Exploratory behaviour of barnacle larvae in field conditions

The aim of this work was to develop techniques for the real time filming of Semibalanus balanoides cyprid larvae in the field in order to describe the exploratory behaviour of S. balanoides cyprids under natural field conditions. The underwater camera system consisted of a high resolution, remotely controlled colour camera attached to a cradle which could be deployed from a pier. A light utilising a battery of far‐red light‐emitting diodes (λ_min = 615 nm) was used for filming at night. Transparent acrylic treated with barnacle settlement factor and smooth green polyester tiles were used as targets. From the films the tracks of the exploring cyprids were digitised from the 30 min trials. During five, 30 min trials a total of 1014 cyprids explored the surface. The numbers of cyprids exploring the surface varied from 49 to 522 cyprids per trial, and a Monte‐Carlo randomisation model showed that larval supply was random or aggregated. The mean exploration time spent on the target was 163 s with a mean distance travelled on the target of 8.5 cm. Exploratory track length was related to water current velocity by a negative curvilinear regression (R² = 0.92, F = 45.65, p < 0.05). There were significant differences in exploratory behaviour between day and night for total distance travelled (p < 0.001), straight line distance (p < 0.001) and velocity (p < 0.001) and the mean heading angle of the exploratory track (p < 0.01). The results are discussed in relation to settlement patterns and the screening of antifouling surfaces.     

Field-based video observations of wild barnacle cyprid behaviour in response to textural and chemical settlement cues

Many marine invertebrate larvae respond behaviourally to environmental settlement cues, yet behaviours are often only inferred from settlement patterns or are limited to laboratory studies. The behaviour of wild cypris larvae of Semibalanus balanoides L. was filmed on settlement tiles in the field. Tiles were of five different textures with a nested treatment of crude conspecific adult extract (AE). The effects of texture and AE on eleven defined behaviours were analysed. Texture affected the gross and net exploratory distances, velocity, acceleration and time spent exploring. AE attracted more cyprids during the first minute of immersion and increased the time spent on surfaces. Relatively few arrivals that either travel far and fast, or exit the surface rapidly, may indicate a lower chance of settlement. An increase in time spent on a surface may increase the probability of being in contact with the surface when the sign stimulus to settle occurs.     

Selective settlement of the barnacle Semibalanus balanoides (L.) facilitates its growth and reproduction on mussel beds in the Wadden Sea

On the unstable sedimentary tidal flats of the Wadden Sea, a suitable attachment substrate for sessile organisms is generally lacking. Epibenthic mussel beds (Mytilus edulis L.) provide the only and strongly limited settlement sites available for the barnacle, Semibalanus balanoides (L.). Field investigations showed that barnacles were non-randomly distributed within a mussel bed. They preferentially occurred near the siphonal apertures of living mussels but rarely grew on dead mussels or shell fragments. Field experiments revealed that this was due to selective settlement of barnacle cyprid larvae. Growth of barnacles was significantly higher upon living mussels than on empty mussel shells. Moreover, a higher reproductive output was obtained by individuals on living mussels which produced twice as many nauplii larvae than barnacles attached to empty shells. This study shows that selective settlement of S. balanoides cyprid larvae on living mussels is adaptive with respect to individual fitness. Received in revised form: 15 January 2001 Electronic Publication     

Adaptive maintenance of genetic polymorphism in an intertidal barnacle: habitat- and life-stage-specific survivorship of Mpi genotypes

In the northern acorn barnacle, Semibalanus balanoides, genotype frequencies of three genetic markers were tracked over time in four types of intertidal habitats. These habitats were selected to represent natural variation in several environmental parameters, specifically the degree of physical stress experienced by barnacles. Frequencies for one allozyme locus (Gpi) and a presumably neutral mtDNA marker were homogeneous among habitats in each temporal sample. Similarly, no temporal stratification in genotype frequencies was evident across the five sampling intervals: from planktonic larvae sampled in March to juveniles collected at the end of June. In contrast to the Gpi and mtDNA loci, Mpi genotypes significantly changed in frequency in two habitats in the high intertidal zone. On exposed substrate, the Mpi-FF homozygote increased in frequency, whereas the alternative homozygote, Mpi-SS, significantly decreased in frequency. Barnacles that were protected from environmental stress at high intertidal heights by the Ascophyllum nodosum algal canopy demonstrated the opposite pattern. In both habitats, the change in frequency of the heterozygote was intermediate to that of the homozygous genotypes. Furthermore, these patterns of genotype-by-environment association reflected a pulse of genotype-specific mortality that occurred over a two-week interval subsequent to metamorphosis from the larval to the adult form. These data indicate that each Mpi homozygote is the highest fitness genotype in some portion of the intertidal environment. Using the Levene (1953) model to evaluate the spatial variation in genotypic fitness, the stable maintenance of the Mpi polymorphism is predicted under certain subsets of conditions. Environmental heterogeneity in the intertidal zone translates to spatial variation in selection pressures, which may result in the active maintenance of the Mpi polymorphism in this species.     

