New diagnostic characters for the order Sporolithales (Corallinophycidae, Rhodophyta).

The diagnosis of the order Sporolithales is currently restricted to tetrasporangial anatomy. Until recently, there were few reports about gametangial, and more specifically carposporangial material for the Sporolithales. This study provides the first detailed observations of the anatomy of the mature carposporophyte phase from three species of Sporolithales commonly found in rhodolith beds from Brazil: Sporolithon episporum, S. ptychoides, and Sporolithon sp. Using these observations, along with previously published descriptions and illustrations from other representative species in the order, a comparison was made with the other three orders (Corallinales, Hapalidiales, and Rhodogorgonales) of the Corallinophycidae. We amend the diagnosis of the order Sporolithales to include the anatomy of the mature carposporophyte as follows: carposporangial conceptacles that lack a central fusion cell, but instead with numerous, short, one to two-celled, filaments that bear oblong terminal carposporangia that are distributed across the conceptacle chamber floor and walls.

Long-term changes in macroalgae assemblages reveal a gradual biodiversity loss over the last 200 years in the hypereutrophic Guanabara Bay.

Guanabara Bay, the second largest bay on the Brazilian coast, has tropical to subtropical, hypereutrophic water conditions. A survey of the macroalgae flora conducted over the past 200 years (1800-2013) yielded a list of 245 species, which represents 15.7% of the total macroalgae recorded in the Tropical Western Atlantic, while comprehends 29.9% and 50.2% of the marine flora in Brazil and Rio de Janeiro, respectively. When evaluating the macroalgal list throughout different periods, it is noticeable the gradual loss of biodiversity as anthropization increased over the last two centuries. The use of phytogeographic indexes and beta diversity confirmed the negative changes in macroalgae assemblages as a response to increasing environmental degradation. The use of floristic surveys to establish the reference conditions employing historical data and/or regionally referenced prove to be an efficient tool using macroalgae as an ecological indicator for water quality.

Diterpenes from the Brown Alga Dictyota crenulata.

The crude extract of the Brazilian brown alga Dictyota crenulata was analyzed by NMR spectroscopy and HRGC-MS techniques. Seven diterpenes were identified: pachydictyol A, dictyodial, 4beta-hydroxydictyodial A, 4beta-acetoxydictyodial A, isopachydictyol A, dictyol C and dictyotadiol. Xeniane diterpenes have previously been found in D. crenulata from the Pacific Ocean. The results characterize D. crenulata as a species that provides prenylated guaiane (group I) and xeniane diterpenes (group III), thus making it a new source of potential antiviral products.

The effect of iron-ore particles on the metal content of the brown alga Padina gymnospora (Espírito Santo Bay, Brazil).

The iron-ore particles discharged by a pellet processing plant (Espírito Santo Bay, Brazil) cover the seabed of Camburi Beach and consequently, the epibenthic community. In order to determine the importance of the contribution of the iron-ore deposits to the metal concentration in macroalgae of Espirito Santo Bay, four methods of cleaning particulate material adhered to the surface of thalli were tested prior to metal tissue analysis (Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) of Padina gymnospora. In addition, heavy metal concentrations were determined in individuals of P. gymnospora from a site (Frade Island) not affected by the iron-ore particles. The most efficient cleaning treatment, a combination of scraping and washing with an ethanol-seawater solution (NA+SC+ET) removed a number of particles on the surface of thalli 10 times higher than that observed in the control (C). Using this treatment, the total-metal concentrations were reduced by 78% for Fe and 50% for Al respect to the control. However, Fe, Al and Cu concentrations after treatment NA+SC+ET were significantly higher than those found at Frade Island. It is suggested that the iron-ore deposit might be a source for metal availability to macroalgae exposed to the dumped material at Espirito Santo Bay.

Marine biodiversity in the Atlantic and Pacific coasts of South America: knowledge and gaps.

