Long-term anti-predator learning and memory differ across populations and sexes in an intertidal snail.

Anti-predator behaviours in response to predator cues can be innate, or they can be learned through prior experience and remembered over time. The duration and strength of continued anti-predator behaviour after predator cues are no longer present, and the potential for an enhanced response when re-exposed to predator cues later is less known but could account for the observed variation in anti-predator responses. We measured the carryover effects of past predation exposure and the potential for anti-predator learning and memory in the marine snail Nucella canaliculata from six populations distributed over 1000 km of coastline. We exposed lab-reared snails to cues associated with a common crab predator or seawater control in two serial experiments separated by over seven months. Responses were population- and sex-dependent, with some populations retaining anti-predator behaviours while others showed a capacity for learning and memory. Male snails showed a strong carryover of risk aversion, while females were able to return to normal feeding rates and grow more quickly. These behavioural differences culminated in strong impacts on feeding and growth rates, demonstrating that this variation has implications for the strength of trait-mediated indirect interactions, which can impact entire ecosystems.

An invasive prey and changing climate interact to shape the breeding phenology of an endangered predator.

Changes in phenology are occurring from global climate change, yet the impacts of other types of global change on the phenology of animals remain less appreciated. Understanding the potential for synergistic effects of different types of global change on phenology is needed, because changing climate regimes can have cascading effects, particularly on invasive species that vary in their thermal tolerances. Using 25 years of data from 5963 nests and 4675 marked individuals across the entire US breeding range of an endangered predator, the snail kite (Rostrhamus sociabilis plumbeus), we isolated the effects of an invasion of novel prey and warming temperatures on breeding phenology and its demographic consequences. Over this time period, breeding season length doubled, increasing by approximately 14 weeks. Both temperature and the establishment of invasive prey interacted to explain the timing of nest initiation. Temperature and invasive prey played distinct roles: earlier nest initiation occurred with increasing temperatures, whereas late nesting increased with invasion. Ultimately, both nest survival and juvenile survival declined later in the year, such that effects from invasive prey, but not warming temperatures, have the apparent potential for mistiming in breeding phenology by some individuals. Nonetheless, relatively few nesting events occurred during late fall when nest survival was very low, and seasonal declines in nest survival were weaker and renesting was more frequent in invaded wetlands, such that total reproductive output increased with invasion. Variation in demographic effects illustrate that considering only particular components of demography (e.g., nest survival rates) may be inadequate to infer the overall consequences of changes in phenology, particularly the potential for mistiming of phenological events. These results emphasize that species invasions may profoundly alter phenology of native species, such effects are distinct from climate effects, and both interact to drive population change.

Climate change could fuel urinary schistosomiasis transmission in Africa and Europe.

The freshwater snail Bulinus truncatus is an important intermediate host for trematode parasites causing urogenital schistosomiasis, a tropical disease affecting over 150 million people. Despite its medical importance, uncertainty remains about its global distribution and the potential impacts of climate change on its future spread. Here, we investigate the distribution of B. truncatus, combining the outputs of correlative and mechanistic modelling methods to fully capitalize on both experimental and occurrence data of the species and to create a more reliable distribution forecast than ever constructed. We constructed ensemble correlative species distribution models using 273 occurrence points collected from different sources and a combination of climatic and (bio)physical environmental variables. Additionally, a mechanistic thermal suitability model was constructed, parameterized by recent life-history data obtained through extensive lab-based snail-temperature experiments and supplemented with an extensive literature review. Our findings reveal that the current suitable habitat for B. truncatus encompasses the Sahel region, the Middle East, and the Mediterranean segment of Africa, stretching from Southern Europe to Mozambique. Regions identified as suitable by both methods generally coincide with areas exhibiting high urogenital schistosomiasis prevalence. Model projections into the future suggest an overall net increase in suitable area of up to 17%. New suitable habitat is in Southern Europe, the Middle East, and large parts of Central Africa, while suitable habitat will be lost in the Sahel region. The change in snail habitat suitability may substantially increase the risk of urogenital schistosomiasis transmission in parts of Africa and Southern Europe while reducing it in the Sahel region.

