Evaluation of acetylcholinesterase inhibitory activity of Brazilian red macroalgae organic extracts

Abstract Alzheimer's disease affects nearly 36.5 million people worldwide, and acetylcholinesterase inhibition is currently considered the main therapeutic strategy against it. Seaweed biodiversity in Brazil represents one of the most important sources of biologically active compounds for applications in phytotherapy. Accordingly, this study aimed to carry out a quantitative and qualitative assessment of Hypnea musciformis (Wulfen) J.V. Lamouroux, Ochtodes secundiramea (Montagne) M.A. Howe, and Pterocladiella capillacea (S.G. Gmelin) Santelices & Hommersand (Rhodophyta) in order to determine the AChE effects from their extracts. As a matter of fact, the O. secundiramea extract showed 48% acetylcholinesterase inhibition at 400 μg/ml. The chemical composition of the bioactive fraction was determined by gas chromatography–mass spectrometry (GC–MS); this fraction is solely composed of halogenated monoterpenes, therefore allowing assignment of acetylcholinesterase inhibition activity to them.

Antibacterial, antifungal and cytotoxic activities exhibited by endophytic fungi from the Brazilian marine red alga Bostrychia tenella (Ceramiales)

Abstract Marine environment is one of the most important sources regarding natural products research. Besides, marine microorganisms have been denominated as a talented natural source for discovery of new leads. Although the association of macroalgae and fungi has been described regarding ecological issues, there is a lack of studies about marine seaweed endophytic fungi. In this context, the goal of this study was to evaluate cytotoxic, antifungal and antibacterial activities of endophytic fungi isolated from the Brazilian marine seaweed Bostrychia tenella (J.V. Lamouroux) J. Agardh (Ceramiales, Rhodophyta). Forty-five endophytic microorganism strains were isolated from B. tenella. Crude extracts and organic fractions of ten selected strains were obtained after growth in rice medium. Samples were evaluated for cytotoxicity, antifungal and antibacterial assays. Penicillium strains showed positive results in a diversity of assays, and other five strains were active in at least one test. In addition, cytochalasin D was isolated from Xylaria sp. This alga is composed of a microbiological potential, since its endophytic strains exhibited remarkable biological properties. Moreover, cytochalasin D isolation has confirmed chemical potential of marine endophytic strains. This is the first study in which cultured fungi isolates from the Brazilian macroalga B. tenella were evaluated concerning biological properties. Results corroborated that this species could be a pharmaceutical source from marine environment. Furthermore, Acremonium implicatum is being firstly described as marine endophyte and Xylaria sp., Trichoderma atroviride and Nigrospora oryzae as marine seaweed endophytes. Thus, this work reports the first study relating detailed isolation, cultivation and biological evaluation (cytotoxic, antifungal and antibacterial) of endophytes Penicillium decaturense and P. waksmanii from the Brazilian marine red alga B. tenella. We are also reporting the isolation of cytochalasin D, a known antitumor and antibiotic compound, from Xylaria sp. strain. Despite widespread prevalence in terrestrial and marine habitats, this present work describes the first occurrence of cytochalasin D as a metabolite from marine seaweed endophyte.

Benthic marine algae from the insular areas of Paraná, Brazil: new database to support the conservation of marine ecosystems / Algas marinhas bentônicas de áreas insulares do Paraná, sul do Brasil: checklist, novas ocorrências para o Estado e implicações para a conservação

