An extract from the brown alga Bifurcaria bifurcata induces irreversible arrest of cell proliferation in a non-small-cell bronchopulmonary carcinoma line

In this study, the antiproliferative activity of a crude extract isolated from Bifurcaria bifurcata (Velley) Ross, a common marine brown alga of the Atlantic shores of Europe, was investigated. Its antiproliferative effect on asynchronous cells of a human non-small-cell bronchopulmonary carcinoma line (NSCLC-N6), particularly chemo-resistant, was expected in vitro. Cell growth appeared to be inhibited in the G1 phase of the cell cycle, and kinetic studies in pretreated cells showed that this growth arrest was irreversible. These events are related to a terminal differentiation induction.     

Screening of Marine Algal Extracts for Anti-settlement Activities against Microalgae and Macroalgae

The ban on the use of TBT-based antifouling paints for boats under 25 m in length has lead to a search for new non-toxic antifoulants. One of the most promising alternative technologies to heavy metal based antifouling paint is the development of antifouling coatings whose active ingredients are naturally occurring compounds from marine organisms. This is based on the principle that marine organisms also face the problem of the presence of epibionts on their own surfaces. In this study, the antifouling activity of a series of aqueous, ethanolic and dichloromethane extracts from thirty algae from the North East Atlantic coast was investigated. The extracts were tested in laboratory assays against species representative of two major groups of fouling organisms, viz . macroalgae and microalgae. The activity of several extracts was comparable to that of heavy metals and biocides (such as TBTO and CuSO 4 ) currently used in antifouling paints and their lack of toxicity with respect to the larvae of oysters and sea urchins suggests a potential for novel active ingredients.     

Seasonal variation in antifouling activity of crude extracts of the brown alga Bifurcaria bifurcata (Cystoseiraceae) against cyprids of Balanus amphitrite and the marine bacteria Cobetia marina and Pseudoalteromonas haloplanktis

Previous studies have demonstrated that macroalgae from Brittany (France) contain products with antifouling activity against marine bacteria, fungi, diatoms, seaweeds and mussels. Little is known regarding the ecological function of these compounds and insufficient attention has been paid to evaluating the possible temporal variation in antifouling activity. Studies of chemical defenses in both terrestrial and marine organisms suggest that organisms vary widely in the production of chemical defenses associated with physical (temperature, light) and biological (e.g. grazing pressure) factors, season and geographical location. The present study aimed to investigate the antifouling activity of crude extracts of monthly collections of the brown alga, Bifurcaria bifurcata, against two marine bacteria, Cobetia marina and Pseudoalteromonas haloplanktis, and cypris larvae of the barnacle, Balanus amphitrite. The toxicity of the extracts was determined with a B. amphitrite nauplius assay.The antimicrobial activity of the extracts was found to be subject to seasonal variation, with the highest level of activity recorded from samples collected between April and September. Results of the anti-settlement experiments showed that the extracts of B. bifurcata (when tested from 0 to 100 μg/ml) can be divided into three groups on the basis of their minimum inhibitory concentrations (MICs): (1) extracts from plants collected from September to March reduced settlement at nontoxic concentrations (50-100 μg/ml); (2) extracts from plants collected from April to July (which were the most active extracts) reduced settlement significantly when tested at >5 μg/ml, but were toxic at 100 μg/ml; (3) the extract prepared from plants harvested in August was inhibitory at >25 μg/ml, but was toxic at 100 μg/ml. Toxicity tests on nauplii showed that LC₅₀ values of samples from the September to March collections were >100 μg/ml, demonstrating that they were nontoxic to nauplii. In contrast, samples obtained from the April to August collections were toxic to nauplii; the most toxic ones being from algae collected in May (LC₅₀=55.6 μg/ml) and in June (LC₅₀=38.3 μg/ml).The antifouling activity of extracts thus reached a peak in summer corresponding to maximal values for water temperature, light intensity and fouling pressure. It remains to be investigated whether this activity has an ecological role in the alga.     

