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.

Marine bacteria from the Gulf of California with antimicrofouling activity against colonizing bacteria and microalgae / Bacterias marinas del Golfo de California con actividad antimicrobiana frente a bacterias colonizadoras y microalgas

Abstract One way of reducing the input of pollutants into the marine environment is to enforce the use of non-toxic antifouling paints in marine protected areas. Thus, the purpose of this study was to detect marine microorganisms that secrete inhibitory substances against bacteria and microalgae to avoid biofouling on manmade structures in La Paz bay, B.C.S., Mexico. The inhibitory potential of 125 bacteria was evaluated against biofilm-forming bacteria. Crude extracts were obtained with methanol and ethyl acetate from 16 bacterial strains that exhibited antagonistic and antibacterial activity in a preliminary screening. Antibacterial and antimicroalgal assays were performed using crude extracts, the minimum inhibitory concentration (MIC) was determined. The highest activity against bacteria and microalgae was found in two strains, Shewanella algae and Staphylococcus sp. The results of this study suggest that extracts of bacteria from the Gulf of California with antimicrobial properties against biofilm-forming bacteria can also prevent the adhesion of microalgae, which may control the development of biofilm formation and, as a consequence, biofouling.(AU)

Resumen Una alternativa para reducir la contaminación en el medio marino es el uso de pinturas anti-incrustantes no tóxicas en áreas marinas protegidas. En el presente estudio se propuso encontrar microorganismos marinos que secreten sustancias capaces de inhibir la adhesión de bacterias y microalgas, de esta manera evitar la bioincrustación en estructuras marinas en la bahía de La Paz, B.C.S., México. Un total de 125 bacterias fueron evaluadas por su capacidad para inhibir el desarrollo de bacterias formadoras de biopelículas. En una selección preliminar de actividad antagónica y antibacteriana, 16 cepas bacterianas mostraron potencial actividad inhibitoria, de las que se obtuvieron los extractos crudos con metanol y acetato de etilo. Se realizaron ensayos antibacterianos y anti-microalgales utilizando los extractos crudos, se determinó la concentración mínima inhibitoria (MIC). Dos cepas mostraron la mayor actividad contra bacterias y microalgas: Shewanella algae y Staphylococcus sp. Los resultados de este estudio sugieren que los extractos de bacterias aisladas en el Golfo de California que poseen propiedades antimicrobianas contra las bacterias formadoras de biofilm y también pueden prevenir la adhesión de microalgas, con lo que se podría controlar el desarrollo de la formación de biopelículas y como consecuencia, el biofouling.(AU)

Inhibition of Nitzschia ovalis biofilm settlement by a bacterial bioactive compound through alteration of EPS and epiphytic bacteria

Background: Marine ecosystems contain benthic microalgae and bacterial species that are capable of secreting extracellular polymeric substances (EPS), suggesting that settlement of these microorganisms can occur on submerged surfaces, a key part of the first stage of biofouling. Currently, anti-fouling treatments that help control this phenomenon involve the use of biocides or antifouling paints that contain heavy metals, which over a long period of exposure can spread to the environment. The bacterium Alteromonas sp. Ni1-LEM has an inhibitory effect on the adhesion of Nitzschia ovalis, an abundant diatom found on submerged surfaces. Results: We evaluated the effect of the bioactive compound secreted by this bacterium on the EPS of biofilms and associated epiphytic bacteria. Three methods of EPS extraction were evaluated to determine the most appropriate and efficient methodology based on the presence of soluble EPS and the total protein and carbohydrate concentrations. Microalgae were cultured with the bacterial compound to evaluate its effect on EPS secretion and variations in its protein and carbohydrate concentrations. An effect of the bacterial supernatant on EPS was observed by assessing biofilm formation and changes in the concentration of proteins and carbohydrates present in the biofilm. Conclusions: These results indicate that a possible mechanism for regulating biofouling could be through alteration of biofilm EPS and alteration of the epiphytic bacterial community associated with the microalga.

