Interspecific variation in patterns of adhesion of marine fouling to silicone surfaces

Abstract

The adhesion of six fouling organisms: the barnacle Balanus eburneus, the gastropod mollusc Crepidula fornicata, the bivalve molluscs Crassostrea virginica and Ostrea/Dendrostrea spp., and the serpulid tubeworms Hydroides dianthus and H. elegans, to 12 silicone fouling-release surfaces was examined. Removal stress (adhesion strength) varied among the fouling species and among the surfaces. Principal component analysis of the removal stress data revealed that the fouling species fell into two distinct groups, one comprising the bivalve molluscs and tubeworms, and the other the barnacle and the gastropod mollusc. None of the silicone materials generated a minimum in removal stress for all the organisms tested, although several surfaces produced low adhesion strengths for both groups of species. These results suggest that fouling-release materials do not rank (in terms of adhesion strength) identically for all fouling organisms, and thus development of a globally-effective hull coating will continue to require testing against a diversity of encrusting species.     

Sediment challenge to promising ultra-low fouling hydrophilic surfaces in the marine environment

Hydrophilic coatings exhibit ultra-low fouling properties in numerous laboratory experiments. In stark contrast, the antifouling effect of such coatings in vitro failed when performing field tests in the marine environment. The fouling release performance of nonionic and zwitterionic hydrophilic polymers was substantially reduced compared to the controlled laboratory environment. Microscopy and spectroscopy revealed that a large proportion of the accumulated material in field tests contains inorganic compounds and diatomaceous soil. Diatoms adhered to the accumulated material on the coating, but not to the pristine polymer. Simulating field tests in the laboratory using sediment samples collected from the test sites showed that incorporated sand and diatomaceous earth impairs the fouling release characteristics of the coatings. When exposed to marine sediment from multiple locations, particulate matter accumulated on these coatings and served as attachment points for diatom adhesion and enhanced fouling. Future developments of hydrophilic coatings should consider accumulated sediment and its potential impact on the antifouling performance.     

Recent developments in superhydrophobic surfaces and their relevance to marine fouling: a review

In this review, a brief synopsis of superhydrophobicity (i.e. extreme non-wettability) and its implications on marine fouling are presented. A short overview of wettability and recent experimental developments aimed at fabricating superhydrophobic surfaces by tailoring their chemical nature and physical appearance (i.e. substratum texture) are reviewed. The formation of responsive/"smart" surfaces, which adjust their physico-chemical properties to variations in some outside physical stimulus, including light, temperature, electric field, or solvent, is also described. Finally, implications of tailoring the surface chemistry, texture, and responsiveness of surfaces on the design of effective marine fouling coatings are considered and discussed.     

Recent developments in superhydrophobic surfaces and their relevance to marine fouling: a review

Abstract

In this review, a brief synopsis of superhydrophobicity (i.e. extreme non-wettability) and its implications on marine fouling are presented. A short overview of wettability and recent experimental developments aimed at fabricating superhydrophobic surfaces by tailoring their chemical nature and physical appearance (i.e. substratum texture) are reviewed. The formation of responsive/“smart” surfaces, which adjust their physico-chemical properties to variations in some outside physical stimulus, including light, temperature, electric field, or solvent, is also described. Finally, implications of tailoring the surface chemistry, texture, and responsiveness of surfaces on the design of effective marine fouling coatings are considered and discussed.     

Temporal and spatial variation in the fouling of silicone coatings in Pearl Harbor, Hawaii

An antifouling or foul-release coating cannot be globally effective if it does not perform well in a range of environmental conditions, against a diversity of fouling organisms. From 1996 to 1998, the field test sites participating in the United States Navy's Office of Naval Research 6.2 Biofouling program examined global variation in the performance of 3 silicone foul-release coatings, viz. GE RTV11, Dow Corning RTV 3140, and Intersleek (International Coatings Ltd), together with a control anticorrosive coating (Ameron Protective Coatings F-150 series). At the University of Hawaii's test site in Pearl Harbor, significant differences were observed among the coatings in the rate of accumulation of fouling. The control coating failed rapidly; after 180-220 d immersion a community dominated by molluscs and sponges developed that persisted for the remainder of the experiment. Fouling of the GE and Dow Corning silicone coatings was slower, but eventually reached a similar community structure and coverage as the control coatings. The Intersleek coating remained lightly fouled throughout the experiment. Spatial variation in the structure of the community fouling the coatings was observed, but not in the extent of fouling. The rate of accumulation of fouling reflected differences among the coatings in adhesion of the tubeworm Hydroides elegans. The surface properties of these coatings may have affected the rate of fouling and the structure of the fouling community through their influence on larval settlement and subsequent interactions with other residents, predators, and the physical environment.     

