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- Nutraceutical potential of Asparagopsis taxiformis (Delile) Trevisan extracts and assessment of a downstream purification strategyPublication . Nunes, N.; Valente, S.; Ferraz, S.; Barreto, Maria Carmo; Carvalho, M. A. A. Pinheiro deThe main goal of the present work was to determine the nutraceutical potential of Asparagopsis taxiformis D. extracts from Madeira Archipelago south coast. Extraction methodologies consisted either/or in 72 hours stirring, at room temperature (M1), or 6 cycles of Soxhlet extraction (M2), both with re-extraction. Solvents used were distilled water, ethanol, methanol and ethyl acetate. M1 allowed to obtain the highest values for extraction yield (31.65 g.100g 1 dw) using water, whereas iodine content (3.37 g.100g 1 dw), TPC (1.71 g GAE.100g 1 dw) and chlorophyll a (45.96 mg.100g 1 dw) were obtained using ethanol, and TCC (36.23 mg.100g 1 dw) with methanol. Extracts that showed higher reduction activity in M1 were derived from ethanol extraction (1,908 mg AAE.100g 1 dw). Water and ethanol were the best solvents for higher DPPH scavenging activity in M2, both with same result (IC50 1.37 mg.mL 1 ). The lowest value of IC50 for chelating activity (1.57 mg.mL 1 ) was determined in M1, using ethyl acetate. The remaining residue was used to obtain other products, i.e. lipid extraction (M1, 2.05 g.100g 1 dw), carrageenans (M2, 21.18 g.100g 1 dw) and cellulose (M1, 23.81 g.100g 1 dw) with subsequent FTIR ATR analysis. Our results show that A. taxiformis is a valuable source of bioactive compounds. The M1 extraction methodology using ethanol is the most effective solvent to produce an iodine rich bioactive extract with potential of being used as a nutraceutical supplement. Also, we have demonstrated a possible downstream strategy that could be implemented for multiple compound extraction from A. taxiformis residue. This has a vital importance for future feasibility, when using this biomass as an industrial feedstock for multiple products production. Statistical analysis, using SPSS 24.0, was also performed and important correlations were found between assays and methods.
- Monitoring system and in situ conservation of endemic and threatened Beta patula Aiton populations in Madeira RegionPublication . Nóbrega, H.; Freitas, G.; Zavattieri, M. A.; Ragonezi, C.; Frese, L.; Carvalho, M. A. A. Pinheiro det Madeira Archipelago is a hotspot for crop wild relatives (CWR) of crop cultures. Some of these CWR are present in very specific environments, such as, in Ponta de Sa˜o Lourenc¸o or Desertas Islands. One such species is Beta patula Aiton, a Critically Endan gered species which belongs to the Gene Pool 1b of cultivated beets. A continuous effort has been devel oped for its ex situ conservation through the storage of accessions (ISOP2512 and ISOP1911) in the ISO Plexis GeneBank at the University of Madeira Simultaneously, a series of studies have been carried out to understand the species’ ecogeographic and ecological requirements, to validate populations’ boundaries and sizes, and to establish population dynamics. This study includes a complete floristic survey at the B. patula locations of, Desembar cadouro islet (DI) with 12 (DI1–DI12) sampling sites, and Cha˜o islet (CI) with 3 (CI1–CI3). Several Biodiversity indices were calculated for these loca tions. Plot DI3 exhibited the highest values for Corrected Evenness (E0 = 0.77 ± 0.07), Shannon– Weaver Diversity Index (H0 = 2.48 ± 0.12), and Hill’s Index (N2 = 4.47 ± 0.72), with a total sum of 306 individuals of B. patula. The demographic status of B. patula populations in DI and CI was determined yearly between 2014 and 2018. The results show an average population size of 16,906 and 2917 plants, respectively. These data will be used for the estab lishment of a protocol to monitor and manage a genetic reserve for B. patula and other CWR. By doing so, our work will contribute to the implemen tation of the European genetic reserve network.
