Influence of five organic antifouling candidates on spore attachment and germination of a fouling alga Ulva pertusa

Screening of test chemicals or formulations for antifouling (AF) activity is important to get first hand information on their nontoxic repelling activities. Especially spores of a fouling alga, Ulva pertusa were used in this study to test the AF efficiency of five organic chemicals. Coatings made with 100 microg cm2 of citral and eugenol significantly inhibited the spore attachment. A low concentration (1 microg cm2) of solanesol exhibited effective AF activity against spore attachment. Spore germination was sensitive to different AF candidates screened in this study. Based on the attachment and germination response of Ulva pertusa spores, AF efficiency of five organic AF candidates is discussed.     

Evaluation of antifouling activity of eight commercially available organic chemicals against the early foulers marine bacteria and Ulva spores

Environmental impacts caused by tin and copper based commercial antifouling (AF) paints were proved to be detrimental to aquatic ecosystems. Therefore, a search of environmental friendly AF compounds to be used in marine paint to protect the surface of maritime developmental structures from the unwanted biofouling is a burning issue of the present time. Commercially available eight organic chemicals--allyl isothiocyanate, beta-myrecene, cis-3-hexenyl acetate, citral, ethyl heptanoate, eugenol, methyl caproate, and octyl alcohol were evaluated forAF activities using both laboratory and field assays. The test chemicals were found to repel the target motile marine bacteria--Alteromonas marina, Bacillus atrophaeus, Roseobactergallaeciensis and Shewanella oneidensis and motile spores of the green alga, Ulva pertusa. The bacterial and Ulva spore repulsion activities of the test chemicals were measured by chemotaxis and agar diffusion methods respectively interestingly these test chemicals were less toxic to the test fouling species. The toxicity of the test chemicals was measured by using antibiotic assay disks against the bacteria and motility test against Ulva spores. Moreover, in field assay, all test chemicals showed a perfect performance ofAF activity showing no fouling during the experimental period of one year Such results and commercial as well as technical feasibility of the test chemicals firmly showed the possibility of using as alternatives of the existing toxic AF agents.     

Biofouling attractants from a brown marine alga Ecklonia cava

In recent years, industrial pollutants and the mountain forest fire ashes released into seawater cause damage to the marine environment, mainly it reduces the algal productivity in the inter tidal region. To get recover from the stress due to pollutants and to increase the growth and development of biofouling algae (benthic organisms), Ecklonia cava extract was investigated for its biofouling attracting efficiency. Bioactive guided fractions of E. cava extract derived from column chromatography were tested against spore attachment of a fouling alga, Ulva pertusa. Fraction B showed increased spore attachment rate with a maximum of 92 +/- 5%. This fraction was further analysed on HPLC, GC-Mass and NMR, deduced as pentadecanoic acid.     

Antifouling activity of six nontoxic chemicals on spore attachment of Ulva fasciata

Marine biofouling cause severe damage to all marine technologies. Indiscriminate use of toxic metallic antifouling (AF) chemicals (cuprous oxide and TBT) to control the biofouling in turn greatly affects the flora and fauna of the marine ecosystem. In search of environmentally safe and effective AF agents, six nontoxic AF candidates were studied against the spore attachment of Ulva fasciata. Spore attachment was inhibited at 100 microg of trans-cinnamic acid (TCA) and benzoic acid (BA). Sulfate-coumaric acid (SCA) had no significant effect on the spore attachment. The release rate of BA exhibited a constant release flow. Varied response of U. fasciata spores to AF chemicals and release rate to the surrounding medium are discussed.     

Inhibitory effects of red algal extracts on larval settlement of the barnacle Balanus improvisus

