Sterols compositions, antibacterial, and antifouling properties from two Malaysian seaweeds: Dictyota dichotoma and Sargassum granuliferum

Seaweeds or macroalgae are the primary producers of an oceanic food source, widely distributed across the globe andknown for their excellent defensive properties against numerous biotic and abiotic factors. These defensive traits comefrom their secondary metabolites that act as protective barriers against pathogens and harmful organisms where amongthese compounds some have been found possessing the antifouling characteristic. In this study, Dictyota dichotomaand Sargassum granuliferum collected from Pulau Nunuyan Laut, Sabah, Malaysia, were studied to determine its sterolcomposition, and isolation was carried out to isolate their pure sterol compounds. Two assays consist of disk diffusionmethod for antibacterial activity and crystal violet assay were carried out to study its antifouling activity. Campesterol,stigmasterol, and β-sitosterol were the dominant sterol compound detected in both samples and six pure sterols wereisolated (compounds 1–6). Results from the antibacterial and antifouling analysis showed better inhibition for Gramnegative compared to Gram-positive bacteria. Fucosterol (4) and epicoprostanol (5) gave the best antibacterial activitywith two bacteria inhibited compared to other compound. Meanwhile, coprostanol (1), campesterol (2), stigmasterol(3), epicoprostanol (5), and 5β-cholestan-3-one (6) showed strong antifouling activity towards the selected bacterialstrains with IC50 values ranges from 266.3 to 425.8 µg/ml.

Notas sobre las algas marinas de la Reserva Internacional de Biosfera Seaflower, Caribe colombiano VI: nuevos registros de algas pardas (Phaeophyceae) / Notes on the marine algae of the international biosphere reserve seaflower, caribbean colombia vi: new records of brown algae (phaeophyceae)

ABSTRACT Two species ofbrown algae (Phaeophyceae), Bachelotia antillarum and Dictyota humifusa are reported for the first time for the Archipelago of San Andrés, Providencia and Santa Catalina, part of the International Biosphere Reserve Seaflower.

RESUMEN Dos nuevos registros de algas pardas (Phaoephyceae), Bachelotia antillarum y Dictyota humifusa son registradas por primera vez para el Archipiélago de San Andrés, Providencia y Santa Catalina, Reserva Internacional de la Biosfera Seaflower.

Prospecção de bioativos farmacológicos em algas marinhas Rhodophyta e Heterokontophyta e avaliação de citotoxicidade / Prospection of pharmaceutical bioactive compounds in marine algae Rhodophyta and Heterokontophyta and cytotoxicity evaluation

Muitas drogas terapêuticas produzidas pela indústria farmacêutica são estruturas químicas isoladas de organismos encontrados na natureza ou moléculas baseadas nelas. Podem ser incluídas nesse grupo drogas isoladas de organismos marinhos, como corais, esponjas e algas marinhas, conhecidos como produtores de grandes quantidades de metabólitos secundários. Com base neste fato o presente estudo teve como objetivo realizar a prospecção de moléculas bioativas com propósito farmacológico, em extratos de algas marinhas vermelhas (Rhodophyta) e pardas (Heterokontophyta) coletadas no litoral brasileiro. A prospecção foi realizada por meio de avaliação de seus potenciais antioxidante, antibacteriano, antifúngico, anticancerígeno, e antiparasitário contra organismos causadores de leishmaniose e esquistossomose. Para as avaliações foram empregadas os extratos supercríticos de 5 espécies diferentes, sendo 2 pardas: Dictyota dichotoma e D. menstrualis e três vermelhas: Chondria littoralis, Spyridia hypnoides e Plocamium brasiliense. Os extratos foram avaliados quanto aos seus potenciais bioativos e os resultados mais promissores foram selecionados para as etapas seguintes do fracionamento. Em uma avaliação geral os extratos apresentaram bons resultados e representam uma potencial fonte de bioativos. Os extratos das espécies de D. dichotoma e D. menstrualis foram então submetidos a um procedimento de fracionamento bioguiado pela atividade esquistossomicida. Incorporou-se ainda um terceiro extrato de D. mertensii aos estudos e todas as etapas do fracionamento foram monitoradas por LC-MS. Comparando-se as massas detectadas em todas as frações que apresentaram atividade, para os 3 extratos, foi verificado que a substância de m/z 271,24 estava presente em todas elas, portanto os procedimentos de isolamento foram direcionados a esta molécula para a qual foi possível isolar 7 mg. Diferentemente do que era esperado a molécula quando avaliada isoladamente não apresentou atividade esquistossomicida, levando a hipótese de que a atividade seja decorrente de uma molécula diferente para cada espécie ou ainda que a mesma seja decorrente de uma interação com outras substâncias por um mecanismo de ação aditivo ou sinérgico. O trabalho avaliado de forma geral apresentou resultados promissores e representa um grande embasamento para servir como base para posteriores trabalho de fracionamento

