AbstractCold acclimation is a biological process that allows animals to survive at low temperatures. The freshwater invertebrate is subject to broad changes in environmental temperature and does not have the required motility in order to move to warmer environments during the winter. For this reason, had to develop robust mechanisms to achieve cold acclimation at the onset of winter. How detects the onset of winter and activates its acclimation mechanism is unknown. Here, we used thermocyclers to induce cold acclimation in and study its properties. We found that cultured at room temperature does not survive an abrupt transition from 22 to 4 °C. However, it can be treated to become cold acclimated and survive at 4 °C by exposure to intermediate temperatures such as 12 °C if the treatment duration exceeds more than a week. Once cold acclimated, is considerably more robust to thermal changes. It survives repeated abrupt transitions from 4 to 22 °C and from 22 to 4 °C. However, acclimation is reversible, and if a cold-acclimated stays at room temperature for more than a week, it will gradually lose its cold acclimation. We developed a mathematical model representing the dynamics of this process and used it to predict survival according to temperature data recorded in one of their natural habitats. The results of these simulations provide an explanation for how survives winter under natural conditions. Accordingly, daily fluctuations are too short to cause injury, and seasonal fluctuations, which are long enough to be lethal, allow acclimation to incrementally build up and protect the animal. Cold acclimation in is therefore an example of a strategy that has adapted during evolution to match the animal's needs for survival.

AbstractIndividuals with similar biological requirements frequently compete for resources. Males and females have evolved different reproductive strategies in which females invest more in fecundity and males in intrasexual competition for mates. Although less common than within-sex competition, intersexual contests may occur to obtain resources. Interindividual differences in fighting ability bias the benefits and costs between opponents, and those differences are expected to be greater in intersexual contests. We compared the chela size, muscular strength, metabolic rate, and relative boldness of males and females of Bouvier, 1898. We further investigated how these traits influence intra- and intersexual contests for shells, with both sexes assuming roles as attackers and defenders. Males and females do not differ in chela size, muscular strength, or boldness. While males exhibited higher metabolic rates, this did not explain contest initiation or outcomes. Surprisingly, females initiated fewer contests against males than against females, and those that did often lost. However, this outcome was not attributed to sexually dimorphic traits. Instead, fighting success correlated with individual boldness and rapping frequency. Interestingly, rapping performance did not correlate with boldness, metabolic rate, or muscular strength, contrary to expectations. Our result reaffirms the common disadvantage of females as attackers in intersexual contests. However, they proved adept at defending their shells in both intra- and intersexual scenarios. This result, in addition to their typically efficient exploratory ability, raises questions about resource acquisition and distribution pathways in each sex.

AbstractSexual dimorphism typically arises as a result of sexual selection or sex-specific natural selection. Species that exhibit cryptic coloration provide an excellent system for studying sex-specific selection for sexual dichromatism. In this study, we examined the sexually dichromatic use of chromatophores in the seawhip shrimp, (Borradaile, 1920), which commonly resides on colonies of (Lamarck, 1815), a gorgonian octocoral that occurs in multiple color morphs. We documented the frequency of cryptic coloration in and tested colony color preference in relation to their current color. In the field, individuals always matched the color of the colony they were found on or were largely translucent. However, when given a choice, individuals chose colonies at random, independent of color. Additionally, we quantified locations across the bodies of males and females that differ most in chromatophore usage and tested the ability of both sexes to change color over 7 days. Females housed with colonies that differed in color from their starting color changed their coloration over the 7 days to better match the colony, while males remained translucent regardless of the color of colony they were housed with. individuals differed most in their coloration in regions corresponding to the saddle region and areas of the abdomen, locations in which females carry their eggs during development. These findings provide insight into the ecological factors underlying sexual dichromatism and shed light on the potential sex-specific selective mechanisms responsible for sexually dimorphic traits.

