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Abstract Mollusca is a morphologically diverse phylum, exhibiting an immense variety of calcium carbonate structures. Proteomic studies of adult shells often report high levels of rapidly-evolving, ‘novel’ shell matrix proteins (SMPs), which are hypothesized to drive shell diversification. However, relatively little is known about the phylogenetic distribution of SMPs, or about the function of individual SMPs in shell construction. To understand how SMPs contribute to shell diversification a thorough characterization of SMPs is required. Here, we build tools and a foundational understanding of SMPs in the marine gastropod species Crepidula fornicata and Crepidula atrasolea because they are genetically-enabled mollusc model organisms. First, we established a staging system of shell development in C. atrasolea for the first time. Next, we leveraged previous findings in C. fornicata combined with phylogenomic analyses of 95 metazoan species to determine the evolutionary lineage of its adult SMP repertoire. We found that 55% of C. fornicata’s SMPs belong to molluscan orthogroups, with 27% restricted to Gastropoda, and only 5% restricted at the species level. The low percentage of species-restricted SMPs underscores the importance of broad-taxon sampling and orthology inference approaches when determining homology of SMPs. From our transcriptome analysis, we found that the majority of C. fornicata SMPs that were found conserved in C. atrasolea were expressed in both larval and adult stages. We then selected a subset of SMPs of varying evolutionary ages for spatial-temporal analysis using in situ hybridization chain reaction (HCR) during larval shell development in C. atrasolea. Out of the 18 SMPs analyzed, 12 were detected in the larval shell field. These results suggest overlapping larval vs. adult SMP repertoires. Using multiplexed HCR, we observed five SMP expression patterns and three distinct cell populations within the shell field. These patterns support the idea that modular expression of SMPs could facilitate divergence of shell morphological characteristics. Collectively, these data establish an evolutionary and developmental framework in Crepidula that enables future comparisons of molluscan biomineralization to reveal mechanisms of shell diversification.

In order to investigate the serendipitous find of a gastropod encrusted by the symbiont-bearing colonial coral Oculina patagonica, we examined several specimens of cnidarian-encrusted gastropods, ranging in age from the Pliocene to the Recent, and characterized in detail their sclerobiont cover. The results of our analysis suggest that gastropod shells can be encrusted by symbiont-bearing colonial corals at various times: (1) when the gastropod is alive; (2) when the shell is being used by a secondary inhabitant (e.g., hermit crabs or sipunculid worms); (3) when the shell is discarded but yet to be buried. The relationship between the symbiont-bearing coral and the inhabitant(s) of the encrusted shell is an example of facultative mutualism, i.e., it is non-obligate yet beneficial for both ends as the former obtains the capability to move, and the latter improves the resistance and resilience of its armor, thus obtaining extra protection from predators. Being able to move could prove particularly useful for a symbiont-bearing coral because, in addition to removing the risk of being smothered by sediment, it would also favor the photosynthetic activity of its algal endosymbionts by allowing the coral to be always clean of sedimentary particles. Although the resulting epibiotic association would be limited in size by the ability of either the gastropod or the secondary inhabitant of the shell to move at the seafloor, these small and easy-to-miss benthic islands might become the seeds that allow sessile carbonate producers such as hermatypic colonial corals to colonize unconsolidated substrates. [ABSTRACT FROM AUTHOR]

Abstract Background One hundred years ago, marine organisms were the dominant systems for the study of developmental biology. The challenges in rearing these organisms outside of a marine setting ultimately contributed to a shift towards work on a smaller number of so-called model systems. Those animals are typically non-marine organisms with advantages afforded by short life cycles, high fecundity, and relative ease in laboratory culture. However, a full understanding of biodiversity, evolution, and anthropogenic effects on biological systems requires a broader survey of development in the animal kingdom. To this day, marine organisms remain relatively understudied, particularly the members of the Lophotrochozoa (Spiralia), which include well over one third of the metazoan phyla (such as the annelids, mollusks, flatworms) and exhibit a tremendous diversity of body plans and developmental modes. To facilitate studies of this group, we have previously described the development and culture of one lophotrochozoan representative, the slipper snail Crepidula atrasolea, which is easy to rear in recirculating marine aquaria. Lab-based culture and rearing of larger populations of animals remain a general challenge for many marine organisms, particularly for inland laboratories. Results Here, we describe the development of an automated marine aquatic rack system for the high-density culture of marine species, which is particularly well suited for rearing filter-feeding animals. Based on existing freshwater recirculating aquatic rack systems, our system is specific to the needs of marine organisms and incorporates robust filtration measures to eliminate wastes, reducing the need for regular water changes. In addition, this system incorporates sensors and associated equipment for automated assessment and adjustment of water quality. An automated feeding system permits precise delivery of liquid food (e.g., phytoplankton) throughout the day, mimicking real-life feeding conditions that contribute to increased growth rates and fecundity. Conclusion This automated system makes laboratory culture of marine animals feasible for both large and small research groups, significantly reducing the time, labor, and overall costs needed to rear these organisms.

