Predators Induce Phenotypic Plasticity in Echinoderms across Life History Stages.

AbstractMarine invertebrates with biphasic life cycles feature life history transitions that coincide with habitat changes from benthic adults to planktonic embryos and larvae, then a return to the benthos as a juvenile at metamorphosis. The metamorphic transition exposes animals to a new suite of benthic predators, and high mortality often occurs in the hours and days following settlement. Juvenile invertebrates may produce phenotypically plastic morphological defenses when predator cues are detected. However, time lags inherent to phenotypic plasticity may delay the production of defenses until after the period of highest vulnerability. It should, therefore, be beneficial for planktonic larvae approaching settlement to detect waterborne cues from benthic predators and produce juvenile phenotypes appropriate for postmetamorphic survival. Echinoderms are useful models for testing transhabitat and trans-life history stage phenotypic plasticity because many species have larvae that construct their juvenile phenotype while still in the water column. In this study, we tested whether planktonic echinoderm larvae exposed to cues from benthic predators modified their juvenile phenotypes at settlement. Green urchin (Strongylocentrotus droebachiensis) and Pacific sand dollar (Dendraster excentricus) larvae were exposed to predatory green crab (Carcinus maenus) or red rock crab (Cancer productus) cues, respectively, from their early-stage juvenile rudiment formation through settlement. Green urchin larvae exposed to predator cues settled with significantly more juvenile spines compared to unexposed controls. Sand dollars exhibited earlier settlement, larger disk area, fewer spines, and shorter spines when exposed to benthic predator cues. Sand dollar larvae were also exposed to cues from planktonic crab larvae and in response settled sooner and larger, with even fewer and shorter spines than those exposed to benthic predator cues. These results suggest that echinoderm larvae alter their juvenile phenotype in response to predator cues, but the response varies between species, and responses to planktonic threats may be prioritized over benthic ones.

Cloning and Selfing Affect Population Genetic Variation in Simulations of Outcrossing, Sexual Sea Stars.

AbstractMany sea stars are well known for facultative or obligate asexual reproduction in both the adult and larval life-cycle stages. Some species and lineages are also capable of facultative or obligate hermaphroditic reproduction with self-fertilization. However, models of population genetic variation and empirical analyses of genetic data typically assume only sexual reproduction and outcrossing. A recent reanalysis of previously published empirical data (microsatellite genotypes) from two studies of one of the most well-known sea star species (the crown-of-thorns sea star; Acanthaster sp.) concluded that cloning and self-fertilization in that species are rare and contribute little to patterns of population genetic variation. Here we reconsider that conclusion by simulating the contribution of cloning and selfing to genetic variation in a series of models of sea star demography. Simulated variation in two simple models (analogous to previous analyses of empirical data) was consistent with high rates of cloning or selfing or both. More realistic scenarios that characterize population flux in sea stars of ecological significance, including outbreaks of crown-of-thorns sea stars that devastate coral reefs, invasions by Asterias amurensis, and epizootics of sea star wasting disease that kill Pisaster ochraceus, also showed significant but smaller effects of cloning and selfing on variation within subpopulations and differentiation between subpopulations. Future models or analyses of genetic variation in similar study systems might benefit from simulation modeling to characterize possible contributions of cloning or selfing to genetic variation in population samples or to understand the limits on inferring the effects of cloning or selfing in nature.

Effects of embryo energy, egg size, and larval food supply on the development of asteroid echinoderms.

Organisms have limited resources available to invest in reproduction, causing a trade-off between the number and size of offspring. One consequence of this trade-off is the evolution of disparate egg sizes and, by extension, developmental modes. In particular, echinoid echinoderms (sea urchins and sand dollars) have been widely used to experimentally manipulate how changes in egg size affect development. Here, we test the generality of the echinoid results by (a) using laser ablations of blastomeres to experimentally reduce embryo energy in the asteroid echinoderms (sea stars), Pisaster ochraceus and Asterias forbesi and (b) comparing naturally produced, variably sized eggs (1.7-fold volume difference between large and small eggs) in A. forbesi. In P. ochraceus and A. forbesi, there were no significant differences between juveniles from both experimentally reduced embryos and naturally produced eggs of variable size. However, in both embryo reduction and egg size variation experiments, simultaneous reductions in larval food had a significant and large effect on larval and juvenile development. These results indicate that (a) food levels are more important than embryo energy or egg size in determining larval and juvenile quality in sea stars and (b) the relative importance of embryo energy or egg size to fundamental life history parameters (time to and size at metamorphosis) does not appear to be consistent within echinoderms.