In situ autofluorescence detection of a fouling marine diatom on different surfaces

A new technique for quantifying fouling diatoms adhering to different surfaces was developed and tested. The method is based on recording the in vivo chlorophyll autofluorescence of diatom cells in situ. The enhanced signal obtained after addition of DCMU was used as a biomass estimate, and the enhancement itself as an indicator of the photosynthetic capacity. A fluorescence spectrometer equipped with a “well plate reader”; accessory was programmed to scan predetermined positions on the microscope slide based test surfaces. In standard tests, a matrix of 7 × 25 equidistant locations was applied to record the central 17 × 67 mm² area of the test surface. Thus, both spatial distributions and the mean value of chlorophyll associated with the specified area could be obtained directly. Microscopical counting was performed for calibration on transparent glass surfaces as well as PVA‐SbQ based hydrogels. There was a good correlation between counting and fluorescence recordings, with a linear range up to 1100 cells mm^?2. Due to the inherent inaccuracies of background estimates, the detection limit on glass and gel was approximately 200 cells mm^?2. The method was also applied successfully to test non‐transparent surfaces. In addition to standard mass screening of different test surfaces, the method may be found useful in studies of algal physiology related to cell adhesion, photosynthesis, growth, detachment and spatial migration.     

Interspecific variation in life history relates to antipredator decisions by marine mesopredators on temperate reefs

As upper-level predatory fishes become overfished, mesopredators rise to become the new 'top' predators of over-exploited marine communities. To gain insight into ensuing mechanisms that might alter indirect species interactions, we examined how behavioural responses to an upper-level predatory fish might differ between mesopredator species with different life histories. In rocky reefs of the northeast Pacific Ocean, adult lingcod (Ophiodon elongatus) are upper-level predators that use a sit-and-wait hunting mode. Reef mesopredators that are prey to adult lingcod include kelp greenling (Hexagrammos decagrammus), younger lingcod, copper rockfish (Sebastes caurinus) and quillback rockfish (S. maliger). Across these mesopredators species, longevity and age at maturity increases and, consequently, the annual proportion of lifetime reproductive output decreases in the order just listed. Therefore, we hypothesized that the level of risk taken to acquire resources would vary interspecifically in that same order. During field experiments we manipulated predation risk with a model adult lingcod and used fixed video cameras to quantify interactions between mesopredators and tethered prey (Pandalus shrimps). We predicted that the probabilities of inspecting and attacking tethered prey would rank from highest to lowest and the timing of these behaviours would rank from earliest to latest as follows: kelp greenling, lingcod, copper rockfish, and quillback rockfish. We also predicted that responses to the model lingcod, such as avoidance of interactions with tethered prey, would rank from weakest to strongest in the same order. Results were consistent with our predictions suggesting that, despite occupying similar trophic levels, longer-lived mesopredators with late maturity have stronger antipredator responses and therefore experience lower foraging rates in the presence of predators than mesopredators with faster life histories. The corollary is that the fishery removal of top predators, which relaxes predation risk, could potentially lead to stronger increases in foraging rates for mesopredators with slower life histories.     

Settlement patterns of species in a marine fouling community and some mechanisms of succession

The succession of fouling organisms that settled on asbestos plates immersed at Garden Island. Western Australia, was partially caused by the settlement patterns of the species involved. Both temporal variations in the abundance and continuity of settlement of each species and the age of the communities in which each species settled abundantly contributed to producing the succession. Additionally, temporal variations in the settlement of each species probably resulted in variability in the pattern of succession.Balanus and Spirorbis, the two earliest species in the succession, settled continuously and abundantly so that they immediately occupied a substantial area. The species which predominated later in the succession, Anomia, Ostrea, encrusting bryozoans, and Mytilus, settled discontinuously so that they were unlikely to settle immediately on a newly immersed surface. Additionally, Anomia, Ostrea, and the encrusting bryozoans did not settle abundantly and did not occupy a substantial area while small. Thus, there was also a delay between the time when these species settled and the time when they had grown sufficiently large to occupy a substantial area.Mytilus was the only species that settled abundantly in established communities and thus could persist indefinitely in a community. Settlement of the other species which occurred earlier in the succession was largely restricted to newly immersed plates and consequently these species were limited in their duration in the succession by the life-spans of the individual organisms.     