We examined the chemical antifouling properties of four sublittoral red algae, Chondrus crispus, Delesseria sanguinea, Osmundea ramosissima, and Polyides rotundus, which are all rarely fouled in the field. Two different approaches were used. Firstly, we tested the effects of lipophilic crude extracts on the settlement behaviour of cyprid larvae of the co-existing barnacle Balanus improvisus. Secondly, in a settlement preference experiment, we tested whether B. improvisus cyprid larvae settle on living algae when given a choice between natural algal surfaces and control surfaces. With this procedure, we were able to test both if the algae inhibit recruitment of cyprids, and if this inhibition is a result of chemistry. The settlement of B. improvisus larvae was strongly inhibited at concentrations estimated to be potentially ecologically relevant for all of the tested extracts. However, only C. crispus significantly inhibited settlement in the preference experiment, even though there was also a tendency for settlement inhibition on P. rotundus and O. ramosissima. In contrast, D. sanguinea seemed to stimulate settlement. This contradiction probably resulted from an extraction of metabolites that naturally occur only inside the alga. However, as this study shows, a combination of settlement assays with whole-cell extracts and preference tests of ecologically relevant fouling organisms on natural algal and control surfaces may be a useful procedure to avoid erroneous conclusions regarding natural antifouling roles of compounds based on settlement assays with only whole-cell extracts. Furthermore, this study also shows that production of inhibitory metabolites may explain the low degree of fouling, especially by B. improvisus, on C. crispus.Communicated by L. Hagerman, Helsingør     

Marine succession of fouling organisms in Hong Kong,with a comparison of woody substrates and common,locally-available,antifouling paints

Successional stages of macrofaunal fouling organisms of test blocks were exposed in the subtropical waters of Hong Kong. A comparison of 5 types of wood commonly used to build boats, and 5 types of locally available antifouling paints is made. Rafts holding the test blocks were exposed at 1 m depth from 24 March, 1970, to 23 October, 1970. The initial macrofoulers on the control (chinese fir) were Balanus and Bugula, and the final dominance was a Styela-Mytilus community. On painted surfaces, the Balanus-Elasmopus community was dominant. Temperature, salinity, dissolved oxygen, and turbidity were recorded weekly during the test. A preliminary chemical analysis of percentage pigment and cuprous oxide content in the antifouling paints is presented.     

Phylogeography of the seaweed Ishige okamurae (Phaeophyceae): evidence for glacial refugia in the northwest Pacific region

Although benthic marine algae are essential components of marine coastal systems that have been influenced profoundly by past and present climate change, our knowledge of seaweed phylogeography is limited. The brown alga Ishige okamurae Yendo occurs in the northwest Pacific, where it occupies a characteristic belt in the exposed intertidal zone. To understand the patterns of genetic diversity and the evolutionary history of this species, we analyzed mitochondrial cox3 from 14 populations (221 individuals) throughout its range. The 17 haplotypes found in this study formed five distinct clades, indicating significant genetic structure. The high differentiation and number of unique (private) haplotypes may result from the recolonization of the species from glacial refugia. Three putative refugia, each with high genetic diversity, were identified: southern Korea (including Jeju Island), northern Taiwan, and central Pacific Japan. Recolonization of I. okamurae was probably determined by ocean currents and changes in sea level during the last glacial period.     

Bacteria on the surface of crustose coralline algae induce metamorphosis of the crown-of-thorns starfish Acanthaster planci

The crustose coralline alga Lithothamnium pseudosorum induces high rates of settlement and metamorphosis of larvae of the coral-eating crown-of-thorns starfish (Acanthaster planci). In cases where crustose coralline algae (CCA) induce metamorphosis of marine invertebrate larvae it is normally assumed that the inductive molecules are produced by the alga, but an alternative is that they originate from bacteria on the plant surface. Bioassays using shards of L. pseudosorum treated with several antibiotics, whereby some shards were reinfected with bacteria from the alga, showed that if bacteria populations are depleted then settlement and metamorphosis of larvae of A. planci are inhibited. This demonstrates that bacteria are necessary for induction and suggests that morphogenic substances are produced by bacteria on the surface of the alga and not directly by the alga itself. However, surface bacteria are not inductive if they are isolated from soluble algal compounds, suggesting either that they require a substrate from the alga to produce the inductive agents or, alternatively but less likely, that compounds from both the alga and bacteria are required. There is no evidence that inductive compounds derive from the alga, since algal cell debris and soluble extracts prepared from the alga do not induce metamorphosis of A. planci. This is the first time that induction of metamorphosis in a marine invertebrate by CCA has been shown to be mediated by bacteria associated with the alga.     