In order to investigate the serendipitous find of a gastropod encrusted by the symbiont-bearing colonial coral Oculina patagonica, we examined several specimens of cnidarian-encrusted gastropods, ranging in age from the Pliocene to the Recent, and characterized in detail their sclerobiont cover. The results of our analysis suggest that gastropod shells can be encrusted by symbiont-bearing colonial corals at various times: (1) when the gastropod is alive; (2) when the shell is being used by a secondary inhabitant (e.g., hermit crabs or sipunculid worms); (3) when the shell is discarded but yet to be buried. The relationship between the symbiont-bearing coral and the inhabitant(s) of the encrusted shell is an example of facultative mutualism, i.e., it is non-obligate yet beneficial for both ends as the former obtains the capability to move, and the latter improves the resistance and resilience of its armor, thus obtaining extra protection from predators. Being able to move could prove particularly useful for a symbiont-bearing coral because, in addition to removing the risk of being smothered by sediment, it would also favor the photosynthetic activity of its algal endosymbionts by allowing the coral to be always clean of sedimentary particles. Although the resulting epibiotic association would be limited in size by the ability of either the gastropod or the secondary inhabitant of the shell to move at the seafloor, these small and easy-to-miss benthic islands might become the seeds that allow sessile carbonate producers such as hermatypic colonial corals to colonize unconsolidated substrates.

Sexual systems are surprisingly diverse, considering the ubiquity of sexual reproduction. Sequential hermaphroditism, the ability of an individual to change sex, has emerged multiple times independently across the animal kingdom. In molluscs, repeated shifts between ancestrally separate sexes and hermaphroditism are generally found at the level of family and above, suggesting recruitment of deeply conserved mechanisms. Despite this, molecular mechanisms of sexual development are poorly known. In molluscs with separate sexes, endocrine disrupting toxins bind the retinoid X receptor (RXR), activating ectopic male development in females, suggesting the retinoid pathway as a candidate controlling sexual transitions in sequential hermaphrodites. We therefore tested the role of retinoic acid signaling in sequentially hermaphroditic Crepidula snails, which develop first into males, then change sex, maturing into females. We show that retinoid agonists induce precocious penis growth in juveniles and superimposition of male development in females. Combining RXR antagonists with retinoid agonists significantly reduces penis length in induced juveniles, while similar treatments using retinoic acid receptor (RAR) antagonists increase penis length. Transcripts of both receptors are expressed in the induced penis. Our findings therefore show that retinoid signaling can initiate molluscan male genital development, and regulate penis length. Further, we show that retinoids induce ectopic male development in multiple Crepidula species. Species-specific influence of conspecific induction of sexual transitions correlates with responsiveness to retinoids. We propose that retinoid signaling plays a conserved role in molluscan male development, and that shifts in the timing of retinoid signaling may have been important for the origins of sequential hermaphroditism within molluscs. [Display omitted] • Retinoid agonists induce penis growth in sequentially hermaphroditic snails. • Blocking RXR or retinoic acid synthesis decreases penis length. • RXR , RAR , and CYP2 6 mRNA are expressed in the induced penis. • Shifts in retinoid timing may be involved in repeated evolution of hermaphroditism. [ABSTRACT FROM AUTHOR]

BMP signaling is involved in many aspects of metazoan development, with two of the most conserved functions being to pattern the dorsal-ventral axis and to specify neural versus epidermal fates. An active area of research within developmental biology asks how BMP signaling was modified over evolution to build disparate body plans. Animals belonging to the superclade Spiralia/Lophotrochozoa are excellent experimental subjects for studying the evolution of BMP signaling because a highly conserved, stereotyped early cleavage program precedes the emergence of distinct body plans. In this study we examine the role of BMP signaling in one representative, the slipper snail Crepidula fornicata. We find that mRNAs encoding BMP pathway components (including the BMP ligand decapentaplegic, and BMP antagonists chordin and noggin-like proteins) are not asymmetrically localized along the dorsal-ventral axis in the early embryo, as they are in other species. Furthermore, when BMP signaling is perturbed by adding ectopic recombinant BMP4 protein, or by treating embryos with the selective Activin receptor-like kinase-2 (ALK-2) inhibitor Dorsomorphin Homolog 1 (DMH1), we observe no obvious effects on dorsal-ventral patterning within the posterior (post-trochal) region of the embryo. Instead, we see effects on head development and the balance between neural and epidermal fates specifically within the anterior, pre-trochal tissue derived from the 1q1 lineage. Our experiments define a window of BMP signaling sensitivity that ends at approximately 44–48 ​hours post fertilization, which occurs well after organizer activity has ended and after the dorsal-ventral axis has been determined. When embryos were exposed to BMP4 protein during this window, we observed morphogenetic defects leading to the separation of the anterior, 1q lineage from the rest of the embryo. The 1q-derived organoid remained largely undifferentiated and was radialized, while the post-trochal portion of the embryo developed relatively normally and exhibited clear signs of dorsal-ventral patterning. When embryos were exposed to DMH1 during the same time interval, we observed defects in the head, including protrusion of the apical plate, enlarged cerebral ganglia and ectopic ocelli, but otherwise the larvae appeared normal. No defects in shell development were noted following DMH1 treatments. The varied roles of BMP signaling in the development of several other spiralians have recently been examined. We discuss our results in this context, and highlight the diversity of developmental mechanisms within spiral-cleaving animals. • BMP signaling was examined during development in the snail Crepidula fornicata. • Perturbing BMP signaling does not affect the primary organizer or dorso-ventral axis. • BMP signaling regulates the patterning and morphogenesis of 1st quartet micromeres. • BMP signaling appears to control the balance between neural and epidermal fates. • Comparisons show that BMP signaling differs in different Lophotrochozoan species. [ABSTRACT FROM AUTHOR]