Assessment of tributyltin contamination based on imposex in Stramonita rustica (Mollusca: Gastropoda) along southern Bahia coast, northeastern Brazil / Avaliação da contaminação por tributil-estanho baseada em imposex em Stramonita rustica (Mollusca: Gastropoda) ao longo do Litoral Sul da Bahia

Abstract This study investigated the effects of tributyltin (TBT) on the morphology of the genital system of the gastropod Stramonita rustica in southern Bahia, Brazil. For this, 330 specimens were collected during the summer of 2014 at eight sampling points to ascertain whether male sex organs had developed in addition to the complete female genital tract in females (= imposex). The analyses were made under a stereoscopic microscope. Imposex and their associated indexes, and the sterile females, exhibited the highest rates in harbors and shipyards areas. Despite the total ban of TBT in anti-fouling paints on a global scale since 2003, the results of this and other studies indicate the continued use of those paints on the Brazilian coast. This shows the inefficiency of existing legislation and the need to strengthen enforcement of the ban.

Resumo Este estudo investigou os efeitos de tributil-estanho (TBT) sobre a morfologia do sistema genital do gastrópode Stramonita rustica (Lamarck, 1822) no Litoral Sul da Bahia, Brasil. Para isso, 330 espécimens foram coletados durante o verão de 2014 em oito pontos de amostragem, para verificar se os órgãos sexuais masculinos tinham se desenvolvido para além do trato genital feminino completo (= imposex). As análises foram feitas sob um microscópio estereoscópico. O imposex e seus índices associados, assim como as fêmeas estéreis, apresentaram as maiores taxas em áreas portuárias e estaleiros. Apesar da proibição total do TBT em tintas anti-incrustantes em escala global desde 2003, os resultados deste e de outros estudos indicam a continuidade de uso dessas tintas na costa brasileira. Isso mostra a ineficácia da legislação vigente e reforça a necessidade de se cumprir integralmente o banimento.

Antifouling evaluation of extracts from Red Sea soft corals against primary biofilm and biofouling

Objectives:To evaluate antifouling property of extracts from Red Sea soft corals against primary biofilm and biofouling.Methods:Seven species of soft corals Sarcophyton glaucum (a),Sinularia compressa,Sinularia cruciata (a),Heteroxenia fuscescens (a),Sarcophyton glaucum (b),Heteroxenia fuscescens (b) and Sinularia cruciata (b) were chosen to test their extracts as antibacterial and antifouling agents in Eastern Harbour of Alexandria,Mediterranean Sea.Bioactive compounds of soft corals were extracted by using methanol and concentrated under vacuum.The residues of extracts were mixed in formulation of inert paint which consisted of rosin,chlorinated robber and ferrous oxide against micro and macro fouling organisms.The formulated paints were then applied on PVC panels twice by brush,hanged in a steel frame and immersed in Eastern Harbour of Alexandria Mediterranean Sea followed by visual inspection and photographic recordings.Results:After 185 days of immersion in seawater,the antifouling results agreed with the antibactedal results where extracts of Sinularia compressa and Heteroxenia fuscescens (b) gave the best activity against marine fouling tubeworms and barnacles.The inhibition activity was correlated with the major functional groups (hydroxyl,amino,carbonyl,aliphatic (fatty acids),C=C of alkene or aromatic rings and C—Cl of aryl halites) of the extracts.Conclusions:The strong antifouling activity makes them promising candidates for new antifouling additives.After the screening and application of natural organic compounds from soft corals,marine organisms show activity against micro and macro fouling organisms.

