Fouling resistance investigations of seaweed products / Investigações de resistência à incrustação de produtos de algas marinhas

Any solid, unprotected, and undefended surface in the aquatic environment will be fouled. Fouling, on the other hand, can affect a wide range of species that can tolerate some epibiosis. Several others, on the other hand, aggressively keep the epibionts off their body surface (antifouling). Antifouling defenses are built into marine plants like seaweed and seagrass. They do have a distinctive surface structure with tightly packed needle-like peaks and antifouling coverings, which may hinder settling bacteria's ability to cling. Chemical antifouling resistance is most probably a biological reaction to epibiosis' ecological drawbacks, especially for organisms capable of performing photosynthesis. The goal of this study was to see how effective natural compounds derived from littoral seaweeds were in preventing fouling. The brown mussel, an important fouling organism, was evaluated in laboratory bioassays against fifty-one populations' crude organic extracts including fort-two macroalgae species. Antifouling activity, exhibited a distinct phylogenetic pattern, with red macroalgae having the largest share of active species, subsequently brown macroalgae. Antifouling action in green seaweeds has never been significant. Seven species showed some level of induced antifouling defense. Our findings appear to back up previous findings about secondary metabolite synthesis in seaweeds, indicating that in the hunt for novel antifoulants, researchers should concentrate their efforts on tropical red macroalgae.

Resumo contaminada. A incrustação, por outro lado, pode afetar uma ampla gama de espécies que podem tolerar alguma epibiose. Vários outros, por outro lado, mantêm agressivamente os epibiontes fora de sua superfície corporal (anti-incrustante). As defesas anti-incrustantes são construídas em plantas marinhas como algas marinhas e ervas marinhas. Elas têm uma estrutura de superfície distinta com picos semelhantes a agulhas bem compactadas e coberturas anti-incrustantes, o que pode dificultar a capacidade de fixação das bactérias. A resistência química anti-incrustante é provavelmente uma reação biológica às desvantagens ecológicas da epibiose, especialmente para organismos capazes de realizar fotossíntese. O objetivo deste estudo foi verificar a eficácia dos compostos naturais derivados de algas marinhas do litoral na prevenção da incrustação. O mexilhão-marrom, importante organismo incrustante, foi avaliado em bioensaios de laboratório contra extratos orgânicos brutos de 51 populações, incluindo duas espécies de macroalgas. A atividade anti-incrustante exibiu um padrão filogenético distinto, com macroalgas vermelhas tendo a maior participação de espécies ativas, posteriormente macroalgas marrons. A ação anti-incrustante em algas verdes nunca foi significativa. Sete espécies apresentaram algum nível de defesa anti-incrustante induzida. Nossas descobertas parecem corroborar descobertas anteriores sobre a síntese de metabólitos secundários em algas marinhas, indicando que, na busca por novos anti-incrustantes, os pesquisadores devem concentrar seus esforços em macroalgas vermelhas tropicais.

Identification and Characterization of the Larval Settlement Pheromone Protein Components in Adult Shells of Crassostrea gigas: A Novel Function of Shell Matrix Proteins.

The global decline of natural oyster populations emphasizes the need to improve our understanding of their biology. Understanding the role of chemical cues from conspecifics on how oysters occupy appropriate substrata is crucial to learning about their evolution, population dynamics, and chemical communication. Here, a novel role of a macromolecular assembly of shell matrix proteins which act as Crassostrea gigas Settlement Pheromone Protein Components in adult shells is demonstrated as the biological cue responsible for gregarious settlement on conspecifics. A bioassay-guided fractionation approach aided by biochemical and molecular analyses reveals that Gigasin-6 isoform X1 and/or X2 isolated from adult shells is the major inducing cue for larval settlement and may also play a role in postlarva-larva settlement interactions. Other isolated Stains-all-stainable acidic proteins may function as a co-factor and a scaffold/structural framework for other matrix proteins to anchor within this assembly and provide protection. Notably, conspecific cue-mediated larval settlement induction in C. gigas presents a complex system that requires an interplay of different glycans, disulfide bonds, amino acid groups, and phosphorylation crosstalk for recognition. These results may find application in the development of oyster aquacultures which could help recover declining marine species and as targets of anti-fouling agents.

Metabolomics on the study of marine organisms.