Site-specific variation in Antarctic marine biofilms established on artificial surfaces

The community structure and composition of marine microbial biofilms established on glass surfaces was investigated across three differentially contaminated Antarctic sites within McMurdo Sound. Diverse microbial communities were revealed at all sites using fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) techniques. Sequencing of excised DGGE bands demonstrated close affiliation with known psychrophiles or undescribed bacteria also recovered from the Antarctic environment. The majority of bacterial sequences were affiliated to the Gammaproteobacteria, Cytophaga/Flavobacteria of Bacteroidetes (CFB), Verrucomicrobia and Planctomycetales. Principal components analysis of quantitative FISH data revealed distinct differences in community composition between sites. Each of the sites were dominated by different bacterial groups: Alphaproteobacteria, Gammaproteobacteria and CFB at the least impacted site, Cape Armitage; green sulfur and sulfate reducing bacteria near the semi-impacted Scott Base and Planctomycetales and sulfate reducing bacteria near the highly impacted McMurdo Station. The highest abundance of archaea was detected near Scott Base (2.5% of total bacteria). Multivariate analyses (non-metric multidimensional scaling and analysis of similarities) of DGGE patterns revealed greater variability in community composition between sites than within sites. This is the first investigation of Antarctic biofilm structure and FISH results suggest that anthropogenic impacts may influence the complex composition of microbial communities.     

Interspecific variation in the response of fish to anthropogenic noise

1. Elevated levels of anthropogenic noise, especially those observed through boating activity, can negatively impact fish species, but it remains unclear which species are most affected and which behavioural metrics are best used in assessing fish responses to underwater noise. The effects of boat sounds on freshwater species are of particular interest because freshwater environments are less studied than the marine realm despite comparably high levels of biodiversity.
2. In the current study, we examine the behavioural responses to boat noise in two freshwater species that differ in their hypothesised response to sound inputs: the spottail shiner (Notropis hudsonius), a species with known hearing specialisations, and the bluegill sunfish (Lepomis macrochirus), a species with more generalised hearing capabilities. Fish were presented with boat noise in a laboratory setting, and their swimming, escape and foraging behaviours were assessed to examine differential responses in relation to hypothesised hearing abilities.
3. Both species showed a decrease in general swimming behaviours but an increase in erratic movements in response to boat noise, indicative of stress responses for both species. Despite the similarities in response based on swimming behaviours however, only spottail shiners exhibited true escape responses to the onset of the noise stimulus, suggesting a more extreme reaction in the species with a more refined hearing ability.
4. Taken together, these results show that freshwater fish can respond to increased levels of anthropogenic noise, but that the severity of the response may differ based on auditory structures and therefore presumed hearing ability. The differences seen between behavioural metrics used (swimming vs. escape responses) also demonstrate how care must be taken in choosing a metric when developing exposure guidelines for underwater sound exposures, as different metrics could lead to differential impact assessments.

     

Photoreversible surfaces to regulate cell adhesion

We report the development of a photoreversible cell culture substrate. We demonstrate the capacity to modify the adhesivity of the substrate using light, altering its capacity to support cell growth. Polyelectrolyte multilayers (PEMs) were used to produce tunable substrates of different thickness and matrix stiffness, which have different intrinsic capacities to support cell adhesion and survival. Surfaces were top-coated with a poly(acrylic acid)-poly(allylamine hydrochloride) polyelectrolyte bilayer functionalized with a small fraction (<1%) of an azobenzene-based photoswitchable sidegroup, which included the cell-adhesive three-amino-acid peptide RGD. Irradiation with light-induced geometric switching of the azo bond, resulting in changes to RGD exposure and consequently to cell adhesion and survival, was investigated on a variety of surfaces of different thickness and stiffness. Substrate stiffness, as modified by the thickness, had a significant influence on the adhesion of NIH 3T3 cells, consistent with previous studies. However, by disrupting the isomerization state of the azobenzene-linked RGD and exposing it to the surface, cell adhesion and survival could be enhanced up to 40% when the positioning of the RGD peptide was manipulated on the softest substrates. These findings identify permissive, yet less-than-optimal, cell culture substrate conditions that can be substantially enhanced using noninvasive modification of the substrate triggered by light. Indeed, where cell adhesion was tuned to be suboptimal under baseline conditions, the light-induced triggers displayed the most enhanced effect, and identification of this 'Goldilocks zone' was key to enabling light triggering.     