Background: One hundred years ago, marine organisms were the dominant systems for the study of developmental biology. The challenges in rearing these organisms outside of a marine setting ultimately contributed to a shift towards work on a smaller number of so-called model systems. Those animals are typically non-marine organisms with advantages afforded by short life cycles, high fecundity, and relative ease in laboratory culture. However, a full understanding of biodiversity, evolution, and anthropogenic effects on biological systems requires a broader survey of development in the animal kingdom. To this day, marine organisms remain relatively understudied, particularly the members of the Lophotrochozoa (Spiralia), which include well over one third of the metazoan phyla (such as the annelids, mollusks, flatworms) and exhibit a tremendous diversity of body plans and developmental modes. To facilitate studies of this group, we have previously described the development and culture of one lophotrochozoan representative, the slipper snail Crepidula atrasolea, which is easy to rear in recirculating marine aquaria. Lab-based culture and rearing of larger populations of animals remain a general challenge for many marine organisms, particularly for inland laboratories. Results: Here, we describe the development of an automated marine aquatic rack system for the high-density culture of marine species, which is particularly well suited for rearing filter-feeding animals. Based on existing freshwater recirculating aquatic rack systems, our system is specific to the needs of marine organisms and incorporates robust filtration measures to eliminate wastes, reducing the need for regular water changes. In addition, this system incorporates sensors and associated equipment for automated assessment and adjustment of water quality. An automated feeding system permits precise delivery of liquid food (e.g., phytoplankton) throughout the day, mimicking real-life feeding conditions that contribute to increased growth rates and fecundity. Conclusion: This automated system makes laboratory culture of marine animals feasible for both large and small research groups, significantly reducing the time, labor, and overall costs needed to rear these organisms. [ABSTRACT FROM AUTHOR]

The anthropogenic introduction in U.K. waters of the north-American marine gastropod Crepidula fornicata (Linné, 1758), commonly called slipper limpet, and its consecutive spreading has led in less than a century to the invasion of a part of benthic grounds along the North-European coasts. Competition for space has hampered the maintenance of the native scallop Pecten maximus , whereas dredge clogging has drastically limited scallop fishing, especially in the Western English Channel. In order to assess the possible future distribution and abundance of both species ( Crepidula and Pecten ), an original model of slipper limpet chains joined to a simple year-class model of the scallop (Ménesguen and Grégoris, 2017) has been coupled to a connectivity matrix summarizing the annual dispersion of emitted larvae in a realistic marine domain. This distributed model can explore the dynamics of both populations at the century scale and has been applied at two different geographical scales: a local one (the bays of Brest and Douarnenez, Western Brittany, France) and a regional one (Bay of Biscay and English Channel). First eigenvectors of the connectivity matrix are used to delineate the main retention areas. For each species taken alone, simulations starting with a few animals in different spots do converge in less than a century towards the same geographic distribution, compatible with the actual field distribution. This suggests that hydrodynamic patterns of larval drift create a strong attractor for these populations; it is reached by different routes of colonization, depending on the initial inoculation. The non-linear interaction created by the lack of space when the populations have filled the benthic area seems to be stronger for scallop populations than for slipper limpets, because of the permanent capacity of Crepidula beds to fix some larvae on the top of existing colonies. Whereas the scallop abundance experiences a rather large limit-cycle, with a dominant 11 years period and a lot of harmonics, the slipper limpet has commonly a stable steady state abundance. Competition between both populations tends to lower the mean scallop abundance and to lengthen and damp its fundamental period of oscillations. Simulated distributions can be locally improved by linking the larval survival to a distributed environmental stressor, as the salinity for scallop or a metallic contamination for the slipper limpet in the case of the bay of Brest. [ABSTRACT FROM AUTHOR]