BACKGROUND: Marine ecosystems are hosts to a vast array of organisms, being among the most richly biodiverse locations on the planet. The study of these ecosystems is very important, as they are not only a significant source of food for the world but also have, in recent years, become a prolific source of compounds with therapeutic potential. Studies of aspects of marine life have involved diverse fields of marine science, and the use of metabolomics as an experimental approach has increased in recent years. As part of the "omics" technologies, metabolomics has been used to deepen the understanding of interactions between marine organisms and their environment at a metabolic level and to discover new metabolites produced by these organisms. AIM OF REVIEW: This review provides an overview of the use of metabolomics in the study of marine organisms. It also explores the use of metabolomics tools common to other fields such as plants and human metabolomics that could potentially contribute to marine organism studies. It deals with the entire process of a metabolomic study, from sample collection considerations, metabolite extraction, analytical techniques, and data analysis. It also includes an overview of recent applications of metabolomics in fields such as marine ecology and drug discovery and future perspectives of its use in the study of marine organisms. KEY SCIENTIFIC CONCEPTS OF REVIEW: The review covers all the steps involved in metabolomic studies of marine organisms including, collection, extraction methods, analytical tools, statistical analysis, and dereplication. It aims to provide insight into all aspects that a newcomer to the field should consider when undertaking marine metabolomics.

Chemical defense in developmental stages and adult of the sea star Echinaster (Othilia) brasiliensis.

To date, evidence regarding the performance of secondary metabolites from larval stages of sea stars as an anti-predation defense relates only to a few species/specimens from a few geographic ranges. Unfortunately, this hinders a comprehensive global understanding of this inter-specific predator-prey interaction. Here, we present laboratory experimental evidence of chemical defense action in the early developmental stages and adults of the sea star Echinaster (Othilia) brasiliensis from Brazil against sympatric and allopatric invertebrate consumers. Blastulae, early and late brachiolarias of E. (O.) brasiliensis were not consumed by the sympatric and allopatric crabs Mithraculus forceps. Blastulae were also avoided by the sympatric and allopatric individuals of the anemone Anemonia sargassensis, but not the larval stages. Extracts from embryos (blastula) and brachiolarias of E. (O.) brasiliensis from one sampled population (João Fernandes beach) significantly inhibited the consumption by sympatric M. forceps, but not by allopatric crabs and A. sargassensi anemone. In this same site, extracts from adults E. (O.) brasiliensis significantly inhibited the consumption by sympatric and allopatric specimens of the crab in a range of concentrations. Whereas equivalent extract concentrations of E. (O.) brasiliensis from other population (Itaipu beach)inhibited the predation by allopatric M. forceps, while sympatric individuals of this crab avoided the only the higher level tested. Then, early stages and adult specimens of E. (O.) brasiliensis can be chemically defended against consumers, but this action is quite variable, depending on the type (anemone or crab) and the origin of the consumer (sympatric or allopatric).

From Marine Origin to Therapeutics: The Antitumor Potential of Marine Algae-Derived Compounds.

Marine environment has demonstrated to be an interesting source of compounds with uncommon and unique chemical features on which the molecular modeling and chemical synthesis of new drugs can be based with greater efficacy and specificity for the therapeutics. Cancer is a growing public health threat, and despite the advances in biomedical research and technology, there is an urgent need for the development of new anticancer drugs. In this field, it is estimated that more than 60% of commercially available anticancer drugs are natural biomimetic inspired. Among the marine organisms, algae have revealed to be one of the major sources of new compounds of marine origin, including those exhibiting antitumor and cytotoxic potential. These compounds demonstrated ability to mediate specific inhibitory activities on a number of key cellular processes, including apoptosis pathways, angiogenesis, migration and invasion, in both in vitro and in vivo models, revealing their potential to be used as anticancer drugs. This review will focus on the bioactive molecules from algae with antitumor potential, from their origin to their potential uses, with special emphasis to the alga Sphaerococcus coronopifolius as a producer of cytotoxic compounds.

A Review of "Polychaeta" Chemicals and their Possible Ecological Role.

Despite the many publications concerning the isolation of substances and the many reviews of marine natural products, some groups of organisms remain poorly studied, including "Polychaeta". In response, this review covers articles published through December 2016 that address marine natural products produced from polychaetes, with a focus on antipredatory strategies, competitors, fouling, and pathogens. A total of 121 compounds were isolated from 1934 to 2016, which includes halogenated aromatics, proteins, amino acids and Lumazine derivatives most notably-with a defensive function were found in the literature, most frequently in the families Sabellidae, Terebellidae, Glyceridae, and Nereididae. The period of highest discovery of natural products in defensive actions for the group was the 2000s. Polychaetes were addressed in 26 revisions of the total 51 articles analyzed and are less reported than other marine invertebrates such as sponges, cnidarians, mollusks, and tunicates. In sum, the present review provides a basis for future research on the marine chemical ecology of polychaetes.

Structuring effects of chemicals from the sea fan Phyllogorgia dilatata on benthic communities.