Plasticity of circadian and circatidal rhythms in activity and transcriptomic dynamics in a freshwater snail.

Organisms have diverse biological clocks synchronised with environmental cycles depending on their habitats. Anticipation of tidal changes has driven the evolution of circatidal rhythms in some marine species. In the freshwater snail, Semisulcospira reiniana, individuals in nontidal areas exhibit circadian rhythms, whereas those in tidal areas exhibit both circadian and circatidal rhythms. We investigated whether the circatidal rhythms are genetically determined or induced by environmental cycles. The exposure to a simulated tidal cycle did not change the intensity of circatidal rhythm in individuals in the nontidal population. However, snails in the tidal population showed different activity rhythms depending on the presence or absence of the exposure. Transcriptome analysis revealed that genes with circatidal oscillation increased due to entrainment to the tidal cycle in both populations and dominant rhythmicity was consistent with the environmental cycle. These results suggest plasticity in the endogenous rhythm in the gene expression in both populations. Note that circatidal oscillating genes were more abundant in the tidal population than in the nontidal population, suggesting that a greater number of genes are associated with circatidal clocks in the tidal population compared to the nontidal population. This increase of circatidal clock-controlled genes in the tidal population could be caused by genetic changes in the biological clock or the experience of tidal cycle in the early life stage. Our findings suggest that the plasticity of biological rhythms may have contributed to the adaptation to the tidal environment in S. reiniana.

Phylogenetic variations in a novel family of hyperstable apple snail egg proteins: insights into structural stability and functional trends.

The relationship between protein stability and functional evolution is little explored in proteins purified from natural sources. Here, we investigated a novel family of egg proteins (Perivitellin-1, PV1) from Pomacea snails. Their remarkable stability and clade-related functions in most derived clades (Canaliculata and Bridgesii) make them excellent candidates for exploring this issue. To that aim, we studied PV1 (PpaPV1) from the most basal lineage, Flagellata. PpaPV1 displays unparalleled structural and kinetic stability, surpassing PV1s from derived clades, ranking among the most hyperstable proteins documented in nature. Its spectral features contribute to a pale egg coloration, exhibiting a milder glycan binding lectin activity with a narrower specificity than PV1s from the closely related Bridgesii clade. These findings provide evidence for substantial structural and functional changes throughout the genus' PV1 evolution. We observed that structural and kinetic stability decreased in a clade-related fashion and was associated with large variations in defensive traits. For instance, pale PpaPV1 lectin turns potent in the Bridgesii clade, adversely affecting gut morphology, while giving rise to brightly colored PV1s providing eggs with a conspicuous, probably warning signal in the Canaliculata clade. This work provides a comprehensive comparative analysis of PV1s from various apple snail species within a phylogenetic framework, offering insights into the interplay among their structural features, stability profiles and functional roles. More broadly, our work provides one of the first examples from natural evolution showing the crucial link among protein structure, stability and evolution of new functions.

Trade and socioeconomic importance of an invasive giant snail in the endemic-rich island of São Tomé, Central Africa.

Managing invasive species is crucial to mitigate their negative impacts on ecosystems, yet conflicts may arise when their social benefits are disregarded. Human pressure on the endemic-rich forests of São Tomé has been high since the island was discovered by the Portuguese in the 15th century, and numerous species have been introduced. These include the invasive West African giant land snail (Archachatina marginata), which was introduced in the mid-20th century, is now widespread on the island, and is a potential threat to native flora and fauna. We assessed the frequency of consumption of this species and its socioeconomic importance to people across the island with household questionnaires, focus group discussions, and semistructured interviews. We explored the prevalence and potential drivers of use (e.g., wealth, household composition, and diversity of occupations) and characterized the commodity chain to identify demographic groups linked to the snail trade. We interviewed 672 people (1 person per household), conducted 6 focus groups, and interviewed 80 key actors belonging to 5 subcategories. The snail was the most widely consumed bushmeat and an important source of income, particularly for women and unemployed youth. Insecure and scarce livelihood alternatives, mostly in rural areas, were reported as drivers for trade involvement. Snail harvesting was more frequent in poorer households with low occupational diversity. Selling tended to occur in households that were well-established in the community and had a higher proportion of children. Both were stimulated by the proximity of communities to the native forest. Buying snails was common in all demographic groups, but it was linked to wealth and occupational diversity. Interventions to manage the impact of this introduced species on the valuable ecosystems of the island should involve multiple sectors of society to ensure societal support. This requires robust consideration of the welfare of vulnerable demographic groups that benefit from the species.