Antifouling evaluation of extracts from Red Sea soft corals against primary biofilm and biofouling

Objectives To evaluate antifouling property of extracts from Red Sea soft corals against primary biofilm and biofouling. Methods Seven species of soft corals Sarcophyton glaucum (a), Sinularia compressa, Sinularia cruciata (a), Heteroxenia fuscescens (a), Sarcophyton glaucum (b), Heteroxenia fuscescens (b) and Sinularia cruciata (b) were chosen to test their extracts as antibacterial and antifouling agents in Eastern Harbour of Alexandria, Mediterranean Sea. Bioactive compounds of soft corals were extracted by using methanol and concentrated under vacuum. The residues of extracts were mixed in formulation of inert paint which consisted of rosin, chlorinated rubber and ferrous oxide against micro and macro fouling organisms. The formulated paints were then applied on PVC panels twice by brush, hanged in a steel frame and immersed in Eastern Harbour of Alexandria Mediterranean Sea followed by visual inspection and photographic recordings. Results After 185 days of immersion in seawater, the antifouling results agreed with the antibacterial results where extracts of Sinularia compressa and Heteroxenia fuscescens (b) gave the best activity against marine fouling tubeworms and barnacles. The inhibition activity was correlated with the major functional groups (hydroxyl, amino, carbonyl, aliphatic (fatty acids), C[dbnd]C of alkene or aromatic rings and C[sbnd]Cl of aryl halides) of the extracts. Conclusions The strong antifouling activity makes them promising candidates for new antifouling additives. After the screening and application of natural organic compounds from soft corals, marine organisms show activity against micro and macro fouling organisms.

Natural antifouling compound production by microbes associated with marine macroorganisms: A review

In the marine environment, all hard surfaces including marine macroorganims are colonized by microorganisms mainly from the surrounding environment. The microorganisms associated with marine macroorganisms offer tremendous potential for exploitation of bioactive metabolites. Biofouling is a continuous problem in marine sectors which needs huge economy for control and cleaning processes. Biotechnological way for searching natural product antifouling compounds gained momentum in recent years because of the environmental pollution associated with the use of toxic chemicals to control biofouling. While, natural product based antifoulants from marine organisms particularly sponges and corals attained significance due to their activities in field assays, collection of larger amount of organisms from the sea is not a viable one. The microorganisms associated with sponges, corals, ascidians, seaweeds and seagrasses showed strong antimicrobial and also antifouling activities. This review highlights the advances in natural product antifoulants research from microbes associated with marine organisms.

Imposex en Voluta musica (Caenogastropoda: Volutidae) en el Noreste de la Península de Araya, Venezuela / Imposex in Voluta musica (Caenogastropoda: Volutidae) from Northeastern Península de Araya, Venezuela

Voluta musica is a dioecious marine gastropod endemic of the South Caribbean. Tributyltin (TBT) and copper (Cu) are potential inducers of imposex, an endocrine disorder by which females develop a penis and/or vas deferens. The goal of this work was to determine the imposex incidence in V. musica populations from Northeastern Península de Araya. For this, we selected three sites (Isla Caribe, Isla Lobos and Bajo Cuspe) and made monthly samplings of 15 snails in each site, during one year, and determined: (1) sizes; (2) sex and imposex incidence and (3) the Relative Penis Length Index (RPLI). We also performed histological analysis of the gonads, and measured TBT and Cu concentrations in sediments from the studied localities. Our results showed that the total number of sampled females affected by imposex was 24.5% at Isla Caribe, 12% at Isla Lobos, and none at Bajo Cuspe. In sediments, Cu was detected mostly in Isla Lobos. The female gonads with imposex did not show any development of male cells in any of the sampled sites. The higher percentage of females with imposex matched with the higher boat traffic locality, and higher TBT level (Isla Caribe). No esterilization was evident in this work, nevertheless, the presence of TBT and Cu in the sediments and females with imposex were considered as a potential threat to V. musica populations in this region. In Venezuela there is no control over this particular issue, possibly because of the lack of information and research in this topic, but certainly, this information will be useful in biodiversity conservation policies.

