Sessile Crustaceans Research Articles

Gene co-option, duplication and divergence of cement proteins underpin the evolution of bioadhesives across barnacle life histories.

Acquisition of new genes often results in the emergence of novel functions and is a key step in lineage-specific adaptation. As a group of sessile crustaceans, barnacles establish permanent attachment through initial cement secretion at the larval phase followed by continuous cement secretion in juveniles and adults. However, the origins and evolution of barnacle larval and adult cement proteins remain poorly understood. By performing microdissection of larval cement glands, transcriptome and shotgun proteomics and immunohistochemistry validation, we identified 30 larval and 27 adult cement proteins of the epibiotic turtle barnacle Chelonibia testudinaria, of which the majority are stage- and barnacle-specific. While only two proteins, SIPC and CP100K, were expressed in both larvae and adults, detection of protease inhibitors and the cross-linking enzyme lysyl oxidase paralogs in larvae and adult cement. Other barnacle-specific cement proteins such as CP100k and CP52k likely share a common origin dating back at least to the divergence of Rhizocephala and Thoracica. Different CP52k paralogues could be detected in larval and adult cement, suggesting stage-specific cement proteins may arise from duplication followed by changes in expression timing of the duplicates. Interestingly, the biochemical properties of larval- and adult-specific CP52k paralogues exhibited remarkable differences. We conclude that barnacle larval and adult cement systems evolved independently, and both emerged from co-option of existing genes and de novo formation, duplication and functional divergence of lineage-specific cement protein genes. Our findings provide important insights into the evolutionary mechanisms of bioadhesives in sessile marine invertebrates.

Structure and Composition of the Cuticle of the Goose Barnacle Pollicipes pollicipes: A Flexible Composite Biomaterial.

Arthropods, the largest animal phylum, including insects, spiders and crustaceans, are characterized by their bodies being covered primarily in chitin. Besides being a source of this biopolymer, crustaceans have also attracted attention from biotechnology given their cuticles' remarkable and diverse mechanical properties. The goose barnacle, Pollicipes pollicipes, is a sessile crustacean characterized by their body parts covered with calcified plates and a peduncle attached to a substrate covered with a cuticle. In this work, the composition and structure of these plates and cuticle were characterized. The morphology of the tergum plate revealed a compact homogeneous structure of calcium carbonate, a typical composition among marine invertebrate hard structures. The cuticle consisted of an outer zone covered with scales and an inner homogenous zone, predominantly organic, composed of successive layers parallel to the surface. The scales are similar to the tergum plate and are arranged in parallel and oriented semi-vertically. Structural and biochemical characterization confirmed a bulk composition of ɑ-chitin and suggested the presence of elastin-based proteins and collagen. The mechanical properties of the cuticle showed that the stiffness values are within the range of values described in elastomers and soft crustacean cuticles resulting from molting. The removal of calcified components exposed round holes, detailed the structure of the lamina, and changed the protein properties, increasing the rigidity of the material. This flexible cuticle, predominantly inorganic, can provide bioinspiration for developing biocompatible and mechanically suitable biomaterials for diverse applications, including in tissue engineering approaches.

Histology and transcriptomic analyses of barnacles with different base materials and habitats shed lights on the duplication and chemical diversification of barnacle cement proteins

BackgroundBarnacles are sessile crustaceans that attach to underwater surfaces using barnacle cement proteins. Barnacles have a calcareous or chitinous membranous base, and their substratum varies from biotic (e.g. corals/sponges) to abiotic surfaces. In this study, we tested the hypothesis that the cement protein (CP) composition and chemical properties of different species vary according to the attachment substrate and/or the basal structure. We examined the histological structure of cement glands and explored the variations in cement protein homologs of 12 barnacle species with different attachment habitats and base materials.ResultsCement gland cells in the rocky shore barnacles Tetraclita japonica formosana and Amphibalanus amphitrite are eosinophilic, while others are basophilic. Transcriptome analyses recovered CP homologs from all species except the scleractinian coral barnacle Galkinia sp. A phylogenomic analysis based on sequences of CP homologs did not reflect a clear phylogenetic pattern in attachment substrates. In some species, certain CPs have a remarkable number of paralogous sequences, suggesting that major duplication events occurred in CP genes. The examined CPs across taxa show consistent bias toward particular sets of amino acid. However, the predicted isoelectric point (pI) and hydropathy are highly divergent. In some species, conserved regions are highly repetitive.ConclusionsInstead of developing specific cement proteins for different attachment substrata, barnacles attached to different substrata rely on a highly duplicated cementation genetic toolkit to generate paralogous CP sequences with diverse chemical and biochemical properties. This general CP cocktail might be the key genetic feature enabling barnacles to adapt to a wide variety of substrata.