Importancia socioeconómica y comercial de un caracol invasor en la isla rica en endemismos de Santo Tomé, África Central Resumen La gestión de las especies invasoras es crucial para mitigar sus efectos negativos en los ecosistemas, aunque pueden surgir conflictos cuando no se tienen en cuenta sus beneficios sociales. La presión humana sobre los bosques de Santo Tomé, ricos en endemismos, ha sido alta desde que la isla fue colonizada por los portugueses en el siglo XV y desde entonces se han introducido numerosas especies. Una de ellas es el caracol terrestre gigante de África Occidental (Archachatina marginata), introducido a mediados del siglo XX y que ahora tiene una distribución amplia en la isla y es una amenaza potencial para la flora y la fauna autóctonas al ser invasor. Evaluamos la frecuencia de consumo de esta especie y su importancia socioeconómica para la población de toda la isla mediante encuestas en hogares, grupos de discusión y entrevistas semiestructuradas a actores clave de cinco categorías. Exploramos la prevalencia y los posibles factores impulsores del consumo (por ejemplo, la riqueza, la composición de los hogares y la diversidad de ocupaciones) y caracterizamos la cadena comercial para identificar los grupos demográficos vinculados al comercio de caracoles. Entrevistamos a 672 personas (una persona por hogar), realizamos seis grupos de discusión y entrevistamos a 80 actores clave. El caracol fue la carne de caza más consumida y una importante fuente de ingresos, sobre todo para las mujeres y los jóvenes desempleados. La inseguridad y la escasez de medios de subsistencia, sobre todo en las zonas rurales, fueron los factores que impulsaron la participación en el mercado. La colecta de caracoles fue más frecuente en los hogares más pobres y con escasa diversidad ocupacional. La venta tendía a producirse en hogares bien establecidos en la comunidad y con una mayor proporción de niños. Ambos factores se vieron estimulados por la proximidad de las comunidades al bosque nativo. La compra de caracoles fue común en todos los grupos demográficos, pero estuvo vinculada a la riqueza y a la diversidad ocupacional. Las intervenciones para gestionar el impacto de esta especie introducida en los valiosos ecosistemas de la isla deben implicar a múltiples sectores de la sociedad para garantizar su apoyo. Para ello es necesario tener muy en cuenta el bienestar de los grupos demográficos vulnerables que se benefician de la especie.

The physical basis of mollusk shell chiral coiling.

Snails are model organisms for studying the genetic, molecular, and developmental bases of left-right asymmetry in Bilateria. However, the development of their typical helicospiral shell, present for the last 540 million years in environments as different as the abyss or our gardens, remains poorly understood. Conversely, ammonites typically have a bilaterally symmetric, planispiraly coiled shell, with only 1% of 3,000 genera displaying either a helicospiral or a meandering asymmetric shell. A comparative analysis suggests that the development of chiral shells in these mollusks is different and that, unlike snails, ammonites with asymmetric shells probably had a bilaterally symmetric body diagnostic of cephalopods. We propose a mathematical model for the growth of shells, taking into account the physical interaction during development between the soft mollusk body and its hard shell. Our model shows that a growth mismatch between the secreted shell tube and a bilaterally symmetric body in ammonites can generate mechanical forces that are balanced by a twist of the body, breaking shell symmetry. In gastropods, where a twist is intrinsic to the body, the same model predicts that helicospiral shells are the most likely shell forms. Our model explains a large diversity of forms and shows that, although molluscan shells are incrementally secreted at their opening, the path followed by the shell edge and the resulting form are partly governed by the mechanics of the body inside the shell, a perspective that explains many aspects of their development and evolution.

