Invasive slipper limpets Crepidula fornicata act like a sink, rather than source, of Vibrio spp.

A large knowledge gap exists regarding the disease profile and pathologic condition of the invasive, non-native, slipper limpet Crepidula fornicata. To help address this, we performed a yearlong health survey across two sites in South Wales UK, subtidal Swansea Bay and intertidal Milford Haven. In total, 1,800 limpets were screened systematically for haemolymph bacterial burdens using both general and vibrio-selective growth media (TSA +2% NaCl and TCBS, respectively), haemolymph (blood) inspection using microscopy, a PCR-based assay targeting Vibrio spp., and multi-tissue histology. Over 99% of haemolymph samples contained cultivable bacterial colony forming units, and 83% of limpets tested positive for the presence of vibrios via PCR (confirmed via Sanger sequencing). Vibrio presence did not vary greatly across sites, yet a strong temporal (seasonal) effect was observed - significantly higher bacterial loads during the summer. Binomial logistic regression models revealed larger (older) limpets were more likely to harbour vibrios, and the growth of bacteria on TCBS was a key predictor for PCR-based vibrio detection. Histological assessment of >340 animals revealed little evidence of inflammation, sepsis, or immune reactivity despite the gross bacterial numbers. We contend that slipper limpets are not susceptible to bacteriosis at either site surveyed, or do not to harbour vibrios known to be pathogenic to humans. The lack of susceptibility to local pathogenic bacteria may explain, in part, the invasion success of C. fornicata across this region.

Mud-Based Construction Material: Promising Properties of French Gravel Wash Mud Mixed with Byproducts, Seashells and Fly Ash as a Binder

The French gravel industry produces approximatively 6.5 million tons of gravel wash mud each year. This material offers very promising properties which require an in-depth characterization study before its use as a construction material, otherwise it is removed from value cycles by disposal in landfills. We examined the suitability of gravel wash mud and seashells, with fly ash as a binder, as an unfired earth construction material. Thermal and mechanical characterizations of the smart mixture composed of gravel wash mud, Crepidula fornicata shells and fly ash are performed. The new specimens exhibit high compressive strengths compared to usual earth construction materials, which appears as a good opportunity for a reduction in the thickness of walls. The use of fly ash and Crepidula shells in addition to gravel wash mud provides high silica and calcium contents, which both react with clay, leading to the formation of tobermorite and Al-tobermorite as a result of a pozzolanic reaction. Considering the reduction in porosity and improvements in strength, these new materials are good candidates to contribute significantly to the Sustainable Development Goals (SDGs) and reduce carbon emissions.

The Marine Gastropod Crepidula fornicata Remains Resilient to Ocean Acidification Across Two Life History Stages

Rising atmospheric CO2 reduces seawater pH causing ocean acidification (OA). Understanding how resilient marine organisms respond to OA may help predict how community dynamics will shift as CO2 continues rising. The common slipper shell snail Crepidula fornicata is a marine gastropod native to eastern North America that has been a successful invader along the western European coastline and elsewhere. It has also been previously shown to be resilient to global change stressors. To examine the mechanisms underlying C. fornicata’s resilience to OA, we conducted two controlled laboratory experiments. First, we examined several phenotypes and genome-wide gene expression of C. fornicata in response to pH treatments (7.5, 7.6, and 8.0) throughout the larval stage and then tested how conditions experienced as larvae influenced juvenile stages (i.e., carry-over effects). Second, we examined genome-wide gene expression patterns of C. fornicata larvae in response to acute (4, 10, 24, and 48 h) pH treatment (7.5 and 8.0). Both C. fornicata larvae and juveniles exhibited resilience to OA and their gene expression responses highlight the role of transcriptome plasticity in this resilience. Larvae did not exhibit reduced growth under OA until they were at least 8 days old. These phenotypic effects were preceded by broad transcriptomic changes, which likely served as an acclimation mechanism for combating reduced pH conditions frequently experienced in littoral zones. Larvae reared in reduced pH conditions also took longer to become competent to metamorphose. In addition, while juvenile sizes at metamorphosis reflected larval rearing pH conditions, no carry-over effects on juvenile growth rates were observed. Transcriptomic analyses suggest increased metabolism under OA, which may indicate compensation in reduced pH environments. Transcriptomic analyses through time suggest that these energetic burdens experienced under OA eventually dissipate, allowing C. fornicata to reduce metabolic demands and acclimate to reduced pH. Carry-over effects from larval OA conditions were observed in juveniles; however, these effects were larger for more severe OA conditions and larvae reared in those conditions also demonstrated less transcriptome elasticity. This study highlights the importance of assessing the effects of OA across life history stages and demonstrates how transcriptomic plasticity may allow highly resilient organisms, like C. fornicata, to acclimate to reduced pH environments.