Trace elements (Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) in Mytilus galloprovincialis and Tapes decussatus from Faro and Ganzirri Lakes (Sicily, Italy): Flow cytometry applied for hemocytes analysis

BackgroundTrace elements present in sessile molluscs, are important because they are used in human consumption and it has significantly increased in recent years. While their filtering of the water can lead to their build-up of organic and inorganic materials that can be sampled and analyzed, this can also lead to bioaccumulation of harmful substances, such as essential and non-essential elements, that can harm the human health if in taken in high concentrations or for a long period of time. MethodsIn the present study, the trace metal content (Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) of two sessile crustaceans, 20 Mytilus galloprovincialis (mussel) and 20 Tapes decussatus (clam) in Faro and Ganzirri Lakes (Messina, Sicily, Italy) were analyzed. Haemolymph samples were taken on both molluscs in order to analyze the haemocyte population by flow cytometric analysis. Unpaired t-tests were used to determine significant differences for the essential and non-essential metallic elements concentrations in the lake waters and in the tissues of M. galloprovincialis and T. decussatus and for hemocyte populations R1 (halinocytes) and R2 (granulocytes). ResultsThe results suggested that that in Faro Lake, the tissue Al, Cr and Pb levels in M. galloprovincialis were higher than those for T. decussatus, in contrast to Mn, Fe, Ni, Cu, Zn and Cd, which were higher in T. decussatus. Unpaired t-tests showed that there were significantly higher proportions of halinocytes in M. galloprovincialis versus T. decussatus for both Faro Lake (41.8 % vs. 24.3 %; P < 0.001) and Ganzirri Lake (43.0 % vs. 22.4 %; P < 0.001). In contrast, while there were significantly higher proportions of granulocytes in Faro Lake (21.2 % vs. 9.1 %; P < 0.001), this difference was not seen for the granulocytes of M. galloprovincialis versus T. decussatus in Ganzirri Lake (9.6 % vs. 13.0 %). ConclusionThis study shows that M. galloprovincialis and T. decussatus can indeed bioaccumulate some of these metal, such that activation of the immune responses is specific to certain cell types. Future research must focus on the balance of trace elements in the consumption of these shellfish, and analyzes with more sophisticated tools can be used to diagnose the increased concentration of trace elements and the quantification of trace metals from shellfish to clams.

Slow crabs - fast genomes: Locomotory capacity predicts skew magnitude in crustacean mitogenomes.

Base composition skews (G-C/G+C) of mitochondrial genomes are believed to be primarily driven by mutational pressure, which is positively correlated with metabolic rate. In marine animals, metabolic rate is also positively correlated with locomotory capacity. Given the central role of mitochondria in energy metabolism, we hypothesised that selection for locomotory capacity should be positively correlated with the strength of purifying selection (dN/dS), and thus be negatively correlated with the skew magnitude. Therefore, these two models assume diametrically opposite associations between the metabolic rate and skew magnitude: positive correlation in the prevailing paradigm, and negative in our working hypothesis. We examined correlations between the skew magnitude, metabolic rate, locomotory capacity, and several other variables previously associated with mitochondrial evolution on 287 crustacean mitogenomes. Weakly locomotory taxa had higher skew magnitude and ω (dN/dS) values, but not the gene order rearrangement rate. Skew and ω magnitudes were correlated. Multilevel regression analyses indicated that three competing variables, body size, gene order rearrangement rate, and effective population size, had negligible impacts on the skew magnitude. In most crustacean lineages selection for locomotory capacity appears to be the primary factor determining the skew magnitude. Contrary to the prevailing paradigm, this implies that adaptive selection outweighs nonadaptive selection (mutation pressure) in crustaceans. However, we found indications that effective population size (nonadaptive factor) may outweigh the impact of locomotory capacity in sessile crustaceans (Thecostraca). In conclusion, skew magnitude is a product of the interplay between adaptive and nonadaptive factors, the balance of which varies among lineages.