AbstractBleeding of horseshoe crabs () for the biomedical industry can have both sublethal and lethal impacts. Bleeding induces a significant drop in the concentration of hemolymph hemocyanin, as well as decreased levels of activity. Furthermore, horseshoe crabs with low hemocyanin prior to being bled have been found to be more likely to die after the procedure. The goal of this project was to determine whether feeding horseshoe crabs after bleeding them could enhance the recovery of their hemocyanin levels and, in doing so, improve their physiological status. The feeds tested in separate experiments included (1) natural forage items, blue mussels () or softshell clams (); (2) a formulated diet containing green crabs () and hemolymph; and (3) a modified commercially available shrimp () broodstock aquafeed. Horseshoe crabs ( = 63) were bled and then either fed or not fed, and their hemolymph hemocyanin concentrations were measured before they were bled and for the following 6-14 days. An additional 25 horseshoe crabs were treated in the same manner but not bled. In three experiments, horseshoe crabs that were fed consistently showed significantly higher hemolymph hemocyanin concentrations compared to those that were not fed. These data suggest that relatively simple modifications of the industrial bleeding procedure, such as feeding horseshoe crabs after bleeding them, may improve their physiological status prior to release.

AbstractThe distance that offspring disperse from their parents affects how a species responds to habitat disturbance, climate change, and interspecific interactions. For many benthic species, this dispersal is planktonic larvae, but the distance these larvae disperse is difficult to observe directly. Dispersal distance has usually been estimated indirectly by combining an observed quantity (, the rate of spread of an invasive organism or genetic similarity between locations) with a model that links that quantity to the dispersal of larvae. The estimates of dispersal distance based on the speed of spread of invasive organisms have led many researchers to conclude that the larvae of most of these organisms disperse much less than would be expected if they were being passively transported by the expected ocean currents (Shanks ; Shanks). I argue that the discrepancy is instead caused by the choice of model linking dispersal distance to invasion speed. Their model neglected the impact of life history, population growth, and oceanographic parameters on invasion speed. When dispersal distance is estimated from a more complete model of invasion speed, it is found that larval dispersal distance is not much less than would be expected for larvae drifting in the observed ocean currents.

AbstractIntegrated chemo- and mechanosensory pathways, along with activated calcium influxes, regulate nematocyst discharge from sea anemone tentacles. Discharge from vibration-sensitive Type A cnidocyte supporting cell complexes use calcium-conducting transient receptor potential V4-like channels. Because calcium influxes often couple with calcium-activated, large-conductance potassium (BK) channels, we hypothesized that BK channels function in nematocyst discharge. To verify this hypothesis, we first tested five selective BK channel blockers on nematocyst-mediated prey killing in (aka ). All tested BK channel blockers inhibited prey killing at concentrations comparable to their inhibition of vertebrate BK channels. In addition, the BK channel blocker paxilline selectively inhibited prey killing mediated by vibration-sensitive Type A cnidocyte supporting cell complexes. We queried a mammalian BK amino acid sequence to the database, from which we identified a putative anemone, pore-forming BK subunit sequence. Using the BK sequence as a template, we assembled a BK transcript from our assembled transcriptome. In addition, the hydra homolog of BK localizes to nematocytes on the hydra single-cell RNA sequencing map. Our findings suggest that expresses BK channels that play a role in vibration-sensitive nematocyst discharge from Type A cnidocyte supporting cell complexes. We believe this is the first functional demonstration of BK channels in nonbilaterians. Because stimulated chemoreceptors frequency tune Type A cnidocyte supporting cell complexes to frequencies matching swimming movements of prey a protein kinase A signaling pathway and protein kinase A generally activates BK channels, we suggest that BK channels may participate in protein kinase A-mediated frequency tuning.

