Seasonal Variation of Antifouling Activities of Marine Algae from the Brittany Coast (France)

The antifouling activity of extracts (aqueous, ethanol, and dichloromethane) of 9 marine macroalgae against bacteria, fungi, diatoms, macroalgal spores, mussel phenoloxidase activity, and barnacle cypris larvae has been investigated in relation to season in bimonthly samples from the Bay of Concarneau (France). Of the extracts tested, 48.2% were active against at least one of the fouling organisms, and of these extracts, 31.2% were seasonally active with a peak of activity in summer corresponding to maximal values for water temperature, light intensity, and fouling pressure, and 17% were active throughout the year. This seasonal activity may be adaptive as it coincides with maximal fouling pressure in the Bay of Concarneau. Dichloromethane extracts of Rhodophyceae were the most active in the antifouling assays.     

Antifouling activities of octocorals on some marine microfoulers

The antifouling activities of vacuum-dried 70% aqueous alcohol extracts of four gorgonian and five soft corals against four dominant marine fouling diatoms (Navicula subinflata Grun, N. crucicula Smith, Amphora sp., Nitzschia sp.) are described. Of the 36 possible combinations (9 corals x 4 diatoms) 23 of the interactions (64%) showed 100% activity. Extracts of the gorgonian coral Echinogorgia complexa Nutting and the soft coral Dendronephthya (Morchellana) sp. showed 100% growth inhibition against all four fouling diatoms, implying the presence of potent broad spectrum antifouling compounds; other extracts showed more limited species specificity. Exposed cells when transferred to extract-free media, resumed normal growth indicating a nontoxic way of action. The effective concentration for 50% growth inhibition of the attached cells (EC50) varied for each extract and test organism used. The EC50 values for the extract of the gorgonian coral E. complexa against the fouling diatoms ranged from 86 microg/ml (N. subinflata) to 505 microg/ml (Amphora sp.) whereas the EC50 values for extracts of the soft coral Dendronephthya (Morchellana) sp. varied from 28 microg/ml (N. crucicula) to 415 microg/ml (Amphora sp.). The results support the hypothesis that octocorals contain antifouling agents, which could be exploited for the development of nontoxic natural antifouling technology.     

IMPACTS OF MESOGRAZERS ON EPIPHYTE AND ENDOPHYTE GROWTH ASSOCIATED WITH CHEMICALLY DEFENDED MACROALGE FROM THE WESTERN ANTARCTIC PENINSULA: A MESOCOSM EXPERIMENT1

It has been hypothesized that the extensive mesograzer community along the western Antarctic Peninsula regulates epiphytic algae as well as emergent filaments from endophytic species. Should grazing limit growth of fouling or potentially pathogenic microphytes, then Antarctic macrophytes may actually benefit from the remarkably high densities of mesograzer amphipods that occur in these waters. Although initially counterintuitive, the negative impacts of epi/endophyte fouling may outweigh stresses caused by limited amphipod grazing on chemically defended macrophytes by reducing stress from endo/epiphyte biomass. If so, then alleviating mesograzing stress should result in significant increases in endo/epiphytic biomass. To test this hypothesis, a mesocosm experiment was conducted. Individuals representing four common species of Antarctic macroalgae were placed in flow‐through seawater mesocosms. Amphipods were added to five mesocosms at simulated natural densities, while the other five remained herbivore free. At the end of 7 weeks, endo/epiphytic growth on individual macrophytes was quantified. Most species of macroalgae demonstrated noticeably higher instances of endophyte coverage, epiphytic diversity, and diatom colonization in consumer‐free mesocosms than in the presence of amphipods. These data suggest that macroalgae along the western Antarctic Peninsula rely on grazers to control populations of potentially harmful epiphytes. We hypothesize that the chemically defended macroalgal flora lives in mutualism with high densities of mesograzers, providing amphipods with shelter from predation while continually being cleaned of potentially harmful endo/epiphytes.     