Capturing shell-crushing by large mobile predators using passive acoustics technology

Durophagy (shell-crushing) is a predatory mode that has evolved across numerous marine taxa from relatively sessile crustaceans to large and highly mobile fishes and mammals. Despite its preponderance in the marine environment, the ecology of durophagy (i.e., dynamics and spatiotemporal distribution) remains poorly understood especially for highly mobile species, limiting our ability to quantify their predatory effects on benthic communities. Here, we used passive acoustics (i.e., remote monitoring of underwater sounds with acoustic recorders) to characterize consumption of hard-shelled mollusk prey by a model predator, the whitespotted eagle ray (Aetobatus narinari). Acoustic recordings were made in captivity for 434 total prey items, spanning eight species of hard-shelled mollusks (1 bivalve, 7 gastropods). For all prey types, consumption sequences were generally characterized by an initial high-energy signal (Sound Pressure Level > 160 dB re 1μPa), presumably associated with shell failure, followed by numerous additional signals of lesser energy as prey was further fractured and winnowed by the predator. Fracture events were short-lived (<0.1 s) with peak frequencies ranging from 3.1 to 5.0 kHz, depending on prey type. Statistical analyses showed capacity to distinguish between the two dominant prey types offered (hard clam, Mercenaria mercenaria and banded tulip, Cinctura lilium) based on processing time, the number of fractures, as well as using a suite of energy and spectral features associated with these acoustic signals. Importantly, we noted that the directionality of these relationships (i.e., relative differences in signals between prey types) can change depending on the chronological location within a consumption sequence and the amount of data analyzed (e.g., first fracture event vs. all fracture event), which may present analytical challenges. Additionally, in situ simulation of fracture events in the target environment suggested events could be detected above ambient noise out to 100 s of m. To our knowledge, this is the first attempt to both quantify and classify durophagy using passive acoustics. We recommend that future studies conduct extensive testing in controlled and target environments to build robust data sets capable of supporting feature extraction as well as detection-classification schemes via machine-learning. Lastly, collaborations with biomechanical scientists are suggested to facilitate a better understanding of the mechanisms driving acoustic variation of shell fracture across prey taxa.

Comparative transcriptomic analysis of deep- and shallow-water barnacle species (Cirripedia, Poecilasmatidae) provides insights into deep-sea adaptation of sessile crustaceans

BackgroundBarnacles are specialized marine organisms that differ from other crustaceans in possession of a calcareous shell, which is attached to submerged surfaces. Barnacles have a wide distribution, mostly in the intertidal zone and shallow waters, but a few species inhabit the deep-sea floor. It is of interest to investigate how such sessile crustaceans became adapted to extreme deep-sea environments. We sequenced the transcriptomes of a deep-sea barnacle, Glyptelasma gigas collected at a depth of 731 m from the northern area of the Zhongjiannan Basin, and a shallow-water coordinal relative, Octolasmis warwicki. The purpose of this study was to provide genetic resources for investigating adaptation mechanisms of deep-sea barnacles.ResultsTotals of 62,470 and 51,585 unigenes were assembled for G. gigas and O. warwicki, respectively, and functional annotation of these unigenes was made using public databases. Comparison of the protein-coding genes between the deep- and shallow-water barnacles, and with those of four other shallow-water crustaceans, revealed 26 gene families that had experienced significant expansion in G. gigas. Functional annotation showed that these expanded genes were predominately related to DNA repair, signal transduction and carbohydrate metabolism. Base substitution analysis on the 11,611 single-copy orthologs between G. gigas and O. warwicki indicated that 25 of them were distinctly positive selected in the deep-sea barnacle, including genes related to transcription, DNA repair, ligand binding, ion channels and energy metabolism, potentially indicating their importance for survival of G. gigas in the deep-sea environment.ConclusionsThe barnacle G. gigas has adopted strategies of expansion of specific gene families and of positive selection of key genes to counteract the negative effects of high hydrostatic pressure, hypoxia, low temperature and food limitation on the deep-sea floor. These expanded gene families and genes under positive selection would tend to enhance the capacities of G. gigas for signal transduction, genetic information processing and energy metabolism, and facilitate networks for perceiving and responding physiologically to the environmental conditions in deep-sea habitats. In short, our results provide genomic evidence relating to deep-sea adaptation of G. gigas, which provide a basis for further biological studies of sessile crustaceans in the deep sea.