Climate change and historical biogeography of the barnacle Semibalanus balanoides

Aim Marine sessile inter‐tidal invertebrates are expected to undergo shifts in distribution due to climate change. Using a combination of survey and transplant data with thermal modelling, we investigated the role of climate on the poleward contraction of the southern range edge of the north temperate barnacle Semibalanus balanoides. Location Western Atlantic of the United States. Methods Barnacle surveys were conducted along the east coast of the United States in 1963 and 2007. Presence, absence and abundance data were collected and the time periods were compared. Transplant experiments monitoring survival with relation to temperature were conducted upon S. balanoides along the more southerly portion of their range, and modelling predicting barnacle survival with relation to biogeography was completed. Results The southern limit of S. balanoides has contracted approximately 350 km to the north. Main conclusions The changes thus far observed in climate along the east coast of the United States have contributed to the southern limit range contraction of S. balanoides. Further changes in the biogeography of S. balanoides are expected with continued climate warming.     

Soil organic matter turnover is governed by accessibility not recalcitrance

Mechanisms to mitigate global climate change by sequestering carbon (C) in different ‘sinks' have been proposed as at least temporary measures. Of the major global C pools, terrestrial ecosystems hold the potential to capture and store substantially increased volumes of C in soil organic matter (SOM) through changes in management that are also of benefit to the multitude of ecosystem services that soils provide. This potential can only be realized by determining the amount of SOM stored in soils now, with subsequent quantification of how this is affected by management strategies intended to increase SOM concentrations, and used in soil C models for the prediction of the roles of soils in future climate change. An apparently obvious method to increase C stocks in soils is to augment the soil C pools with the longest mean residence times (MRT). Computer simulation models of soil C dynamics, e.g. RothC and Century, partition these refractory constituents into slow and passive pools with MRTs of centuries to millennia. This partitioning is assumed to reflect: (i) the average biomolecular properties of SOM in the pools with reference to their source in plant litter, (ii) the accessibility of the SOM to decomposer organisms or catalytic enzymes, or (iii) constraints imposed on decomposition by environmental conditions, including soil moisture and temperature. However, contemporary analytical approaches suggest that the chemical composition of these pools is not necessarily predictable because, despite considerable progress with understanding decomposition processes and the role of decomposer organisms, along with refinements in simulation models, little progress has been made in reconciling biochemical properties with the kinetically defined pools. In this review, we will explore how advances in quantitative analytical techniques have redefined the new understanding of SOM dynamics and how this is affecting the development and application of new modelling approaches to soil C.     

Possible molecular mechanisms of species recognition by barnacle larvae inferred from multi-specific sequencing analysis of proteinaceous settlement-inducing pheromone

Gregarious settlement is essential for reproduction and survival of many barnacles. A glycoprotein, settlement-inducing protein complex (SIPC) has been recognized as a signal for settlement and it is expressed in both conspecific adults and larvae. Although the settlement-inducing activities of SIPC are species-specific, the molecular-based mechanism by which larvae distinguish conspecific SIPC from the SIPC of other species is still unknown. Here, the complete primary structure of the SIPC of Megabalanus coccopoma, as well as the partial structure of the SIPCs of Balanus improvisus, Megabalanus rosa, and Elminius modestus are reported. These SIPCs contain highly variable regions that possibly modulate the affinity for the receptor, resulting in the species specificity of SIPC. In addition, the distribution patterns of potential N-glycosylation sites were seen to be different among the various species. Differences in such post-translational modifications may contribute to the species specificity of SIPC.     