The marine areas of South America (SA) include almost 30,000 km of coastline and encompass three different oceanic domains--the Caribbean, the Pacific, and the Atlantic--ranging in latitude from 12∘N to 55∘S. The 10 countries that border these coasts have different research capabilities and taxonomic traditions that affect taxonomic knowledge. This paper analyzes the status of knowledge of marine biodiversity in five subregions along the Atlantic and Pacific coasts of South America (SA): the Tropical East Pacific, the Humboldt Current,the Patagonian Shelf, the Brazilian Shelves, and the Tropical West Atlantic, and it provides a review of ecosystem threats and regional marine conservation strategies. South American marine biodiversity is least well known in the tropical subregions (with the exception of Costa Rica and Panama). Differences in total biodiversity were observed between the Atlantic and Pacific oceans at the same latitude. In the north of the continent, the Tropical East Pacific is richer in species than the Tropical West Atlantic, however, when standardized by coastal length, there is very little difference among them. In the south, the Humboldt Current system is much richer than the Patagonian Shelf. An analysis of endemism shows that 75% of the species are reported within only one of the SA regions, while about 22% of the species of SA are not reported elsewhere in the world. National and regional initiatives focusing on new exploration, especially to unknown areas and ecosystems, as well as collaboration among countries are fundamental to achieving the goal of completing inventories of species diversity and distribution.These inventories will allow accurate interpretation of the biogeography of its two oceanic coasts and latitudinal trends,and will also provide relevant information for science based policies.

Antiviral activity of the green marine alga Ulva fasciata on the replication of human metapneumovirus.

We evaluated the antiviral activity of the marine alga, Ulva fasciata, collected from Rasa beach and Forno beach, Búzios, Rio de Janeiro, Brazil on the replication of human metapneumovirus (HMPV). The algae extracts were prepared using three different methodologies to compare the activity of different groups of chemical composites obtained through these different methodologies. Four out of the six extracts inhibited nearly 100% of viral replication. The results demonstrated that the majority of the extracts (five out of six) possess virucidal activity and therefore have the ability to interact with the extracellular viral particles and prevent the infection. On the other hand, only two extracts (from Forno beach, obtained by maceration and maceration of the decoction) were able to interact with cell receptors, hindering the viral entry. Finally, only the extract of algae collected at Forno beach, obtained by maceration presented intracellular activity. To our knowledge, this is a pioneer study on antiviral activity of marine algae against HMPV. It is also the first on antiviral activity against HMPV ever done in Brazil. The study also shows the effect of different environment factors and different chemical procedures used to obtain the extract on its biological properties.

Characterization of the photosynthetic conditions and pigment profiles of the colour strains of Hypnea musciformis from field-collected and in vitro cultured samples

Hypnea musciformis (Wulfen) JV Lamour. is a species of great economic interest as it produces Κ-carrageenan and has shown biological activities against HIV and HSV viruses. This species displays different colour strains in its natural habitat, which may have implications for the biotechnological potential of the species. The aim of this study was to characterize the photosynthetic apparatus and pigment profile of three colour strains of H. musciformis (green, brown and red) in their natural habitat and in culture. Chlorophyll a fluorescence of photosystem II was measured with a pulse-amplitude modulated fluorometer and pigments were quantified by spectrofluorimetry (chlorophyll a) and spectrophotometry (phycobiliproteins). In the natural habitat, we detected significant differences between the colour strains for the following photochemical parameters: the green strain had a higher effective quantum yield (ΦPSII) than the red strain and a higher maximum relative electron transport rate (rETRmax) than the brown and red strains. Saturation irradiances were 1000 µE.m-2.s-1 (green) and 500 µE.m-2.s-1 (brown and red). Concerning in vitro culture, the green strain presented the lowest ΦPSII, rETRmax, and α rETR, while the brown strain presented the highest values for these same parameters. The chlorophyll a content of the cultured green strain was the lowest. The phycoerythrin contents of the three colour strains were unchanged by either natural of in vitro conditions: lower in green, intermediate in brown and higher in the red strain, ensuring the chromatic identity of the strains. Our results suggest that the green strain has a better performance when exposed to high irradiance, but a lower efficiency under low irradiance compared to the brown and red strains.

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.