Microplastics exposure could be detrimental to marine organisms especially under high concentrations. However, few studies have considered the multiphasic nature of marine invertebrates' life history and investigated the impact of experiencing microplastics during early development on post-metamorphic stages (legacy effect). Many planktonic larvae can feed selectively and it is unclear whether such selectivity could modulate the impact of algal food-sized microplastic. In this two-stage experiment, veligers of Crepidula onyx were first exposed to additions of algae-sized micro-polystyrene (micro-PS) beads at different concentrations, including ones that were comparable their algal diet. These additions were then either halted or continued after settlement. At environmentally relevant concentration (ten 2-μm microplastic beads ml −1 ), larval and juvenile C. onyx was not affected. At higher concentrations, these micro-PS fed larvae consumed a similar amount of algae compared to those in control but grew relatively slower than those in the control suggesting that ingestion and/or removal of microplastic was/were energetically costly. These larvae also settled earlier at a smaller size compared to the control, which could negatively affect post-settlement success. Juvenile C. onyx receiving continuous micro-PS addition had slower growth rates. Individuals only exposed to micro-PS during their larval stage continued to have slower growth rates than those in the control even if micro-PS had been absent in their surroundings for 65 days highlighting a legacy effect of microplastic exposure. [ABSTRACT FROM AUTHOR]

During development in metazoan embryos, the fundamental embryonic axes are established by organizing centers that influence the fates of nearby cells. Among the spiralians, a large and diverse branch of protostome metazoans, studies have shown that an organizer sets up the dorsal-ventral axis, which arises from one of the four basic cell quadrants during development (the dorsal, D quadrant). Studies in a few species have also revealed variation in terms of how and when the D quadrant and the organizer are established. In some species the D quadrant is specified conditionally, via cell-cell interactions, while in others it is specified autonomously, via asymmetric cell divisions (such as those involving the formation of polar lobes). The third quartet macromere (3D) typically serves as the spiralian organizer; however, other cells born earlier or later in the D quadrant lineage can serve as the organizer, such as the 2d micromere in the annelid Capitella teleta or the 4d micromere in the mollusc Crepidula fornicata . Here we present work carried out in the snail C. fornicata to show that establishment of a single D quadrant appears to rely on a combination of both autonomous (via inheritance of the polar lobe) and conditional mechanisms (involving induction via the progeny of the first quartet micromeres). Through systematic ablation of cells, we show that D quadrant identity is established between 5th and 6th cleavage stages, as it is in other spiralians that use conditional specification. Subsequently, following the next cell cycle, organizer activity takes place soon after the birth of the 4d micromere. Therefore, unlike the case in other spiralians that use conditional specification, the specification of the D quadrant and the activity of the dorso-ventral organizer are temporally and spatially uncoupled. We also present data on organizer function in naturally-occurring and experimentally-induced twin embryos, which possess multiple D quadrants. We show that supernumerary D quadrants can arise in C. fornicata (either spontaneously or following polar lobe removal); when multiple D quadrants are present these do not exhibit effective organizer activity. We conclude that the polar lobe is not required for D quadrant specification, though it could play a role in effective organizer activity. We also tested whether the inheritance of the small polar lobe by the D quadrant is associated with the ability to laterally inhibit neighboring quadrants by direct contact in order to normally prevent supernumerary organizers from arising. Finally, we discuss the variation of spiralian organizers in a phylogenetic context. [ABSTRACT FROM AUTHOR]

The slipper limpet Crepidula fornicata Linnaeus, 1758 is a major exotic invader of East North Atlantic coasts. Individuals live on top of each other and form stacks with the youngest on the top. Earlier studies reported that one individual typically settles per year. If true, it is a simplified means to provide a “shell length-age” relationship for population dynamics studies, especially Production/Biomass ratio (P/B) assessment. However, estimated P/B ranges between 0.15 and 0.45 yr-1 seem low compared to those of closely-related marine invertebrates and considering the invasiveness of C. fornicata in coastal habitats. In this study, we placed artificial substrates (“tiles”) for one year in the middle of a C. fornicata colony and measured settlement. We sometimes observed more than one individual settling per stack in a year: 4% of stacks were composed by 3 individuals, 27% by 2 individuals and 69% were single specimen. On this basis, we formulated a model to better link the position of each C. fornicata within a stack to its age. In addition, the C. fornicata population was annually sampled for 5 years. Then, population dynamics parameters, density at recruitment, mortality rate, growth performance, production and P/B were estimated. We compared two cases: (i) individual age was defined by its position in stacks; (ii) individual age was corrected by the model. Recalculation moderately increased growth performance expectation (+2 to 8% per year) but greatly enhanced production and P/B (ca. 1.2 to 2.6-fold per year). Recalculated P/B values ranged from 0.55 to 0.72 yr-1. While still low, they were more consistent with published values for similar large marine invertebrates, in particular for invasive species. [ABSTRACT FROM AUTHOR]