Chitin is a functional component of the larval adhesive of barnacles

Barnacles are the only sessile crustaceans, and their larva, the cyprid, is supremely adapted for attachment to surfaces. Barnacles have a universal requirement for strong adhesion at the point of larval attachment. Selective pressure on the cyprid adhesive has been intense and led to evolution of a tenacious and versatile natural glue. Here we provide evidence that carbohydrate polymers in the form of chitin provide stability to the cyprid adhesive of Balanus amphitrite. Chitin was identified surrounding lipid-rich vesicles in the cyprid cement glands. The functional role of chitin was demonstrated via removal of freshly attached cyprids from surfaces using a chitinase. Proteomic analysis identified a single cement gland-specific protein via its association with chitin and localized this protein to the same vesicles. The role of chitin in cyprid adhesion raises intriguing questions about the evolution of barnacle adhesion, as well as providing a new target for antifouling technologies.

Phylogenetic Relationship of Barnacles (Fam:Balanidae) Sampled on Artificial Substrata of Kuala Selangor and Morib

Barnacles are marine sessile crustacean inhabiting intertidal areas of the Selangor coastline. They are seen attaching themselves to rocks and artificial structures such as jetty, piers, boats and sea walls. Being the most successful biofoulers, barnacles cause economic losses to some extent. Most of barnacles study focused on morphological identification only. Since molecular method gave more accurate results by sequence comparison, species identification was done on samples of obviously different species inhabiting artificial substrata by using mitochondrial 16S rDNA identification. In Kuala Selangor, there was only one species found on artificial substrata in Bagan Pasir and Pasir Penambang which was identified as Amphibalanus cirratus. Two species that differed in their morphological characteristics found on Morib sea walls were identified as Amphibalanus cirratus and Chthamalus malayensis. Phylogenetic tree based on the 16S rDNA showed that all the samples were in the same cluster reflecting that they are in the same clade.

Osmoregulation in Barnacles: An Evolutionary Perspective of Potential Mechanisms and Future Research Directions.

Barnacles form a globally ubiquitous group of sessile crustaceans that are particularly common in the coastal intertidal. Several barnacle species are described as highly euryhaline and a few species even have the ability to colonize estuarine and brackish habitats below 5 PSU. However, the physiological and/or morphological adaptations that allow barnacles to live at low salinities are poorly understood and current knowledge is largely based on classical eco-physiological studies offering limited insight into the molecular mechanisms. This review provides an overview of available knowledge of salinity tolerance in barnacles and what is currently known about their osmoregulatory strategies. To stimulate future studies on barnacle euryhalinity, we briefly review and compare barnacles to other marine invertebrates with known mechanisms of osmoregulation with focus on crustaceans. Different mechanisms are described based on the current understanding of molecular biology and integrative physiology of osmoregulation. We focus on ion and water transport across epithelial cell layers, including transport mechanisms across cell membranes and paracellular transfer across tight junctions as well as on the use of intra- and extracellular osmolytes. Based on this current knowledge, we discuss the osmoregulatory mechanisms possibly present in barnacles. We further discuss evolutionary consequences of barnacle osmoregulation including invasion-success in new habitats and life-history evolution. Tolerance to low salinities may play a crucial role in determining future distributions of barnacles since forthcoming climate-change scenarios predict decreased salinity in shallow coastal areas. Finally, we outline future research directions to identify osmoregulatory tissues, characterize physiological and molecular mechanisms, and explore ecological and evolutionary implications of osmoregulation in barnacles.