Diterpenes from marine brown alga Dictyota guineensis (Dictyotaceae, Phaeophyceae)

The crude extract of the marine brown alga Dictyota guineensis was analyzed by high-resolution gas chromatography-mass spectrometry (HRGC-MS). Five diterpenes were identified: dictyol E (the most abundant diterpene), dictyotadiol, dictyoxide, isopachydictyol A and pachydictyol A, all diterpenes from the chemical group I, i.e., mainly prenylated derivatives of known sesquiterpene skeletons that result from a first cyclization of geranyl-geraniol between positions 1 and 10. These diterpenes are known for their activity against bacteria, fungi and other activities. The results characterize D. guineensis as a species that yields exclusively diterpenes from group I, with low oxidation and low structural complexity. On Brazilian coasts, only D. mertensii provides exclusively prenylated guaiane diterpenes. Although D. guineensis presents alternate branches and fixing by rhizoidal branches, it is easily distinguishable from D. mertensii by the much narrower stem, short stature and flabelliform habit of the former species. On the other hand, both species have been characterized as producers of diterpenes of group I, in particular, prenylated guaiane. However, D. guineensis has a majority dictyol E in the lipophilic extract, while D. mertensii produces more complex prenylated guaianes, like dictyol H.

Antiviral activity of extracts from Brazilian seaweeds against herpes simplex virus

Organic extracts of 36 species of marine algae (sixteen species of Rhodophyta, eight species of Ochrophyta and twelve species of Chlorophyta) from seven locations on the Brazilian coast were evaluated for their anti-HSV-1 and anti-HSV-2 activity resistant to Acyclovir (ACV). Activity tests in crude extracts, followed by the identification of the major compounds present, were performed for all species. The chemical profiles of all crude extracts were obtained by ¹H-NMR and 13C-NMR spectroscopy. The percentage of extracts with antiviral activity was higher for HSV-1 (86.1%) than for HSV-2 (55.5%). The green algae Ulva fasciata and Codium decorticatum both showed the highest activity (99.9%) against HSV-1, with triacylglycerols and fatty acids as the major components. The red alga Laurencia dendroidea showed good activity against HSV-1 (97.5%) and the halogenated sesquiterpenes obtusol and (-)-elatol were identified as the major components in the extract. Against HSV-2, the green alga Penicillus capitatus (Chlorophyta) and Stypopodium zonale (Ochrophyta) were the most active (96.0 and 95.8%). Atomaric acid, a meroditerpene, was identified as the major secondary metabolite in the S. zonale extract. These results reinforce the role of seaweeds as important sources of compounds with the potential to enter into the pipeline for development of new drugs against herpes simplex.

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.

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.

Sulfated fucan from marine alga inhibits HeLa cells infection by HTLV-1 free particles: semi-quantitative analysis

A sulfated fucan from Laminaria abyssalis marine alga prevented the interaction of HTLV-1 particles, purified from the MT-2 cell line, with HeLa cells. The infection obtained using a concentrated virus suspension was detected only by amplification of the newly synthesized HTLV-1 proviral cDNA by the nested-polymerase chain reaction (PCR). The sulfated polysaccharide was not toxic to the cells at a concentration of 100 µg/mL and prevented infection by the viral particles when added to the cell monolayers. The proviral cDNA was only detected when the sulfated polysaccharide was added to the cells three hours post-infection, indicating that the inhibitory activity occurred in the initial stages of virus-cell interaction. Our results demonstrate, for the first time, the ability of a sulfated fucan from marine algae to inhibit virus transmission through free virus particles.

Antiviral activity of the green marine alga Ulva fasciata on the replication of human metapneumovirus / Atividade antiviral da alga verde marinha Ulva fasciata na replicação do metapneumovírus humano

We evaluated the antiviral activity of the marine alga, Ulva fasciata, collected from Rasa beach and Forno beach, Búzios, Rio de Janeiro, Brazil on the replication of human metapneumovirus (HMPV). The algae extracts were prepared using three different methodologies to compare the activity of different groups of chemical composites obtained through these different methodologies. Four out of the six extracts inhibited nearly 100 percent of viral replication. The results demonstrated that the majority of the extracts (five out of six) possess virucidal activity and therefore have the ability to interact with the extracellular viral particles and prevent the infection. On the other hand, only two extracts (from Forno beach, obtained by maceration and maceration of the decoction) were able to interact with cell receptors, hindering the viral entry. Finally, only the extract of algae collected at Forno beach, obtained by maceration presented intracellular activity. To our knowledge, this is a pioneer study on antiviral activity of marine algae against HMPV. It is also the first on antiviral activity against HMPV ever done in Brazil. The study also shows the effect of different environment factors and different chemical procedures used to obtain the extract on its biological properties.