Several therapeutic drugs manufactured by the pharmaceutical industry are chemical structures isolated from organisms that are found in nature or molecules based on that. May be included at this group drugs isolated from marine organisms, like corals, sponges and seaweeds, known as great secondary metabolites producers. Based on this facts the objective of the present study is to perform a prospection study to achieve bioactive molecules with pharmaceutical purposes, on extracts made from red (Rhodophyta) and brown (Heterokontophyta) seaweed collected in the Brazilian shore. The prospection studies was performed by means of evaluation of the antioxidant, antibacterial, antifungal, anticancer and antiparasitic (against Leishmania and Schistosoma) potential. In the evaluation were tested the supercritical extracts of 5 different species, including 2 brown species: Dictyota dichotoma and D. menstrualis and 3 red species: Chondria littoralis, Spyridia hypnoides and Plocamium brasiliense. The extracts were evaluated by their potential bioactive compounds and the most promising results were selected for the following fractionation steps. Overall the extracts have shown good results and may be represent a potential source of bioactive molecules. The extracts of both D. dichotoma and D. menstrualis were submitted to a bioguided fractionation process by their antischistosomal activities. It was still included a third extract from D. mertensii to the studies and every step was monitored by LC-MS techniques. Comparing the detected mass for each active fraction, it was observed the presence of a substance with m/z 271,24 in all of the extracts, so the isolating procedures were directed to obtain that specific molecule, which was obtained in a biomass of 7 mg. Differently than expected the molecule when evaluated isolated do not show the antischistosomal activity, leading to the hypothesis that the activity was related to different molecules for each species or even the observed effect is resulted by an interaction mechanism with another substances by an additive or synergist mechanism. The overall evaluation of the whole work show some promising results and it represent a great support for future fractionation works

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.

Marine natural seaweed products as potential antiviral drugs against Bovine viral diarrhea virus

Bovine viral diarrhea virus (BVDV) is an etiologic agent that causes important economic losses in the world. It is endemic in cattle herds in most parts of the world. The purpose of this study was to evaluate the in vitro cytotoxic effect and antiviral properties of several marine natural products obtained from seaweeds: the indole alkaloid caulerpin (CAV, 1) and three diterpenes: 6-hydroxydichotoma-3,14-diene-1,17-dial (DA, 2), 10,18-diacetoxy-8-hydroxy-2,6-dolabelladiene (DB1, 3) and 8,10,18-trihydroxy-2,6-dolabelladiene (DB3, 4). The screening to evaluate the cytotoxicity of compounds did not show toxic effects to MDBK cells. The antiviral activity of the compounds was measured by the inhibition of the cytopathic effect on infected cells by plaque assay (PA) and EC50 values were calculated for CAV (EC=2,0± 5.8), DA (EC 2,8± 7.7), DB1 (EC 2,0±9.7), and DB3 (EC 2,3±7.4). Acyclovir (EC50 322± 5.9) was used in all experiments as the control standard. Although the results of the antiviral activity suggest that all compounds are promising as antiviral agents against BVDV, the Selectivity Index suggests that DB1 is the safest of the compounds tested.

UV- VIS Spectroscopic and HPLC Studies on Dictyota bartayresiana Lamour

Objective: The present study was aimed to explore phytochemical constituents present in Dictyota bartayresiana Lamour and produce the UV-VIS and HPLC spectrum profile for Dictyotabartayresiana. Methods: Phytochemical screening of the extracts was carried out according to the standard methods. For the HPLC analysis, the methanol: water (45:55) was used as mobile phase. Results: The phytochemical results showed the presence of alkaloids, steroids, phenolic groups, saponins, tannins, glycosides and sugars. The UV- VIS profile of methanolic, petroleum ether, chloroform, isopropanol of D. bartayresiana extract showed various peaks with different functional groups. The HPLC profile of D. bartayresiana petroleum ether, chloroform and benzene extracts showed some prominent and moderate peaks with different retention time. Conclusions:The results of the present study showed that Dictyota bartayresiana Lamour may be rich sources of phytoconstituents which can be isolated and further screened for different kinds of biological activities, depending on their reported therapeutic uses.