Palaeoenvironmental analysis of the Miocene barnacle facies: case studies from Europe and South America

Abstract Acorn barnacles are sessile crustaceans common in shallow-water settings, both in modern oceans and in the Miocene geological record. Barnacle-rich facies occur from polar to equatorial latitudes, generally associated with shallow-water, high-energy, hard substrates. The aim of this work is to investigate this type of facies by analysing, from the palaeontological, sedimentological and petrographical points of view, early Miocene examples from Northern Italy, Southern France and South-western Peru. Our results are then compared with the existing information on both modern and fossil barnacle-rich deposits. The studied facies can be divided into two groups. The first one consists of very shallow, nearshore assemblages where barnacles are associated with an abundant hard-substrate biota (e.g., barnamol). The second one includes a barnacle-coralline algae association, here named “barnalgal” (= barnacle / red algal dominated), related to a deeper setting. The same pattern occurs in the distribution of both fossil and recent barnacle facies. The majority of them are related to very shallow, high-energy, hard-substrate, a setting that represents the environmental optimum for the development of barnacle facies, but exceptions do occur. These atypical facies can be identified through a complete analysis of both the skeletal assemblage and the barnacle association, showing that barnacle palaeontology can be a powerful tool for palaeoenvironmental reconstruction.

The diversity of acorn barnacles (Cirripedia, Balanomorpha) across Thailand’s coasts: The Andaman Sea and the Gulf of Thailand

The acorn barnacle is a sessile crustacean, inhabiting the intertidal areas of tropical and temperate regions worldwide. According to current practices on Cirripedia morphology, shell, opercular valves, and arthropodal characters including cirri and mouthparts are used as a tool for taxonomic classification, and using these characteristics the present study aimed to provide better resolution for the barnacle diversity and geographical distribution within coastlines of Thailand: the Andaman Sea and the Gulf of Thailand. A total of ten species belonging to three families (Chthamalidae, Tetraclitidae, and Balanidae) were identified in this study. Subsequently, five species were newly recorded for the first time from Thailand’s coasts: Newmanella spinosus Chan &amp; Cheang, 2016, Euraphia hembeli Conrad, 1837, Euraphia depressa (Poli, 1795), Tetraclita kuroshioensis Chan, Tsang &amp; Chu, 2007, and Tetraclita singaporensis Chan, Tsang &amp; Chu, 2007. The others, already mentioned in previous records, include: Tetraclita squamosa (Bruguière, 1789), Chthamalus malayensis Pilsbry, 1916, Amphibalanus amphitrite (Darwin, 1854), Amphibalanus reticulatus (Utinomi, 1967), and Megabalanus tintinnabulum (Linnaeus, 1758). Interestingly, acorn barnacles along the Andaman Sea occur abundantly, and are much higher in number of species (up to 8 species) than those found in the Gulf of Thailand’s coast (up to 6 species). This biased trend of species’ preferences is possibly due to the differences in oceanographic nature between two coastlines and the history of barnacle colonization.

Structural specialties, curiosities, and record-breaking features of crustacean reproduction.

Crustaceans are a morphologically, physiologically, and ecologically highly diverse animal group and correspondingly diverse are their reproductive characteristics. They have evolved structural specialties with respect to penis construction, sperm form, sperm storage, fertilization, and brood care. Unique in the animal kingdom are safety lines that safeguard hatching and first molting. Further curiosities are dwarf males in parasitic and sessile crustaceans and bacteria-induced feminization and gigantism of crustacean hosts. Record-breaking features are relative penis length, sperm size, clutch size, chromosome number, viability of dormant eggs, and fossil ages of penis, sperm, and brooded embryos. These examples from a single invertebrate subphylum and a single life history aspect illustrate that morphological solutions to functional requirements can be as spectacular as behavioral adaptations. They may provide valuable sources for comparative morphologists, ecologists, evolutionary biologists, and applied biologists to advance topical issues such as sperm competition, posthumous paternity, evolution of brood care, adaptation to freshwater, infectious feminization, sustainable male-based fishery, maintenance of genetic diversity under conditions of limited mating opportunity, and long-term impact of pollution on genotype and phenotype. J. Morphol. 277:1399-1422, 2016. © 2016 Wiley Periodicals, Inc.