Seaweed secondary metabolites as antifoulants: effects of Dictyota spp. diterpenes on survivorship, settlement, and development of marine invertebrate larvae

Summary. A recent investigation showed that the brown seaweed Dictyota menstrualis was unfouled relative to co-occurring seaweeds, and that larvae of fouling invertebrates avoided settling on D. menstrualis due to chemicals on its surface. The secondary metabolites dictyol E and pachydictyol A are among the compounds found on this alga's surface. In the present study, we tested the effects of specific diterpenes from Dictyota on the survivorship, growth, and development of invertebrate larvae and developing juveniles that could foul seaweeds. Exposure to dictyol E, dictyol B acetate, pachydictyol A, and dictyodial from Dictyota menstrualis and D. ciliolata caused significant larval mortality, abnormal development, and reduce growth rates for three species of co-occurring invertebrates when their larvae were forced into contact with these metabolites. Larvae were damaged at metabolite concentrations as low as 5% of maximum possible surface concentrations of these compounds for the populations of Dictyota we studied. The negative effects of these secondary metabolites on potential foulers, in conjunction with data demonstrating larval avoidance of dictyol-covered surfaces, suggest that these compounds could function as chemical defenses against fouling, and could select for larvae that avoid hosts producing these metabolites. Received 25 May 1998; accepted 22 June 1998.     

防污漆中活性物质海洋环境风险评估关键技术探讨

防污漆中的活性物质对海洋生态环境和人类健康造成的潜在风险受到日益广泛的关注,一些发达国家已建立了针对活性物质海洋环境风险评估的技术体系,但我国相关研究目前尚属空白。综述了防污漆活性物质海洋环境风险评估的研究背景、相关法规、技术标准和发展现状,针对环境风险评估的2个重要组成部分(危害性评估和暴露评估)中的关键技术进行了探讨。在危害性评估中,重点分析和比较了受试生物物种的选择原则、生态毒理数据的要求以及预测无效应浓度的推导方法和应用范围;在暴露评估中,系统阐述了活性物质在水环境中释放速率的计算及修正方法、环境浓度的预测模型、现有的暴露场景及其局限性等。本文以期为我国开展防污漆活性物质海洋环境风险评估提供研究基础和科学依据,并提出了今后的研究重点和方向。     

Effects of copper (I) oxide on growth and biochemical compositions of two marine microalgae

In copper based antifouling (AF) paints Cu (I) oxide was largely used as booster biocide. In this study effect of Cu (I) oxide on two marine microalgae, Tetraselmis suecica and Dunaliella tertiolecta was demonstrated. EC50 (96 hr) concentrations estimated for T. suecica and D. tertiolecta were 1.3 mg l(-1) and 1.34 mg l(-1), respectively. Copper (I) oxide induced changes in growth, chlorophyll, carbohydrate and protein contents were observed in T. suecica and D. tertiolecta. At low concentration of 0.0625 mg l(-1), 3-26% and 1-16% growth stimulation was observed in T. suecica and T. tertiolecta respectively. Increasing Cu (I) oxide concentrations proportionately decreased the carbohydrate and protein contents. This study clearly indicates the toxicity of excessive Cu (I) oxide on growth and biochemical compositions of T. suecica and D. tertiolecta.     

Identification of marine bacteria affecting lithium adsorbents in seawater

Lithium manganese oxide–based adsorbents have been developed for the recovery of lithium from seawater. To maximize the recovery efficiency, it is important to prevent microfouling of lithium adsorbents by marine bacteria. To identify the marine bacteria that cause biofouling against the lithium adsorbents, lithium adsorbents were installed into a non-coated frame or a frame coated with an antifouling agent soaked in seawater. Microorganisms from the surface of lithium adsorbents were collected for 30 days at 10-day intervals, cultured in marine broth, isolated, and identified by 16S rDNA sequencing. Pseudoalteromonas and Vibrio were constituted to 35.6 and 28.8 % of total isolates, respectively, and were predominant in the non-coated frame, whereas Vibrio was poorly isolated (2.3 %) from the antifouling agent–coated frame. In this study, antifouling strategy for maximum lithium recovery efficiency in the marine area takes account of Pseudoalteromonas and Vibrio.     