AbstractThe order Fecampiida, a group of parasitic turbellarians, has been poorly studied in terms of its species diversity, morphology, and ecology. Fecampiida is positioned within the monophyletic clade Adiaphanida, along with Tricladida and Prolecithophora, but their phylogenetic relationships are not well understood. Although the nervous and muscular systems of only two species in Fecampiida have been studied, recent research inferred morphological similarities between Fecampiida and Prolecithophora. In this study, we collected fecampiid cocoons and juveniles at depths of 1861-4438 m in Japanese waters. We identified the species on the basis of swimming juvenile specimens and by using histological and molecular methods, while we also examined its musculature and nervous system. Our study revealed a more complex nervous system than previously reported, with dorsal, lateral, and ventral pairs of longitudinal nerve cords connected through an anterior neuropile and posterior transverse commissures. While the nervous and muscular morphology suggested similarities with Prolecithophora, our phylogenetic analysis did not support a close relationship between Fecampiida and Prolecithophora.

AbstractThe insulin-like androgenic gland hormone is a crucial sexual regulator that is involved in the masculine sexual differentiation of crustaceans. As an insulin-like peptide, the insulin-like androgenic gland hormone has been proposed to act through the insulin receptor-mediated pathway. The present study cloned and characterized two insulin receptors ( and ) from the swimming crab hallmarked with a conserved intracellular tyrosine kinase catalytic domain and several other typical insulin receptor domains in their deduced amino acid sequences. Both insulin receptors were predominately expressed in the testis and the insulin-like androgenic gland hormone-producing organ androgenic gland. Their testicular expression during the annual cycle suggested that they may play critical roles in spermatogenesis. By using the protein colocalization analysis in HEK293 cells, interactions of PtIAG with the two PtIRs were further confirmed. In addition, the insulin receptor antagonist was found to attenuate the stimulatory effects of androgenic gland homogenate on the phosphorylated MAPK levels in testis explants, suggesting that the insulin receptor-dependent MAPK pathway may be essential for insulin-like androgenic gland hormone functions.

AbstractThe Pacific razor clam, (Sugpiaq: Cingtaataq, Dixon, 1789), is vital to commercial, recreational, and subsistence fisheries across the Pacific Northwest Coast of North America. Despite the species' status as one of the most popular shellfish species harvested in the Pacific Northwest, British Columbia, and Alaska, its larval development has never been fully characterized. Generating a developmental times series, and describing development fully, is crucial for guiding targeted management, developing a mariculture strategy for the species, and providing a more pointed avenue for studies examining the response of to ocean change. This study presents the first photographic documentation of larval development in , including the timing of key transitions during embryogenesis and early larval development. Scanning electron microscopy revealed that the larval shell forms a concretion, a process typically documented in early gastropod development. This novel characterization is pertinent, as it conveys the need for the inclusion of alternative bivalve development processes, such as a concretion, in bivalve research. This study also compared development in to a global assortment of bivalve species, including two other members of the Pharidae family, determining that the timing to D-veliger and trochophore stages was similar to the majority of bivalves surveyed. While bivalve response to climate change is a topic of great interest, not all species of concern have undergone comprehensive developmental assessments, a requisite benchmark for designing climate change studies that examine early life history sensitivity to such changes. This research supports the use of comprehensive developmental studies as prerequisites for designing climate change experimentation, establishes the necessity of high-magnification and high-resolution scanning electron microscopy within developmental assessments, and provides information about the development of a cornerstone bivalve species.

AbstractGulf of Maine waters are warming rapidly, prompting a reevaluation of how commercially important marine species will respond. The goal of this study was to determine the respiratory, cardiac, and locomotory responses of American lobsters () to increasing water temperatures and to compare these to similar published studies. First, we measured the heart rate and ventilation rate of 10 lobsters that were confined in a temperature-controlled chamber while exposing them to gradually warming temperatures from 16 to 30 °C over 7 h. Both heart rate and ventilation rate increased along with the temperature up to a break point, with the mean heart rate peaking at 26.5 ± 1.6 °C, while the ventilation rate peaked at 27.4 ± 0.8 °C. In a subset of these trials ( = 5), oxygen consumption was also monitored and peaked at similar temperatures. In a second experiment, both the heart rate and activity of five lobsters were monitored with custom-built dataloggers while they moved freely in a large tank, while the temperature was increased from 18 to 29 °C over 24 h. The heart rate of these lobsters also increased with temperature, but their initial heart rates were lower than we recorded from confined lobsters. Finally, we confirmed that the low heart rates of the freely moving lobsters were due to the methods used by comparing heart rate data from eight lobsters collected using both methods with each individual animal. Thus, while our overall results are consistent with data from previous studies, they also show that the methods used in studies of physiological and behavioral responses to warming temperatures can impact the results obtained.