Organisms inhabiting coastal waters naturally experience diel and seasonal physico-chemical variations. According to various assumptions, coastal species are either considered to be highly tolerant to environmental changes or, conversely, living at the thresholds of their physiological performance. Therefore, these species are either more resistant or more sensitive, respectively, to ocean acidification and warming. Here, we focused on Crepidula fornicata, an invasive gastropod that colonized bays and estuaries on northwestern European coasts during the 20th century. Small (<3 cm in length) and large (>4.5 cm in length), sexually mature individuals of C. fornicata were raised for 6 months in three different pCO2 conditions (390 μatm, 750 μatm, and 1400 μatm) at four successive temperature levels (10°C, 13°C, 16°C, and 19°C). At each temperature level and in each pCO2 condition, we assessed the physiological rates of respiration, ammonia excretion, filtration and calcification on small and large individuals. Results show that, in general, temperature positively influenced respiration, excretion and filtration rates in both small and large individuals. Conversely, increasing pCO2 negatively affected calcification rates, leading to net dissolution in the most drastic pCO2 condition (1400 μatm) but did not affect the other physiological rates. Overall, our results indicate that C. fornicata can tolerate ocean acidification, particularly in the intermediate pCO2 scenario. Moreover, in this eurythermal species, moderate warming may play a buffering role in the future responses of organisms to ocean acidification. [ABSTRACT FROM AUTHOR]

In the current context of environmental change, ocean acidification is predicted to affect the cellular processes, physiology and behaviour of all marine organisms, impacting survival, growth and reproduction. In relation to thermal tolerance limits, the effects of elevated pCO2 could be expected to be more pronounced at the upper limits of the thermal tolerance window. Our study focused on Crepidulafornicata, an invasive gastropod which colonized shallow waters around European coasts during the 20th century. We investigated the effects of 10 weeks' exposure to current (380 μatm) and elevated (550, 750, 1,000 μatm) pCO2 on this engineer species using an acute temperature increase (1 °C 12 h -1) as the test. Respiration rates were measured on both males (small individuals) and females (large individuals). Mortality increased suddenly from 34 °C, particularly in females. Respiration rate in C.fornicata increased linearly with temperature between 18 and 34 °C, but no differences were detected between the different pCO2 conditions either in the regressions between respiration rate and temperature or in Q10 values. In the same way, condition indices were similar in all the pCO2 treatments at the end of the experiment, but decreased from the beginning of the experiment. This species was highly resistant to acute exposure to high temperature regardless of pCO2 levels, even though food was limited during the experiment. Crepidulafornicata appears to have either developed resistance mechanisms or a strong phenotypic plasticity to deal with fluctuations of physicochemical parameters in its habitat. This suggests that invasive species may be more resistant to future environmental changes than its native competitors. [ABSTRACT FROM AUTHOR]

Food quality and quantity, as well as temperature, are all factors that are expected to affect rates of development, and are likely to be affected by expected climatic change. We tested the effect of a mixed diet versus a single-food diet on metamorphic competence in the emerging model species Crepidula fornicata. We then compared our results with other published studies on this species that examined time to metamorphic competence across a range of food concentrations and rearing temperatures. Ours was the only study to test the effects of single food versus a mixed diet on metamorphic competence for this species. Diet composition did not affect metamorphic competence or survivorship. Comparing results across studies, we found that the shortest time to metamorphic competence was typically found when the food availability per larva was the greatest, independent of rearing temperature. Unfortunately, some published studies did not include important metadata needed for comparison with other studies; these data included larval rearing density, food density, frequency of feeding, and rearing temperature. Mortality rates were not always reported and when reported were often measured in different ways, preventing comparison. Such metadata are essential for comparisons among studies as well as among taxa, and for the determination of generalizable patterns and evolutionary trends. Increased reporting of all such metadata is essential if we are to use scientific studies performed to their fullest potential. [ABSTRACT FROM AUTHOR]

In sex-changing animals with internal fertilization, gregarious behavior may increase mating opportunities and the frequency of multiple paternity, thus increasing maternal reproductive success. Crepidula convexa is a direct-developing protandrous gastropod characterized by only modest gregarious behavior compared with previously studied members of the genus: females are frequently found isolated. Using 6 microsatellite markers, we analyzed paternity profiles in 10 broods (25 embryos per mother). The number of assigned fathers varied among families from 1 to 4 fathers per brood. Interestingly, polyandry was not detected in solitary females but only in females grouped with conspecific individuals. Overall, we found an average of 1.8 fathers per brood, but this increased to 2.6 fathers per brood when considering only the nonisolated females. Among 18 unambiguously identified fathers, only 5 were collected in our samples, suggesting substantial male mobility. Comparison with previous paternity analyses in Crepidula fornicata and Crepidula coquimbensis revealed that polyandry appears as a common trait of these sex-changing gastropods despite their different grouping behaviors and life histories. As expected, the level of polyandry was nevertheless lower in the modestly gregarious C. convexa. [ABSTRACT FROM AUTHOR]