Characterization of EGF coupling to aminated silicone rubber surfaces

Tethering of growth factors to biomaterial substrates via a polyethylene glycol (PEG) spacer has been established as a means of controlling dosage and conformation of the protein at the material surface, while retaining biological activity. However, the extent of modification through a comparison of bound versus unbound protein has not generally been characterized. In this work, covalent tethering of epidermal growth factor (EGF) to allylamine plasma modified polydimethylsiloxane (PDMS) substrates is characterized to determine the nature of the bound growth factor and to optimize the conditions for the reaction. Tethering is achieved via conjugation of EGF with homobifunctional N-hydroxysuccinimide (NHS) ester of PEG-butanoic acid (SBA2-PEG) in solution, followed by exposure of the pegylated EGF to the aminated surfaces (solution first reaction). SDS-PAGE analysis indicates that a low ratio of EGF:PEG is required to maximize the yield of the EGF-PEG reaction; a relatively short reaction time is needed to limit hydrolysis of the NHS ester. With increasing amounts of PEG and a higher reaction time, a higher fraction of the EGF can be covalently tethered to the surfaces, as shown by binding of 125I-labeled EGF and subsequent washing with sodium dodecyl sulfate (SDS) to remove adsorbed protein. However, even under the optimal reaction conditions established by the SDS-PAGE analysis, higher molecular weight EGF-PEG complexes are observed by SDS-PAGE and matrix-assisted laser desorption/ionization (MALDI). The presence of these complexes, as well as unreacted growth factor, can lead to a surface of heterogeneous composition. While these surfaces were found to have biological activity, stimulating the adhesion and growth of corneal epithelial cells versus PDMS controls, further optimization of reaction conditions, including the use of a homobifunctional PEG linker and possibly separation of reaction species are required to achieve a uniformly active and well-defined biomaterial surface.     

Interspecific and temporal variation in herbivore responses to hybrid willows

We studied herbivory of two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids to test alternative hypotheses concerning the effects of hybridization on plant resistance. Individually marked plants were identified using morphological traits in the field and random amplified polymorphic DNA (RAPD) band analysis was used to verify the genetic status of many parental and hybrid plants. The desities of 12 herbivore species on plants in the field were compared between two parents and their F₂-type hybrids. We found about equal support for the additive, dominance, and hybrid susceptibility hypotheses over 4 years. In one year, one species supported the hybrid resistance hypothesis. Guild membership was not a good predictor of similar responses of species to hybrid versus parental plants. There were marked differences in support for particular hypotheses among years for four herbivore species. This study demonstrates the diversity of responses of phytophages in response to interspecific hybridization, and indicates that year-to-year variation in relative resistance of hybrid plants can be important.     

Interspecific interactions in the marine rotifer microcosm

Copepods and protozoans often co-exist in marinerotifer mass cultures. Interspecific interactionbetween the rotifer Brachionus rotundiformisTschugunoff and eight other zooplankton species,namely Brachionus plicatilis O. F. Müller (rotifer),Diaphanosoma celebensis Stingelin (cladoceran),Tigriopus japonicus Mori, Acartia sp. (copepod),Euplotes sp., Vorticella sp., an unidentifiedprotozoan species (P1 strain) (protozoan) and Artemiasp. (anostracan) at two developmental stages (nauplii –0.95 mm, 0 days old; adults – 3.3 mm, 19 days old) wereinvestigated in the laboratory.There was no contaminating species that contributedto an increase in rotifer population growth during theexperiments. Four types of interspecific interactionswere seen between B. rotundiformis and otherco-existing zooplankton species. These include effectson population growth: (1) both species declined, (2) onespecies is promoted while the other is not influenced,(3) one species is declined while the other is notinfluenced and (4) one species is promoted while theother declined. The first type was exhibited by B. rotundiformis vs B. plicatilis, B. rotundiformis vsD. celebensis and B. rotundiformis vs Artemia sp. The second type was exhibited by B. rotundiformis vsVorticella sp. and the third type by B. rotundiformisvs Euplotes sp. and B. rotundiformis vs T. japonicus. The fourth type was exhibited by B. rotundiformis vs Acartia sp. and B. rotundiformis vs P1 strain.     

