Species specificity of barnacle settlement-inducing proteins

We previously isolated a larval settlement-inducing protein complex (SIPC) from adult extracts of the barnacle, Balanus amphitrite using a nitrocellulose membrane settlement assay. In the present study, we found that the extracts of other adult barnacles, Megabalanus rosa and Balanus eburneus, also induced the settlement of B. amphitrite cyprids although the inductive activity was slightly lower than that of conspecific extracts. Furthermore, we examined reactivity to anti-SIPC antibody in adult extracts from six species of Japanese barnacles other than B. amphitrite, brine shrimp and eight marine sessile organisms besides barnacles. The results showed that all barnacles examined contained SIPC-like proteins with slightly different molecular weight, while the other animals did not react to the antibody by immunoblot analysis. These findings suggest that species specificity in settlement-inducing proteins of barnacles is not so strict, but these proteins are characteristic to barnacle species.     

A comparison of the antifouling/foul-release characteristics of non-biocidal xerogel and commercial coatings toward micro- and macrofouling organisms

Five non-biocidal xerogel coatings were compared to two commercial non-biocidal coatings and a silicone standard with respect to antifouling (AF)/fouling-release (FR) characteristics. The formation and release of biofilm of the marine bacterium Cellulophaga lytica, the attachment and release of the microalga Navicula incerta, and the fraction removal and critical removal stress of reattached adult barnacles of Amphibalanus amphitrite were evaluated in laboratory assays. Correlations of AF/FR performance with surface characteristics such as wettability, surface energy, elastic modulus, and surface roughness were examined. Several of the xerogel coating compositions performed well against both microfouling organisms while the commercial coatings performed less well toward the removal of microalgae. Reattached barnacle adhesion as measured by critical removal stress was significantly lower on the commercial coatings when compared to the xerogel coatings. However, two xerogel compositions showed release of 89-100% of reattached barnacles. These two formulations were also tested in the field and showed similar results.     

Multiple foundation species shape benthic habitat islands

Pattern generation by foundation species (FS) is a primary structuring agent in marine and terrestrial communities. Prior research, focused on single-species or guild-dominated habitats, stressed the role of facilitation in maintaining community structure. However, many habitats are developed by multiple FS from different guilds. Competition between these FS may provide an additional agent potentially responsible for spatial and temporal patterns. In the White Sea, epibenthic patches formed by barnacles (Balanus crenatus) and solitary ascidians (mainly Styela spp. and Molgula spp.) on small stones and empty bivalve shells (mainly Serripes groenlandicus) produce microhabitats for different sessile taxa. We hypothesized that: (1) several FS would provide habitats for most of other species in the community; (2) different FS promote different assemblages of sessile organisms; (3) the interplay of facilitation and competition best explains observed patterns of abundance and demography in FS; and (4) these interactions shape the whole community, increasing the diversity compared to less heterogeneous patches constituted by single FS. We examined 459 patches and the results generally supported this hypothesis. The number of FS in a patch positively affected species diversity. Most sessile species (72% of individuals) resided on barnacles, ascidians and red algae, except barnacles that dominated the primary substrate. The size structure of barnacles (live individuals and empty shells) and ascidians were interrelated, suggesting long-term patch dynamics whereby ascidians regularly replace barnacles. Following this replacement, we expect consequent changes to the entire dependent assemblage. Evidence for these changes exists in the spatial pattern: most sessile and motile taxa demonstrated significant associations with either FS. Our results indicate that the small-scale patterns observed in patches formed by multiple FS are primarily generated by facilitation of dependent taxa by FS, and facilitation and competition between different FS. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Dedicated to E. A. Ninbourg, our late tutor.     