In this work we studied bioerosion and encrustation on dead shells of the shallow benthic clam Venus antiqua from Patagonia Argentina with the aim of understanding biotic interactions (live/live interaction) and postmortem encrustation (live/dead interaction). In this regard, drill hole analysis and epibiont distribution in shells from modern death assemblages were performed. Additionally, we analyzed crushing traces in the shells of the drilling gastropod Trophon geversianus, which were caused by another predator. The analysis of drill hole placement and epibiont distribution on V. antiqua shells show drill holes (75%) and the epibiont Crepidula spp. (70.91%) more concentrated in the upper sector of the valve, which might be explained by the vertical position and a semi-infaunal mode of life in this clam. Nevertheless, the presence of drill holes in the lower sector of the valve indicates that clams spent part of the time reclining on the sediment. There is also evidence that clams with Crepidula spp. as commensals are less frequently attacked by drilling gastropods. Besides, a high percentage of articulated clams (30.97%) show signs of attack by drilling gastropods, but incomplete drill holes (7.67%) also suggest failed attempts, resulting in a minimum of 23.30% of successful predation. These incomplete drill holes may suggest failed attempts due to another predator attack upon the snails consuming the clams thus interrupting the feeding activity: more than 60% of marks of crushing in T. geversianus shells could have been produced by other predators, such as crabs. The postmortem encrustation on V. antiqua shells (35%) was mainly produced by calcareous polychaetes, preferably located on the inner side of the valve indicating that the empty shells of V. antiqua served as cryptic environment before they become exposed on the beach. Finally our data show that drilling frequency is very low in Pleistocene assemblages, suggesting changes in burial depth dynamics of this suspension feeder. [ABSTRACT FROM AUTHOR]

Sibling cannibalism is commonly observed in marine species. For instance, intrabrood cannibalism has been documented in marine gastropods with direct development, suggesting a relationship between embryo behavior and the evolution of life history strategies. However, there has been little effort to document the factors driving sibling cannibalism in marine species. The kin selection theory suggests that the level of relatedness plays an important role in cannibalism patterns. We examined Crepidula coquimbensis, a marine gastropod that broods and encloses its brooded offspring in capsules. Encapsulated embryos show sibling cannibalism and high levels of intracapsular multiple paternity. Given these features, cannibalistic behavior may be driven by kin-relatedness. To test this hypothesis, we constructed artificial aggregations of embryos to mimic three levels of relatedness: high, medium and low. For each category of aggregation, the cannibalism rate and benefits (i.e. size at hatching of surviving offspring) were estimated. In addition, at the end of embryo development, we performed parentage analyses to determine if cannibalism was associated with the relatedness between cannibal and victim embryos. Our results show that the intensity of sibling cannibalism increased in aggregations characterized by the lowest level of relatedness. There were important benefits of cannibalism in terms of hatching cannibal size. In addition, cannibalism between embryos was not random: the variation in reproductive success between males increased over the course of the experiment and the effective number of fathers decreased. Altogether, these results suggest that polyandry may play an important role in the evolution of sibling cannibalism in C. coquimbensis and that kin selection may operate during early embryonic stages in this species. [ABSTRACT FROM AUTHOR]

Sibling cannibalism is commonly observed in marine species. For instance, intrabrood cannibalism has been documented in marine gastropods with direct development, suggesting a relationship between embryo behavior and the evolution of life history strategies. However, there has been little effort to document the factors driving sibling cannibalism in marine species. The kin selection theory suggests that the level of relatedness plays an important role in cannibalism patterns. We examined Crepidula coquimbensis, a marine gastropod that broods and encloses its brooded offspring in capsules. Encapsulated embryos show sibling cannibalism and high levels of intracapsular multiple paternity. Given these features, cannibalistic behavior may be driven by kin-relatedness. To test this hypothesis, we constructed artificial aggregations of embryos to mimic three levels of relatedness: high, medium and low. For each category of aggregation, the cannibalism rate and benefits (i.e. size at hatching of surviving offspring) were estimated. In addition, at the end of embryo development, we performed parentage analyses to determine if cannibalism was associated with the relatedness between cannibal and victim embryos. Our results show that the intensity of sibling cannibalism increased in aggregations characterized by the lowest level of relatedness. There were important benefits of cannibalism in terms of hatching cannibal size. In addition, cannibalism between embryos was not random: the variation in reproductive success between males increased over the course of the experiment and the effective number of fathers decreased. Altogether, these results suggest that polyandry may play an important role in the evolution of sibling cannibalism in C. coquimbensis and that kin selection may operate during early embryonic stages in this species. [ABSTRACT FROM AUTHOR]

