Under pressure: Assessing the ecological boundaries of the epipelagic goose barnacle Lepas anatifera using ocean gliders and laboratory experiments.

Epipelagic barnacles have been considered good bioindicators since they are abundant and broadly distributed but with apparent tolerance restrictions to temperature and salinity, and also bioaccumulate pollutants. However, Lepas anatifera was found attached to the oceanic gliders, thriving through drastic and unreported environmental fluctuations. This study aimed to assess the resistance and oxidative stress responses of L. anatifera collected from gliders and attached to floating litter to temperature, salinity, and pressure. Barnacles withstood all tested pressure, temperature, and salinity ranges, except the extreme salt concentration. The activities of antioxidant enzymes - catalase and superoxide dismutase - were significantly increased under high temperature, high pressure, and low salinity. Malondialdehyde levels significantly increased only under high pressure. In conclusion, L. anatifera can be considered resistant organisms to extreme environmental changes. However, the instauration of oxidative stress under certain circumstances makes them vulnerable to predicted future trends in marine environments.

First record of the cirripede Anelasma squalicola Darwin, 1852 (Pollicipedidae) in the Mediterranean Sea.

Anelasma squalicola Darwin 1852, is a barnacle that occurs on elasmobranchs from the Atlantic, Pacific, Arctic and Indian Ocean. It infects sharks of the families Etmopteridae and Pentanchidae. The barnacle attaches sub-dermally to its hosts, relying on its interstitial fluid for its nourishment. Herein we present the first record of this species in the Mediterranean Sea, from Etmopterus spinax (Linnaeus, 1758) caught during a bottom-trawl survey in the Alboran Sea (western Mediterranean) in 2017. The overall prevalence of infection was very low (0.22%). The calculated HSI index for the parasitized individual was lower compared to overall means of other individuals and could indicate a potential physiological alteration in the host.

Theoretical studies of diverse sexual patterns in marine animals.

Marine animals show diverse and flexible sexual systems. Here, we review several advancements of theoretical studies made in the last decade. (i) Sex change in coral fishes is often accompanied by a long break in reproductive activity. The delay can be shortened by retaining the inactive gonad for the opposite sex. (ii) Barnacles adopt diverse sexual patterns. The game model was analysed assuming that newly settled larvae choose either growth or immediate reproduction and large individuals adjust male-female investments. (iii) Some parasitic barnacles produce larvae with sexual size dimorphism and others produce larvae with the sex determined after settlement on hosts. (iv) In some fish and many reptiles, sex is determined by the temperature experienced as eggs. The dynamics of sex hormones were studied when the enzymatic reaction rates were followed by the Arrhenius equation. The FMF pattern (male at intermediates temperature; female both at high and low temperatures) required some reactions with enhanced temperature dependence at higher temperatures. The game model provides a useful framework for understanding diverse sexual patterns if we incorporate various constraints, such as unpredictability, cost of trait change and social situations. For further developments, we need to consider constraints imposed by physiological and molecular mechanisms.

A Rhizocephalan Parasite Induces Pervasive Effects on Its Shrimp Host.

AbstractRhizocephalan barnacles are parasites of crustaceans that are known for dramatic effects on hosts, including parasitic castration, feminization, molt inhibition, and the facilitation of epibiosis. Most research on rhizocephalans has focused on carcinized hosts, with relatively little research directed to shrimp hosts that may experience distinct consequences of infection. Here, we describe a high-prevalence rhizocephalan-shrimp system in which multiple host changes are associated with infection: the dock shrimp Pandalus danae infected by the rhizocephalan Sylon hippolytes. In field-collected P. danae, infection by Sylon was associated with development of female sex characters at a smaller size and greater probability of epibiosis. Standardized video observations showed that infected P. danae performed grooming activities at higher rates than uninfected shrimp, suggesting that inhibited molting rather than direct behavioral modification is a likely mechanism for higher epibiosis rates. There was no difference in the composition of grooming behavior types or in general activity between infected and uninfected shrimp. Fatty acid compositions differed with infection, but total lipid concentrations did not, suggesting that parasite-driven shifts in host resource allocation were compensated or redirected from unmeasured tissues. Our results show that Sylon alters its host's role by provisioning an epibiotic substrate and also that it influences host physiology, resulting in feminization and fatty acid shifts. This study lays the groundwork for expanding rhizocephalan-shrimp research and encourages recognition of oft-ignored roles of parasitism in ecological communities.