Chemical defense and antifouling activity of three Mediterranean sponges of the genus Ircinia

The defense roles and the antifouling activity of the organic extracts and the major metabolites of the sponges Ircinia oros, I. variabilis and I. spinosula were investigated. The antifeedant activity was tested in experimental aquaria on the generalist predator fish Thalassoma pavo as well as in coastal ecosystems rich in fishes. Some of the major metabolites exhibited high levels of antifeedant activity. The antifouling activity was tested in laboratory assays, against representatives of the major groups of fouling organisms (marine bacteria, marine fungi, diatoms, macroalgae and mussels). All extracts showed promising levels of activity. As was expected, no single extract was active in all tests and some fractions that were effective against one organism showed little or no activity against the others. The high but variable level of antifouling activity in combination with the absence of toxicity (tested on the development of oyster and sea urchin larvae) shows the potential of these metabolites to become ingredients in environmentally friendly antifouling preparations.     

Planktonic bacteria and fungi are selectively eliminated by exposure to marine macroalgae in close proximity

To test whether macroalgae affect microbial colonizers in close proximity in a phylum-specific fashion, the community richness of planktonic bacteria and fungi was analyzed with selective oligonucleotide probes targeting the Cytophaga/Flavobacterium/Bacteroides (CFB), Alphaproteobacteria and Roseobacter group and the ITS1 region of marine fungi. Naturally occuring planktonic microorganisms were incubated in the presence of macroalgae or in seawater previously conditioned with macroalgal metabolites. The red algae Ceramium rubrum and Mastocarpus stellatus as well as seawater conditioned with these algae reduced the community composition of bacteria to a greater extent than the brown alga Laminaria digitata, indicating that metabolites differed among macroalgae or that the susceptibility of planktonic bacteria towards alga-derived antimicrobials correlated with their phylogenetic affiliation. The most affected phylotypes belonged to the CFB and the Roseobacter clade. The planktonic fungal community was only affected in the presence of macroalgae and not in algal-conditioned water, but with a specificity different from that observed for bacteria. The macroalgae L. digitata and M. stellatus exhibited more pronounced antifungal effects than C. rubrum. This study demonstrates macroalgal defenses against epiphytic microorganisms based on natural delivery mechanisms of allelochemicals utilizing a culture-independent approach, thus minimizing the ecological bias inherent to culture-dependent studies based on few microbial isolates.     

Growth inhibition of fouling diatoms by antipatharian colonies

Growth inhibition of four dominant marine fouling diatoms by alcohol extracts of antipatharian colonies, collected off the east coast of India, are reported here. Of the three antipatharian colonies tested one antipatharian inhibited the growth of all the four fouling diatoms on glass surface, implying the presence of potent algal growth inhibitors whilst the other extracts were species specific in their action. The same cells exhibited normal growth, when transferred in extract free media, indicating a nontoxic mode of action. The effective concentration for 50% inhibition (EC50) ranged from 90 microg/ml (Nitzschia sp.) to 756 microg/ml (Amphora sp). The EC50 values for N. subinflata and N. crucicula were 215 microg/ml and 108 microg/ml, respectively. The active extracts totally arrested the silicate uptake by diatoms treated with EC100 values, in relation to controls. The degree of silicate uptake impairment varies with the test organisms. The results show the presence of antifouling agents in this group of organisms. This is the first report of anltifouling properties of antipatharian colonies.     