AbstractSea stars are a major component of the megabenthos in most marine habitats, including those within the deep sea. Being radially symmetric, sea stars have sensory structures that are evenly distributed along the arms, with a compound eye located on each arm tip of most examined species. Surprisingly, eyes with a spatial resolution that rivals the highest acuity known among sea stars so far were recently found in , a member of the deep-sea sea star order Brisingida. Here, we examined 21 species across 11 brisingid genera for the presence of eyes; where eyes were present, we used morphological characteristics to evaluate spatial resolution and sensitivity. This study found that eyes were present within 43% of the examined species. These brisingid eyes were relatively large compared to those of other deep-sea sea stars, with a high number of densely packed ommatidia. One of the examined species, , had more than 600 ommatidia per eye, which is the highest number of ommatidia found in any sea star eye so far. Combined, the results indicate that brisingid eyes are adapted for spatial resolution over sensitivity. Together with results showing that many brisingids are bioluminescent, this relatively high spatial resolution suggests that the group may use their eyes to support visually guided intraspecific communication based on bioluminescent signals. Phylogenetic analysis indicated that the common ancestor of brisingids had eyes ( = 0.72) and that eyes were lost once within the clade.

AbstractThe form of the cyphonautes larva of bryozoans changes little during development. The ciliated band that generates the feeding current increases nearly in proportion to body length, so that the maximum rate of clearing planktonic food from a volume of water becomes increasingly low relative to body protein. This development is unlike the other larvae that produce a feeding current with bands of simple cilia. The cyphonautes' growth rate has therefore been predicted to be unusually low when food is scarce. As predicted, cyphonautes larvae of a species of starved at concentrations of food that supported growth of pluteus larvae. Comparisons between the cyphonautes and plutei of a sand dollar were for growth from first feeding to metamorphosis, with a mix of two algal species. Another comparison was for growth of cyphonautes at an advanced stage and plutei of a regular sea urchin at an early stage, with food in seawater at a reduced concentration. The low maximum clearance rate did not prevent rapid growth and development of some cyphonautes from egg through metamorphosis when food was abundant. Twenty-nine days for development to metamorphosis in the laboratory with abundant food was close to Yoshioka's estimate of larval duration from the time lag between adult zooid density and larval abundance in a population in the Southern California Bight. Despite individual variation in growth rates and other physiological and environmental influences, simple measures of larval form predicted the differences in larval performance: scarce food extended larval duration for the cyphonautes more than for plutei.

AbstractWe describe the cloning and expression of a nonreceptor tyrosine kinase, (), a () gene, identified in a subtractive screen for maternal ascidian cDNAs in , an ascidian species with a tadpole larva. The gene encodes a 4-kb mRNA expressed in gonads, eggs, and embryos in the tailed but is not detected in eggs or embryos of the closely related tailless species . There is a large insertion in in the genome, as shown by transcriptome and genome analyses, resulting in it becoming a pseudogene. The amino acid sequence encodes a nonreceptor tyrosine kinase with an N-terminal region containing two SH2 domains and five ankyrin repeats, similar to the gene found in other ascidians. Thus, the ascidian genes are members of the SHARK (Src-homology ankyrin-repeat containing tyrosine kinase) family of nonreceptor tyrosine kinases, which are found throughout invertebrates and missing from vertebrates. We show that is lacking the tyrosine kinase domain in the tailless , although the truncated mRNA is still expressed in transcriptome data. This maternal and zygotic tyrosine kinase is another described pseudogene from and appears not to be necessary for adult development.