Proteomic analysis of larvae during development, attachment, and metamorphosis in the fouling barnacle, Balanus amphitrite

The barnacle, Balanus amphitrite, is one of the primary model organisms for rocky-shore ecology studies and biofouling research. This barnacle species has a complex life cycle during which the swimming nauplius molts six times and transforms into a cyprid stage. Cyprids must attach to a surface to metamorphose into a juvenile barnacle. To clarify the overall profile of protein expression during larval development and metamorphosis, 2-DE was used to compare the proteome of the nauplius, the swimming cyprid, the attached cyprid, and the metamorphosed cyprid. The proteome of the swimming cyprid was distinctly different from that of other life stages and had about 400 spots. The proteomes of the attached and metamorphosed cyprids were similar with respect to major proteins but had significantly lower numbers of spots compared to that of swimming larval stages. Obviously, synthesis of most proteins from swimming cyprids was switched off after attachment and metamorphosis. Our advanced MS analysis (MALDI-TOF/TOF MS/MS) allowed us to identify the proteins that were differentially and abundantly expressed in the swimming cyprid. These proteins included signal transduction proteins (adenylate cyclase and calmodulin) and juvenile hormone binding proteins. In summary, for the first time, we have analyzed the global protein expression pattern of fouling marine invertebrate larvae during metamorphosis. Our study provides new insights into the mechanisms of barnacle larval metamorphosis and also provides a foundation for exploring novel targets for antifouling treatments.     

Bacterial colonisation and settlement of algal spores and barnacle larvae on low surface energy materials

Melt‐coated films of poly(fluoroalkylacrylate)s and poly(fluoroalkylmethacrylate)s have been exposed respectively to bacterial cultures (Pseudomonas spp., Alteromonas sp. NCIMB 1534 and Desulphovibrio alaskensis NCIMB 13491), Enteromorpha zoospores and cyprid larvae of Balanus amphitrite Darwin. In all experiments, settlement and development was considerably less on the films than on controls of glass, poly‐(methylmethacrylate), poly(tetrafluoroethene) (ptfe) and polyester/glassf ibre composite. The settled bacteria were also much more weakly attached to the fluoropo‐lymer films than to the controls. Enteromorpha zoospores tended to settle at surface faults/cracks and also showed some sensitivity to the fluorine content of the materials. Cyprids did not settle on ptfe and could find only occasional sights for attachment on the films. Although the fluoropolymers show promise as fouling‐resistant coatings, improvements in film quality are required.     

Evaluating the occurrence of cryptic invasions of a rocky shore barnacle,Semibalanus cariosus,between the north-eastern Pacific and Japan

Many coastal barnacles are introduced to non-native regions. However, data are lacking on cryptic invasion, which is defined as an invasion that remains unrecognised because the invader is mistaken for a native or previously introduced species or clade. In this work, cryptic invasions of an intertidal barnacle, Semibalanus cariosus, between Japan and the north-eastern Pacific were evaluated based on population genetic analyses. A significant genetic differentiation was found between the Japanese and north-eastern Pacific populations, suggesting a limited introduction of non-native genotypes between these regions. Haplotype frequencies did not differ significantly between the past (museum samples collected in 1971 from Hokkaido, Japan) and present Japanese populations, implying the rare occurrence of human-mediated migration from the north-eastern Pacific to Japan. Migrate-n analysis revealed a low level of directional gene flow in S. cariosus from the north-eastern Pacific to Japan, possibly by natural stepping-stone dispersal via directional water currents or human-mediated transport.     

On the limits of fractal surface behaviour in silica. A virtual adsorbates simulation

A computer simulation technique, suited to replicate real adsorption experiments, was applied to pure simulated silica in order to gain insight into the fractal regime of its surface. The previously reported experimental fractal dimension was closely approached and the hitherto uncharted lower limit of fractal surface behaviour is reported herein.     

Effects of surface charge and Gibbs surface energy on the settlement behaviour of barnacle cyprids (Balanus amphitrite)

Gibbs surface energy has long been considered to be an important parameter in the design of fouling-resistant surfaces for marine applications. Rigorous testing of the hypothesis that settlement is related to Gibbs surface energy however has never been accomplished, due mainly to practical limitations imposed by the necessary combination of surface engineering and biological evaluation methods. In this article, the effects of surface charge and Gibbs surface energy on the settlement of cyprids of an important fouling barnacle, Balanus amphitrite, were evaluated. Settlement assays were conducted on a range of self-assembled monolayers (SAMs) (CH₃-, OH-, COOH-, N(CH₃)₃ ⁺-, NH₂-terminated), presented in gold-coated polystyrene well plates, varying in terms of their surface charge and Gibbs surface energy. Contrary to contemporary theory, settlement was not increased by high-energy surfaces, rather the opposite was found to be the case with cyprids settling in greater numbers on a low-energy CH₃- SAM compared to a high-energy OH- SAM. Settlement was also greater on negatively-charged SAMs, compared to neutral and positively-charged SAMs. These findings are discussed in the context of data drawn from surfaces that varied in multiple characteristics simultaneously, as have been used previously for such experiments. The finding that surface charge, rather than total surface energy, may be responsible for surface selection by cyprids, will have significant implications for the design of future fouling-resistant materials.     

