietary fiber (33.7% to 41.0%) that presented a good balance of both soluble and insoluble fibers (15.5% to 19.2% and 18.2% to 21.8%, respectively) was the most abundant component in these macroalgae. Protein and ash (ranged from 15.7% to 23.4% and 10.7% to 20.7%, respectively) were the 2nd most abundant components in red and green macroalgae, respectively. Moderate soluble carbohydrate contents (14.5% to 19.8%) were found in all species. Lipid contents were low (1.0% to 4.3%), particularly in Ulva species (1.0% to 1.7%), but contained higher unsaturated fatty acids (FAs) (7.5% to 32.9% and 29.6% to 69.2% of total FA for monounsaturated fatty acid and polyunsaturated fatty acid groups, respectively) than saturated fatty acid (23.3% to 46.8% of total FA) contents. All the macroalgal proteins had high digestibility in vitro (82.2% to 89.4%, relatively to sodium caseinate), contained high quantity of essential amino acids (45.3% to 58.1% of total amino acids), but in different proportions, and were rich in aspartic and glutamic acids that together account for 17.2% to 36.2% of the total amino acids. These results suggested that regular consumption of the selected macroalgae may improve human health and revealed that they can be used for producing food supplements for human and animal nutrition and/or pharmaceuticals with potential effect on the regional economy. Furthermore, the ACE-inhibitory IC50 values of 0.095 to 0.695 mg/mL for the <1 kDa protein hydrolysate fraction revealed a potential impact on hypertension disorder.

Angiotensin I-converting enzyme (ACE) inhibitory activity of Fucus spiralis macroalgae and influence of the extracts storage temperature-A short report.

Recently, increasing attention has been paid to the marine algae as a natural source of novel angiotensin-I converting enzyme (ACE) inhibitors, such as the phlorotannins that are the predominant polyphenols in brown algae. This study reports, for the first time, the ACE inhibition of methanol extract/fractions from Azorean brown algae Fucus spiralis (Fs) determined by HPLC-UV method, their total phenolic content (TPC) quantified as phloroglucinol equivalents (PE) and the effect of the Fs dry powder methanol extracts (Fs-DME) storage temperature on ACE inhibition. The results indicate that the ACE inhibition of Fs-DME decreased by 28.8% and 78.2% when stored during 15days at -80°C and -13°C, respectively, as compared with the activity of Fs-DME at a refrigerated temperature of 6°C and assayed immediately after extraction that showed a value of 80.1±2.1%. This Fs-DME sample was fractionated by ultrafiltration membranes into three molecular weight ranges (<1kDa, 1-3kDa and >3kDa), presenting the fraction>3kDa remarkably high ACE inhibition (88.8±2.4%), TPC value (156.6±1.4mg PE/g of dry weight fraction) and yield. Furthermore, chromatographic and spectrophotometric analyses corroborate that phenolic compounds were present in Fs methanol extract/fractions, and also revealed that phloroglucinol occurs in Fs. The results seem to suggest that Azorean Fs can be a source of powerful ACE-inhibitory phlorotannins with potential impact on public health, particularly on hypertensive patients.

Health-promoting ingredients from four selected Azorean macroalgae.

This study presents, for the first time, the nutritional and health promoting aspects of four selected Azorean macroalgae (Ulva compressa, Ulva rigida, Gelidium microdon and Pterocladiella capillacea) in terms of total lipids, fatty acids (FA) profile, n6/n3 and hypocholesterolemic (h)/hypercholesterolemic (H) FA ratios, minerals, total essential amino acids (AA), anti-ageing and anti-phenylketonuria AA content, coenzyme Q10, α-tocopherol, total phenolics, antioxidant properties and energy value, on a dry weight basis. The results revealed low lipid content (1.02-4.32%) but significant PUFA content (29.57-69.22% of total FA), suitable FA ratios (0.48-9.49 for n6/n3 and 1.26-4.22 for h/H), balanced macromineral ratios (0.27-1.91 for Na/K and 0.15-1.07 for Ca/Mg), appreciable amount of essential AA (45.27-58.13% of total AA), high amount of anti-ageing AA, low Phe content, coenzyme Q10 (1.25-8.27µg/g), α-tocopherol (2.61-9.14mg/100g), high total phenolic content (27.70-55.07mg of gallic acid equivalents/g extract), significant free radical scavenging activity (29.32-47.73%) and valuable energy content (6.80-9.80kJ/g). A regular consumption of these algae either directly or through food supplements may improve human health or may have a protective effect on some diseases and ageing process. They can also be used for producing pharmaceuticals and cosmeceuticals with potential economic value.

Edible Azorean macroalgae as source of rich nutrients with impact on human health.

Fucus spiralis, Porphyra sp. and Osmundea pinnatifida are macroalgae consumed as food in some of the Azorean Islands, but little information is available on their nutritional value. This paper presents information on the nutritional aspects in terms of proteins (9.71-24.82%), lipids (5.23-8.88%), soluble carbohydrates (17.59-25.37%), ash (22.31-38.55%), minerals (0.96-1.82 for Na/K ratios) and fat soluble vitamins A (1.20-1.41%) and E (4.86-51.14%) contents on dry weight basis. Fatty acid (SFA, MUFA, PUFA and n6/n3 and h/H ratios) and amino acid profiles, and antioxidant properties were also reported. All the three species contained measurable quantities of 9 of 10 essential amino acids (41.62-63.50% of the total amino acids). The regular consumption of these algae allied with the low level of seawater pollution around Azores Islands will improve human health conditions and will have a protective effect on the most common degenerative diseases.