Unravelling the trophic interaction between a parasitic barnacle (Anelasma squalicola) and its host Southern lanternshark (Etmopterus granulosus) using stable isotopes.

The parasitic barnacle, Anelasma squalicola, is a rare and evolutionary fascinating organism. Unlike most other filter-feeding barnacles, A. squalicola has evolved the capability to uptake nutrient from its host, exclusively parasitizing deepwater sharks of the families Etmopteridae and Pentanchidae. The physiological mechanisms involved in the uptake of nutrients from its host are not yet known. Using stable isotopes and elemental compositions, we followed the fate of nitrogen, carbon and sulphur through various tissues of A. squalicola and its host, the Southern lanternshark Etmopterus granulosus, to better understand the trophic relationship between parasite and host. Like most marine parasites, A. squalicola is lipid-rich and clear differences were found in the stable isotope ratios between barnacle organs. It is evident that the deployment of a system of 'rootlets', which merge with host tissues, allows A. squalicola to draw nutrients from its host. Through this system, proteins are then rerouted to the exterior structural tissues of A. squalicola while lipids are used for maintenance and egg synthesis. The nutrient requirement of A. squalicola was found to change from protein-rich to lipid-rich between its early development stage and its definitive size.

Functional morphology of cirri in the barnacle Amphibalanus improvisus (crustacea: Balanidae).

Barnacles rely heavily on their mobile cirri for food capture because of the sessile lifestyle. These filamentous food capturing devices are extended into the water current and perform undulating movements. Cuticular structures with corresponding musculature work together, to allow these highly repetitive movements. This paper studies the interplay between structure and function of the cirri using scanning electron microscopy (SEM), microcomputed tomography (µCT) and high-speed video recordings (HSV) in Amphibalanus improvisus. Barnacles use the external cuticle structures (denticles and setae) for efficient grooming, food capturing, and providing support for the muscular system responsible for movement control. Entanglement of the cirri during extension is probably avoided through an interlocking of the serrate setae on cirri IV-VI, creating a "zipper-like" effect, which was recorded here using the HSV. We analyzed the muscular arrangement using µCT and found a new flexor muscle in both the endo- and exopod of cirrus II. Supported by the intrinsic cirral muscles, the new flexor muscles may provide variable movements of the anterior cirri (cirri I-III), which is important for further food handling. Our results provide a foundation for further comparative studies of the feeding apparatus of barnacles and for possible implications in the area of bio-inspired robotics.

Amphiphilic Nitroxide-Bearing Siloxane-Based Block Copolymer Coatings for Enhanced Marine Fouling Release.

The buildup of organic matter and organisms on surfaces exposed to marine environments, known as biofouling, is a disruptive and costly process affecting maritime operations. Previous research has identified some of the surface characteristics particularly suited to the creation of antifouling and fouling-release surfaces, but there remains room for improvement against both macrofouling and microfouling organisms. Characterization of their adhesives has shown that many rely on oxidative chemistries. In this work, we explore the incorporation of the stable radical 2,2,6,6-tetramethylpipiderin-1-oxyl (TEMPO) as a component in an amphiphilic block copolymer system to act as an inhibitor for marine cements, disrupting adhesion of macrofouling organisms. Using polystyrene-b-poly(dimethylsiloxane-r-vinylmethysiloxane) block copolymers, pendent vinyl groups were functionalized with TEMPO and poly(ethylene glycol) to construct an amphiphilic material with redox active character. The antifouling and fouling-release performance of these materials was investigated through settlement and removal assays of three model fouling organisms and correlated to surface structure and chemistry. Surfaces showed significant antifouling character and fouling-release performance was increased substantially toward barnacles by the incorporation of stable radicals, indicating their potential for marine antifouling applications.

Larval tolerance to food limitation is stronger in an exotic barnacle than in its native competitor.