This study describes the diversity of benthic marine algae from insular areas of the southern Brazilian coast. Algal samples were collected between 2006 and 2010 during the winter and the summer seasons at three sites in the coastal waters of Paraná, Brazil: Ilha do Mel, Currais Archipelago and Ilha do Farol. The samples were collected along parallel transects on the coast. In this survey, Paraná marine phycoflora comprised 139 taxa (90 Rhodophyta, 27 Chlorophyta and 22 Phaeophyceae). Fifty-two species represent new records for the state, and 14 taxa are present at all sampling sites and in both seasons. Higher diversities of rhodophytes and chlorophytes were observed at Ilha do Mel, while phaeophytes were more diverse at Currais Archipelago. Lower algal diversity was observed at Ilha do Farol, a sampling station which is near an urban area. Ceramiaceae, Rhodomelaceae and Corallinaceae were dominant among Rhodophyta, Cladophoraceae and Ulvaceae among Chlorophyta, and Dictyotaceae and Sargassaceae among Phaeophyceae. Seasonal and spatial differences in species composition could be explained by the availability of consolidated substrate, water transparency and proximity to an urban area. Seaweed biodiversity from the Paraná coast also presents low species richness compared to other Brazilian states as a result of the shorter coastline, lower availability of rocky shores, and the location between estuarine systems (Paranaguá and Guaratuba Bays). These bays input a large amount of continental water, resulting in decreased salinity, high concentrations of suspended particulate matter and low transparency in the water column. Knowledge of seaweed diversity is essential for conservation studies. In addition, environmental monitoring programs undertaken during medium- to long-term seasonal changes could be improved to reflect changes detected through new records, the introduction of alien species in the area of interest, or even dominance of opportunistic species over other taxa. Therefore, a database able to support the monitoring of biodiversity is a fundamental step in detecting environmental impacts that could change seaweed biogeography, mainly in urbanized and harbor areas.

O presente estudo descreve a diversidade de algas marinhas bentônicas em áreas insulares da costa do Paraná, sul do Brasil. As amostras de algas foram coletadas entre 2006 e 2010 durante o inverno e o verão, em três locais: Ilha do Mel, Arquipélago de Currais e Ilha do Farol. A amostragem foi realizada ao longo de transectos paralelos à linha de costa e, na Ilha dos Currais, em diferentes profundidades. A ficoflora marinha paranaense é composta por 139 táxons (90 Rhodophyta, 27 Chlorophyta e 22 Phaeophyceae). Cinquenta e duas espécies são novas citações para o Estado do Paraná, e 14 táxons foram registrados em todos os locais estudados durante o verão e o inverno. Diferenças sazonais e espaciais na composição específica podem ser explicadas pela disponibilidade de substratos rochosos, transparência da água e proximidade com a área urbana. A maior diversidade de rodófitas e clorófitas foi observada na Ilha do Mel e a de feofíceas no Arquipélago de Currais. A menor diversidade de algas foi observada na Ilha do Farol, ponto amostral mais próximo a uma área urbana. Ceramiaceae, Rhodomelaceae e Corallinaceae foram dominantes entre Rhodophyta, Cladophoraceae e Ulvaceae entre Chlorophyta, e Dictyotaceae e Sargassaceae entre Phaeophyceae. A diversidade de algas marinhas bentônicas na costa do Paraná é menor quando comparada a outros estados brasileiros e isto está associado à menor extensão de sua linha de costa, menor disponibilidade de substratos consolidados para recrutamento e localização entre dois sistemas estuarinos (baías de Paranaguá e de Guaratuba). Os estuários aumentam consideravelmente o aporte de água continental, diminuindo a salinidade, elevando as concentrações de material particulado em suspensão e reduzindo a transparência da água. O conhecimento da diversidade algácea é fundamental para estudos de conservação, sendo que câmbios sazonais em médio e longo prazo podem estar relacionados a alterações ambientais. O indicador destas mudanças pode ser a detecção de novas ocorrências, espécies novas ou ainda introdução de espécies exóticas na área em questão, ou mesmo pela dominância de táxons oportunistas. Portanto, uma base de dados que sustente um monitoramento da biodiversidade é relevante por detectar impactos que podem alterar padrões biogeográficos das comunidades de macroalgas, principalmente em regiões com influência de atividades antrópicas, como as zonas urbanas ou áreas portuárias.

Anti-HSV activity of Hypnea musciformis cultured with different phytohormones

Four extracts from the seaweed Hypnea musciformis (Wulfen in Jacq.) J.V. Lamour. (Rhodophyta), collected directly from its natural habitat or cultivated in the presence of phytohormones, were evaluated for their ability to inhibit the replication of acyclovir-resistant herpes simplex viruses types 1 (HSV-1) and 2 (HSV-2) strains. The main purpose was to determinate whether these growth conditions would affect the antiviral activity. Our results showed the possibility of improving the anti-HSV activity by using extracts from algae cultured in the presence of phytohormones.