Marine antifoulants from bifurcaria bifurcata (phaeophyceae,cystoseiraceae) and other brown macroalgae

In this study, the antifouling activity of a series of extracts and linear diterpenes isolated from Bifurcaria bifurcata (Velley) Ross, a common brown alga of the Atlantic shores of Europe, and derivatives of these compounds was investigated. Antifouling assays with crude extracts from other brown algae, found abundantly along the coast of South Africa (Bifurcaria brassicaeformis, Bifurcariopsis capensis), the Atlantic shores of Europe (Halidrys siliquosa) and the coast of Mediterranean sea (Cladostephus verticillatus, Halopteris scoparia), are also reported. The fractions were tested in laboratory assays against representative species of the major groups of fouling organisms, viz. bacteria, fungi, diatoms, spores and zygotes of macroalgae and the blue mussel Mytilus edulis. Several components showed promising levels of activity. The high, albeit variable, level of antifouling activity suggests a potential for novel active ingredients in antifouling preparations.     

Adhesion and motility of fouling diatoms on a silicone elastomer

Recent demands for non-toxic antifouling technologies have led to increased interest in coatings based on silicone elastomers that 'release' macrofouling organisms when hydrodynamic conditions are sufficiently robust. However, these types of coatings accumulate diatom slimes, which are not released even from vessels operating at high speeds (>30 knots). In this study, adhesion strength and motility of three common fouling diatoms (Amphora coffeaeformis var. perpusilla (Grunow) Cleve, Craspedostauros australis Cox and Navicula perminuta Grunow) were measured on a poly-dimethylsiloxane elastomer (PDMSE) and acid-washed glass. Adhesion of the three species was stronger to PDMSE than to glass but the adhesion strengths varied. The wall shear stress required to remove 50% of cells from PDMSE was 17 Pa for Craspedostauros, 24 Pa for Amphora and >53 Pa for Navicula; the corresponding values for glass were 3, 10 and 25 Pa. In contrast, the motility of the three species showed little or no correlation between the two surfaces. Craspedostauros moved equally well on glass and PDMSE, Amphora moved more on glass initially before movement ceased and Navicula moved more on PDMSE before movement ceased. The results show that fouling diatoms adhere more strongly to a hydrophobic PDMSE surface, and this feature may contribute to their successful colonization of low surface energy, foul-release coatings. The results also indicate that diatom motility is not related to adhesion strength, and motility does not appear to be a useful indicator of surface preference by diatoms.     

Effect of grazing by isopods and amphipods on growth of Ulva spp. (Chlorophyta)

Eutrophication of shallow coastal waters often leads to blooms of macroalgae. Grazing by crustaceans, such as amphipods and isopods, can reduce macroalgal biomass accumulation. At the same time, growth of the macroalgae can be stimulated by epiphyte removal. The role of grazing by isopods and amphipods on Ulva spp. biomass development was investigated in the Veerse Meer, a brackish lagoon situated in the southwest Netherlands. Exclusion of grazing in the field did not stimulate Ulva spp. growth. In fact, growth rates were higher in exclosures that allowed grazers to enter. Edibility tests identified the amphipod Gammarus locusta, and the isopods Idotea chelipes and Sphaeroma hookeri as potential grazers on Ulva spp. However, when epiphytic diatoms were present on the Ulva spp. thalli, Gammarus and Sphaeroma grazed on ephiphytes and not on Ulva tissue. Only Idotea continued to graze on Ulva spp. A laboratory growth experiment revealed a positive effect of Gammarus presence on Ulva spp. growth, probably caused by preferential removal of epiphytic diatoms from the Ulva spp. thalli. The growth stimulation by epiphyte removing grazers such as Gammarus may explain the higher growth rates in the presence of grazers observed in the field. When determining the potential role of invertebrate grazers in controlling macroalgal biomass accumulation, it is important to include an assessment of the epiphyte abundance on the macroalgae, as preferential removal of epiphytes may stimulate growth and thus have the opposite effect.     

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.     