Within-thallus variation in polyphenolic content and antifouling activity in Sargassum vulgare

The brown seaweed Sargassum vulgare C. Agardh abounds along the southeast coast of Rio de Janeiro State, Brazil, creating unique habitats, and its polyphenols play an important ecological role as antifouling agents. In order to understand more precisely this defensive strategy of S. vulgare, we collected this macroalga at Ilha de Itacuruçá in May and October 2009. Thalli were separated according to tissue type: pneumatocysts, receptacles, leaflets, and axis. Phenolic extracts from each specific part of the plant were obtained and the associated antifouling activity was tested in order to assess whether a tissue specialisation/antifouling activity pattern exists. Preliminary separation process was carried out on a phenolic extract to separate the polyphenols from the remaining compounds. Such fractionation allowed us to study the involvement of strictly phenolic compounds in antifouling activity. Within-thallus variation in both phenolic content and antifouling activity were highlighted, and seasonal variation in both those characters was also observed. For both experiments, pneumatocyst and leaflet phenolic extracts were the most active against the attachment of Perna perna, but no correlation was observed between phenolic concentration and antifouling activity. On the contrary, the extract containing only phenolic compounds was twice less active than the extract from which 80 % of the polyphenols were removed. These results led us to hypothesise that polar compounds other than polyphenols are involved in the antifouling activity.     

NORTH CAROLINA MARINE ALGAE. IX. ONSLOWIA ENDOPHYTICA GEN. ET SP. NOV. (PHAEOPHYTA,SPHACELARIALES) AND NOTES ON OTHER NEW RECORDS FOR NORTH CAROLINA1

Four benthic algae are reported here for the first time in the North Carolina flora. The new brown algal genus and species, Onslowia endophytica Searles, is described as an endophyte of Halymenia floridana from the North Carolina continental shelf. New records of Boodleopsis pusilla and Naccaria corymbosa from North Carolina constitute range extensions of these tropical species on the American coast north from Florida. Blastophysa rhizopus, an endophyte and epiphyte known from the North Atlantic coast of Europe and America as well as the Caribbean is reported from North Carolina for the first time and in a new host, Predaea feldmannii.     

Invasion or invisibility: using genetic and distributional data to investigate the alien or indigenous status of the Atlantic populations of the peracarid isopod,Stenosoma nadejda (Rezig 1989)

The peracarid isopod, Stenosoma nadejda (Rezig 1989), until recently considered to be endemic of the Mediterranean region, was first reported in the Atlantic coast of southern Spain in 2001, and in 2006 abundant populations were discovered throughout the southwestern Portuguese coast. This fast expansion was intriguing because, as a direct brooder, this species has limited mechanisms for dispersal, such as rafting on seaweeds. Did S. nadejda recently extend its range into the Atlantic or was it overlooked in the past? We examined the patterns of genetic diversity and population differentiation accordingly by sequencing the cytochrome c oxidase subunit I mitochondrial gene from 75 individuals collected in five locations in Atlantic Iberia and one in the Mediterranean. Our results indicate that the newly discovered Atlantic populations of S. nadejda appear to be old and have long persisted on Atlantic shores rather than being a recent introduction. High levels of genetic diversity and geographic structure were uncovered in what was initially suspected to be an ‘invasive’ species. Recent changes in population dynamics may have made S. nadejda more conspicuous in the Atlantic shores, or a more comprehensive survey led to the recognition of this species where it was not expected.     

A long-term comparison of the benthic algal flora of Clare Island, County Mayo, western Ireland

The marine benthic algal flora of Clare Island, off County Mayo, western shore of Ireland, was investigated; collections of intertidal and subtidal marine algae were made at 16 sites along the eastern and southern shores in the years 1990, 1993 and 2000–2002. The data and observations obtained were compared with the results of a similar survey conducted by Arthur Disbrowe Cotton in 1910–1911. Considering the results of the original survey and the new survey together, the marine algal flora of the island currently totals 293 species; 224 species were recorded by Cotton in the original survey, whereas 223 species were identified in the present study. Most species are common to the original and the new list and the main differences are easily explainable; the new survey used SCUBA diving, which allowed the collection of several subtidal species not collected in 1910, and Cotton reported several microscopic green and brown algae, usually difficult to recognise in the field, which were not rediscovered. The most remarkable differences consist in the current presence of some large intertidal brown algae (Bifurcaria bifurcata, Cystoseira foeniculacea and Cystoseira nodicaulis) that were not reported in the survey of 1910. Two algae, Codium fragile subsp. tomentosoides and Asparagopsis armata, were introduced in Europe after the original survey. At present, the benthic algal assemblages of Clare Island still have basically the same structure and distribution as in 1910 and, if compared with other coastal areas of Europe, the intertidal marine environment of Clare Island appears remarkably well conserved.     