Lifestyles and feeding biology

Chapter 1. The Role of Natural History in Understanding the Diversity of Lifestyles in Crustaceans Les Watling and Martin Thiel Chapter 2. Diversity of Lifestyles, Sexual Systems and Larval Development Pattern in Sessile Crustaceans Benny K.K. Chan and Jens T. Hoeg Chapter 3. The Tube-dwelling Lifestyle in Crustaceans and its Relation to Feeding P. Geoffrey Moore and Lucas B. Eastman Chapter 4. Burrow-dwelling in Crustacea R. James A. Atkinson and Lucas B. Eastman Chapter 5. Crustaceans Inhabiting Domiciles Excavated from Macrophytes and Stone Barbara A. Mejaes, Alistair G.B. Poore and Martin Thiel Chapter 6. Crustaceans in Mobile Homes Patsy A. McLaughlin Chapter 7. Crustaceans as Symbionts: An Overview of their Diversity, Host Use and Lifestyles J. Antonio Baeza Chapter 8. Predator Adaptations of Decapods Kari L. Lavalli and Ehud Spanier Chapter 9. Small Free-living Crustaceans Richard B. Taylor Chapter 10. Planktonic Crustaceans: Lifestyles in the Water Column Ruben Escribano and Ramiro Riquelme-Bugueno Chapter 11. Oxygen Lifestyles of the Species-rich and Fabulous: the Deep-sea Crustaceans George D.F. Wilson and Shane T. Ahyong Chapter 12. Lifestyles of Terrestrial Crustaceans Alastair Richardson and Paula Araujo Chapter 13. Freshwater Crustaceans: Adaptations to Complex Inland Habitats and Species Interactions Alan P. Covich Chapter 14. Crustaceans of Extreme Environments Chiara Benvenuto, Brenton Knott, and Stephen C. Weeks Chapter 15. Filter-feeding Mechanisms in Crustaceans Hans Ulrik Riisgard Chapter 16. Deposit-Feeding: Obtaining Nutrition from Sediment Les Watling Chapter 17. Lifestyle of Detritus-Feeding Crustaceans Sven Hammann and Martin Zimmer Chapter 18. Grazers of Macroalgae and Higher Plants Veijo Jormalainen Chapter 19. Foraging Behavior of Crustacean Predators and Scavengers Lucas B. Eastman and Martin Thiel

Amyloid-like conformation and interaction for the self-assembly in barnacle underwater cement.

Barnacles are unique marine sessile crustaceans and permanently attach to various foreign surfaces during most of their lifespan. The protein complex secreted from their body and used to attach their calcareous shell to almost all surfaces in water has long fascinated us because we have limited technology with which to attach materials in water. Unraveling the mechanism of underwater attachment by barnacles is thus important for interface science, for the understanding of the biology and physiology of barnacles, and for the development of technology to prevent fouling. Previous studies have indicated that the intermolecular interactions optimized by conformations of the adhesive proteins are crucial in the self-assembly and/or curing of the adhesive. This study aimed to identify the possible structural determinants responsible for the self-assembly. Thioflavin T binding screening of peptides designed on the basis of the primary structure of a bulk 52 kDa cement protein indicated the presence of some amyloidogenic motifs in the protein. The conformation of the peptide was transformed to a β-sheet by an increase in either pH or ionic strength, resulting in its self-assembly. Thioflavin T binding was inhibited by small polyphenolic molecules, suggesting the contribution of aromatic interactions during self-assembly. The occurrence of amyloid-like units in the protein implies that the protein conformation is an important factor contributing to the self-assembly of the cement, the first event of the curing, as the adhesive material is secreted into the seawater out of the animal's body.