The role of macroalgae in epiphytism of the toxic dinoflagellate Gambierdiscus toxicus (Dinophyceae)

Populations of the toxic, epiphytic dinoflagellate Gambierdiscus toxicus Adachi et Fukuyo are asSociated closely with Jania sp. on Hitiaa and Papara fringing reefs in Tahiti. Small populations were also observed to be asSociated with Amphiroa sp. and Halimeda opuntia (L.) Lamouroux. The cells attached themselves to the thallus by means of a short thread. When the thalli were irradiated, the cells began to detach from them and swim around the branches. The swimming cells stopped and attached to substrata when a disturbance occurred. The attached cells began to swim within a short time under light conditions when the thallus of Jania sp. were placed near the attached cells. Amphiroa sp. and H. opuntia also induced this re-commencement of swimming of the attached cells. These observations suggest that G. toxicus usually swims around macroalgal thalli on coral reefs. When sudden disturbance or strong water motion occurs, they attach to the surface of macroalgae and are not dispersed. Soon after water motion becomes slow, the cells begin to swim into the water around the thalli. The epiphytism of G. toxicus is different from epiphytic pennate diatoms, most of which adhere to the thallus all the time. The population of G. toxicus is maintained as an asSociation to a limited number of species of macroalgae which support the re-commencement of swimming after disturbance.     

ASSOCIATION OF A FREE-LIVING EUGLENA WITH A COLONIAL ROTIFER

Large epiphytic and epilithic diatom species hosting other diatoms were observed in several fresh- and brackish water sites in Southern California. The most commonly encountered hosts were species forming long filaments attached to rocks or macroalgae, Hydrosera whampoensis (Schwarz) Hendey, Melosira varians Agardh, Pleurosira laevis (Ehrenberg) Compere and Terpsinoe musica Ehrenberg. These large diatoms often had smaller diatoms attached, usually to the girdle bands and occasionally to the mucilage pads connecting the cells. For example, cells of T. musica were observed supporting growth of a diverse diatom assemblage composed of species of the genera Achnanthes , Achnanthidium , Amphora , Cocconeis and Tabularia; Synedra sp. was attached to M. varians and B. paxillifer; and Cocconeis placentula was seen on H. whampoensis. Thus, large epiphytic and epilithic diatoms seem to provide suitable sites for attachment of small epiphytic diatom species, and it appears that this phenomenon is more common than previously thought.     

Poly(ethylene glycol)-containing hydrogel surfaces for antifouling applications in marine and freshwater environments

This work describes the fabrication, characterization, and biological evaluation of a thin protein-resistant poly(ethylene glycol) (PEG)-based hydrogel coating for antifouling applications. The coating was fabricated by free-radical polymerization on silanized glass and silicon and on polystyrene-covered silicon and gold. The physicochemical properties of the coating were characterized by infrared spectroscopy, ellipsometry, and contact angle measurements. In particular, the chemical stability of the coating in artificial seawater was evaluated over a six-month period. These measurements indicated that the degradation process was slow under the test conditions chosen, with the coating thickness and composition changing only marginally over the period. The settlement behavior of a broad and diverse group of marine and freshwater fouling organisms was evaluated. The tested organisms were barnacle larvae (Balanus amphitrite), algal zoospores (Ulva linza), diatoms (Navicula perminuta), and three bacteria species (Cobetia marina, Marinobacter hydrocarbonoclasticus, and Pseudomonas fluorescens). The biological results showed that the hydrogel coating exhibited excellent antifouling properties with respect to settlement and removal.     

Isolation and structural determination of the antifouling diketopiperazines from marine-derived Streptomyces praecox 291-11