Benefit from an invader: American slipper limpet <Emphasis Type="Italic">Crepidula fornicata</Emphasis>reduces star fish predation on basibiont European mussels

Introduced species have recently become a major concern in ecological research and aquatic conservation. This is due to an increasing appearance of introduced species at a global scale and a multitude of negative impacts on native biota. However, impacts of introduced species are not necessarily only negative. The epizootic American slipper limpet Crepidula fornicata, native at North American Atlantic shores, was introduced to Europe in the 1870s and is now widespread along the Atlantic coast of Europe. Negative effects like trophic and spatial competition have been reported. In its major basibiont in the Wadden Sea, the blue mussel Mytilus edulis, attached limpets reduce survival and growth. However, a laboratory experiment also showed sea star (Asterias rubens) predation on mussels with limpet epigrowth to be three times lower than in unfouled mussels. Hence, although negatively affected by C. fornicatain one way, this epigrowth is beneficial for fouled mussels in another. This indicates that the actual impact of an introduced species is a complex interplay of positive and negative effects which may only be revealed experimentally.     

Fouling deterrent chemical defence in three muricid gastropod egg masses from the Southeast coast of India

Abstract

The egg masses of the marine muricid gastropod molluscs Chicoreus virgineus, Chicoreus ramosus and Rapana rapiformis were studied for antifouling activities. The minimum inhibitory concentrations of crude extracts for the inhibition of byssal production and attachment of the brown mussel Perna indica were 650 μg ml^?1, 1150 μg ml^?1 and 925 μg ml^?1 from the three muricid gastropods, respectively. Higher LC₅₀ values than EC₅₀ values and 100% recovery of the mussels in the toxicity assay indicated the non-toxic nature of the extracts. The gradient partitioning of the egg mass extracts and subsequent antimicrofouling screening against 40 biofilm bacteria showed wide-spectrum antibacterial activity of the medium polar fraction from C. virgineus; the non-polar fraction from R. rapiformis and both non-polar and medium polar fractions from C. ramosus. The antimicrofouling activity from extracts of the three egg masses was found to be more prominent than antimacrofouling activity. This may be attributed to the targeting of a defence strategy against microbes in order to protect the developing mollusc embryos.     

Trihydroxylated linear diterpenes from the brown alga Bifurcaria bifurcata

Two novel polar diterpenes were isolated from the brown alga Bifurcaria bifurcata collected off the Atlantic coast of Morocco, and their structures established by spectral methods. Both compounds are trihydroxylated acyclic diterpenes derived from 12-hydroxygeranylgeraniol. They were tested in vitro for their cytotoxicity and proved to be active against the NSCLC-N6 cell line. Their absolute configuration at the C-12 position has been determined with a modified Mosher's method [J. Am. Chem. Soc. 113 (1991) 4092] and that of the 12-hydroxygeranylgeraniol (bifurcadiol) has been revised.     

A provisional classification of algal-characterised rocky shore biotopes in the Azores

Recent studies of the rocky shores of the Azores archipelago have provided information on community structure allowing provisional identification of plant-characterised biotopes (habitats and their associated communities). Although the Azores share some littoral and sublittoral biotopes with the Atlantic coast of mainland Europe, shores in the archipelago mostly lack the functionally important leathery macrophyte communities of fucoids and laminarians widespread in the North Atlantic. Intertidal biotopes are mainly turfs typical of warm-temperate and tropical regions, and characterised by articulated Corallinaceae or by non-coralline algae such as Cladophora spp., Gelidium spp., Pterocladiella capillacea, Stypocaulon scoparia, and Valonia utricularis. Subtidal algal biotopes are characterised by Dictyota spp., Halopteris filicina, Sphaerococcus coronopifolius and, most commonly, Zonaria tournefortii.     