Despite advances in understanding the ecological functions of secondary metabolites from marine organisms, there has been little focus on the influence of chemically-defended species at the community level. Several compounds have been isolated from the gorgonian octocoral Phyllogorgia dilatata, a conspicuous species that forms dense canopies on rocky reefs of northern Rio de Janeiro State, Brazil. Manipulative experiments were performed to study: (1) the effects of live colonies of P. dilatata (physical presence and chemistry) on recruitment of sympatric benthic organisms; (2) the allelopathic effects of its chemicals on competitors; and (3) chemotactic responses of the non-indigenous brittle star, Ophiothela mirabilis. Early establishment of benthic species was influenced on substrates around live P. dilatata colonies and some effects could be attributed to the gorgonian's secondary metabolites.In addition, the gorgonian chemicals also exerted an allelopathic effect on the sympatric zoanthid Palythoa caribaeorum, and positive chemotaxis upon O. mirabilis. These results indicate multiple ecological roles of a chemically-defended gorgonian on settlement, sympatric competitors, and non-indigenous species.

Volatile secondary metabolites as aposematic olfactory signals and defensive weapons in aquatic environments.

Olfaction is considered a distance sense; hence, aquatic olfaction is thought to be mediated only by molecules dissolved in water. Here, we challenge this view by showing that shrimp and fish can recognize the presence of hydrophobic olfactory cues by a "tactile" form of chemoreception. We found that odiferous furanosesquiterpenes protect both the Mediterranean octocoral Maasella edwardsi and its specialist predator, the nudibranch gastropod Tritonia striata, from potential predators. Food treated with the terpenes elicited avoidance responses in the cooccurring shrimp Palaemon elegans Rejection was also induced in the shrimp by the memory recall of postingestive aversive effects (vomiting), evoked by repeatedly touching the food with chemosensory mouthparts. Consistent with their emetic properties once ingested, the compounds were highly toxic to brine shrimp. Further experiments on the zebrafish showed that this vertebrate aquatic model also avoids food treated with one of the terpenes, after having experienced gastrointestinal malaise. The fish refused the food after repeatedly touching it with their mouths. The compounds studied thus act simultaneously as (i) toxins, (ii) avoidance-learning inducers, and (iii) aposematic odorant cues. Although they produce a characteristic smell when exposed to air, the compounds are detected by direct contact with the emitter in aquatic environments and are perceived at high doses that are not compatible with their transport in water. The mouthparts of both the shrimp and the fish have thus been shown to act as "aquatic noses," supporting a substantial revision of the current definition of the chemical senses based upon spatial criteria.

Sulfated phenolic acids from Dasycladales siphonous green algae.

Sulfated aromatic acids play a central role as mediators of chemical interactions and physiological processes in marine algae and seagrass. Among others, Dasycladus vermicularis (Scopoli) Krasser 1898 uses a sulfated hydroxylated coumarin derivative as storage metabolite for a protein cross linker that can be activated upon mechanical disruption of the alga. We introduce a comprehensive monitoring technique for sulfated metabolites based on fragmentation patterns in liquid chromatography/mass spectrometry and applied it to Dasycladales. This allowed the identification of two new aromatic sulfate esters 4-(sulfooxy)phenylacetic acid and 4-(sulfooxy)benzoic acid. The two metabolites were synthesized to prove the mass spectrometry-based structure elucidation in co-injections. We show that both metabolites are transformed to the corresponding desulfated phenols by sulfatases of bacteria. In biofouling experiments with Escherichia coli and Vibrio natriegens the desulfated forms were more active than the sulfated ones. Sulfatation might thus represent a measure of detoxification that enables the algae to store inactive forms of metabolites that are activated by settling organisms and then act as defense.

Estudio de prefrrmulación para el desarrollo deun gel a base de polímeros con capacidad mucoadhesiva para ensayos de alelopatía en invertebrados mriinos. / Preformulation study for the development of a polymer-based gel with mucoadhesive propierties for its use in allelopathy assays in marine invertebrates

La investigación se desarrolló con el fin de implementar una metodología que permitiera la realización de ensayos de interacciones alelopáticas entre corales y esponjas, minimizando la interacción física provocada por el uso de dispositivos que producen roce y efecto abrasivo sobre los pólipos del coral. Se llevó a cabo un estudio de preformulación utilizando polímeros con características mucoadhesivas sobre mucus que recubre los pólipos de coral, con el fin de obtener un gel, incorporando en las formulaciones un extracto de la esponja Cliona delitrix. Se caracterizaron propiedades como extensibilidad y adherencia, así como la capacidad bioadhesiva de las formulaciones propuestas, considerando su comportamiento reológico. Estas mostraron una buena estabilidad física frente a las condiciones del medio marino tanto in vitro como in situ. De igual manera, se diseñó un dispositivo que facilitó la aplicación del gel sobre la superficie de los corales por parte de los buzos en el arrecife coralino. Finalmente se estudió el comportamiento de liberación al medio acuoso simulado del gel con el extracto de la esponja objeto de estudio.