Prokaryotic Expression and Functional Characterization of the 19 kDa Protein in &lt;i&gt;Balanus albicostatus&lt;/i&gt; Cement

Barnacle is a unique sessile crustacean, which produces a multi-protein complex historically called barnacle cement to attach to diverse immersed materials permanently. The proteinaceous cement exhibits powerful adhesive property and special waterproof capability to cure at solid-liquid boundaries, which makes it ideal biomaterial for technical, medical and dental applications. It has been proved that a 19 kDa protein component, termed cp-19k in the cement plays a key role in surface coupling during underwater attachment. To verify whether the bacterial recombinant 19 kDa protein retains the adhesive ability, we cloned and sequenced the Bacp-19k gene in Balanus albicostatus. It encodes 173 amino acid residues, with seven biased ones, Thr, Lys, Gly, Ala, Val, Ser and Leu, comprising about 80% of the total. Two amino acid substitutions (F69L, I106L) were discovered in Bacp-19k due to the polymorphisms in barnacle cp-19ks, compared with the submitted one (GenBank: AB242295.1). Recombinant Bacp-19k was highly expressed in host strain Escherichia coli BL21 (DE3) and purified by affinity chromatography. Adsorption of recombinant Bacp-19k to glass substrata was examined by Coomassie brilliant blue staining. Future study will reveal the relationship between specific structures and functions for molecular design of novel biomimetic underwater adhesives.

Significance of the conformation of building blocks in curing of barnacle underwater adhesive

Barnacles are a unique sessile crustacean that attach irreversibly and firmly to foreign underwater surfaces. Its biological underwater adhesive is a peculiar extracellular multi-protein complex. Here we characterize one of the two major proteins, a 52 kDa protein found in the barnacle cement complex. Cloning of the cDNA revealed that the protein has no homolog in the nonredundant database. The primary structure consists of four long sequence repeats. The process of dissolving the protein at the adhesive joint of the animal by various treatments was monitored in order to obtain insight into the molecular mechanism involved in curing of the adhesive bulk. Treatments with protein denaturant, reducing agents and/or chemical-specific proteolysis in combination with 2D diagonal PAGE indicated no involvement of the protein in intermolecular cross-linkage/polymerization, including formation of intermolecular disulfide bonds. As solubilization of the proteins required high concentrations of denaturing agents, it appears that both the conformation of the protein as building blocks and non-covalent molecular interactions between the building blocks, possibly hydrophobic interactions and hydrogen bonds, are crucial for curing of the cement. It was also suggested that the protein contributes to surface coupling by an anchoring effect to micro- to nanoscopic roughness of surfaces.

Charles Darwin and Joseph de Bosquet – Brothers in barnacles: How diminutive crustaceans helped shape a theory

Ten years after his return to England on board ‘The Beagle’ in 1836, Charles Robert Darwin (1809–1882) started collecting, dissecting, describing and interpreting both extant and fossil acorn and goose barnacles (cirripedes). In all, he spent eight years on this animal group and, between 1851 and 1855, published four authoritative and admirably illustrated volumes on these sessile crustaceans; a highly valuable source of information, even to the present day. The Maastricht pharmacist, Joseph Augustin Hubert de Bosquet (1814–1880), who also was a well-versed collector and student of Late Cretaceous fossils from the nearby St Pietersberg, corresponded with Darwin on the subject of cirripedes between 17 December 1852 and early November 1856. In addition, these gentlemen exchanged monographs, manuscripts, engravings and specimens. At a time when Darwin was finally trying to come to terms with his ‘wretched’ barnacles, he much appreciated the support and understanding coming from Maastricht and his letters clearly show the respect he had for his fellow barnacle worker. In fact, Darwin’s barnacle work strengthened his resolve to publish his 1859 masterpiece, On the Origin of Species. Current studies of cirripedes in the Maastricht area seek to complement de Bosquet’s pioneering work and document in more detail the stratigraphic ranges of the various taxa as well as their relationships with species elsewhere in Europe as well as overseas.

Molecular Design of Barnacle Cement in Comparison with Those of Mussel and Tubeworm

Various organisms are known to attach themselves to a wide range of foreign materials in water as one of their essential physiological functions. To accomplish this, these organisms have acquired their specialized molecular systems in the process of their evolution. The molecular systems of sessile organisms are excellently designed for the purpose of underwater adhesion from the macroscopic scale to the molecular level. This review focuses on the unique sessile crustacean, the barnacle, in which a molecular system called cement was found for the underwater adhesion, which is completely different from the molecular system found in the mussel and tubeworm. The components, properties, and unique functions of the cement proteins from barnacles in comparison with those of the mussel and tubeworm are discussed.

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

AbstractThoracic 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.