Marine derived actinomycetes constituting 185 strains were screened for their antifouling activity against the marine seaweed, Ulva pertusa, and fouling diatom, Navicula annexa. Strain 291-11 isolated from the seaweed, Undaria pinnatifida, rhizosphere showed the highest antifouling activity and was identified as Streptomyces praecox based on a 16S rDNA sequence analysis. Strain 291-11 was therefore named S. praecox 291-11. The antifouling compounds from S. praecox 291-11 were isolated, and their structures were analyzed. The chemical constituents representing the antifouling activity were identified as (6S,3S)-6-benzyl-3-methyl-2,5-diketopiperazine (bmDKP) and (6S,3S)-6-isobutyl-3-methyl-2,5-diketopiperazine (imDKP) by interpreting the nuclear magnetic resonance and high-resolution mass spectroscopy data. Approximately 4.8 mg of bmDKP and 3.1 mg of imDKP were isolated from 1.2 g of the S. praecox 291-11 crude extract. Eight different compositions of culture media were investigated for culture, the TBFeC medium being best for bmDKP and TCGC being the optimum for imDKP production. Two compounds respectively showed a 17.7 and 21 therapeutic ratio (LC50/EC50) to inhibit zoospores, and two compounds respectively showed a 263 and 120.2 therapeutic ratio to inhibit diatoms.     

The development of marine biofilms on two commercial non-biocidal coatings: a comparison between silicone and fluoropolymer technologies

The antimicrobial performance of two fouling-release coating systems, Intersleek 700? (IS700; silicone technology), Intersleek 900? (IS900; fluoropolymer technology) and a tie coat (TC, control surface) was investigated in a short term (10 days) field experiment conducted at a depth of ca 0.5 m in the Marina Bandar Rawdha (Muscat, Oman). Microfouling on coated glass slides was analyzed using epifluorescence microscopy and adenosine-5'-triphosphate (ATP) luminometry. All the coatings developed biofilms composed of heterotrophic bacteria, cyanobacteria, seven species of diatoms (2 species of Navicula, Cylindrotheca sp., Nitzschia sp., Amphora sp., Diploneis sp., and Bacillaria sp.) and algal spores (Ulva sp.). IS900 had significantly thinner biofilms with fewer diatom species, no algal spores and the least number of bacteria in comparison with IS700 and the TC. The ATP readings did not correspond to the numbers of bacteria and diatoms in the biofilms. The density of diatoms was negatively correlated with the density of the bacteria in biofilms on the IS900 coating, and, conversely, diatom density was positively correlated in biofilms on the TC. The higher antifouling efficacy of IS900 over IS700 may lead to lower roughness and thus lower fuel consumption for those vessels that utilise the IS900 fouling-release coating.     

Epiphytic diatom communities on intertidal seaweeds from Iceland

Epiphytic diatom communities on macroalgae from Iceland coastal waters were investigated during July 2005. Ten species of seaweeds have been collected belonging to brown, red and green algae. The analysis of epiphytic diatom community was carried out under scanning electron microscopy. The epiphytic diatom abundances varied from 7 ± 5 to 7524 ± 3491 cells mm⁻². Erect growth forms were the most abundant, representing on average 50% of the total diatoms (Achnanthes cf. brevipes var. parvula, Tabularia investiens, T. fasciculata, Hyalosira cf. delicatula, Gomphoseptatum aestuarii, Pseudogomphonema plinskii), followed by adnate (29%) (Cocconeis stauroneiformis, C. scutellum) and motile forms (21%) (Nitzschia cf. amphibia and Navicula perminuta). Highly branched seaweeds with articulated thallus surface, offering a number of microenvironments to be occupied by the epiphytes, showed a high level of colonization, mainly due to erect and motile diatoms. Flat thalli with smooth surface allowed for the growth of mainly erect diatoms.     

Growth form analysis of epiphytic diatom communities of Terra Nova Bay (Ross Sea, Antarctica)

Diatoms have been long collected from the Southern Ocean but almost no data exist for epiphytic communities, despite their high ecological significance as an important food source in Antarctic coastal food chains. Here, we present a first growth form analysis of diatoms associated with rhodophyte hosts from Terra Nova Bay, Ross Sea, Antarctica. We performed this study to gather baseline information on the species composition of epiphytic diatom communities, determine the influence of some environmental variables on the diatom distribution patterns, and assess the caveats that must be taken into account in terms of sampling design. Macroalgal material was collected during the Italian Antarctic expeditions between 1990 and 2004. Epiphytic diatoms were studied by means of scanning electron microscopy. In terms of growth forms, there were no significant differences between the diatom communities on the different macroalgal host species. Motile (mainly small-celled Navicula perminuta and other Navicula spp.) and adnate (Cocconeis spp.) diatoms dominated the community throughout the study period. Many of the macroalgal blades examined were also covered by epiphytic animals (calcareous bryozoans, hydroids) over most of their surface, with a significant effect on the associated diatom community structure. Our findings suggest that the bio-physicochemical characteristics of each sampling site affected the epiphytic diatom communities more than the substrate type provided by the macroalgal host or the sampling depth.     