- Biochemical composition, nutritional value, and antioxidant properties of seven seaweed species from the Madeira ArchipelagoPublication . Nunes, N.; Ferraz, S.; Valente, S.; Barreto, Maria Carmo; Carvalho, M. A. A. Pinheiro deBiochemical composition and antioxidant activity were assessed in seven seaweeds from Madeira Archipelago, namely, the chlorophyte (Ulva lactuca), the rhodophytes (Asparagopsis taxiformis, Chondrus crispus, Galaxaura rugosa, Grateloupia lanceola and Nemalion elminthoides), and the phaeophyte (Zonaria tournefortii). Seaweed mineral content varied from 16.60 to 84.16 g (100 g)-1 dry weight (dw). Organic matter, composed by fiber and matrix polysaccharides (8.33 to 54.04 g (100 g)-1 dw), starch (1.95 to 25.41 g (100 g)-1 dw), protein (2.80 to 17.55 g (100 g)-1 dw), and fat (1.46 to 12.04 g (100 g)-1 dw), was also determined. Asparagopsis taxiformis was found to have substantial quantities of protein, fat, fiber, and matricial polysaccharides, compared to the other analyzed seaweeds. Analysis of antioxidant components included the measurement of chlorophyll a (28.81 to 244.3 g (100 g)-1 dw), total carotenoids (0 to 297.8 g (100 g)-1 dw), total phenolic compounds (0 to 2154 mg GAE (100 g)-1 dw), and total flavonols (7.27 to 604.8 mg QE (100 g)-1 dw). Zonaria tournefortii was found to possess the highest contents of chlorophyll a, total carotenoids, total phenolic content (TPC), and antioxidant activity, determined through ferric reduction antioxidant potential (FRAP), ferrous ion chelating (FIC), free radical-scavenging assay (FRSA), and β-carotene bleaching (β-CB). Statistical analysis showed 38 significant correlations between various biochemical and antioxidant parameters or activity and determined that fat content showed the highest number of correlations. Overall, this study gives a better understanding of Madeira autochthonous seaweeds in their potential of being introduced as a raw material for nutrient supplementation in various food products or to produce functional foods using seaweed natural properties.
- Propagation of nine endemic plant species from Madeira Island (Portugal)Publication . Henriques, Dina; Fontinha, Susana; Neves, Margarida Costa; Nóbrega, Humberto; Ferro, António; Carvalho, M. A. A. Pinheiro deEfficient propagation of endangered plant species is a critical factor in successful ecological restoration and conscientious habitat management. Hence, propagation trials of nine endemic plant species of Madeira (Anthyllis lemanniana Lowe, Armeria maderensis Lowe, Cedronella canariensis (L.) Webb & Berthel., Erica maderensis (Benth.) Bornm., Genista tenera (Jacq. ex Murray) Kuntze, Helichrysum melaleucum Rchb. ex Holl, Pericallis aurita (L'Her.) B. Nord., Sideritis candicans Aiton and Teline maderensis Webb & Berthel.) were carried out. Plant propagation requirements and their sexual and vegetative propagation methods were studied. Seed germination success varied between species. Germination rate exceeded 70% in six out of nine species, being lower than 30% in Pericallis aurita, while H. melaleucum seeds did not germinate. Vegetative propagation yielded lower success rates, with three species (Erica maderensis, Genista tenera and Teline maderensis) unable to establish roots, and three species (Helichrysum melaleucum, Pericallis aurita and Sideritis candicans) exceeding 60% of the rooting success. Establishment of the propagation requirements of these species could be regarded an important tool for supporting Madeira’s flora conservation programs.