Combining Traits and Density to Model Recruitment of Sessile Organisms

We propose an integrative approach that explains patterns of recruitment to adult populations in sessile organisms by considering the numbers of individuals and their body size. A recruitment model, based on a small number of parameters, was developed for sessile organisms and tested using the barnacle Semibalanus balanoides, a marine invertebrate inhabiting North Atlantic intertidal shores. Incorporating barnacle body size improved model fit beyond that based on density alone, showing that growth played an important role in how resource limitation affected survival. Our approach uncovered the following: First, changes in the shape of the recruitment curve resulted from the balance between individual growth and mortality. Second, recruitment was limited by the least plastic trait used to characterise body size, operculum area. Basal area, a trait that responded to increases in barnacle density, did not contribute significantly to explain patterns of recruitment. Third, some temporal variation is explained by changes in the amount of space occupied by shells of dead barnacles: at high cover barnacles are densely packed and these shells remain long after death. Fourth, seasonal variation and spatial variation in survival can be separated from that resulting from resource limitation; survival was predicted for two different shores and four sampling times using a single recruitment model. We conclude that applying this integrative approach to recruitment will lead to a considerable advance in understanding patterns of mortality of early stages of sessile organisms.     

Antifouling activity against barnacle cypris larvae: Do target species matter (Amphibalanus amphitrite versus Semibalanus balanoides)?

Larvae of many benthic invertebrates settle on surfaces where they metamorphose into juveniles if suitable substrata are available, and are responsible for the major costs of biofouling. When assessing new formulations or compounds for potential antifouling (AF) application, constraints such as seasonal availability may restrict most bioassays to relatively few taxa and species. For example, amongst barnacles, Amphibalanus amphitrite is popular as a test organism but is it really representative of other barnacle species? In order to test this hypothesis, we have chosen to work with marine natural extracts as a probe. Indeed, one substitution technology to toxic metal-based coatings to control fouling is the development of AF coatings with active compounds derived from marine organisms or analogues of the lead compounds. In this study, the AF activity and toxicity of extracts from 30 algae from the North East Atlantic coast were investigated for their potential anti-settlement activities against larvae of two species of barnacle, A. amphitrite and Semibalanus balanoides. As a trend, most of the active extracts displayed activity towards S. balanoides, only few displayed targeted activity against A. amphitrite, or against both species. In order to better understand if this tendency could be linked to chemical ecology, surface extracts were prepared on a selection of species. The results highlight that surface extracts of algae all displayed highest levels of activity than total extracts when tested on S. balanoides. This difference illustrates that specific compounds in their ecological context can have potentially a better efficacy on target species.     

Smelly feet are not always a bad thing: the relationship between cyprid footprint protein and the barnacle settlement pheromone

A critical phase in the life cycle of sessile benthic marine invertebrates is locating a suitable substratum for settlement. For barnacles, it is the lecithotrophic cypris larva that makes this plankto-benthic transition. In exploring possible substrata for settlement, the cyprid leaves behind 'footprints' of a proteinaceous secretion that reportedly functions as a temporary adhesive, and also acts as a secondary cue in larval-larval interactions at settlement. Here, we show that two polyclonal antibodies raised against peptides localized at the N- and C-terminal regions of the adult settlement cue--the settlement-inducing protein complex (SIPC)--could both detect 'temporary adhesive' indicating that the SIPC is either a component of this secretion or that they are the same protein.     

Prolonged morphometric study of barnacles grown on soft substrata of hydrogels and elastomers

A long-term investigation of the shell shape and the basal morphology of barnacles grown on tough, double-network (DN) hydrogels and polydimethylsiloxane (PDMS) elastomer was conducted in a laboratory environment. The elastic modulus of these soft substrata varied between 0.01 and 0.47 MPa. Polystyrene (PS) (elastic modulus, 3 GPa) was used as a hard substratum control. It was found that the shell shape and the basal plate morphology of barnacles were different on the rigid PS substratum compared to the soft substrata of PDMS and DN hydrogels. Barnacles on the PS substratum had a truncated cone shape with a flat basal plate while on soft PDMS and DN gels, barnacles had a pseudo-cylindrical shape and their basal plates showed curvature. In addition, a large adhesive layer was observed under barnacles on PDMS, but not on DN gels. The effect of substratum stiffness is discussed in terms of barnacle muscle contraction, whereby the relative stiffness of the substratum compared to that of the muscle is considered as the key parameter.     