AbstractExtracellular calcium has been known to be required for nematocyst discharge for more than 60 years, yet calcium's role in nematocyst discharge is poorly understood. Currently, we know that extracellular calcium plays at least two distinct roles in nematocyst discharge. First, calcium plays a role in the triggering of discharge by physical contact, most likely involving transient receptor potential channels. Second, activated L-type calcium channels desensitize nematocyst discharge predisposed to discharge by stimulated chemoreceptors for -acetylated sugars, such as -acetylneuraminic acid (NANA). It is not known whether the stimulated NANA signaling pathway activates L-type channels electrogenically through membrane depolarization or directly by phosphorylation of the channel. We hypothesize that the activated NANA signaling pathway initiates desensitization by depolarizing cell membrane potentials to activate voltage-gated L-type calcium channels. Consistent with our hypothesis, we show that depolarization induced by blocking voltage-gated potassium channels with 4-aminopyridine selectively activates Ca influx into tentacle ectodermal cells L-type channels and inhibits nematocyst discharge from chemosensitized anemones. Furthermore, preventing membrane depolarization with valinomycin or hyperpolarizing resting membrane potentials with low-potassium seawater suppresses NANA-induced Ca influx, prevents desensitization of nematocyst discharge, and enhances NANA sensitivity. Thus, changing resting membrane potentials modulates NANA sensitivity, and NANA-induced depolarization drives desensitization. We suggest that desensitization of the NANA signaling pathway occurs by a feedback pathway involving calcium channels that are activated by NANA-induced depolarization. Elucidating the desensitization pathway may suggest methods to protect or prevent public health cases of nematocyst stinging.

AbstractColonial animals are composed of clonal individuals that remain physically connected and physiologically integrated. Salps are tunicates with a dual life cycle, including an asexual solitary stage that buds sexual colonies composed of jet-propelling zooids that efficiently swim together as a single unit by multijet propulsion. Colonies from different species develop distinct architectures characterized by their zooid arrangement patterns, but this diversity has received little attention. Thus, these architectures have never been formally defined using a framework of variables and axes that would allow comparative analyses. We set out to define an ontology of the salp colony architecture morphospace and describe the developmental pathways that build the different architectures. To inform these definitions, we collected and photographed live specimens of adult and developing colonies through offshore scuba diving. Since all salp colonies begin their development as a transversal double chain, we characterized each adult colonial architecture as a series of developmental transitions, such as rotations and translations of zooids, relative to their orientation at this early shared stage. We hypothesize that all adult architectures are either final or intermediate stages within three developmental pathways toward bipinnate, cluster, or helical forms. This framework will enable comparative studies on the biomechanical implications, ecological functions, evolutionary history, and engineering applications of the diversity of salp colony architectures.

AbstractUnderstanding the evolution of development (evo-devo) in the Ophiuroidea and the pathways in the switch from a feeding to a nonfeeding larva is complicated by the variability in the phenotype of the metamorphic larva, being a reduced yolky ophiopluteus in some species (type I development) and a vitellaria larva in others (type II development). We investigated evo-devo in the family Ophionereididae, a group dominated by lecithotrophic development through a vitellaria larva. We reared the planktotrophic larvae of to settlement to determine the metamorphic phenotype. Counter to expectations, did not exhibit type II metamorphosis through a vitellaria, although it did exhibit transient vitellaria-like features. Resorption of the larval arms in the same interradial positions where the ciliary bands form in vitellariae gave them a fleeting vitellaria-like appearance. Development of exhibits heterochronic features in early formation of the skeletal primordium of the third pair (postoral) of larval arms and in the presettlement juvenile early appearance of the juvenile terminal arm plates on external view in parallel with larval arm resorption. Development of the fourth pair (posterodorsal) of larval arms, the last pair to be formed, is plastic, with 44% of larvae exhibiting partial arm growth. Heterochronic traits in development, as seen in , may have facilitated evolution of a lecithotrophic mode of development in . Comparison of the ophiopluteus of and the vestigial pluteus of provided insights into the simplification of larval form from the ancestral (feeding larva) state in . The diverse metamorphic phenotypes in ophiuroids indicate that type I and type II development may not be completely divergent lines of evo-devo and point to selective pressure in the pelagic-benthic transition in the evolution of ophiuroid development.