A critical question in marine ecology is understanding how organisms will cope with environmental conditions under climate change. Increasing temperatures not only have a direct effect on marine organisms but may also lead to food limitation through for example trophic mismatches, or by the increased metabolic demands imposed by developing at high temperatures. Using barnacles from a population of North Wales, we studied the combined effect of temperature and food density on the survival, settlement success, developmental time and body size of larvae of the native barnacle Semibalanus balanoides and its exotic competitor, the barnacle Austrominius modestus. Larvae were reared at similar food levels but at temperature ranges which varied among species reflecting their different phenology and tolerances. For S. balanoides (spring larval release) we used a lower temperature of 9 °C, reflecting spring temperatures from N Wales to SW England, and 15 °C representing warmer conditions; for A. modestus (summer larval release) a typical summer temperature for this geographic range of 15 °C was used with a raised temperature of 18 °C. Larvae were reared under controlled conditions in automated, computer programmable incubators and fed diatoms (Skeletonema costatum) at three food levels. We found stress effects of food limitation on larval performance of S. balanoides. While survival during naupliar development was little affected by food and temperature, low food levels strongly depressed survival and settlement during the cyprid stage of S. balanoides at both tested temperatures, but especially at 15 °C. By contrast, at the tested temperatures little effects were found on survival and settlement success in the exotic A. modestus. Both species delayed development in response to low food levels while S. balanoides cyprids showed decreased body size at the high tested temperature. The main impact occurred as a delayed effect, at the time when cyprids attempt to settle, rather than as an effect on naupliar survival or metamorphosis to the cyprid stage. Response in body size and developmental time may have costs at the time of metamorphosis (delayed settlement) or after metamorphosis. Overall, our experiments suggest that as temperature increases, settlement success of S. balanoides larvae (but not that of its competitor A. modestus) will become more sensitive to conditions of food limitation, imposed for instance by phenological mismatches with periods of phytoplankton peak.

Differences in the parasitic effects of a bopyrid isopod and rhizocephalan barnacle on the portunid crab, Charybdis bimaculata.

Bopyrid isopods and rhizocephalan barnacles are obligate parasite crustaceans which harm their decapod hosts. However, to the best of our knowledge, studies have not compared which of these parasites has a greater parasitic effect on its hosts. Here, the parasitic effect of the bopyrid isopod, Allokepon hendersoni, and an unidentified sacculinid rhizocephalan species, infesting the same population of portunid crabs, Charybdis bimaculata, was investigated and compared for the first time. Samples were collected from the bycatch of a trawl fishery in Tosa Bay, Japan. A total of 2601 crabs were collected, of which 14 (0.55%) were parasitized by the bopyrid and 21 (0.82%) by the rhizocephalan. One of the two female crabs parasitized by the bopyrid was ovigerous (with much fewer eggs than unparasitized females). No ovigerous crab was found from the eight females parasitized by the rhizocephalan. Because only two female crabs were parasitized by the bopyrid, the following analyses were made using the male crabs. Both parasites reduced the wet weight (crab condition) and the cheliped size (secondary growth) of C. bimaculata, but the impact of the parasitism did not differ between the parasite species. The size of the abdominal flap of male hosts was reduced by the bopyrid infestation; however, rhizocephalan infestation caused enlargement of the abdominal flap, which is an indication of feminization. The present study provides information on how the effect of these two parasitic castrators on the same host crab varies. A moderate decrease in crab condition and cheliped development was common among the parasites, suggesting that the degree of impact might be favorable for the survival of the two parasites species.

Porous silicone substrates inhibit permanent barnacle attachment under natural conditions.

Barnacles are able to effectively adhere to most surfaces underwater. Dewetting of the corresponding surface prior to the release of their permanent adhesive plays an important role in the attachment process. Possibly, a surface that is able to interfere with this process may have exceptional fouling repellence and fouling release abilities. Therefore, open-pored foams made from polydimethylsiloxane (PDMS) were tested together with flat PDMS samples as controls in a 13-week-long field experiment in the Baltic Sea. On a weekly basis, both settlement and fouling density development of the bay barnacle Balanus (=Amphibalanus) improvisus were monitored. The overall settlement was close to zero on PDMS foams and the few attached barnacles were not able to stay on the PDMS foams longer than 1 week after initial settlement. Changes in the stiffness of the PDMS foams did not affect these results. Open-pored PDMS foam systems may be a promising tool in the development of new, innovative antifouling strategies.

Swimming kinematics and hydrodynamics of barnacle larvae throughout development.