Antifouling activity as a function of population variation in <Emphasis Type="Italic">Sargassum vulgare</Emphasis> from the littoral of Rio de Janeiro (Brazil)

The brown seaweed Sargassum vulgare is abundant along the coast of Rio de Janeiro state. An investigation of the spatial variation of antifouling activity was conducted, in which algae were collected at five locations along the coast of Rio de Janeiro during October 2008. Hexane, dichloromethane, methanol and acetone/water extracts were prepared and screened for their bioactivities against the growth of five strains of marine fouling bacteria, five biofilm-associated microphytobenthic strains and attachment of the mussel Perna perna. The most active were the hexane extracts from Bananal algae that inhibited the growth of all microalgae tested; the methanol and dichloromethane extracts from Mar do Norte, which inhibited Vibrio aestuarianus and Pseudoalteromonas elyakovii and the polyphenol extracts from Ilha de Itacuruçá and Bananal that inhibited mussel attachment, respectively, by 64% and 71% compared to controls.     

Woodlands at the edge: A European perspective on the Atlantic Oakwood Plant Communities

Summary

The Atlantic Oakwoods can be seen as ‘at the edge’ in two ways. First, they lie at the extreme oceanic end of a series of Quercus robur and Q. petraea woodlands which stretches eastwards from the North Atlantic coast to the continental heartland of Europe. Second, they are at the northern limit of a range of oceanic oakwoods which extends southwards to Iberia where Q. pyrenaica is an important canopy companion. This paper outlines the main variations in trees, shrubs, herbs and cryptogams that can be seen along these two climatic gradients, so as to highlight what is truly distinctive about the Atlantic Oakwoods and how we might rightly value them.     

Antifouling activity of seaweed extracts on the green alga Enteromorpha prolifera and the mussel Mytilus edulis

Twenty-seven species of common seaweeds from the coast of Korea havebeen screened for antifouling activity. The seaweed extracts were tested inlaboratory assays against the marine fouling green alga Enteromorphaprolifera and the blue mussel Mytilus edulis. Tissue growth, sporesettlement, zygote formation and germlings of the E. prolifera wereinhibited by methanol extracts of the seaweed Ishige sinicola (= I. foliacea) and Sargassum horneri. Spore settlement was stronglyinhibited by using extract concentrations as low as 30 g mL^-1with I. sinicola and 120 g mL^-1 with S. horneri. The repulsive activity of the foot of the mussel was completely inhibited bymethanol extracts of I. sinicola and Scytosiphon lomentaria atconcentrations of 40 g per 10 L drop supplied to eachmussel. These extracts also showed strong antifouling activities onlarval settlement with, respectively, no or only 6% of the spat settlingwhen a test concentration of 0.8 mg mL^-1 was used. This work isthe first stage towards the development of novel antifouling agents frommarine macroalgae.     

(S)-12-hydroxygeranylgeraniol-derived diterpenes from the brown alga Bifurcaria bifurcata

Four novel diterpenes were isolated from the brown alga Bifurcaria bifurcata collected off the Atlantic coast from Morocco, and their structures established by spectral and chemical methods. These compounds are acyclic diterpenes derived from (S)-12-hydroxygeranylgeraniol. One of them is its dehydration product at C-12, while the others are its oxidation derivatives: the methyl ester of the acid at C-1 and two stereoisomers (Z and E) of the aldehyde at C-1. These results are discussed from a chemotaxonomic point of view.     

Antifouling substances of natural origin: Activity of benzoquinone compounds from Maesa Lanceolata against marine crustaceans

A screening of antifouling activity from plants extracts led to selection and further study of Maesa lanceolata Forssk. Two p‐benzoquinone compounds were isolated from the fruits and found to be active against Artemia salina. The anti‐crustacean activity of both p‐benzoquinones is reported for the first time.     