Adelphophagic development, where embryos consume sibling embryos or nurse eggs, is particularly common in marine caenogastropods and some families of polychaetes. When exogenous nutrition is provided before hatching, egg size and hatching size can be uncoupled, but advantages and constraints of adelphophagic development compared to development from large eggs are unknown. Here we examine temperature-mediated plasticity in offspring size, brooding duration, and fecundity in the adelphophagic marine gastropod Crepidula cf. onyx. We use these data combined with previously published data on two planktotrophic Crepidula and two Crepidula species that develop from large eggs to test hypotheses about the consequences of adelphophagic development and patterns of variation in offspring size. In Crepidula cf. onyx, egg size shows no significant effect of temperature. Hatching size is significantly larger at 28 °C than at 23°C but proceeds from fewer eggs per capsule at 28 °C. Hatching size is therefore decoupled from both egg size and the number of eggs per capsule. Although development is faster at the higher temperature, broods are produced roughly every 26-27 days at both temperatures. Increased rate of development has been cited as a potential advantage of adelphophagic development in muricids, but the adelphophagic C. cf. onyx did not develop more quickly than C. atrasolea or C. ustulatulina, species that produce similarly sized hatchlings from large eggs. Comparisons across calyptraeid species support the role of adelphophagy in increasing variance in offspring size. This increased variability is primarily expressed within broods or among broods from the same female, not among females. [ABSTRACT FROM AUTHOR]

Many marine gastropod species brood their embryos in thin-walled capsules to protect them during development. Despite its beneficial effects, encapsulation has two major constraints, nutrition and oxygen supply, which affect embryo development and larval survival. Developing embryos usually rely on intracapsular food sources provided by the mother, in the form of yolk, nurse eggs and intracapsular fluid. However, it is still not clear if they are able to feed on extracapsular sources that may cross the capsule wall. We investigated this possibility in the calyptraeid species Crepidula fornicata. In this species, the internal capsule wall thickness sharply decreases during embryonic development, which might change wall permeability to small organic molecules, thus providing embryos with external dissolved organic matter. To test this hypothesis, encapsulated and excapsulated embryos of C. fornicata were placed for 48 h in a 13C-enriched amino acid (l-alanine) solution. Excapsulated embryos were enriched in 13C (+5.75‰), which suggested that they were able to assimilate the labelled amino acid. In contrast, encapsulated embryos were weakly enriched (+0.75‰), suggesting that encapsulation greatly reduces the potential for the use of extracapsular amino acids and that encapsulated embryos mainly rely on maternal food. [ABSTRACT FROM PUBLISHER]

Background: Animals with a spiral cleavage program, such as mollusks and annelids, make up the majority of the superphylum Lophotrochozoa. The great diversity of larval and adult body plans in this group emerges from this highly conserved developmental program. The 4d micromere is one of the most conserved aspects of spiralian development. Unlike the preceding pattern of spiral divisions, cleavages within the 4d teloblastic sublineages are bilateral, representing a critical transition towards constructing the bilaterian body plan. These cells give rise to the visceral mesoderm in virtually all spiralians examined and in many species they also contribute to the endodermal intestine. Hence, the 4d lineage is an ideal one for studying the evolution and diversification of the bipotential endomesodermal germ layer in protostomes at the level of individual cells. Little is known of how division patterns are controlled or how mesodermal and endodermal sublineages diverge in spiralians. Detailed modern fate maps for 4d exist in only a few species of clitellate annelids, specifically in glossiphoniid leeches and the sludge worm Tubifex. We investigated the 4d lineage in the gastropod Crepidula fornicata, an established model system for spiralian biology, and in a closely related direct-developing species, C. convexa. Results: High-resolution cell lineage tracing techniques were used to study the 4d lineage of C. fornicata and C. convexa. We present a new nomenclature to name the progeny of 4d, and report the fate map for the sublineages up through the birth of the first five pairs of teloblast daughter cells (when 28 cells are present in the 4d sublineage), and describe each clone's behavior during gastrulation and later stages as these undergo differentiation. We identify the precise origin of the intestine, two cells of the larval kidney complex, the larval retractor muscles and the presumptive germ cells, among others. Other tissues that arise later in the 4d lineage include the adult heart, internal foot tissues, and additional muscle and mesenchymal cells derived from later-bornprogeny of the left and right teloblasts. To test whether other cells can compensate for the loss of these tissues (that is, undergo regulation), specific cells were ablated in C. fornicata. Conclusions: Our results present the first fate map of the 4d micromere sublineages in a mollusk. The fate mapreveals that endodermal and mesodermal fates segregate much later than previously thought. We observed little evidence of regulation between sublineages, consistent with a lineage-driven cell specification process. Our results provide a framework for comparisons with other spiralians and lay the groundwork for investigation of the molecular mechanisms of endomesoderm formation, germ line segregation and bilateral differentiation in Crepidula. [ABSTRACT FROM AUTHOR]

The article focuses on a study on the impact of rising coastal seawater temperatures on the reproductive output of Crepidula (C.) fornicata, an invasive gastropod. It notes that the nature and the structure of a habitat get to be modified by the abundance of C. fornicata species to their advantage. Findings show extended periods of brood presence and greater intensity in the reproductive activity of C. fornicata population in Bourneuf Bay, France during the 2006-2007 period than the 2000-2001 period in which the former posted higher water temperature. Results also indicate that the species exhibited greater intensity of gametogenesis amid higher water temperatures and chlorophyll (chl) α values.