Novel whole cell adhesion assays of three isolates of the fouling diatom Amphora coffeaeformis reveal diverse responses to surfaces of different wettability

Whole cell, strength of adhesion assays of three different isolates of the fouling diatom Amphora coffeaeformis were compared using a hydrophilic surface viz. acid washed glass (AWG), and a hydrophobic surface viz. a self assembled monolayer (SAM) of undecanethiol (UDT). Assays were performed using a newly designed turbulent flow channel that permits direct observation and recording of cell populations on a test surface. Exposure to continuous shear stress over 3?h revealed that the more motile isolate, WIL2, adhered much more strongly to both test surfaces compared to the other two strains. When the response of the isolates to shear stress after 3?h was compared, there was no significant difference in the percentage of cells removed, irrespective of surface wettability. Cells of the three isolates of A. coffeaeformis varied significantly in their response to different surfaces during initial adhesion, indicating the presence of a wide range of 'physiological races' within this species.     

Novel whole cell adhesion assays of three isolates of the fouling diatom Amphora coffeaeformis reveal diverse responses to surfaces of different wettability

Whole cell, strength of adhesion assays of three different isolates of the fouling diatom Amphora coffeaeformis were compared using a hydrophilic surface viz. acid washed glass (AWG), and a hydrophobic surface viz. a self assembled monolayer (SAM) of undecanethiol (UDT). Assays were performed using a newly designed turbulent flow channel that permits direct observation and recording of cell populations on a test surface. Exposure to continuous shear stress over 3 h revealed that the more motile isolate, WIL2, adhered much more strongly to both test surfaces compared to the other two strains. When the response of the isolates to shear stress after 3 h was compared, there was no significant difference in the percentage of cells removed, irrespective of surface wettability. Cells of the three isolates of A. coffeaeformis varied significantly in their response to different surfaces during initial adhesion, indicating the presence of a wide range of ‘physiological races’ within this species.     

Latitudinal variation in diet and patterns of human interaction in the marine otter

The marine otter (Lontra felina) inhabits patches of rocky coastline from central Peru to southern Chile and is classified as Endangered by the IUCN. Given the limited information available about the species, we set out to assess marine otter diet with a view to detecting latitudinal differences, and to assess marine otter activity budgets and interspecific interactions (including anthropogenic) at Peruvian fishing villages and to compare results with similar Chilean studies. Nine study sites from central Chile to southern Peru were sampled for otter spraints to assess relative frequency of prey types and two fishing ports in southern Peru were monitored through focal and scan observations to assess activity patterns, interspecific interactions, habitat use patterns, and dive durations. Results indicate that toward the northern part of its range, crustaceans become less important and fish more important in the diet. Interactions were observed between marine otters and other species, including stray dogs and cats. The strong dependence of marine otters on the availability of safe rocky shelters, and the species’ apparent tolerance to living alongside humans raise conservation concerns about vulnerability to anthropogenic threats. These factors, if not correctly managed, could turn some of these rocky seashore patches into population sinks.     

Interspecific variation in plant responses to mycorrhizal colonization in tallgrass prairie

Symbiotic associations between plants and arbuscular mycorrhizal fungi are ubiquitous and ecologically important in many grasslands. Differences in species responses to mycorrhizal colonization can have a significant influence on plant community structure. The growth responses of 36 species of warm- and cool-season tallgrass prairie grasses and 59 tallgrass prairie forbs to arbuscular mycorrhizal (AM) fungal colonization were assessed in greenhouse studies to examine the extent of interspecific variation in host-plant benefit from the symbiosis and patterns of mycorrhizal dependence among host plant life history (e.g., annual, perennial) and taxonomic (e.g., grass, forb, legume, nonlegume) groups and phenological guilds. There was a strong and significant relationship between phenology of prairie grasses and mycorrhizal responsiveness, however this relationship was less apparent in forbs. Perennial warm-season C(4) grasses and forbs generally benefited significantly from the mycorrhizal symbiosis, whereas biomass production of the cool-season C(3) grasses was not affected. The root systems of the cool-season grasses were also less highly colonized by the AM fungi, as compared to the warm-season grasses or forbs. Unlike the native perennials, annuals were generally not responsive to mycorrhizal colonization and were lower in percentage root colonization than the perennial species. Plant growth responsiveness and AM root colonization were positively correlated for the nonleguminous species, with this relationship being strongest for the cool-season grasses. In contrast, root colonization of prairie legumes showed a significant, but negative, relationship to mycorrhizal growth responsiveness.