Instant Ocean Versus Natural Seawater: Impacts on Aspects of Reproduction and Development in Three Marine Invertebrates.

Marine invertebrate larvae have often been reared in artificial rather than natural seawater, either for convenience or to avoid potentially confounding effects of unknown contaminants. This study sought to determine the impact of artificial seawater on various aspects of development for three marine invertebrate species. We examined the impact of Instant Ocean on growth, survival, and fecundity of the deposit-feeding polychaete Capitella teleta at 2 salinities: 24 and 34 ppt; the impact on survival, growth rate, and time to metamorphic competence for the slipper limpet Crepidula fornicata; and the impact on larval growth for the sea star Asterias forbesi. Juveniles of C. teleta survived better in natural seawater than in Instant Ocean at both salinities but at the higher salinity grew more quickly in Instant Ocean; fecundity was not significantly affected by the type of seawater used at either salinity. Using Instant Ocean in place of natural seawater had no pronounced impact on the survival of C. fornicata larvae or on how long it took them to become competent to metamorphose; however, larvae grew somewhat more quickly in Instant Ocean than in natural seawater for the first 4 days of development, but by day 7 they were about 4.5% larger if they had been reared in seawater. The type of seawater used affected the growth of A. forbesi larvae, with larvae growing significantly more slowly in Instant Ocean than in natural seawater, no matter how growth was measured. In conclusion, our results suggest that although Instant Ocean may be a reasonable substitute for natural seawater for work with some species, using it may affect experimental outcomes in some aspects of work with other species.

Culturing echinoderm larvae through metamorphosis.

Echinoderms are favored study organisms not only in cell and developmental biology, but also physiology, larval biology, benthic ecology, population biology and paleontology, among other fields. However, many echinoderm embryology labs are not well-equipped to continue to rear the post-embryonic stages that result. This is unfortunate, as such labs are thus unable to address many intriguing biological phenomena, related to their own cell and developmental biology studies, that emerge during larval and juvenile stages. To facilitate broader studies of post-embryonic echinoderms, we provide here our collective experience rearing these organisms, with suggestions to try and pitfalls to avoid. Furthermore, we present information on rearing larvae from small laboratory to large aquaculture scales. Finally, we review taxon-specific approaches to larval rearing through metamorphosis in each of the four most commonly-studied echinoderm classes-asteroids, echinoids, holothuroids and ophiuroids.

Developmental Consequences of Temperature and Salinity Stress in the Sand Dollar Dendraster excentricus.

Animals that reside, reproduce, and develop in nearshore habitats are often exposed to strong fluctuations in abiotic conditions, including temperature and salinity. We studied the developmental response of the sand dollar Dendraster excentricus to increased temperature and reduced salinity at levels comparable to those induced by summer freshwater input into the San Juan Archipelago, Washington. We observed that embryos exposed to temperature and salinity stress exhibited polyembryony (the splitting of one embryo into multiple independent individuals), and we subsequently tested the competency of twin and normal embryos to reach metamorphosis. We found that twin embryos generated from a single egg are each capable of reaching metamorphosis. To begin investigating the mechanisms underlying polyembryony, we tested whether osmotic stress caused swelling of the fertilization envelope, thus allowing embryos the physical space to produce multiples within a single envelope. We also tested whether reduced calcium levels in low-salinity seawater reduced cell-cell adhesion and allowed cells to separate and develop as multiple embryos within a fertilization envelope. However, neither osmotic stress nor reduced calcium levels alone appear sufficient to induce polyembryony. We hypothesize that changes in the properties of the hyaline layer that lies beneath the fertilization envelope facilitate polyembryony.

Environmental Induction of Polyembryony in Echinoid Echinoderms.

Polyembryony, or the production of multiple offspring from a single zygote, is a widespread phenomenon in the animal kingdom. Various types of polyembryony have been described in arthropods, bryozoans, chordates, cnidarians, echinoderms, and platyhelminthes. We describe the induction of polyembryony in embryos of the sand dollar Echinarachnius parma and the pencil urchin Eucidaris tribuloides in response to elevated temperature and reduced salinity. Data on the environmental variation in temperature and salinity that normally occurs during the spawning season, combined with the range of laboratory conditions over which polyembryony was induced, suggest that polyembryony may occur frequently in these species under natural conditions. We tested an additional two species of echinoids for similar responses, but found little evidence for polyembryony in the green urchin Strongylocentrotus droebachiensis or the variegated urchin Lytechinus variegatus, suggesting that polyembryony is not a universal response of echinoids to fluctuations in temperature and salinity. The unexpected developmental changes that we observed in response to present-day fluctuations in temperature and salinity suggest that ongoing and future environmental shifts may drive substantial changes in marine invertebrate developmental patterns, and that these changes will be different across taxa.