Species-specific oxylipins and the effects of ontogeny and predation on their emission from freshwater snails.

Chemical cues play important roles in mediating ecological interactions. Oxylipins, oxygenated metabolites of fatty acids, are one signalling molecule type that influences the physiology and function of species, suggesting their broader significance in chemical communication within aquatic systems. Yet, our current understanding of their function is restricted taxonomically and contextually making it difficult to infer their ecological significance. Snails and leeches are ubiquitous in freshwater ecosystems worldwide, yet little is known about their oxylipin profiles and the factors that cause their profiles to change. As snails and leeches differ taxonomically and represent different trophic groups, we postulated oxylipin profile differences. For snails, we hypothesized that ontogeny (non-reproductive vs reproductive) and predation (non-infested vs leech-infested) would affect oxylipin profiles. Oxylipins were characterized from water conditioned with the snail Planorbella duryi and leech Helobdella lineata, and included three treatment types (snails, leeches, and leech-infested snails) with the snails consisting of three size classes: small (5-6 mm, non-reproductive) and medium and large (13-14 and 19-20 mm, reproductive). The two species differed in the composition of their oxylipin profiles both in diversity and amounts. Further, ontogeny and predation affected the diversity of oxylipins emitted by snails. Our experimental profiles of oxylipins show that chemical cues within freshwater systems vary depending upon the species emitting the signals, the developmental stage of the species, as well as from ecological interactions such as predation. We also identified some candidates, like 9-HETE and PGE2, that could be explored more directly for their physiological and ecological roles in freshwater systems.

Effects of cadmium on the intestinal health of the snail Bradybaena ravida Benson.

The heavy metal cadmium (Cd) is a toxic and bioaccumulative metal that can be enriched in the tissues and organs of living organisms through the digestive tract. However, more research is needed to determine whether food-sourced Cd affects the homeostasis of host gut microflora. In this study, the snail Bradybaena ravida (Benson) was used as a model organism fed with mulberry leaves spiked with different concentrations of Cd (0, 0.052, 0.71, and 1.94 mg kg-1). By combining 16S rRNA high-throughput sequencing with biochemical characterization, it was found that there were increases in the overall microbial diversity and abundances of pathogenic bacteria such as Corynebacterium, Enterococcus, Aeromonas, and Rickettsia in the gut of B. ravida after exposure to Cd. However, the abundances of potential Cd-resistant microbes in the host's gut, including Sphingobacterium, Lactococcus, and Chryseobacterium, decreased with increasing Cd concentrations in the mulberry leaves. In addition, there was a significant reduction in activities of energy, nutrient metabolism, and antioxidant enzymes for gut microbiota of snails treated with high concentrations of Cd compared to those with low ones. These findings highlight the interaction of snail gut microbiota with Cd exposure, indicating the potential role of terrestrial animal gut microbiota in environmental monitoring through rapid recognition and response to environmental pollution.

Maternal effects, paternal effects, and their interactions in the freshwater snail Physa acuta.

Individuals exposed to predation risk can produce offspring with altered phenotypes. Most work on predation-induced parental effects has focused on maternal effects or on generalized parental effects where both parents are exposed to risk. We conducted an experiment to measure and compare maternal and paternal effects on offspring phenotypes and test for interactions in those effects. We exposed 82 snails from 22 lines to control or predator cues and created line dyads with the four possible mating pairings of control and predator cue exposed individuals. We measured the resulting body masses, shell masses, shell shapes, and anti-predator behaviors of the offspring. We found some evidence that offspring were larger and heavier when the mother was exposed to predation cues, but that this effect was negated when the father was also exposed. The mass of offspring shells relative to their total mass was unaffected by parental treatments. Shell shape was marginally affected by maternal treatment, but not paternal treatment. Behavioral responses to cues were not affected by maternal or paternal treatments. Our results suggest potential conflict between male and female parental effects and highlight the importance of examining the interactions of maternal and paternal effects.

Post-exposure Period as a key Factor to Assess Cadmium Toxicity: Lethal vs. Behavioral Responses.

