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.

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.

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.

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.

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.

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.

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.

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.

Parasite and host biomass and reproductive output in barnacle populations in the rocky intertidal zone.

The rocky intertidal zone has a long history of ecological study with barnacles frequently serving as a model system to explore foundational theories. Parasites are often ignored in community ecology studies, and this particularly holds for true for the rocky intertidal zone. We explore the role of the isopod parasite, Hemioniscus balani, on its host, the acorn barnacle, Chthamalus fissus. We use the currencies of biomass and reproduction measured at the individual level, then applied to the population level, to evaluate the importance of this parasite to barnacle populations. We found H. balani can comprise substantial biomass in 'apparent' barnacle populations, sometimes even equaling barnacle biomass. Additionally, parasite reproduction sometimes matched barnacle reproduction. Thus, parasites divert substantial energy flow from the barnacle population and to near-shore communities in the form of parasite larvae. Parasites appeared to decrease barnacle reproduction per area. Potentially, this parasite may control barnacle populations, depending on the extent to which heavily infected barnacle populations contribute to barnacle populations at larger scales. These findings regarding the importance of a particular parasite for host population dynamics in this well studied ecosystem call for the integration of disease dynamics into community ecological studies of the rocky intertidal zone.

Changes in Lipids During the Ovary Maturation Process of Balanus rostratus.

Balanus rostratus is a large cold-water acorn barnacle distributed around the northern coast of the Pacific Ocean. In Mutsu Bay, Aomori, Japan, B. rostratus, which adhere naturally to scallop shells, are cultured as food. However, current culture methods do not generate sufficient supplies to satisfy market demand. Knowledge of the physiology of B. rostratus reproduction is important for the development of more efficient aquaculture methods. Previous studies have suggested that fatty acids and their metabolites play an important role in barnacle reproduction and development; however, few studies have analyzed lipids, particularly during ovary maturation. Here we analyzed lipid content, lipid class, and fatty acid composition of B. rostratus ovary throughout the year. The clutch in the present study was observed once per year at the end of November. The lipid content increased as the ovary underwent maturation. The proportion of triacylglycerol increased with increasing lipid content. The proportions of myristic acid, arachidonic acid, EPA and DHA significantly decreased in December. By contrast, the proportion of these fatty acids in lipid extracted from larvae was high relative to lipid extracted from B. rostratus ovary in December. These findings suggest that these fatty acids are transferred from the ovary to the larvae. Our novel findings on lipid metabolism during ovary maturation in B. rostratus indicate the importance of lipids during reproduction. This information may be useful in establishing methods for the aquaculture of B. rostratus.

Quantitative analysis of the complete larval settlement process confirms Crisp's model of surface selectivity by barnacles.

For barnacle cypris larvae at the point of settlement, selection of an appropriate surface is critical. Since post-settlement relocation is usually impossible, barnacles have evolved finely tuned surface-sensing capabilities to identify suitable substrata, and a temporary adhesion system for extensive surface exploration. The pattern of exploratory behaviour appears complex and may last for several hours, imposing significant barriers to quantitative measurement. Here, we employ a novel tracking system that enables simultaneous analysis of the larval body movement of multiple individuals over their entire planktonic phase. For the first time, to our knowledge, we describe quantitatively the complete settlement process of cyprids as they explore and select surfaces for attachment. We confirm the 'classic' behaviours of wide searching, close searching and inspection that comprise a model originally proposed by Prof. Dennis Crisp FRS. Moreover, a short-term assay of cyprid body movement has identified inspection behaviour as the best indicator of propensity to settle, with more inspection-related movements occurring in conditions that also promote higher settlement. More than half a century after the model was first proposed by Crisp, there exists a precise method for quantifying cyprid settlement behaviour in wide-ranging investigations of barnacle ecology and applied studies of fouling management.

Annual temperature variation as a time machine to understand the effects of long-term climate change on a poleward range shift.

Range shifts due to annual variation in temperature are more tractable than range shifts linked to decadal to century long temperature changes due to climate change, providing natural experiments to determine the mechanisms responsible for driving long-term distributional shifts. In this study we couple physiologically grounded mechanistic models with biogeographic surveys in 2 years with high levels of annual temperature variation to disentangle the drivers of a historical range shift driven by climate change. The distribution of the barnacle Semibalanus balanoides has shifted 350 km poleward in the past half century along the east coast of the United States. Recruits were present throughout the historical range following the 2015 reproductive season, when temperatures were similar to those in the past century, and absent following the 2016 reproductive season when temperatures were warmer than they have been since 1870, the earliest date for temperature records. Our dispersal dependent mechanistic models of reproductive success were highly accurate and predicted patterns of reproduction success documented in field surveys throughout the historical range in 2015 and 2016. Our mechanistic models of reproductive success not only predicted recruitment dynamics near the range edge but also predicted interior range fragmentation in a number of years between 1870 and 2016. All recruits monitored within the historical range following the 2015 colonization died before 2016 suggesting juvenile survival was likely the primary driver of the historical range retraction. However, if 2016 is indicative of future temperatures mechanisms of range limitation will shift and reproductive failure will lead to further range retraction in the future. Mechanistic models are necessary for accurately predicting the effects of climate change on ranges of species.

Should I stay or should I go? The settlement-inducing protein complex guides barnacle settlement decisions.