Gongylonema pulchrum infection in a resident of Williamsburg, Virginia, verified by genetic analysis.

We describe the thirteenth reported case of human infection with Gongylonema spp. in the United States and the first to be confirmed as Gongylonema pulchrum. The parasite described was isolated from the oral cavity of a resident of Williamsburg, Virginia. The identity of the parasite was verified through morphological and genetic approaches, and provided the first genetic confirmation of a Gongylonema sp. in humans.

A novel report of hatching plasticity in the phylum Echinodermata.

Hatching plasticity occurs in response to a wide range of stimuli across many animal taxa, including annelids, arthropods, mollusks, and chordates. Despite the prominence of echinoderms in developmental biology and more than 100 years of detailed examination of their development under a variety of conditions, environmentally cued hatching plasticity has never been reported in the phylum Echinodermata. Here we report plasticity in the timing and stage of hatching of embryos of the sand dollar Echinarachnius parma in response to reductions in salinity. Embryos of E. parma increased their time to hatching more than twofold in response to ecologically relevant salinity reductions, while maintaining an otherwise normal developmental schedule. Embryos that experienced the greatest delay in hatching time emerged from the fertilization envelope as four-arm pluteus larvae rather than hatching as blastulae or early gastrulae. Salinity manipulations across multiple male-female pairs indicated high variability in hatching time both within and among clutches, suggesting significant intraspecific variation in developmental responses to salinity.

Understanding the effects of low salinity on fertilization success and early development in the sand dollar Echinarachnius parma.

Free-spawning marine invertebrates that live near shore or in estuaries may experience reduced fertilization success during low-salinity events. Although several studies have documented reproductive failure at reduced salinity in estuarine animals, few have looked at whether developmental failure is due to a failure of fertilization or to a failure of fertilized eggs to cleave. In this study, we examined the effects of salinities ranging from 18 to 32 psu on fertilization success and early development in the sand dollar Echinarachnius parma. In addition to decoupling the effects of low salinity on fertilization from its effects on early cleavage, we also assessed whether eggs or sperm were the weak link in accounting for reproductive failure. We found that both fertilization and cleavage failed at salinities below about 22 psu but that development could be partially rescued by returning zygotes to full-strength seawater. We also found that sperm remained active and capable of fertilizing eggs even after being exposed to low salinities for 30 min.. Taken together, these results suggest that reproductive failure at low salinities in E. parma is due more to an inability of the fertilized eggs to cleave than to an inability of sperm to fertilize eggs.

The peril of the plankton.

The pelagic environment is characterized by unevenly distributed resources and risks. Such unpredictability presents adaptive challenges to diverse planktonic organisms including the larvae of benthic marine invertebrates. Estimates of mortality during planktonic development are highly variable, ranging from 0% to 100% per day. Predation is considered a significant source of this mortality, but what explains the variability in estimates of the mortality of marine invertebrate larvae? While differential exposure of larval prey to predators may explain these widely variable estimates, adaptations that reduce vulnerability of marine larvae to predators may also be important. Although there are excellent reviews of predation upon larvae and of larval mortality and defenses, nearly 15 years have elapsed since these topics were formally reviewed. Here, we highlight recent advances in understanding the behavioral, chemical, and morphological defenses that larvae possess and assess their effectiveness in reducing the risk of predation. While recent work confirms that larval mortality is generally high, it also demonstrates that larvae can reduce their risk of predation in several ways, including: (1) temporarily escaping the benthos during vulnerable early stages, (2) producing chemical compounds that reduce palatability, (3) possessing morphological defenses such as spines and shells, and (4) exhibiting induced defensive responses whereby larvae can alter their behavior, morphology, and life histories in the presence of predators. Taken together, these studies indicate that marine invertebrate larvae possess a sophisticated suite of defensive phenotypes that have allowed them to persist in the life cycle of benthic invertebrates for eons.