Principles of Biofouling Protection in Marine Sponges: A Model for the Design of Novel Biomimetic and Bio-inspired Coatings in the Marine Environment?

The process of biofouling of marine structures and substrates, such as platforms or ship hulls, proceeds in multiple steps. Soon after the formation of an initial conditioning film, formed via the adsorption of organic particles to natural or man-made substrates, a population of different bacterial taxa associates under the formation of a biofilm. These microorganisms communicate through a complex quorum sensing network. Macro-foulers, e.g., barnacles, then settle and form a fouling layer on the marine surfaces, a process that globally has severe impacts both on the economy and on the environment. Since the ban of tributyltin, an efficient replacement of this antifouling compound by next-generation antifouling coatings that are environmentally more acceptable and also showing longer half-lives has not yet been developed. The sponges, as sessile filter-feeder animals, have evolved antifouling strategies to protect themselves against micro- and subsequent macro-biofouling processes. Experimental data are summarized and suggest that coating of the sponge surface with bio-silica contributes to the inhibition of the formation of a conditioning film. A direct adsorption of the surfaces by microorganisms can be impaired through poisoning the organisms with direct-acting secondary metabolites or toxic peptides. In addition, first, compounds from sponges have been identified that interfere with the anti-quorum sensing network. Sponge secondary metabolites acting selectively on diatom colonization have not yet been identified. Finally, it is outlined that direct-acting secondary metabolites inhibiting the growth of macro-fouling animals and those that poison the multidrug resistance pump are available. It is concluded that rational screening programs for inhibitors of the complex and dynamic problem of biofilm production, based on multidisciplinary studies and using sponges as a model, are required in the future.     

Polysulfone and polyacrylate-based zwitterionic coatings for the prevention and easy removal of marine biofouling

A series of polysulfone and polyacrylate-based zwitterionic coatings were prepared on epoxy-primed aluminum substrata and characterized for their antifouling (AF) and fouling-release (FR) properties towards marine bacteria, microalgae and barnacles. The zwitterionic polymer coatings provided minimal resistance against bacterial biofilm retention and microalgal cell attachment, but facilitated good removal of attached microbial biomass by exposure to water-jet apparatus generated hydrodynamic shearing forces. Increasing the ion content of the coatings improved the AF properties, but required a stronger adhesive bond to the epoxy-primed aluminum substratum to prevent coating swelling and dissolution. Grafted poly(sulfobetaine) (gpSBMA), the most promising zwitterionic coating identified from microfouling evaluations, enabled the removal of four out of five barnacles reattached to its surface without incurring damage to their baseplates. This significant result indicated that gpSBMA relied predominately on its surface chemistry for its FR properties since it was very thin (~1–2 µm) relative to commercial coating standards (>200 µm).     

A new flow-through bioassay for testing low-emission antifouling coatings

Current antifouling (AF) technologies are based on the continuous release of biocides into the water, and consequently discharge into the environment. Major efforts to develop more environmentally friendly coatings require efficient testing in laboratory assays, followed by field studies. Barnacles are important fouling organisms worldwide, increasing hydrodynamic drag on ships and damaging coatings on underwater surfaces, and thus are extensively used as models in AF research, mostly in static, laboratory-based systems. Reliable flow-through test assays for the screening of biocide-containing AF paints, however, are rare. Herein, a flow-through bioassay was developed to screen for diverse low-release biocide paints, and to evaluate their effects on pre- and post-settlement traits in barnacles. The assay distinguishes between the effects from direct surface contact and bulk-water effects, which are crucial when developing low-emission AF coatings. This flow-through bioassay adds a new tool for rapid laboratory-based first-stage screening of candidate compounds and novel AF formulations.     