Evaluation of the acute toxicity of dolabelladienotriol, a potential antiviral from the brown alga Dictyota pfaffii, in BALB/c mice

Dolabelladienotriol is a product extracted from the brown marine alga Dictyota pfaffii from Brazil that has been shown to have antiviral activity and low cytotoxicity. Our studies have evaluated the acute toxicity of dolabelladienotriol in BALB/c mice for ten days after administration of a single dose. Among the parameters considered were behavior, weight, biochemical and histological analyses of blood samples taken at three different times (Bs.0, Bs.1 and Bs.2) and optical microscopic examination of organs like liver, kidney, stomach and small intestine. Mice deaths were not observed at any dose during the ten day period. There were some changes in the biochemical analysis results for urea nitrogen (BUN) and alanine aminotransferase (ALT), but the changes were not significantly different from the reference levels of the animals before administration of the substance. Histological analyses of tissues were very similar for all animals. The alterations in liver and kidney tissues did not affect the animals´ behavior at any concentration, not even at 50 mg/kg, where the most significant changes in tissues were seen. This study indicates that dolabelladienotriol has low toxicity in administered dose range.

Feeding preferences of the endemic gastropod Astraea latispina in relation to chemical defenses of Brazilian tropical seaweeds

Seaweed preference by the Brazilian endemic gastropod Astraea latispina was examined in the laboratory to evaluate the role of secondary metabolites in determining food choice. Of three species of seaweeds examined, Plocamium brasiliense was highly preferred; less so were Sargassum furcatum and Dictyota cervicornis were preferred less. Extracts and/or pure major metabolites of the two potentially chemically-defended seaweeds (P. brasiliense and D. cervicornis) were tested as feeding deterrents against A. latispina. Algal extract assays demonstrated that three concentrations of crude organic extract of the red alga P. brasiliense (50%, 100%: natural concentration, and 200% of dry weight: dw) did not affect feeding of this gastropod. In contrast, the three concentrations of crude organic extract of the brown alga D. cervicornis (50%, 100% and 200% dw) inhibited feeding by A. latispina. The chemical deterrent property of D. cervicornis extract against the gastropod A. latispina occurred due to a mixture of the secodolastane diterpenes isolinearol/linearol (4:1 -- 0.08% dry weight). This is the first report showing that Dictyota cervicornis produces a chemical defense against herbivores using secodolastane diterpenoid. In addition, these results widen the action spectrum of secondary metabolites found in seaweed belonging to this brown algal genus.

A preferência alimentar do gastrópodo endêmico Astraea latispina por macroalgas bentônicas foi avaliada em laboratório com o intuito de evidenciar o papel de metabólitos secundários em sua escolha alimentar. Dentre as macroalgas examinadas, Plocamium brasiliense foi preferencialmente consumida, enquanto Sargassum furcatum e Dictyota cervicornis foram menos consumidas. Os extratos brutos e/ou metabólitos majoritários puros das duas espécies de macroalgas potencialmente produtoras de defesa química (P. brasiliense e D. cervicornis) foram testados em relação à A. latispina. As três concentrações de extrato bruto (50%, 100%: concentração natural, e 200%: peso seco -- ps) de P. brasiliense não inibiram a herbivoria dessa espécie de gastrópodo. Por outro lado, as três concentrações de extrato bruto de D. cervicornis (50%, 100% e 200% ps) inibiram a herbivoria de A. latispina. Constatou-se que a propriedade deterrente do extrato de D. cervicornis em relação ao gastrópodo A. latispina deve-se à mistura dos diterpenos isolinearol/linearol (4:1 -- 0,08% ps). Este é o primeiro relato de que D. cervicornis produz defesa química contra herbívoros, particularmente diterpenos com esqueleto carbônico do tipo secodolastano. Além disso, esses resultados ampliam o espectro de ação dos metabólitos secundários encontrados em espécies desse gênero de alga parda.