Current knowledge on biotechnological interesting seaweeds from the Magellan Region, Chile

This paper is a compilation of data from investigations made with marine benthic algae from the Magellan Region that have biotechnological utilization in human consumption or medicine or as a source of phycolloids or food supplements or animal feed. The most important Rhodophyta species are: Ahnfeltia plicata (Hudson) E.M. Fries for agarose production, Gigartina skottsbergii Setchell & N.L.Gardner for carrageenan production, and Callophyllis variegata (Bory de Saint-Vincent) Kützing for human consumption. The most important Heterokontophyta species are: Macrocystis pyrifera (L.) C. Agardh, and Durvillaea antarctica (Chamisso) Hariot for human consumption, alginate production, and as biofertilizer for agricultural crops. M. pyrifera is also used as a food supplement for salmon, chickens, quails, sheep and bovines and for biofuel production.

Comparison of extraction and transesterification methods on the determination of the fatty acid contents of three Brazilian seaweed species

Seaweeds are photosynthetic organisms important to their ecosystem and constitute a source of compounds with several different applications in the pharmaceutical, cosmetic and biotechnology industries, such as triacylglycerols, which can be converted to fatty acid methyl esters that make up biodiesel, an alternative source of fuel applied in economic important areas. This study evaluates the fatty acid profiles and concentrations of three Brazilian seaweed species, Hypnea musciformis (Wulfen) J.V. Lamouroux (Rhodophya), Sargassum cymosum C. Agardh (Heterokontophyta), and Ulva lactuca L. (Chlorophyta), comparing three extraction methods (Bligh & Dyer - B&D; AOAC Official Methods - AOM; and extraction with methanol and ultrasound - EMU) and two transesterification methods (7% BF3 in methanol - BF3; and 5% HCl in methanol - HCl). The fatty acid contents of the three species of seaweeds were significantly different when extracted and transesterified by the different methods. Moreover, the best method for one species was not the same for the other species. The best extraction and transesterification methods for H. musciformis, S. cymosum and U. lactuca were, respectively, AOM-HCl, B&D-BF3 and B&D-BF3/B&D-HCl. These results point to a matrix effect and the method used for the analysis of the fatty acid content of different organisms should be selected carefully.

Effects of temperature, salinity and irradiance on carposporeling development of Hidropuntia caudata (Gracilariales, Rhodophyta)

The success of seaweed cultivation depends on the scientific control of the tolerance limits and the optimal physiological conditions that affect the spore germination and the early development of algal species. In order to establish cultivation techniques for spores of Hidropuntia caudata (J. Agardh) Gurgel & Fredericq, the effects of irradiance, salinity, and temperature on the carpospore germination and carposporeling development were evaluated under laboratory conditions. Five photon flux densities (PFD, from 18 to 200 µmol photons m-2s-1), six salinity values (from 7 to 55 psu), and four temperatures (from 20 ºC to 35 ºC) were investigated. The level of irradiance caused significant differences in the growth, in the following order: 200±5 > 100±5 ~= 62.5±2.5 > 30±1.5 > 18±1 µmol of photons m-2s-1, but they did not inhibit the carposporeling development. Maximum growth occurred under 35 psu, while at 15 psu the formation of carposporeling erect axis was limited. The optimal temperature for growth was 25 ºC, while at 35 ºC the spores died. These results show the importance of previous knowledge on the tolerance limits and optimal conditions for sporeling development of H. caudata for the implementation of an aquaculture program.

Protocol for seaweed decontamination to isolate unialgal cultures

Decontamination protocols in seaweeds are essential tools for ecophysiological studies in laboratory cultures. These protocols consist of a set of procedures and physical and chemical treatments that must be adjusted for each species. Thus, the effects of explant size and of combinations of physical treatments (brushing and cutting) and chemical treatments (sodium hypochlorite, detergent, seawater, distilled water, germanium dioxide) on the process of obtaining unialgal culture of two pigmentar morphos of Hypnea musciformis were investigated. It was found that thallus segments 50 mm in length, when transported from the field to the laboratory, remained healthier and were less susceptible to epiphytes than those 7 mm in length. The collected material had surfaces contaminated by diatoms, which were weakly attached, as well as surface contamination caused by strongly attached Sahlingia subintegra. The most efficient combination of physical and chemical treatments was explant brushing, cutting and washing with detergent. This combination eliminated the contamination by S. subintegra, but not all of the diatom contamination. The population of the latter was reduced by using physical treatment and by washing with detergent and distilled water and then exterminated by using germanium dioxide (0.003 mg/L). Employing this protocol, unialgal cultures of H. musciformis could be established in approximately eight to ten weeks.