This study was carried out with the purpose of implementing a methodology to assess allelopathic interactions assays between corals and reef sponges reducing the physical interaction caused by the use of devices that involve abrasion and harm over the coral polyps. Was carried out a preformulation study using polymers with mucoadhesive on the mucus that cover the coral polyps, with the purpose of develop a gel, incorporating an extract from Cliona Delitrix into the formulations. Obtained formulations were characterized by properties such as extensibility, adherence and mucoadhesive capacity. These formulations showed great physical stability under prevalent marine conditions both in vitro and in situ. In the same way was designed a device that let the smearing of the gel over the coral surfaces carried out for the divers in the coral reefs. Finally, was studied the releasing behavior of the gel with the sponge´s extract into the sea water conditions.

Seaweed allelopathy degrades the resilience and function of coral reefs.

Coral reefs are in dramatic global decline due to a host of local- and global-scale anthropogenic disturbances that suppress corals and enhance seaweeds. This decline is exacerbated, and recovery made less likely, due to over-fishing of herbivores that normally limit seaweed effects on corals. Seaweeds were known to suppress coral reproduction and recruitment, but in a recent study, we demonstrated that numerous seaweeds also directly poison corals via lipid-soluble allelochemicals transferred during contact. These allelopathic interactions may limit reef recovery once seaweeds proliferate and commonly contact remaining corals. Other recent studies suggest that seaweeds may also damage corals by enhancing coral disease or via release of water-soluble compounds that stimulate damaging microbes. For some of these mechanisms, cause versus effect is not yet clear. Here, we suggest that these different mechanisms are not mutually exclusive, may interact in context-dependent ways, but need to be assessed under ecologically realistic field conditions where flow may limit impacts of some mechanisms.

NOVEL STEROLS OF THE TOXIC DINOFLAGELLATE KARENIA BREVIS (DINOPHYCEAE): A DEFENSIVE FUNCTION FOR UNUSUAL MARINE STEROLS?1.

The "red tide" organism Karenia brevis (Davis) Hansen & Moestrup (=Gymnodinium breve Davis) produces a mixture of brevetoxins, potent neurotoxins responsible for neurotoxic shellfish poisoning in humans and massive fish kills in the Gulf of Mexico and the southern Atlantic coast of the United States. The sterol composition of K. brevis was found to be a mixture of six novel and rare Δ8(14) sterols. The two predominant sterols, (24R)-4α-methylergosta-8(14), 22-dienol and (24R)-4α-methyl-27-norergosta-8(14), 22-dienol, were named gymnodinosterol and brevesterol and represent potentially useful biomarkers for K. brevis. A possible function for such unusual marine sterols is proposed whereby structural modifications render the sterols non-nutritious to marine invertebrates, reducing predation and thereby enhancing the ability of the dinoflagellates to form massive blooms.

Chemical defense of brown algae (Dictyopteris spp.) against the herbivorous amphipod Ampithoe longimana.

Terpenoids, polyphenols, and C11 metabolites are broadly distributed among brown seaweeds. Terpenoids and polyphenols have often been investigated as chemical defenses against herbivores, while there are only few investigations of the fatty-acid-derived C11 hydrocarbons and C11 sulfur compounds as potential defenses. We investigated effects of C11 sulfur metabolites from the cosmopolitan brown alga Dictyopteris membranacea on feeding and fitness of the herbivorous amphipod Ampithoe longimana. In choice tests between freshly collected thalli of D. hoytii (which lacks C11 sulfur compounds) and D. membranacea (which contains C11 sulfur compounds) amphipods consumed about 4 times more of the species lacking the C11 sulfur compounds. The same feeding preference was observed when these plants were finely ground and embedded in an agar matrix to destroy morphological differences. When a diet made from field-collected thalli of D. membranacea containing C11 sulfur compounds was tested against a diet made from a laboratory culture of D. membranacea that had lost the ability to produce C11 sulfur compounds, the same magnitude of preference was observed for the population lacking the sulfur compounds. In addition to the C11 sulfur compounds, a water-soluble C9-oxo acid that appears to be a by-product in the biosynthesis of the C11 metabolites also suppressed amphipod feeding to a comparable extent. Both classes of compound may contribute to the effective chemical protection of D. membranacea. When juvenile amphipods were reared for 28 days on artificial diets containing the above compounds, their survivorship (≤10%) closely resembled that of a starved treatment, but differed dramatically from a control treatment (60%) consisting of the same food, but without the metabolites. Most other classes of brown algal secondary metabolites are defensive against a broad spectrum of larger herbivores, but relatively ineffective against the amphipod studied here. In contrast, the fatty-acid-derived sulfur compounds and the C9-oxo acid strongly deter Ampithoe-like mesograzers but appear less effective against other herbivores, suggesting that these metabolites could be ecologically important in defending zygotes and germlings against these small consumers.