Neste artigo, foi avaliada a atividade antiviral da alga marinha Ulva fasciata, coletada nas Praias do Forno e Rasa, em Búzios, Rio de Janeiro, Brasil, sobre a replicação do metapneumovírus humano (HMPV). Os extratos desta alga foram preparados utilizando três diferentes metodologias, visando a comparação da atividade de diferentes grupos de compostos químicos que são obtidos dependendo da metodologia empregada. Quatro, do total de seis extratos foram capazes de inibir praticamente 100 por cento da replicação viral. Os resultados demonstram também que a maioria dos extratos (cinco, dos seis), possui atividade virucida e, portanto, possuem a habilidade de interagir com a partícula viral extracelularmente impedindo a infecção. Por outro lado, apenas dois extratos (coletado da Praia do Forno e, preparado através de maceração e maceração do decocto) foram capazes de se ligar a receptores celulares, impossibilitando assim a entrada das partículas virais nas células. Finalmente, apenas o extrato que foi preparado por maceração da alga coletada na Praia do Forno, demonstrou atividade intracelular. Até onde sabemos, este é um estudo pioneiro sobre a atividade antiviral de algas marinhas sobre o HMPV. É também o primeiro estudo sobre atividade antiviral sobre HMPV realizado no Brasil. O estudo também mostra o efeito de diferentes condições ambientais e procedimentos químicos utilizados na preparação do extrato sobre suas propriedades biológicas.

Is the mussel test a good indicator of antifouling activity? A comparison between laboratory and field assays.

Current antifouling technologies rely on metal-based paints, but due to their toxicity, an expected worldwide ban of organotin-containing paints is now prompting the quest for safe and effective alternatives. One of these is antifouling coatings whose active components are naturally occurring compounds in marine organisms. A number of laboratory bioassays has been designed to search for antifouling compounds. However, there is no evidence to date that these assays provide results reproducible through ecologically realistic field experiments. Natural concentrations of the extracts from the Brazilian seaweeds Laurencia obtusa and Stypopodium zonale were tested in the laboratory through the 'mussel test' and in the field through the 'phytagel method' in order to compare the efficiency of these methods in assessing antifouling activity. L. obtusa extract significantly inhibited fouling in both the laboratory and field assays, while S. zonale stimulated fouling in both assays. Major compounds from the extracts were identified. The findings suggest that the 'mussel test' is a reliable time and cost-saving screening method for antifouling substances, although field assays are more sensitive for detection of their activity spectrum.

Inhibitory effect of extracts of Brazilian marine algae on human T-cell lymphotropic virus type 1 (HTLV-1)-induced syncytium formation in vitro.

Extracts from four species of Brazilian marine algae collected from the Rio de Janeiro State coast were screened to determine the inhibitory effect on HTLV-1-induced syncytium formation. Before performing the syncytium inhibition assay the 50% cytotoxic dose (CyD50) of the algal extracts was evaluated. The antiviral test was carried out in HeLa cells co-cultured with HTLV-I infected T-cell line (C91/PL cells) in the presence of marine algal extracts in the concentration inferior to that corresponding to the CyD50. It was observed that co-cultured cells exposed to Ulva fasciata extract showed 60.2% syncytium inhibition at a concentration of 2.5%. At 5% concentration, Sargassum vulgare and Vidalia obtusiloba extracts presented 78.8 and 76% syncytium inhibition, respectively. The best inhibitory activity was observed with Laminaria abyssalis that presented 100% syncytium inhibition at a concentration of 2.5%. This work shows that extracts of marine algae, mainly L. abyssalis extract, are able to inhibit the cell-to-cell contact essential for the spreading of the virus and could be useful to prevent the infection.