Marine biofilm bacteria evade eukaryotic predation by targeted chemical defense

Many plants and animals are defended from predation or herbivory by inhibitory secondary metabolites, which in the marine environment are very common among sessile organisms. Among bacteria, where there is the greatest metabolic potential, little is known about chemical defenses against bacterivorous consumers. An emerging hypothesis is that sessile bacterial communities organized as biofilms serve as bacterial refuge from predation. By testing growth and survival of two common bacterivorous nanoflagellates, we find evidence that chemically mediated resistance against protozoan predators is common among biofilm populations in a diverse set of marine bacteria. Using bioassay-guided chemical and genetic analysis, we identified one of the most effective antiprotozoal compounds as violacein, an alkaloid that we demonstrate is produced predominately within biofilm cells. Nanomolar concentrations of violacein inhibit protozoan feeding by inducing a conserved eukaryotic cell death program. Such biofilm-specific chemical defenses could contribute to the successful persistence of biofilm bacteria in various environments and provide the ecological and evolutionary context for a number of eukaryote-targeting bacterial metabolites.     

Bioassays and field immersion tests: a comparison of the antifouling activity of copper-free poly(methacrylic)-based coatings containing tertiary amines and ammonium salt groups

This paper focuses on the activity spectrum of three dimethylalkyl tertiary amines as potential active molecules and the corresponding ammonium salt-based antifouling (AF) paints. Bioassays (using marine bacteria, microalgae and barnacles) and field tests were combined to assess the AF activity of coatings. Bioassay results demonstrated that the ammonium salt-based paints did not inhibit the growth of microorganisms (except the dimethyldodecylammonium-based coatings) and that the tertiary amines were potent towards bacteria, diatoms, and barnacle larvae at non-toxic concentrations (therapeutic ratio, LC₅₀/EC₅₀, < 1). The results from field tests indicated that the ammonium salt-based coatings inhibited the settlement of macrofouling and the dimethylhexadecylammonium-based coatings provided protection against slime in comparison with PVC blank panels. Thus, results from laboratory assays did not fully concur with the AF activity of the paints in the field trial.     

The distribution and abundance of barnacles in a mangrove forest

Abstract The grey mangrove tree, Avicennia marina, forms a hard substratum in an area otherwise dominated by soft-sediment. Various intertidal organisms attach to and move about on the trees. The abundant barnacles Elminius covenus, Hexaminius popeiana and Hexaminius foliorum live on a variety of substrata in mangrove forests. Their patterns of distribution and abundance were recorded and models proposed to explain these patterns. Densities of barnacles on bark, leaves and twigs of A. marina were estimated in a locality near Sydney. Elminius covenus were more abundant on bark than on leaves or twigs of A. marina. Hexaminius popeiana, although less abundant on bark than E. covenus, were not found on twigs or leaves. In contrast, H. foliorum were more abundant on twigs than leaves and were not found on the bark of A. marina. Densities of barnacles were greater in the seaward than in landward parts of the forest. Elminius covenus were the most abundant barnacles, H. foliorum were less abundant and H. popeiana were virtually absent in the landward zone. Barnacles in the seaward zone were most abundant at mid-tidal levels, less at high and low tidal levels of trunks and more abundant on lower than upper surfaces of trunks and leaves. The densities of barnacles differed according to the orientation of the bark or leaf. Models are proposed to explain the patterns of distribution and abundance of these barnacles. Each involves larval supply, settlement and post-settlement mortality. It is concluded that similar processes determining patterns of distribution and abundance for sessile organisms living on rock platforms may be applicable to sessile organisms living in the very different habitats of mangrove forests.     