Changes in size strongly influence organisms' ecological performances. For aquatic organisms, they can transition from viscosity- to inertia-dominated fluid regimes as they grow. Such transitions are often associated with changes in morphology, swimming speed and kinematics. Barnacles do not fit into this norm as they have two morphologically distinct planktonic larval phases that swim differently but are of comparable sizes and operate in the same fluid regime (Reynolds number 100-101). We quantified the hydrodynamics of the rocky intertidal stalked barnacle Capitulum mitella from the nauplius II to cyprid stage and examined how kinematics and size increases affect its swimming performance. Cyprids beat their appendages in a metachronal wave to swim faster, more smoothly, and with less backwards slip per beat cycle than did all naupliar stages. Micro-particle image velocimetry showed that cyprids generated trailing viscous vortex rings that pushed water backwards for propulsion, contrary to the nauplii's forward suction current for particle capture. Our observations highlight that zooplankton swimming performance can shift via morphological and kinematic modifications without a significant size increase. The divergence in ecological functions through ontogeny in barnacles and the removal of feeding requirement likely contributed to the evolution of the specialized, taxonomically unique cyprid phase.

Sponge symbiosis is facilitated by adaptive evolution of larval sensory and attachment structures in barnacles.

Symbiotic relations and range of host usage are prominent in coral reefs and crucial to the stability of such systems. In order to explain how symbiotic relations are established and evolve, we used sponge-associated barnacles to ask three questions. (1) Does larval settlement on sponge hosts require novel adaptations facilitating symbiosis? (2) How do larvae settle and start life on their hosts? (3) How has this remarkable symbiotic lifestyle involving many barnacle species evolved? We found that the larvae (cyprids) of sponge-associated barnacles show a remarkably high level of interspecific variation compared with other barnacles. We document that variation in larval attachment devices are specifically related to properties of the surface on which they attach and metamorphose. Mapping of the larval and sponge surface features onto a molecular-based phylogeny showed that sponge symbiosis evolved separately at least three times within barnacles, with the same adaptive features being found in all larvae irrespective of phylogenetic relatedness. Furthermore, the metamorphosis of two species proceeded very differently, with one species remaining superficially on the host and developing a set of white calcareous structures, the other embedding itself into the live host tissue almost immediately after settlement. We argue that such a high degree of evolutionary flexibility of barnacle larvae played an important role in the successful evolution of complex symbiotic relationships in both coral reefs and other marine systems.

Transcriptome analyses suggest a molecular mechanism for the SIPC response of Amphibalanus amphitrite.

Barnacles are notorious marine fouling organisms. Their successful attachment to a substrate requires that they search for an appropriate habitat during their cyprid stage. A chemical cue called SIPC (Settlement-Inducing Protein Complex) has been shown to play a key role in the induction of cyprid gregarious settlement; however, the underlying biochemical mechanism remains unclear. Here, RNA-seq was used to examine the gene expression profiles of Amphibalanus amphitrite cyprids in response to SIPC and to identify SIPC-activated intracellular signaling pathways. A total of 389 unigenes were differentially expressed in response to SIPC, and cement protein genes were not among them. KEGG enrichment analysis suggested that SNARE interactions in the vesicular transport pathway were significantly influenced by SIPC treatment, indicating a possible role for SIPC in triggering protein transportation and secretion. Several genes with specific functions in metamorphosis were found among the differentially expressed genes (DEGs). GO (Gene Ontology) enrichment analysis revealed that the DEGs were significantly enriched in enamel mineralization pathways, suggesting that SIPC may also be involved in the activation of mineralization.

Food sources for the Ediacara biota communities.

The Ediacara biota represents the first complex macroscopic organisms in the geological record, foreshadowing the radiation of eumetazoan animals in the Cambrian explosion. However, little is known about the contingencies that lead to their emergence, including the possible roles of nutrient availability and the quality of food sources. Here we present information on primary producers in the Ediacaran based on biomarker molecules that were extracted from sediments hosting Ediacaran macrofossils. High relative abundances of algal steranes over bacterial hopanes suggest that the Ediacara biota inhabited nutrient replete environments with an abundance of algal food sources comparable to Phanerozoic ecosystems. Thus, organisms of the Ediacara biota inhabited nutrient-rich environments akin to those that later fuelled the Cambrian explosion.

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

BACKGROUND: Barnacles 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. RESULTS: Totals 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. CONCLUSIONS: The 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.

Effects of polystyrene microplastics on larval development, settlement, and metamorphosis of the intertidal barnacle Amphibalanus amphitrite.