The exposure of animals to pollution in ecosystems is not always chronic. Toxicants can remain in aquatic ecosystems for a short-term. To improve the extrapolation of laboratory results to natural scenarios the inclusion of post-exposure periods is a relevant issue. The present study focuses on the assessment of cadmium toxicity on survival and behavior of the aquatic snail Potamopyrgus antipodarum (Tateidae, Mollusca) during exposure and post-exposure. Animals were exposed for 48 h to cadmium (0.05, 0.14, 0.44 and 1.34 mg Cd/L) and 168 h of post-exposure. During the post-exposure period an increase in mortality in all concentrations was observed. The effects observed during the post-exposure period on the LC50 and EC50 were significant. During the post-exposure, behavior showed a clear recovery in surviving animals exposed to 0.44 mg Cd/L. Animals exposed to 0.05 mg Cd/L did not show differences with control. Therefore, mortality after exposure should be included in the ecotoxicological bioassays for a more realistic estimation of the cadmium effects. To assess the degree of animal recovery after cadmium exposure, behaviour has been shown as an adequate parameter.

Polyethylene Microplastics Affected Survival Rate, Food Intake and Altered Oxidative Stress Parameters in Freshwater Snail Indoplanorbis exustus.

Microplastics have a negative impact on aquatic ecosystems. Gastropod mollusks serve as bioindicators and are good model systems for ecotoxicological studies. To assess oxidative damage, we exposed the ram's horn snail, Indoplanorbis exustus, to various concentrations of low-density polyethylene microplastics (size range 8-100 µm). The main objectives were microplastics preparation, characterization, and examination of their effect on the essential organs of I. exustus. Scanning electron microscopy, fourier transform infrared spectroscopy and x-ray diffraction techniques confirmed the polymer type of laboratory prepared polyethylene microplastics. The LC50 value of microplastics for snails was calculated to be 872 mg/L after 96 h of exposure. We observed a significant elevation in superoxide dismutase, catalase and lipid peroxidation levels with increasing concentrations of microplastics. Microplastics exposure also affected protein content, total food intake and total weights. Moreover, snails failed to recover post-treatment. Snails collected from contaminated source of microplastics served as positive control for the study. Hence, we can conclude that microplastics cause overall impairment in the physiological parameters and show adverse effects on the freshwater snail, I. exustus.

Sexual inactivation induced by the mucus that covers land snail love darts: sexual selection and evolution of allohormones in hermaphrodites.

The transfer of male accessory gland secretions is a well-investigated reproductive strategy for winning in sexual selection. An example of such a strategy is the conspicuous mating behaviour of simultaneously hermaphroditic land snails, the so-called shooting of love darts, whereby a snail drives a love dart(s) into the body of its mating partner. In the land snail Euhadra quaesita, it has been shown that a specific mucus which coats the love dart is transferred into the partner's haemolymph and that it suppresses subsequent matings in the darted individual. However, how the mucus of the love dart suppresses rematings remains unclear. In the present study, we tested the hypothesis that by injection of the dart mucus, love-dart shooters manipulate the physiology of a dart recipient and make the individual sexually inactive. In an experiment in which snails were provided with opportunities to encounter a potential mating partner, we found that the latency period to achieve sexual arousal was longer in snails injected with the dart mucus than in snails of the control treatments. This finding indicates that the dart mucus delays sexual arousal in injected snails. This delay in arousal is a novel example of the effects of the mucus in simultaneously hermaphroditic land snails. The remating suppression effect of the dart mucus is likely to occur through sexual inactivation.

Peptidergic modulation of a multi-functional central pattern generator in the pulmonate snail.