Effects of intensity and seasonal timing of disturbances on a rocky intertidal benthic community on the southern coast of Korea

The effects of intensity and timing of disturbances on recovery of marine benthic organisms were investigated on a rocky intertidal shore in Gwangyang Bay, Korea. We hypothesized that the recovery pattern of the benthic community structure would be affected by disturbance intensity and season. Twenty-eight permanent plots were set up, with disturbance intensity (cleared plots and sterile plots) and seasonal disturbance (fall 1999 and spring 2000) incorporated into the experimental design. To monitor natural seasonal variation in benthic community abundances, we established seven permanent unmanipulated plots. Turf-forming algae were observed in the unmanipulated plots throughout the experimental period, whereas green algae and invertebrate presence varied with season. In the disturbance-intensity experiment, turf-forming and green algae were dominant in cleared plots. The highest coverage of sessile organisms was observed in sterile plots, which exhibited the highest species richness because of their relatively low macroalgal coverage. Seasonal effects of disturbance were an important factor in the recovery pattern of benthic organisms under high disturbance intensity. Coverage of green algae was higher in sterile spring plots than in sterile fall plots; this result was attributed to low spatial competition, as the disturbances occurred just before green algal blooms. On the other hand, the abundances of barnacles and bivalves were highest on sterile fall plots, as these organisms were suppressed by green algal blooms in other periods. These results indicate that the effects of disturbance intensity on benthic community recovery patterns can be influenced by season of disturbance.     

Recolonization of a high latitude hard-bottom nearshore community

Early recolonization of sessile organisms and grazer effects on recolonization was examined in a high latitude nearshore rocky environment. A manipulative experiment using cleared boulders and cages was used to determine the time needed for initial recruitment and composition of early recruits. The hypothesis tested was invertebrate grazing is causing slow recolonization. Four years of monitoring cleared boulders resulted in an initial recruitment of primarily barnacles in year 3 on boulders that were caged to exclude grazers. The total percent cover on these boulders after 4 years was less than 2%. Cage control and uncaged boulders showed less recruitment. Uncleared and uncaged control rocks showed insignificant temporal variation. Concurrent observations revealed that macroalgae were reproductive during the study and that space was limiting in this community. This study strongly suggests that perturbations causing removal of hard substrate sessile communities in the Beaufort Sea will result in very slow community recovery.     

Too close for comfort: spatial patterns in acorn barnacle populations

Spatial patterns in aggregations form as a result of the interplay between costs and benefits experienced by individuals. Such self-organisation of aggregations can be explained using a zonal model in which a short-range zone of repulsion and longer-range zone of attraction surrounding individuals leads to emergent pattern properties. The signal of these processes can be detected using spatial pattern analyses. Furthermore, in sessile organisms, post-settlement mortality reveals the relative costs and benefits of positions within the aggregation. Acorn barnacles are known to require contact with conspecifics for reproduction and are therefore believed to aggregate for this purpose; isolated individuals may also be more susceptible to abiotic stress and predation. At short distances, however, competition for space and resources is likely to occur. In this study spatial patterns of barnacles (Semibalanus balanoides L.) were analysed using pair-correlation functions. Individuals were dispersed at distances below 0.30 cm, but peak relative density occurred at a distance of 0.36 cm from conspecifics. This is much closer than required for reproductive access, implying a strong aggregative drive, up to the point of physical contact with neighbours. Nevertheless, analysis of dead barnacles illustrated that such proximity carries a cost as barnacles with many neighbours were more likely to have died. The inferences obtained from these patterns are that barnacles aggregate as closely as they can, and that local neighbourhood competition is a powerful determinant of mortality. These processes give rise to the observed pattern properties.     