How do changes in parental investment influence development in echinoid echinoderms?

Understanding the relationship between egg size, development time, and juvenile size is critical to explaining patterns of life-history evolution in marine invertebrates. Currently there is conflicting information about the effects of changes in egg size on the life histories of echinoid echinoderms. We sought to resolve this conflict by manipulating egg size and food level during the development of two planktotrophic echinoid echinoderms: the green sea urchin, Strongylocentrotus droebachiensis and the sand dollar, Echinarachnius parma. Based on comparative datasets, we predicted that decreasing food availability and egg size would increase development time and reduce juvenile size. To test our prediction, blastomere separations were performed in both species at the two-cell stage to reduce egg volume by 50%, producing whole- and half-size larvae that were reared to metamorphosis under high or low food levels. Upon settlement, age at metamorphosis, juvenile size, spine number, and spine length were measured. As predicted, reducing egg size and food availability significantly increased age at metamorphosis and reduced juvenile quality. Along with previous egg size manipulations in other echinoids, this study suggests that the relationship between egg size, development time, and juvenile size is strongly dependent upon the initial size of the egg.

Size-specific predation on marine invertebrate larvae.

Predation on planktonic larval stages is frequently a major source of mortality for the offspring of benthic marine invertebrates. Mortality rate likely varies with larval size and developmental stage, but few experiments have measured how these factors affect predation rates. I used experimental reductions in egg size to test how variation in larval size affects the likelihood of predation during planktonic development. Blastomeres of the sand dollar Dendraster excentricus were separated at the two-cell stage to produce half-sized zygotes. Larvae resulting from this manipulation were tested for their susceptibility to predation relative to whole-sized siblings at four ages. Individuals from each size class were simultaneously presented as prey items to five predators (crab zoeae, crab megalopae, chaetognaths, solitary tunicates, and postlarval fish) in the laboratory. Four predators consumed significantly more half-sized larvae than whole-sized larvae, but one predator type (postlarval fish) consumed more whole-sized larvae. Predators that consumed more half-sized larvae also preferentially consumed younger larvae. In contrast, postlarval fish showed no significant prey preference based on larval age. These results suggest that assumptions of constant mortality rates during development should be modified to account for the effects of larval size and age.

Intermediate modes of larval development: bridging the gap between planktotrophy and lecithotrophy.

The extraordinary diversity of larval form and function in marine invertebrates has motivated many studies of development, ecology, and evolution. Among organisms with pelagic development via a larval stage, this diversity is often reduced to a dichotomy between two broad nutritional categories: planktotrophy and lecithotrophy. Despite the clear utility of the planktotrophy-lecithotrophy dichotomy to those interested in the history or consequences of life history patterns, it is also clear that a number of larval forms do not fit neatly into either of these general categories. Here we review studies of these intermediate larval forms, focusing on descriptions of larvae known as facultative feeders. Recent descriptions of larval development suggest that facultative feeders and other intermediate larval forms are not as rare as commonly assumed. We assess the importance of these forms for models of life-history evolution and call for a more-detailed and nuanced view of larval biology to account for their existence. Clearer knowledge of the phylogenetic distribution and frequency of occurrence of larvae that exhibit intermediate nutritional requirements is also essential for evaluating current ideas on evolutionary transitions between planktotrophy and lecithotrophy. Finally, intermediate larval types provide valuable and underutilized opportunities for testing hypotheses in the fields of larval ecology and the evolution of development.

How does metabolic rate scale with egg size? An experimental test with sea urchin embryos.

The consequences of changes in egg size for the development of marine invertebrates have been the subject of much theoretical and experimental work. Models that explore larval developmental modes in the context of maternal investment per offspring are often couched in an energetic framework, but the relationships between egg size and the energetics of larval development are poorly understood. We used blastomere separations to examine how experimental reductions in egg size affected (1) larval metabolic rate and (2) larval resistance to starvation. We found that separating blastomeres at the 2- and 4-cell stage resulted in average reductions of 50% and 75%, respectively, in larval metabolic rate. This suggests that, in an experimental context, mass-specific metabolic rate does not change with egg size. We also found that a 50% reduction in egg volume did not reduce the resistance of larvae to starvation when particulate food was withheld. This suggests that the material supplied to larvae in the egg is used primarily for construction of the larval body, rather than as a buffer against starvation or as a means of reducing reliance on exogenous fuel to sustain maintenance metabolism.