Voluta musica es un gasterópodo dioico endémico del Caribe sur. El TBT y el Cu, son potenciales causantes del imposex, fenómeno donde las hembras desarrollan un pene y/o vaso deferente. El objetivo fue determinar la incidencia de imposex en V. musica en el noreste de la Península de Araya. Se seleccionaron tres localidades y se captura-ron mensualmente 15 individuos durante un año para determinar: (1) talla de los individuos; (2) sexo y presencia de imposex; (3) índice Largo Relativo del Pene (RPLI). Se realizó histología de la gónada de los individuos. Se determinó TBT y Cu en el sedimento de cada localidad. En Isla Caribe, el 24.5% de las hembras presentó imposex, y se halló 3.9ngSn/g de TBT; en Isla Lobos, el 12% de las hembras desarrollaron imposex; en Bajo del Cuspe no se observó imposex. Se halló Cu en mayor concentración en Isla Lobos. Las gónadas femeninas con imposex no demos-traron masculinización. El mayor porcentaje de imposex coincide con la localidad de mayor tráfico de embarcacio-nes y con mayor nivel de TBT (Isla Caribe). No se eviden-ció esterilización, sin embargo la presencia de TBT, Cu e imposex son potenciales amenazas para las poblaciones de V. musica en la región. Hasta ahora, en Venezuela no se está tomando ninguna medida de control sobre este tema en particular, posiblemente por la escasez de información y orientación de las investigaciones hacia este tema, pero que sin duda se debería tomar en cuenta en las políticas para la conservación de la biodiversidad.

Antifouling activity of twelve demosponges from Brazil / Atividade anti-incrustante de doze demosponjas do Brasil

Benthic marine organisms are constantly exposed to fouling, which is harmful to most host species. Thus, the production of secondary metabolites containing antifouling properties is an important ecological advantage for sessile organisms and may also provide leading compounds for the development of antifouling paints. High antifouling potential of sponges has been demonstrated in the Indian and Pacific oceans and in the Caribbean and Mediterranean seas. Brazilian sponges remain understudied concerning antifouling activities. Only two scientific articles reported this activity in sponges of Brazil. The objective of this study was to test crude extracts of twelve species of sponges from Brazil against the attachment of the mussel Perna perna through laboratorial assays, and highlight promising species for future studies. The species Petromica citrina, Amphimedon viridis, Desmapsamma anchorata, Chondrosia sp., Polymastia janeirensis, Tedania ignis, Aplysina fulva, Mycale angulosa, Hymeniacidon heliophila, Dysidea etheria, Tethya rubra, and Tethya maza were frozen and freeze-dried before extraction with acetone or dichloromethane. The crude extract of four species significantly inhibited the attachment of byssus: Tethya rubra (p = 0.0009), Tethya maza (p = 0.0039), Petromica citrina (p = 0.0277), and Hymeniacidon heliophila (p = 0.00003). These species, specially, should be the target of future studies to detail the substances involved in the ability antifouling well as to define its amplitude of action.

Organismos bentônicos marinhos estão expostos constantemente à incrustação, que pode ser danosa para a maioria das espécies. Assim, a produção de metabólitos secundários com propriedades anti-incrustantes é uma vantagem ecológica importante para organismos sésseis e pode também orientar o estudo de substâncias para o desenvolvimento de tintas anti-incrustantes. O alto potencial anti-incrustante de esponjas tem sido demonstrado nos oceanos Índico e Pacífico, nos mares Mediterrâneo e Caribenho. Esponjas brasileiras permanecem pouco estudadas em relação à atividade anti-incrustante. Apenas dois artigos científicos registraram essa atividade em esponjas do Brasil. O objetivo desse estudo foi testar os extratos brutos de doze espécies de esponjas do Brasil contra a fixação do molusco Perna perna através de ensaios laboratoriais e também destacar espécies promissoras para estudos futuros. As espécies Petromica citrina, Amphimedon viridis, Desmapsamma anchorata, Chondrosia sp., Polymastia janeirensis, Tedania ignis, Aplysina fulva, Mycale angulosa, Hymeniacidon heliophila, Dysidea etheria, Tethya rubra e Tethya maza foram congeladas e liofilizadas logo após a coleta e posteriormente procedeu-se a extração com acetona ou diclorometano. O extrato bruto de quatro espécies inibiu significativamente a fixação de bissos: Tethya rubra (p = 0.0009), Tethya maza (p = 0.0039), Petromica citrina (p = 0.0277), e Hymeniacidon heliophila (p = 0.00003). Essas espécies, especialmente, devem ser priorizadas em estudos futuros para detalhamento das substâncias envolvidas na capacidade anti-incrustante, bem como para definir sua amplitude de ação.