- Fatty acid composition, TLC screening, ATR-FTIR analysis, anti-cholinesterase activity, and in vitro cytotoxicity to A549 tumor cell line of extracts of 3 macroalgae collected in MadeiraPublication . Nunes, N.; Rosa, G. P.; Ferraz, S.; Barreto, Maria Carmo; Carvalho, M. A. A. Pinheiro deThree macroalgae collected at Madeira Island were included in this study to determine their potential for drug, nutraceutical, food, or supplement application. Fatty acid content was higher in Zonaria tournefortii (12.32 mg g−1 dw) with 16.58% of PUFAs, eicosapentaenoic acid (C20:5ω3), and arachidonic acid (20:4ω6) having concentrations of 2.59 and 1.17%, respectively. The anti-thrombogenic and anti-atherogenicity potential was higher for Z. tournefortii due to relevant fatty acids in the biochemical composition this macroalgae. Lipid classes were assessed in the lipid extract and neutral lipids (NL) were in higher yield in Asparagopsis taxiformis (51.16%) and lower in Z. tournefortii (26.96%). The glycolipids (GL) were between 36.03 and 16.11% in Z. tournefortii and Ulva lactuca. Phospholipids (PL) fraction varied from 35.91 and 31.60% in A. taxiformis and Z. tournefortii. TLC screening identified that U. lactuca contains phytol and cholesterol in its NL, digalactosyldiacylglycerol in its GL, and cardiolipin and L-α-phosphatidylcholine in its PL. Zonaria tournefortii contains phytol and cholesterol in its NL classes, and the PL classes contain L-α-phosphatidylethanolamine and 1-(3-sn-phosphatidyl)-rac-glycerol. The macroalgae A. taxiformis revealed cholesterol in its NL fraction and the same phospholipids as Z. tournefortii in its PL fraction. ATRFTIR analysis enabled a “fingerprint” spectra and important sulfation absorption bands were identified, revealing the functional polysaccharides within these macroalgae. Anti-cholinesterasic activity was assessed in A. taxiformis, with a low IC50 for AChE (8.92 ± 0.43 μg mL−1 ) and BuChE (13.96 ± 0.32 μg mL−1 ), demonstrating dual inhibitory activity, justifying the interest to identify the active principle which may be the scaffold of a novel drug.
- Evaluation of maize germplasm based on zein polymorphism from the archipelago of MadeiraPublication . Freitas, I. R. A. de; Ganança, F.; Santos, T. M. dos; Carvalho, M. A. A. Pinheiro de; Motto, M.; Vieira, M. ClementeZein polypeptides are a group of proteins that accumulate in maize endosperm during seed development, representing more than 60% of the total endosperm proteins in the mature seeds. To evaluate genotype variability of Madeiran maize germplasm, a biochemical study was conducted based on the prolamins of maize, zeins, extracted from endosperm meal of 43 populations of Zea mays L. maintained in the Germplasm bank of Madeira University along with the inbreed W64A which was used as a polymorphic zein polypeptide standard profile. The zein polymorphism of these 44 maize populations were compared using two different electrophoresis techniques, SDS-PAGE in 15% discontinuous polyacrylamide gel and Acid-PAGE in 10% continuous polyacrylamide gel. SDS-PAGE allowed up to 16 polypeptides to be identified with apparent molecular mass ranging from 28-kDa to 10-kDa. Acid-PAGE allowed up to 20 zein fractions to be identified. The data was submitted to principal component analysis (PCA) and canonical discriminate and similarity analysis. The SDS-PAGE zein polymorphism allowed us to detect 6 groups, assembling all maize populations and explaining 55.32% of all variability. The similarity analysis of zein patterns obtained by Acid-PAGE showed that among regional maize germplasm, 22.5 % of all population seems to be related and have a common ancestor. The ISOP71 seems to be the population more closed to the common ancestor and appears related with the remaining maize populations, excluding the ISOP125.The obtained results and the importance of zein polymorphism in the evaluation of maize germplasm from Madeiran Archipelago are discussed
- Palynological study of the endemic woody sonchus fron the flora of Madeira: a morphological and molecular approachPublication . Carvalho, M. A. A. Pinheiro de; Câmara, I. G.; Santos, T. M. M. dos; Correia, R. S.