The effects of microplastic on mortality and sublethal responses on larval development of meroplankton are still largely unknown. Present study investigated the effects of four sizes of virgin spherical polystyrene microplastics (diameter 1.7, 6.8, 10.4, 19.0 µm) on naupliar (stage II-VI) and cypris larvae of barnacle Amphibalanus amphitrite at environmentally relevant concentrations (1, 10, 100, 1000 beads mL-1). Essential life-history traits, including mortality, development time and rates of growth, settling, and metamorphosis were measured throughout the entire larval development. Feeding experiments were conducted to evaluate if microplastics decreased naupliar feeding due to physical impacts or selective feeding of nauplii. The results showed that A. amphitrite stage II nauplii were able to ingest and efficiently egest all sizes of microplastics. All the life-history endpoints measured were not significantly affected by all sizes of microplastics at any concentration tested. Presence of all sizes of microplastics did not cause physical interference on naupliar feeding and all stages of nauplius larvae (stage III-VI) did not selectively feed on microplastics. However, the feeding ability of stage III nauplius appeared to be affected by 1.7 µm at 1000 beads mL-1 which was possibly due to individual variations rather than microplastics' impacts. Overall, the full larval development of barnacle A. amphitrite was not affected by microplastics at environmentally relevant concentrations under laboratory condition.

Prevalence and histopathology of the parasitic barnacle, Sacculina carcini in shore crabs, Carcinus maenas.

Sacculina carcini is a common parasite of the European shore crab, Carcinus maenas. Following successful penetration of the host, numerous rootlets are formed that permeate through the hosts' tissues. Ultimately, these form an externa that houses the developing nauplii larvae of the parasite. Most studies have quantified levels of infection by counting the presence of reproductive externae and their breakdown structures, called scars. However, the diagnosis of the disease based only on external features may lead to underreporting the prevalence of the parasite. In the current study, we examined the presence and severity of S. carcini in C. maenas (n = 221) in the Prince of Wales Dock, South Wales, U.K. using a range of diagnostic approaches to give an accurate representation of temporal dynamics of infection. Parasitized crabs were found with a mean prevalence of 24% as determined by histological examination of the hepatopancreas. However, the prevalence of S. carcini based on the presence of externae and scars was only 6.3% and 1.8%, respectively. Overall, parasitism was associated with smaller crabs, crabs later in the moulting cycle that were orange in colour (as opposed to green or yellow), and those with a higher number of bacteria in the haemolymph. Interestingly, only 7.5% of infected crabs showed evidence of distinct host (cellular) response to the presence of rootlets in the hepatopancreas.

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.

Specialized structures on the border between rhizocephalan parasites and their host's nervous system reveal potential sites for host-parasite interactions.

Rhizocephalan barnacles are a unique group of endoparasitic crustaceans. In their extreme adaptation to endoparasitism, rhizocephalan adults have lost almost all features of their free-living relatives but acquired an outstanding degree of control over the body of their hosts (mostly decapods). The subtle influence exercised by rhizocephalans on the physiology, morphology and behaviour of their hosts is a vivid example of the most intimate host-parasite interactions but their mechanisms are very poorly known. In this study we examined the morphology and the adaptive ultrastructure of the organs invading the nervous system of the host in two rhizocephalan species from the families Peltogastridae, (Peltogaster paguri) and Peltogasterellidae (Peltogasterella gracilis). We found two essentially different types of structures involved in interactions of these two rhizocephalans with the nervous system of their hosts: modified rhizocephalan rootlets lying inside the ganglia and the neural fibres of the host enlacing the trophic rootlets of the parasites. We suggest that both these structures may be highly specialized tools allowing the parasite to interact with the host on the humoral level via neuromediators, hormones, attractants and trophic factors.

Competing barnacle species with a time dependent reproduction rate.

A simple competition model with time varying periodic coefficients, in which two species use different reproduction strategies, is explored in this paper. The two species considered comprise a native species which reproduces once a year over a short time period and an invasive species which is capable of reproducing throughout the entire year. A monotonicity property of the model is instrumental for its analysis. The model reveals that the time difference between the peak of reproduction for the two species is a critical factor in determining the outcome of competition between these species. The impact of climate change and an anthropogenic disturbance, comprising the creation of additional substrate, is also investigated using a modified model. The results of this paper describe how climate change will favour the invasive species by reducing the time period between the reproductive peaks of the two species and how the addition of new substrates is likely to endanger a small population of either of the two species, depending on the timing of the introduction of the substrates.