Demography and phenology of the intertidal green algaCodium setchellii: The enigma of local scarcity on sand-influenced rocky shores

Demographic and phenological attributes that characterize rare and scarce species have not been extensively investigated for marine benthic organisms. From 1984 to 1987, 1 investigated the population ecology of the low intertidal, psammophytic green algaCodium setchellii Gardner that forms low-density populations on sand-influenced rocky benches on N.E. Pacific shores. To address why the alga is not more common in the habitat in which it is most abundant, I quantified sand fluctuations and population dynamics ofC. setchellii at several sites along the central coast of Oregon, USA. Sand movement was highly unpredictable both spatially and temporally on a local scale. AlthoughC. setchellii became fertile in late autumn to winter, the alga and rocky surfaces were often deeply buried by sand in these seasons. Algal recruitment was also spatially and temporally variable, and mortality of small thalli was high. Thallus growth was generally slow, and the alga had the capacity to persist at a given size for long periods. On sand-influenced rocky shores, mortality of largethalli was low, with some thalli persisting for several years. In laboratory experiments,C. setchellii lost substantial biomass during 4 to 5 wk sand burials, whereas two sympatric species of psammophytic algae lost little to no biomass. The within-site scarcity ofC. setchellii may be a consequence of (1) largely unpredictable environmental conditions that reduce algal recruitment and survival, and (2) apparent lack of specialized anatomical or reproductive adaptations to persist on sand-scoured rocky shores. The alga's persistence may be due to its slow growth and long lifespan.     

Comparative effects of indole derivatives as antifouling agents on the growth of two marine diatom species

Antifouling agents, used to prevent biofouling, need to be assessed for their impacts on marine organisms and environment before the application. Diatoms are one of the main components of fouling biofilms, which play important roles in the formation of biofouling. Particularly, diatoms are also the important ingredients of primary production and present interest as ecotoxicological models in marine environment. In this study, two benthic diatoms Nitzschia closterium f. minutissima and Navicula climacospheniae, widely distributed in fouling biofilm, were used as models for screening the activities of potential antifoulants. Nine indole derivatives were tested and CuSO₄ was used as a reference. Indole derivatives showed significant anti-algal activities and the EC₅₀ values of most indole derivatives were lower than that of CuSO₄. Halogen substituent enhanced the anti-algal activities of compounds, and the most efficient compounds for N. closterium f. minutissima were gramine and 7-chloroindole with the EC₅₀ values of 1.94 and 2.1?mg/L, while for N. climacospheniae, 7-chloroindole and 6-bromoindole were the most efficient and the EC₅₀ values were 3.91 and 4.25?mg/L, respectively. In conclusion, indole derivatives would be one of the promising candidates as antifoulants and our results strengthened the need to perform antifouling activity assays and environment-friendly evaluations.     

Offshore marine cheilostome Bryozoa from Fort Lauderdale,Florida

Cheilostome Bryozoa were collected upon marine fouling test panels exposed offshore from Fort Lauderdale, Florida, USA between 1961 and 1967. Panels were exposed at certain depths, ranging from the surface to the bottom, in several arrays extending to a distance of 4 miles offshore. Twenty-three species were identified from the 114 panels recovered and examined. Most species were found on panels in shallow water, or on panels near the bottom in deeper, offshore water. A notable exception was a large, arborescent colony of Biflustra savartii which engulfed a panel 50 feet (15 m) deep, 4 miles from shore, in the Florida current. The lack of cheilostome Bryozoa on shallow and mid-water panels at the offshore, deepwater stations was apparently due to the low number of larvae reaching them from the distant inshore colonies.     

Mechanisms of copper tolerance in the marine fouling alga Ectocarpus siliculosus — Evidence for an exclusion mechanism

Copper-tolerant strains of the marine fouling alga Ectocarpus siliculosus (Dillw) Lyngb. have been recorded. The mechanisms of tolerance which may operate in a copper-tolerant strain were investigated by comparing the growth characteristics, copper content and extracellular organic production of tolerant and non-tolerant strains. Growth was measured by a wet-weight method. The copper content of dryashed cells was measured spectrophotometrically using tetraethylthiuram disulphide. The extracellular material is ultra-violet absorbing, and the production was measured by recording absorption at 265 nm. Evidence suggests that an exclusion mechanism is operating in the tolerant strain under investigation. The extracellular material, the cell wall or an associated organism make little or no contribution no this exclusion. Membrane and intracellular changes are believed to account for the tolerance in this strain of E. siliculosus.     