AbstractAlthough the gut and the brain vastly differ in physiological function, they have been interlinked in a variety of different neurological and behavioral disorders. The bacteria that comprise the gut microbiome communicate and influence the function of various physiological processes within the body, including nervous system function. However, the effects of social experience in the context of dominance and social stress on gut microbiome remain poorly understood. Here, we examined whether social experience impacts the host zebrafish () gut microbiome. We studied how social dominance during the first 2 weeks of social interactions changed the composition of zebrafish gut microbiome by comparing gut bacterial composition, diversity, and relative abundance between socially dominant, submissive, social isolates and control group-housed communal fish. Using amplicon sequencing of the rRNA gene, we report that social dominance significantly affects host gut bacterial community composition but not bacterial diversity. At the genus level, and unclassified Enterobacteriaceae relative abundance decreased in dominant individuals while commensal bacteria (, and ) increased in relative abundance. Conversely, the relative abundance of and was increased in subordinates, isolates, and communal fish compared to dominant fish. The shift in commensal and pathogenic bacteria highlights the impact of social experience and the accompanying stress on gut microbiome, with potentially similar effects in other social organisms.

AbstractSpecies undergoing postreproductive death experience great changes in their reproductive organs, which are driven by numerous physiological processes. To assess whether apoptotic processes are involved in the dynamics of the reproductive organs of , the gonadal structure of this semelparous side-gilled sea slug was studied using light and scanning electron microscopy. Apoptotic cells at different gonadal developmental stages were detected by TUNEL assay. Apoptosis was primarily focused on spermatogonia during gonadal cell proliferation, probably as a regulatory mechanism that maintains homeostasis in reproductive cells. Visible gonadal degeneration at the end of the reproductive period is accompanied by apoptosis of the basal lamina cells of the acini, suggesting that apoptotic processes are involved in the gonadal degeneration observed in .

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 infected by the rhizocephalan . In field-collected , infection by was associated with development of female sex characters at a smaller size and greater probability of epibiosis. Standardized video observations showed that infected 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 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.

AbstractMass mortality events provide valuable insight into biological extremes and also ecological interactions more generally. The sea star wasting epidemic that began in 2013 catalyzed study of the microbiome, genetics, population dynamics, and community ecology of several high-profile species inhabiting the northeastern Pacific but exposed a dearth of information on the diversity, distributions, and impacts of sea star wasting for many lesser-known sea stars and a need for integration across scales. Here, we combine datasets from single-site to coast-wide studies, across time lines from weeks to decades, for 65 species. We evaluated the impacts of abiotic characteristics hypothetically associated with sea star wasting (sea surface temperature, pelagic primary productivity, upwelling wind forcing, wave exposure, freshwater runoff) and species characteristics (depth distribution, developmental mode, diet, habitat, reproductive period). We find that the 2010s sea star wasting outbreak clearly affected a little over a dozen species, primarily intertidal and shallow subtidal taxa, causing instantaneous wasting prevalence rates of 5%-80%. Despite the collapse of some populations within weeks, environmental and species variation protracted the outbreak, which lasted 2-3 years from onset until declining to chronic background rates of ∼2% sea star wasting prevalence. Recruitment began immediately in many species, and in general, sea star assemblages trended toward recovery; however, recovery was heterogeneous, and a marine heatwave in 2019 raised concerns of a second decline. The abiotic stressors most associated with the 2010s sea star wasting outbreak were elevated sea surface temperature and low wave exposure, as well as freshwater discharge in the north. However, detailed data speaking directly to the biological, ecological, and environmental cause(s) and consequences of the sea star wasting outbreak remain limited in scope, unavoidably retrospective, and perhaps always indeterminate. Redressing this shortfall for the future will require a broad spectrum of monitoring studies not less than the taxonomically broad cross-scale framework we have modeled in this synthesis.