Antifouling activity of symbiotic bacteria from sponge Aplysina gerardogreeni

A key area in marine antifoulant research is the discovery of new environmentally friendly solutions that prevent biofilm formation and associated biocorrosion. Taking into consideration the natural mechanisms of marine organisms to protect against epibiosis, new biomimetic solutions can be utilised against biofouling, and marine bacteria are promising agents. Therefore, the goal of this study was to identify cultivable bacteria with antifouling (AF) activity associated with the sponge Aplysina gerardogreeni. A collection of 63 bacteria was isolated, and the organic extracts were assayed against various microfouler strains (16 bacteria and five microalgae). The results showed that 87% of bacterial extracts were active against the microfoulers tested. Sixteen of them can be considered to possess AF potential and belong to the genera Bacillus, Micrococcus, Paracoccus, Pseudobacter, Pseudovibrio, Psychrobacter, Staphylocuccus and Terribacillus. Bioactivity showed temporal variations; the highest activity was in February and June and the lowest in October. Bacillus bacteria were dominant and showed AF activities throughout the year. The results revealed those marine bacteria sponge-associated and the genus Bacillus in particular, are promising AF agents.     

Inbreeding shapes the evolution of marine invertebrates

Inbreeding is a potent evolutionary force shaping the distribution of genetic variation within and among populations of plants and animals. Yet, our understanding of the forces shaping the expression and evolution of nonrandom mating in general, and inbreeding in particular, remains remarkably incomplete. Most research on plant mating systems focuses on self-fertilization and its consequences for automatic selection, inbreeding depression, purging, and reproductive assurance, whereas studies of animal mating systems have often assumed that inbreeding is rare, and that natural selection favors traits that promote outbreeding. Given that many sessile and sedentary marine invertebrates and marine macroalgae share key life history features with seed plants (e.g., low mobility, modular construction, and the release of gametes into the environment), their mating systems may be similar. Here, we show that published estimates of inbreeding coefficients (F_IS) for sessile and sedentary marine organisms are similar and at least as high as noted in terrestrial seed plants. We also found that variation in F_IS within invertebrates is related to the potential to self-fertilize, disperse, and choose mates. The similarity of F_IS for these organismal groups suggests that inbreeding could play a larger role in the evolution of sessile and sedentary marine organisms than is currently recognized. Specifically, associations between traits of marine invertebrates and F_IS suggest that inbreeding could drive evolutionary transitions between hermaphroditism and separate sexes, direct development and multiphasic life cycles, and external and internal fertilization.     

Setal morphology and cirral setation of thoracican barnacle cirri: adaptations and implications for thoracican evolution

Thoracic cirripedes are sessile crustaceans that use six pairs of thoracic appendages (the cirri) to catch and handle food. We used scanning electron microscopy to examine the cirri, which include one to three pairs of maxillipeds in six species of thoracican barnacles, in search of correlations between cirral setation and feeding mode. The species studied comprise both pedunculate and sessile forms and represent a wide range of marine habitats as well as morphologies, viz., Ibla cumingi , Octolasmis warwickii , Capitulum mitella , Pollicipes polymerus , Tetraclita japonica japonica and Megabalanus volcano . Of the pedunculates, I. cumingi has the least complex setation pattern consisting of only serrulate types. This is consistent with its very simplified feeding mode and an apparent inability to discriminate between food items. Octolasmis warwickii is slightly more modified, while both P. polymerus and C. mitella have a more diversified setation. The balanomorphan species exhibit by far the most complex cirral setation. This is consistent with the several types of suspension feeding seen in these species, their ability to identify and sort captured food items and even to perform microfiltration in the mantle cavity using the setae on their three pairs of maxillipeds. Our results indicate that in thoracican barnacles, adaptations in feeding behaviour are associated with changes in the setation pattern of the cirri. In addition, the setal types and their distribution on the cirri are potential new characters in future morphology-based analyses of the phylogeny of the Cirripedia Thoracica.     

海洋真菌抗污损活性天然产物研究

黏附于海洋船舶或人工设施表面的污损生物给人类海洋生产活动与生态环境带来诸多不利影响.将具有抗污损活性的化合物开发成防污涂料是目前防治海洋生物污损的最常用手段之一.而大量传统有机金属防污剂因其严重毒副作用被禁用,亟须开发高效、环境友好型抗污损涂料.海洋真菌能够产生大量结构新颖、作用机制独特的高效、低毒/无毒抗污损活性次级...