Diatoms: unicellular surrogates for macroalgal community structure in streams?

As wholesale biodiversity assessment is often impractical, the use of surrogates that reflect the assemblage structure and diversity of other taxa has attracted increased attention. We sampled 47 boreal streams for diatoms and macroalgae and examined their assemblage patterns along major environmental gradients. Our main intention was to examine whether diatoms might be useful surrogates for macroalgae in boreal streams. We also assessed whether taxon richness and community composition provided similar insights into the patterns of cross-taxon concordance. According to canonical correspondence analysis, diatom distribution was most strongly related to water pH, conductivity, latitude and longitude, and macroalgal distribution to water pH and iron content, latitude and bed instability. In Mantel’s test, diatoms and macroalgae showed significant cross-taxon concordance. However, there was no significant correlation between taxon richness of the two algal groups, likely reflecting their differing responses to environmental variables. We found evidence that although diatoms and macroalgae are partly controlled by different environmental factors, they are segregated rather similarly along latitude and a few environmental gradients such as water pH and iron content. We conclude that, at least at broad geographical extents and in small streams, diatoms reflect the structure of the macroalgal community and are therefore useful surrogates for cost-effective biomonitoring of algal communities in streams.     

Larvae and postlarvae production of the green-white sea urchin Lytechinus variegatus (Echinodermata: Echinoidea) in culture conditions

We evaluated the biological feasibility of massive larvae and postlarvae production of the Caribbean green-white urchin Lytechinus variegatus. Experiments were designed to choose the initial larval density and microalgae diets under culture, to study metamorphosis, postlarval and juvenile growth. Massive production of competent larvae 650 microm long at 12-13 days is possible using larval densities of 0.25 to 1 larva/ml. The microalgae Rhodomonas sp. was suitable for the optimization of larval growth and survival. Metamorphosis of 100% of the larvae can be induced with films of bentic diatoms (Navicula sp. and Amphora sp.), after 96 h; however, diatoms are not adequate for postlarval development and a food supply of Ulva lactuta is necessary for proper growth. For juveniles, a diet of macroalgae (U. lactuta) and/or commercial marine shrimp culture pellet food is enough for growth, but the best results were obtained with shrimp or U. lactuta used alone (85-86%, against 46% with the mixed diet). We recommend future experiments on nutritional requirements to optimize growth of these and subsequent stages.     

Seasonal variations in the fouling diatom community structure from a monsoon influenced tropical estuary

Seasonal variations in the fouling diatom community from a monsoon influenced tropical estuary were investigated. The community composition did not differ significantly between stainless steel and polystyrene substrata due to dominance by Navicula spp. The experimental evidence suggests that Amphora, which is a dominant fouling diatom in temperate waters, ultimately dominates the community in tropical regions when conditions are favourable. These observations reveal that a faster onset of macrofouling interferes with the microfouling community wherein the faster recruiters that have a higher abundance in ambient waters, dominate the community. Seasonal variations were observed in the fouling diatom community. Navicula delicatula dominated during the post-monsoon and tychopelagic diatoms (Melosira and Odontella) were observed during the monsoon. Low diatom abundance was recorded during the pre-monsoon season. The results indicate that although the fouling diatom community composition does not vary between substrata, there is a seasonal change in the community depending on the physical, chemical and biological interactions.