Trends in abundance of rocky intertidal seaweeds and filter feeders across gradients of elevation,wave exposure,and ice scour in eastern Canada

On NW Atlantic rocky shores, the main basal organisms in intertidal communities are seaweeds (Ascophyllum nodosum, Fucus spp. and Chondrus crispus) and filter feeders (barnacles, Semibalanus balanoides, and mussels, Mytilus spp.). Their ecology has been extensively studied in New England (United States), but knowledge gaps exist for northern shores, which are subjected to stronger environmental stress. Therefore, we studied the above organisms on Canadian shores. We quantified the summer abundance of these seaweeds and filter feeders across full vertical (intertidal elevation) and horizontal (wave exposure and winter ice scour) environmental gradients on the Gulf of St. Lawrence and open Atlantic coasts of Nova Scotia. At the regional scale along the open Atlantic coast, seaweeds showed similar abundances in Nova Scotia than values reported for New England. However, both filter feeders were considerably less abundant in Nova Scotia. At the local scale in Nova Scotia, intense winter ice scour (which only occurs on the Gulf of St. Lawrence coast) was associated with a very low abundance of all species except barnacles. Spatial trends in Nova Scotia were similar to patterns known for certain species elsewhere, such as A. nodosum being almost restricted to sheltered habitats, regardless of elevation, and C. crispus being almost restricted to low elevations, regardless of exposure. Other trends were, however, characteristic of Nova Scotia, such as C. crispus being frequent at low elevations in exposed habitats, unlike in New England, where mussels often predominate there because of competitive advantages. In Nova Scotia, mussels were always restricted to cracks and crevices, unlike in New England, where they form extensive intertidal beds on exposed shores. The direct effects of increased environmental stress and indirect effects through altered interspecific interactions might explain the regional differences in local species distribution, which will require experimental validation. Handling editor: K. Martens     

Marine microbial natural products in antifouling coatings

Ecological problems associated with current antifouling technologies have increased interest in the natural strategies that marine organisms use to keep their surfaces clean and free from fouling. Bacteria isolated from living surfaces in the marine environment have been shown to produce chemicals that are potential antifoulants. Active compounds from the cells and culture supernatant of two bacterial strains, FS‐55 and NudMB50–11, isolated from surface of the seaweed, Fucus serratus, and the nudibranch, Archidoris pseudoargus, respectively, were extracted using solid phase extraction. The extracts were combined with acrylic base paint resin and assayed for antifouling activity by measuring their ability to inhibit the growth of fouling bacteria. These formulations were found to be active against fouling bacteria isolated from marine surfaces. The formulation of antifouling paints that incorporate marine microbial natural products is reported here for the first time. This is a significant advance towards the production of an environmentally friendly antifouling paint that utilises a sustainable supply of natural biodegradable compounds.     

Isolation and cross‐species amplification of microsatellite loci from the fucoid seaweeds Fucus vesiculosus,F. serratus and Ascophyllum nodosum (Heterokontophyta,Fucaceae)

The Fucaceae is a family of brown seaweeds that dominate and frequently co‐occur on North Atlantic rocky shores. We developed nine polymorphic microsatellite markers for the fucoid seaweeds Fucus vesiculosus, F. serratus and Ascophyllum nodosum using a combined, enriched library. Six of these loci were polymorphic in at least two species, showing from two to eight alleles with heterozygosities ranging from 0.41 to 0.85. Loci were also tested on F. spiralis, revealing five polymorphic microsatellite loci in this species.     

Inhibitory Effects of Mediterranean Sponge Extracts and Metabolites on Larval Settlement of the Barnacle Balanus amphitrite

One of the most promising alternative technologies to antifouling paints based on heavy metals is the development of coatings whose active ingredients are compounds naturally occurring in marine organisms. This approach is based on the problem of epibiosis faced by all marine organisms and the fact that a great number of them cope with it successfully. The present study investigated the antifouling activity of a series of extracts and secondary metabolites from the epibiont-free Mediterranean sponges Ircinia oros, I. spinosula, Cacospongia scalaris, Dysidea sp., and Hippospongia communis. Antifouling efficacy was evaluated by the settlement inhibition of laboratory-reared Balanus amphitrite Darwin cyprids. The most promising activity was exhibited by the metabolites 2-[24-acetoxy]-octaprenyl-1-4-hydroquinone (8a), dihydrofurospongin II (10), and the alcoholic extract of Dysidea sp.