Expression Pattern of Nitric Oxide Synthase during Development of the Marine Gastropod Mollusc, Crepidula fornicata

Nitric Oxide (NO) plays a key role in the induction of larval metamorphosis in several invertebrate phyla. The inhibition of the NO synthase in Crepidula fornicata, a molluscan model for evolutionary, developmental, and ecological research, has been demonstrated to block the initiation of metamorphosis highlighting that endogenous NO is crucial in the control of this developmental and morphological process. Nitric Oxide Synthase contributes to the development of shell gland, digestive gland and kidney, being expressed in cells that presumably correspond to FMRF-amide, serotoninergic and catecolaminergic neurons. Here we identified a single Nos gene in embryonic and larval transcriptomes of C. fornicata and studied its localization during development, through whole-mount in situ hybridization, in order to compare its expression pattern with that of other marine invertebrate animal models.

Departures from isotropy: the kinematics of a larval snail in response to food

The swimming behavior of invertebrate larvae can affect their dispersal, survival, and settlement in the ocean. Modelling this behavior accurately poses unique challenges as behavior is controlled both by physiology and environmental cues. Some larvae use cilia to both swim and create feeding currents, resulting in potential trade-offs between the two functions. Food availability is naturally patchy and often occurs in shallow horizontal layers in the ocean. Also, larval swimming motions generally differ in the horizontal and vertical. In order to investigate behavioral response to food by ciliated larvae, we measure their behavioral anisotropy by quantifying deviations from a model based in isotropic diffusion. We hypothesize that larvae will increase horizontal swimming and decrease vertical swimming after encountering food which could lead to aggregation at food layers. We consider Crepidula fornicata larvae which are specifically of interest as they exhibit unsteady and variable swimming behaviors that are difficult to categorize. We tracked the larvae in still water with and without food, with a portion of the larvae starved beforehand. On average, larvae in the presence of food were observed higher in the water column, with higher swimming speeds and higher horizontal swimming velocities when compared to larvae without food. Starved larvae also exhibited higher vertical velocities in food, suggesting no aggregation behavior. While most treatments showed strong anisotropy in larval behavior, we found that starved larvae without food exhibited approximately isotropic kinematics, indicating that behavioral anisotropy can vary with environmental history and conditions to enhance foraging success or mitigate food-poor environments.

The marine gastropod Crepidula fornicata remains resilient to ocean acidification across two life history stages

Rising atmospheric CO2 reduces seawater pH causing ocean acidification (OA). Understanding how resilient marine organisms respond to OA may help predict how community dynamics will shift as CO2 continues rising. The common slipper shell snail Crepidula fornicata is a resilient marine gastropod native to eastern North America, which has been a successful invader along the western European coastline and elsewhere. To examine its potential resilience to OA, we conducted two controlled laboratory experiments. First, we examined several phenotypes and genome-wide gene expression of C. fornicata in response to pH treatments (7.5, 7.6, 8.0) throughout the larval stage and then tested how conditions experienced as larvae influenced juvenile stages (i.e. carryover effects). Second, we examined genome-wide gene expression patterns of C. fornicata larvae in response to acute (4, 10, 24 and 48 hours) pH treatment (7.5, 8.0). Both C. fornicata larvae and juveniles exhibited resilience to OA and gene expression responses highlight the role of transcriptome plasticity in OA resilience. Larvae did not exhibit reduced growth under OA until they were at least 4 days old. These phenotypic effects were preceded by broad transcriptomic changes, which likely serve as an acclimation mechanism for combating reduced pH conditions frequently experienced in littoral zones. Delayed metamorphosis was observed for larvae reared at reduced pH. Although juvenile size reflected larval rearing pH conditions, no carryover effects in juvenile growth rates were observed. Transcriptomic analyses suggest increased metabolism under OA, which may indicate compensation in reduced pH environments. Time course transcriptomic analyses suggest energetic burdens experienced under OA eventually dissipate, allowing C. fornicata to reduce metabolic demands and acclimate to reduced pH. This study highlights the importance of assessing the effects of OA across life history stages and demonstrates how transcriptomic plasticity can allow highly resilient organisms, like C. fornicata, acclimate to reduced pH environments.