Biochemical composition,energy content and chemical antifeedant and antifoulant defenses of the colonial Antarctic ascidian<Emphasis Type="Italic"> Distaplia cylindrica</Emphasis>

The colonial ascidian Distaplia cylindrica occurs both as scattered individual colonies or in gardens of colonies in fine-grained soft substrata below 20 m depths off Anvers Island along the Antarctic Peninsula. Individual colonies, shaped as tall rod-like cylinders and anchored in the sediments by a bulbous base, may measure up to 7 m in height. D. cylindrica represent a considerable source of materials and energy for prospective predators, as well as potential surface area for fouling organisms. Nonetheless, qualitative in situ observations provided no evidence of predation by sympatric predators such as abundant sea stars, nor obvious biofouling of colony surfaces. Mean energy content of whole-colony tissue of D. cylindrica was relatively high for an ascidian (14.7 kJ g^–1 dry wt), with most of this energy attributable to protein (12.7 kJ g^–1 dry wt). The sympatric omnivorous sea star Odontaster validus consistently rejected pieces of D. cylindrica colonies in laboratory feeding assays, while readily ingesting similarly sized alginate food pellets. Feeding deterrence was determined to be attributable to defensive chemistry, as colonies of D. cylindrica are nutritionally attractive and lack physical protection (conspicuous skeletal elements or a tough outer tunic), and O. validus display significant feeding-deterrent responses to alginate food pellets containing tissue-level concentrations of organic extracts. In addition, high acidity measured on outer colony surfaces (pH 1.5) as well as homogenized whole-colony tissues (pH 2.5) are indicative of surface sequestration of inorganic acids. Agar food pellets prepared at tissue levels of acidity resulted in significant feeding deterrence in sea stars. Thus, both inorganic acids and secondary metabolites contribute to chemical feeding defenses. D. cylindrica also possesses potent antifoulant secondary metabolites. Tissue-level concentrations of hydrophilic and lipophilic extracts caused significant mortality in a sympatric pennate diatom. Chemical feeding deterrents and antifoulants are likely to contribute to the abundance of D. cylindrica and, in turn, play a role in regulating energy transfer and community structure in benthic marine environments surrounding Antarctica.Communicated by P.W. Sammarco, Chauvin     

Do plant density, nutrient availability, and herbivore grazing interact to affect phlorotannin plasticity in the brown seaweed Ascophyllum nodosum

Plants have different strategies to cope with herbivory, including induction of chemical defences and compensatory growth. The most favourable strategy for an individual plant may depend on the density at which the plants are growing and on the availability of nutrients, but this has not been tested previously for marine plant–herbivore interactions. We investigated the separate and interactive effects of plant density, nutrient availability, and herbivore grazing on the phlorotannin (polyphenolic) production in the brown seaweed Ascophyllum nodosum. Seaweed plants grown at low or high densities were exposed either to nutrient enrichment, herbivorous littorinid gastropods (Littorina obtusata), or a combination of nutrients and herbivores in an outdoor mesocosm experiment for 2 weeks. Seaweeds grown at a low density tended to have higher tissue nitrogen content compared to plants grown at a high density when exposed to elevated nutrient levels, indicating that there was a density dependent competition for nitrogen. Herbivore grazing induced a higher phlorotannin content in plants grown under ambient, but not enriched, nutrient levels, indicting either that phlorotannin plasticity is more costly when nutrients are abundant or that plants responded to herbivory by compensatory growth. However, there were no significant interactive or main effects of plant density on the seaweed phlorotannin content. The results indicate that plants in both high and low densities induce chemical defence, and that eutrophication may have indirect effects on marine plant–herbivore interactions through alterations of plant chemical defence allocation.     

Alkaloids and ascidian chemical defense: Evidence for the ecological role of natural products fromEudistoma olivaceum

The surface of the colonial ascidianEudistoma olivaceum (Van Name) is almost completely free of fouling organisms. I provide evidence that this epibiont-free surface is maintained by the alkaloids, Eudistomins G and H. These alkaloids were extracted from colonies collected in the shallow subtidal of Indian River Lagoon, Florida. USA, in November 1987. Concentrations of less than one-fifth of those found in the living ascidian inhibited the settlement of the larvae of five invertebrate species relative to appropriate controls in laboratory and field trials. Standard pharmacological assays have revealed that Eudistomins G and H were not the most biologically active of the eudistomins, indicating that these standard assays are not necessarily good predictors of antifouling activity. In an examination of possible alternate roles for these eudistomins, they proved ineffective as a fish antifeedant. PinfishLagadon rhomboides consumed agar pellets to which Eudistomins G and H had been added at concentrations 10- to 100-fold higher than those effective against settling larvae. These findings indicate that biologically active marine natural products may serve specific ecological roles.