The article presents research on the variation and repeatability of egg size among broods from a single female from two gastropods, Crepidula atrasolea and ustulatulina. It says that egg size was measured from three to seven broods per female and females were raised from hatching under under invariant temperature in incubators during the preliminary experiment. The females were raised under changing temperatures on the laboratory bench top in the second experiment. It suggests that there were differences in egg size among broods from each female and in females from the bench top under both conditions. The estimates of repeatability were reduced when among brood variation was included in the calculations. It adds that brood order and female do not contribute to intraspecific variation.

Recent developmental and genomic research focused on "slipper snails" in the genus Crepidula has positioned Crepidula fornicata as a de facto model system for lophotrochozoan development. Here we review recent developments, as well as earlier reports demonstrating the widespread use of this system in studies of development and life history. Recent studies have resulted in a well-resolved fate map of embryonic cell lineage, documented mechanisms for axis determination and D quadrant specification, preliminary gene expression patterns, and the successful application of loss- and gain-of-function assays. The recent development of expressed sequence tags and preliminary genomics work will promote the use of this system, particularly in the area of developmental biology. A wealth of comparative information on phylogenetic relationships, variation in mode of development within the family, and numerous studies on larval biology and metamorphosis, primarily in Crepidula fornicata, make these snails a powerful tool for studies of the evolution of the mechanisms of development in the Mollusca and Lophotrochozoa. By bringing a review of the current state of knowledge of Crepidula life histories and development together with some detailed experimental methods, we hope to encourage further use of this system in various fields of investigation. [ABSTRACT FROM AUTHOR]

Reproductive strategies and parental effects play a major role in shaping early life-history traits. Although polyandry is a common reproductive strategy, its role is still poorly documented in relation to paternal effects. Here, we used as a case study the invasive sessile marine gastropod Crepidula fornicata, a mollusc with polyandry and extreme larval growth variation among sibling larvae. Based on paternity analyses, the relationships between paternal identity and the variations in a major early life-history trait in marine organisms, that is, larval growth, were investigated. Using microsatellite markers, paternities of 437 fast- and slow-growing larvae from 6 broods were reliably assigned to a set of 20 fathers. No particular fathers were found responsible for the specific growth performances of their offspring. However, the range of larval growth rates within a brood was significantly correlated to 1) an index of sire diversity and 2) the degree of larvae relatedness within broods. Multiple paternity could thus play an important role in determining the extent of pelagic larval duration and consequently the range of dispersal distances achieved during larval life. This study also highlighted the usefulness of using indices based on fathers' relative contribution to the progeny in paternity studies. [ABSTRACT FROM AUTHOR]

The article examines the proposed two-dimensional, depth-averaged, hydrodynamic and reactive-transport model that has been specifically developed for the estuaries and the Bay of Brest. The proposed model, accordingly, includes an explicit representation of the benthic-pelagic coupling which is stimulated by the increasing density of the slipper limpet Crepidula fornicata. The testing of proposed model reveals that the benthic nutrient dynamics are highly variable and strongly depend on the local conditions.

Cell lineage studies in mollusk embryos have documented numerous variations on the lophotrochozoan theme of spiral cleavage. In the experimentally tractable embryo of the mud snail Ilyanassa, cell lineage has previously been described only up to the 29-cell stage. Here I provide a chronology of cell divisions in Ilyanassa to the stage of 84 cells (about 16 hours after first cleavage at 23°C), and show spatial arrangements of identified nuclei at stages ranging from 27 to 84 cells. During this period the spiral cleavage pattern gives way to a bilaterally symmetric, dorsoventrally polarized pattern of mitotic timing and geometry. At the same time, the mesentoblast cell 4d rapidly proliferates to form twelve cells lying deep to the dorsal ectoderm. The onset of epiboly coincides with a period of mitotic quiescence throughout the ectoderm. As in other gastropod embryos, cell cycle lengths vary widely and predictably according to cell identity, and many of the longest cell cycles occur in small daughters of highly asymmetric divisions. While Ilyanassa shares many features of embryonic cell lineage with two other caenogastropod genera, Crepidula and Bithynia, it is distinguished by a general tendency toward earlier and more pronounced diversification of cell division pattern along axes of later differential growth. [ABSTRACT FROM AUTHOR]