- Assessing the diversity of sea beet (Beta vulgaris L. ssp. maritima) populationsPublication . Ascarini, F.; Nóbrega, H. G.M.; Leite, I. S.; Freitas, G.; Ragonezi, C.; Zavattieri, M. Amely; Carvalho, M. A. A. Pinheiro deBeta vulgaris L. subsp. maritima (L.) Arcang., sea beet, is a morphologically and genetically variable species, belonging to beet primary gene -pool. This crop wild relative is a valuable genetic resource for resistance improvement in beets and could play an important role in crop yield sustainability. Eleven Madeiran sea beet populations were characterized using morphological descriptors and genetic markers. Our goal was to evaluate these populations as a potential source of valuable genetic material. Morphological characterization showed a high quantitative variation among populations. Plant height and inflorescence height parameters had the highest influence in the separation of populations. Molecular analysis was performed with polymorphic SSRs to determine genetic variability between populations. Both PCA and PCoA revealed three clusters that separated the populations according to morphological and genetic traits, respectively. This study contributes to the knowledge of sea beet diversity in Madeira’s archipelago and to the perception that the islands' specific environmental conditions influence its genetic variability, making these populations a possible gene source for sugar beet breeding programs
- Helminth parasites of the oceanic horse mackerel Trachurus picturatus Bowdich 1825 (Pisces: Carangidae) from Madeira Island, Atlantic Ocean, PortugalPublication . Costa, G.; Moreira, E. Melo; Carvalho, M. A. A. Pinheiro deThe helminth parasite fauna of the oceanic horse mackerel Trachurus picturatus Bowdich 1825, caught off the Madeira Islands was composed of six different taxa. Prevalence and abundance of larval Anisakis sp. (Nematoda: Anisakidae) and Nybelinia lingualis (Trypanorhyncha: Tentaculariidae), the most common parasite taxa, were 24.3%, 0.9 and 37.9%, 0.7, respectively. Bolbosoma vasculosum (Acanthocephala: Polymorphidae) and the monogeneans Heteraxinoides atlanticus (Monogenea: Heteraxinidae) and Pseudaxine trachuri (Monogenea: Gastrocotylidae) were comparatively rare. The depauperate helminth fauna of the oceanic horse mackerel at Madeira compared to other geographical regions of the north-eastern Atlantic, namely the Azores banks and the West African coast, may be attributed to the paucity of nutrients off oceanic islands and to a low density of the fish population.
- Genetic diversification and dispersal of taro (Colocasia esculenta (L.) Schott)Publication . Chaïr, H.; Traore, R. E.; Duval, M. F.; Rivallan, R.; Mukherjee, A.; Aboagye, L. M.; Van Rensburg, W. J.; Andrianavalona, V.; Carvalho, M. A. A. Pinheiro de; Saborio, F.; Sri Prana, M.; Komolong, B.; Lawac, F.; Lebot, V.Taro (Colocasia esculenta (L.) Schott) is widely distributed in tropical and sub-tropical areas. However, its origin, diversification and dispersal remain unclear. While taro genetic diversity has been documented at the country and regional levels in Asia and the Pacific, few reports are available from Americas and Africa where it has been introduced through human migrations. We used eleven microsatellite markers to investigate the diversity and diversification of taro accessions from nineteen countries in Asia, the Pacific, Africa and America. The highest genetic diversity and number of private alleles were observed in Asian accessions, mainly from India. While taro has been diversified in Asia and the Pacific mostly via sexual reproduction, clonal reproduction with mutation appeared predominant in African and American countries investigated. Bayesian clustering revealed a first genetic group of diploids from the Asia-Pacific region and to a second diploid-triploid group mainly from India. Admixed cultivars between the two genetic pools were also found. In West Africa, most cultivars were found to have originated from India. Only one multi-locus lineage was assigned to the Asian pool, while cultivars in Madagascar originated from India and Indonesia. The South African cultivars shared lineages with Japan. The Caribbean Islands cultivars were found to have originated from the Pacific, while in Costa Rica they were from India or admixed between Indian and Asian groups. Taro dispersal in the different areas of Africa and America is thus discussed in the light of available records of voyages and settlements.