Reproduction in barnacles relies on chemical cues that guide their gregarious settlement. These cues have been pinned down to several sources of settlement pheromones, one of which is a protein termed settlement-inducing protein complex (SIPC), a large glycoprotein acting as a pheromone to induce larval settlement and as an adhesive in surface exploration by the cyprids. Settlement assays in laboratory conditions with Amphibalanus (=Balanus) amphitrite cyprids in the presence of SIPC showed that cyprids exhibit settlement preference behaviour at lower concentrations of SIPC [half maximal effective concentration (EC50)=3.73 nmol l-1] and settlement avoidance behaviour at higher concentrations (EC50=101 nmol l-1). By using truncated fragments of SIPC in settlement assays, we identify that domains at the N-terminus of SIPC transduce settlement preference cues that mask the settlement avoidance cues transduced by domains at its C-terminus. Removing the N-terminal 600 amino acids from SIPC resulted in truncated fragments that transduced only settlement avoidance cues to the cyprids. From the sexual reproduction point of view, this bimodal response of barnacles to SIPC suggests that barnacles will settle gregariously when conspecific cues are sparse but will not settle if conspecific cues inform of overcrowding that will increase reproductive competition and diminish their reproductive chances.

Probing the settlement signals of Amphibalanus amphitrite.

To achieve their reproductive potential, barnacles combine tactile exploration of surface structural properties and integration of cellular signals originating from their antennular sensory setae within a developmentally defined, temporally narrow window of settlement opportunity. Behavioural assays with cyprids coupled with biometric analysis of scanning electron microscopy-acquired images in the presence of specific chemical compounds were used to investigate how settlement on a substratum is altered in response to the presence of these compounds. It is shown that impeding tactile exploration, altering cellular signalling and/or inducing malformations of anatomical features of the antennular sensory setae can disrupt the settlement behaviour of the model barnacle species Amphibalanus amphitrite. It is concluded that surface exploration by the cyprids relies on mechanical and nociception-related and calcium-mediated signals while a protein kinase C signalling cascade controls the timely metamorphosis of the cyprids to sessile juveniles.

Species fluctuations sustained by a cyclic succession at the edge of chaos.

Although mathematical models and laboratory experiments have shown that species interactions can generate chaos, field evidence of chaos in natural ecosystems is rare. We report on a pristine rocky intertidal community located in one of the world's oldest marine reserves that has displayed a complex cyclic succession for more than 20 y. Bare rock was colonized by barnacles and crustose algae, they were overgrown by mussels, and the subsequent detachment of the mussels returned bare rock again. These processes generated irregular species fluctuations, such that the species coexisted over many generations without ever approaching a stable equilibrium state. Analysis of the species fluctuations revealed a dominant periodicity of about 2 y, a global Lyapunov exponent statistically indistinguishable from zero, and local Lyapunov exponents that alternated systematically between negative and positive values. This pattern indicates that the community moved back and forth between stabilizing and chaotic dynamics during the cyclic succession. The results are supported by a patch-occupancy model predicting similar patterns when the species interactions were exposed to seasonal variation. Our findings show that natural ecosystems can sustain continued changes in species abundances and that seasonal forcing may push these nonequilibrium dynamics to the edge of chaos.

Two's a crowd? Crowding effect in a parasitic castrator drives differences in reproductive resource allocation in single vs double infections.

The 'crowding effect' is a result of competition by parasites within a host for finite resources. Typically, the severity of this effect increases with increasing numbers of parasites within a host and manifests in reduced body size and thus fitness. Evidence for the crowding effect is mixed - while some have found negative effects, others have found a positive effect of increased parasite load on parasite fitness. Parasites are consumers with diverse trophic strategies reflected in their life history traits. These distinctions are useful to predict the effects of crowding. We studied a parasitic castrator, a parasite that usurps host reproductive energy and renders the host sterile. Parasitic castrators typically occur as single infections within hosts. With multiple parasitic castrators, we expect strong competition and evidence of crowding. We directly assess the effect of crowding on reproductive success in a barnacle population infected by a unique parasitic castrator, Hemioniscus balani, an isopod parasite that infects and blocks reproduction of barnacles. We find (1) strong evidence of crowding in double infections, (2) increased frequency of double infections in larger barnacle hosts with more resources and (3) perfect compensation in egg production, supporting strong space limitation. Our results document that the effects of crowding are particularly severe for this parasitic castrator, and may be applicable to other castrators that are also resource or space limited.

Parasites as prey: the effect of cercarial density and alternative prey on consumption of cercariae by four non-host species.

In parasites with complex life cycles the transmission of free-living infective stages can be influenced by ambient community diversity, in particular via predation. Here, we experimentally investigated whether parasite density and the presence of alternative prey can alter predation rates on free-living cercarial stages of a marine trematode by several non-host predators. All four predator species consumed increasing numbers of cercariae with an increase in cercarial density, indicating that the removal of cercariae by predators is effective over a range of natural densities as well as in the presence of alternative prey for a number of predators typical of marine ecosystems. However, the relative removal rates and the effects of cercarial density and alternative prey differed among predator species. In barnacles and shrimps, significant interactive effects of cercarial density and alternative prey on cercarial predation occurred while in oysters and crabs cercarial removal rates were unaffected by both factors. As changes in cercarial densities directly translate into changes in infection levels in down-stream hosts in this parasite-host system, the observed predator-specific responses suggest that cercarial predation effects on disease risks will depend on the specific species composition of ambient communities and not on non-host biodiversity per se.

Effect of barnacle fouling on ship resistance and powering.

Predictions of added resistance and the effective power of ships were made for varying barnacle fouling conditions. A series of towing tests was carried out using flat plates covered with artificial barnacles. The tests were designed to allow the examination of the effects of barnacle height and percentage coverage on the resistance and effective power of ships. The drag coefficients and roughness function values were evaluated for the flat plates. The roughness effects of the fouling conditions on the ships' frictional resistances were predicted. Added resistance diagrams were then plotted using these predictions, and powering penalties for these ships were calculated using the diagrams generated. The results indicate that the effect of barnacle size is significant, since a 10% coverage of barnacles each 5 mm in height caused a similar level of added power requirements to a 50% coverage of barnacles each 1.25 mm in height.