Effects of nitrate and phosphate availabilities on growth, photosynthesis and pigment and protein contents in colour strains of Hypnea musciformis (Wulfen in Jacqu.) J.V. Lamour. (Gigartinales, Rhodophyta)

In Brazil, Hypnea musciformis is the main raw material for carrageenan production and the knowledge of nitrogen and phosphorus metabolism in algae is critical for the success of cultivation because these elements can limit seaweed productivity. Thus, the objective of this study was to evaluate the effects of nitrate (zero to 100 μM) and nitrate plus phosphate (zero to 25 μM) availabilities on the growth, the contents of photosynthetic pigments (phycobiliproteins and chlorophyll a) and proteins, and the photosynthesis and respiration of the brown (BR) and light green (LG) strains of H. musciformis. The results revealed metabolic differences between the colour strains of H. musciformis for nitrogen metabolism: upon nitrate addition, the LG strain stored nitrogen mainly as proteins, while the BR strain stored it as proteins and pigments. Moreover, the respiration of the LG strain and the photosynthesis of the BR strain increased with nitrate concentrations, indicating that the BR strain fixed more photosynthetic carbon than the LG strain.

Growth and accumulation of carotenoids and nitrogen compounds in Gracilaria domingensis (Kütz.) Sonder ex Dickie (Gracilariales, Rhodophyta) cultured under different irradiance and nutrient levels

Effects of the interaction of irradiance and nutrient levels on growth and contents of photosynthetic pigments, carotenoids and proteins in Gracilaria domingensis (Kütz.) Sonder ex Dickie (Gracilariales, Rhodophyta) were investigated experimentally. Nutrient availability provided by dilutions of the nutrient solution of von Stosch (25 and 50 percent, which corresponded to nitrate concentrations of 125 and 250 μmol, respectively) and two photon flux densities [low PFD (50±5) and high PFD (100±5) μmol photons.m-2.s-1] were tested. Growth rates of G. domingensis were stimulated by high PFD. The interaction between high nutrient availability (50 percent VSES) and high PFD stimulated the accumulation of total soluble protein. Phycobiliprotein concentrations (phycoerythrin, phycocyanin, and allophycocyanin) and carotenoid contents were influenced by irradiance levels. Phycobiliprotein concentrations were higher at low PFD and high irradiances stimulated carotenoid accumulation. These results reflect the function of these pigments in photoprotection and the acclimation of G. domingensis to changes in irradiance levels. Our results indicate that light is a limiting factor for G. domingensis growth, that variations in phycobiliprotein contents under different irradiance levels are related to photoacclimation process, and that higher carotenoid contents at high irradiances are due to a photoprotection mechanism.

Micropropagation as a tool for sustainable utilization and conservation of populations of Rhodophyta

Micropropagation as a tool for sustainable utilization and conservation of populations of Rhodophyta. Micropropagation, or in vitro clonal propagation, allows the production of a large number of individuals within a short period. These micropropagated clones could be used as seedlings for seaweed cultivation, avoiding collection from natural beds. Consequently, there has been an increasing interest in micropropagation as a tool for preservation of populations of marine red algae on the Brazilian coast and for the sustainable production of raw material for commercial exploration. This paper reviews the literature on tissue culture and micropropagation of red algae published during the three last decades. Based on the literature, we can conclude that the regeneration process is complex and diversified in different species of Rhodophyta and that the success of micropropagation depends on this process. Species belonging to the orders Bangiales and Ceramiales showed low potential for regeneration, while Gigartinales species showed the highest potential for regeneration. Micropropagation of commercially important red algae is fundamental for the conservation of natural populations by providing seedlings for cultivation and for germplasm collections, both for the conservation of genetic diversity and for biotechnological applications.