The complex barnacle perfume: identification of waterborne pheromone homologues in Balanus improvisus and their differential expression during settlement

A key question in barnacle biology is the nature of cues that induce gregarious settlement. One of the characterised cues is the waterborne settlement pheromone (WSP). This study aimed to identify WSP homologues in Balanus improvisus and to investigate their expression during settlement. Six WSP homologues were identified, all containing an N-terminal signal peptide, a conserved core region, and a variable C-terminus comprising several -GR- and -HDDH- motifs. The B. improvisus WSP homologues were expressed in all settlement stages but showed different expression patterns. The homologue most similar to the B. amphitrite WSP was the most abundant and was constantly expressed during settlement. In contrast, several of the other WSP homologues showed the greatest expression in the juvenile stage. The presence of several WSP homologues suggests the existence of a pheromone mix, where con-specificity might be determined by a combination of sequence characteristics and the concentration of the individual components.     

线型链支化链网状链生物大分子的分维测量

本文应用光散射技术研究了几种生物大分子在水溶液中的分形性质。当温度高于胶凝临界温度时,直链淀粉（线型链）、明胶（线圈状链）的分维ｄｆ为５／３,支链淀粉（支化链）的分维为２．２￣２．５。在直链淀粉及明胶水溶液胶凝点（网状链）ｄｆ＝２．０。文章还讨论了ｄｆ与样品温度、浓度的关系。     

Enhancing the settlement and attachment strength of pediveligers of Mytilus galloprovincialis bychanging surface wettability and microtopography

Surface wettability and microtopography can either enhance or deter larval settlement of many sessile marine organisms. This study quantifies the effect of these surface properties on the settlement of pediveligers of Mytilus galloprovincialis, using polymers spanning a range of wettability and microtextured polydimethylsiloxane (PDMS). Furthermore, the adhesion strength of settled pediveligers on microtextured PDMS surfaces was quantified using a flow chamber. Settlement was enhanced at the hydrophilic end of the wettability spectrum, where mean settlement on nylon reached 33.5 ± 13.1%. In contrast, mean settlement on the most hydrophobic polymer (PDMS) was 4.2 ± 3.2%. Microtopography had a much stronger effect compared to wettability, where 400 μm textured PDMS enhanced settlement above 90%. Settlement preferences were also positively correlated to adhesion strength at flow rates of 4 knots, with all initially settled pediveligers on smooth PDMS detaching, while 79.9 ± 5.7% of pediveligers remained on the 400 μm texture.     

Correlative assays of barnacle cyprid behaviour for the laboratory evaluation of antifouling coatings: a study of surface energy components

Laboratory evaluation of antifouling coatings is underpinned by settlement studies with specific fouling organisms. Established methods provide insight into the likelihood of failure of a particular coating system, but can neglect the process of surface selection that often precedes attachment. The present approach for quantifying the exploratory behaviour of barnacle cypris larvae suggested that inspection behaviour can be a rapid and predictive proxy for settlement. Two series of xerogels with comparable total surface energy, but different dispersive and polar components, were evaluated. Settlement assays with three-day-old cyprids of Balanus improvisus demonstrated that while attachment was not linked directly to dispersive free energy, the composition of the xerogel was nevertheless significant. Behavioural analysis provided insight into the mechanism of surface rejection. In the case of a 50:50?PH/TEOS (phenyltriethoxysilane-based) xerogel vs a 50:50 TFP/TEOS (3,3,3-trifluoropropyltrimethoxysilane-based) xerogel, wide-searching behaviour was absent on the former.     

Host location by larvae of a parasitic barnacle: larval chemotaxis and plume tracking in flow

Numerous studies describe stimulation and/or enhancement oflarval settlement by distance chemoreception in response tochemical factors emitted by conspecific adults, host and preyspecies and microbial films. However, active upstream trackingof odor plumes, needed in order to locate specific, spatiallylimited settlement sites, has thus far received little scientificattention. This study examines host location in flow and stillwater by larvae of the parasitic barnacle Heterosaccus dollfusi,which inhabits the brachyuran crab Charybdis longicollis. Experimentsincluded analysis of larval motion patterns under four conditions:still water, in flow, in still water with waterborne host metabolitesand in flow with host metabolites. Our results show that H.dollfusi larvae are capable of actively and effectively locatingtheir host in still water and in flow, using chemotaxis andrheotaxis and modifying their swimming pattern, direction, velocity,determination and turning rate to accommodate efficient navigationin changing environmental conditions.