Structural diversity of the secondary metabolites derived from marine corals in the south China sea / 国际药学研究杂志

This review reported the structural diversity and novelty of the secondary metabolites derived from eight marine corals in the south China sea. The structural patterns from eight coral species were summarized, in which 130 new compounds of a total of 155 derivatives were uncovered. Some of the secondary metabolites showed potent anti-fouling and anti-cancer activities. These findings ndicated that marine organisms can provide structurally unique compounds for further pharmacological evaluation.

COMPOSICIÓN QUÍMICA Y ACTIVIDAD ANTIFOULING DE LA FRACCIÓN LIPÍDICA DE LA ESPONJA MARINA Cliona tenuis (Clionidae) / CHEMICAL COMPOSITION AND ANTIFOULING ACTIVITY OF THE LIPID FRACTION OF THE CARIBBEAN MARINE SPONGE Cliona tenuis (Clionidae) / COMPOSIÇÃO QUÍMICA E PROPRIEDADES ANTIFOULING DA FRAÇÃO LIPÍDICA DA ESPONJA MARINHA Cliona tenuis (Clionidae)

Del extracto orgánico de la esponja marina Cliona tenuis, recolectada en las Islas del Rosario (Colombia, mar Caribe), fue obtenida la fracción lipídica, la cual presentó propiedades antifouling en pruebas en campo. Esta fracción fue separada por CC sobre gel de sílice hasta obtener fracciones de ésteres metílicos, glicéridos, glicolípidos, fosfolípidos y ácidos grasos libres, las cuales fueron identificadas por CCD y técnicas de de-replicación (RMN 1H y 13C). Posteriormente, las fracciones de glicéridos, glicolípidos y fosfolípidos fueron hidrolizadas, y los ácidos obtenidos, junto con los provenientes de la fracción de ácidos grasos libres, fueron transformados en ésteres metílicos, y todos se analizaron por CGAR-EM. Para ubicar las insaturaciones y ramificaciones, los ésteres metílicos se transformaron luego en sus correspondientes pirrolididas, las cuales también se analizaron por CGAR-EM. El estudio cromatográfico (valores de ECL) y de los espectros de masas de los ésteres metílicos y de sus derivados pirrolididas permitió identificar 81 ácidos grasos diferentes, de los cuales no habían sido previamente reportados: los ácidos 4,8-hexadecadienoico, 11-metil- 4,10-octadecadienoico, 6,9,12,14- icosatetraenoico, y 6,9,12,14,17-icosapentanoico.

The lipid fraction obtained from the marine sponge Cliona tenuis, collected at the Islas del Rosario (Colombia, Caribbean Sea), showed antifouling activity using field bioassays. This fraction was separated by CC on silica gel obtaining a mixture of methyl esters, and enriched fractions of glycerides, glycolipids, phospholipids and free fatty acids. All of them were identified by TLC and dereplication techniques (NMR). The glyceride, glycolipid, and phospholipid fractions were hydrolyzed and the fatty acids obtained, together with the initial fatty acids fraction, were converted into their methyl esters and analyzed by HRGC-MS. To locate unsaturations and alkyl branches in fatty acids, their methyl esters derivates were transformed to pyrrolidides and subsequently analyzed by HRGC-MS. The identity of the fatty acids was carried out using their ECL value as methyl esters, and particularly the study of their pirrolidide spectra was used to locate unsaturations and alkyl branch positions. This study allowed us to identify 81 fatty acids, four of them never reported before: 4,8-hexadecadienoic acid, 11-methy l-4, 10-octadecadienoic acid, 6,9,12,14-icosatetraenoic acid, and 6,9,12,14,17-icosapentanoic acid.

A partir do extrato orgânico da esponja marinha Cliona tenuis, coletada nas ilhas do Rosario (Colombia, Mar Caribe), foi obtido uma fração lipídica, que apresentou propriedades antifouling in situ. Esta fração foi separada através de CC em gel de sílica para obter as frações de ésteres metílicos, glicerídeos, glicolipídeos, folfolipídeos e ácidos libres, as quais foram identificadas por CCD e técnicas espectroscópicas de RMN 1H e 13C. Posteriormente, as frações contendo glicerídeos, glicolipídeos e folfolipídeos foram hidrolizadas e os ácidos obtidos, juntamente com os ácidos obtidos das frações que continham ácidos livres foram transformadas em ésteres metílicos e analisados por CGAR-EM. Para identificar as insaturações e ramificações, os ésteres metílicos foram transformados em derivados pirrolidínicos para serem analisadas por CG-EM. Através da análise cromatográfica (valores de ECL) e dos espectros de massas dos ésteres metílicos e seus derivados pirrolidínicos foi possível identificar 81 ácidos graxos diferentes, sendo que quatro são compostos ainda não relatados: os ácidos 4,8-hexadecadienóico, 11-metil-4,10-octadecadienóico, 6,9, 12, 14- icosatetraenóico, e 6,9,12,14,17-icosapentanóico.

Research progress on mangrove symbiotic microorganisms and their metabolites / 中华中医药杂志

The individuality of mangrove ecotope determined the diversity and specificity of symbiotic microorganisms and their metabolites. However, being confined by the knowledge and cultivation means, the field on mangrove symbiotic microorganisms was on starting. In this paper, the progresses on symbiotic microorganisms of mangrove were reviewed, which hint the essentiality and trends of that field.

Field experimental evaluation of secondary metabolites from marine invertebrates as antifoulants

The crude organic extracts of the endemic gorgonian Phyllogorgia dilatata and two sponge species Aplysina fulva and Mycale microsigmatosa were evaluated for anti-fouling properties through field experiments. To investigate this property in ecologically meaningful conditions, crude extracts from these invertebrates were incorporated at concentrations naturally found in these marine organisms into a stable gel used as a substratum for fouling settlement. Crude extract from A. fulva showed no significant anti-fouling property at the natural concentrations used in the field experiments. In fact, fouling organisms settled significantly more on gels treated with A. fulva extract than on the control gel. On the other hand, both M. microsigmatosa and P. dilatata yielded crude extracts that exhibited a selective action inhibiting only the settlement of barnacles. The evidences obtained here by means of field experiments can provide a basis for future development of one kind of natural antifoulant technology to prevent marine biofouling.

As propriedades antiincrustantes dos extratos brutos da gorgônia endêmica Phyllogorgia dilatata e de duas espécies de esponja, Aplysina fulva e Mycale microsigmatosa, foram avaliadas por intermédio de ensaios de campo. Para investigar essas propriedades sob condições ecologicamente relevantes, as concentrações naturais dos extratos brutos encontrados nesses invertebrados foram incorporadas a um gel estável que serviu de substrato para o assentamento de organismos incrustantes. O extrato bruto de A. fulva não mostrou efeito antiincrustante na concentração natural usada nos experimentos. De fato, os organismos incrustantes colonizaram significativamente mais o gel contendo extrato de A. fulva do que o gel-controle. Por outro lado, ambos os invertebrados, M. microsigmatosa e P. dilatata, produzem extratos brutos que foram seletivamente ativos contra o estabelecimento de balanídeos. As evidências obtidas aqui, por meio de experimentos em campo, podem ser importantes subsídios para futuras investigações visando à aplicação industrial de produtos naturais antiincrustantes em tecnologias de prevenção da bioincrustação marinha.