Egg laying in pulmonate snails is a well-orchestrated process that involves a period of reduced locomotion, followed by substrate cleaning with rhythmic rasping of the surface to make tiny grooves, into which eggs are deposited. Although the neurohormonal control of initiating egg laying has been well established, the signals that modulate the buccal central pattern generator to substrate cleaning during egg laying are not known. Neuropeptides of the invertebrate gonadotropin-releasing hormone/corazonin family (invGnRH/CRZ) have been shown to be involved in reproduction and allied behaviors in many vertebrates and invertebrates. Here, we show that the buccal motor pattern underlying substrate cleaning during egg laying is altered by a vertebrate GnRH agonist. Signals from the intestinal nerve innervating reproductive structures, previously shown to be both necessary and sufficient for egg-laying behaviors, are blocked by a vertebrate GnRH antagonist. Further, the vertebrate GnRH-triggered response elicits rhythmic, phase 2 and non-phase 2 activity in the buccal motor pattern, with a shutdown of phase 3, indicative of repetitive rasping without accompanied swallowing behavior. Using immunohistochemistry, intracellular electrophysiology and extracellular nerve stimulation, we show that a member of the invGnRH/CRZ family of neuropeptides could be the signal that contextually switches the multifunctional buccal CPG to a biphasic rasping rhythm that underlies substrate cleaning behavior during egg laying in the pulmonate snail Planorbella (Helisoma) trivolvis.

The making of natural iron sulfide nanoparticles in a hot vent snail.

Biomineralization in animals exclusively features oxygen-based minerals with a single exception of the scaly-foot gastropod Chrysomallon squamiferum, the only metazoan with an iron sulfide skeleton. This unique snail inhabits deep-sea hot vents and possesses scales infused with iron sulfide nanoparticles, including pyrite, giving it a characteristic metallic black sheen. Since the scaly-foot is capable of making iron sulfide nanoparticles in its natural habitat at a relatively low temperature (∼15 °C) and in a chemically dynamic vent environment, elucidating its biomineralization pathways is expected to have significant industrial applications for the production of metal chalcogenide nanoparticles. Nevertheless, this biomineralization has remained a mystery for decades since the snail's discovery, except that it requires the environment to be rich in iron, with a white population lacking in iron sulfide known from a naturally iron-poor locality. Here, we reveal a biologically controlled mineralization mechanism employed by the scaly-foot snail to achieve this nanoparticle biomineralization, through δ34 S measurements and detailed electron-microscopic investigations of both natural scales and scales from the white population artificially incubated in an iron-rich environment. We show that the scaly-foot snail mediates biomineralization in its scales by supplying sulfur through channel-like columns in which reaction with iron ions diffusing inward from the surrounding vent fluid mineralizes iron sulfides.

Evolution of reproductive strategies in the species-rich land snail subfamily Phaedusinae (Stylommatophora: Clausiliidae).

Most of the present knowledge on animal reproductive mode evolution, and possible factors driving transitions between oviparity and viviparity is based on studies on vertebrates. The species rich door snail (Clausiliidae) subfamily Phaedusinae represents a suitable and unique model for further examining parity evolution, as three different strategies, oviparity, viviparity, and the intermediate mode of embryo-retention, occur in this group. The present study reconstructs the evolution of reproductive strategies in Phaedusinae based on time-calibrated molecular phylogenetics, reproductive mode examinations and ancestral state reconstruction. Our phylogenetic analysis employing multiple mitochondrial and nuclear markers identified a well-supported clade (including the tribes Phaedusini and Serrulinini) that contains species exhibiting various reproductive strategies. This clade evolved from an oviparous most recent common ancestor according to our reconstruction. All non-oviparous taxa are confined to a highly supported subclade, coinciding with the tribe Phaedusini. Both oviparity and viviparity occur frequently in different lineages of this subclade that are not closely related. During Phaedusini diversification, multiple transitions in reproductive strategy must have taken place, which could have been promoted by a high fitness of embryo-retaining species. The evolutionary success of this group might result from the maintenance of various strategies.

Structural and functional insights into macrophage migration inhibitory factor from Oncomelania hupensis, the intermediate host of Schistosoma japonicum.

Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum. As an irreplaceable prerequisite in the transmission and prevalence of schistosomiasis japonica, an in-depth study of this obligate host-parasite interaction can provide glimpse into the molecular events in the competition between schistosome infectivity and snail immune resistance. In previous studies, we identified a macrophage migration inhibitory factor (MIF) from O. hupensis (OhMIF), and showed that it was involved in the snail host immune response to the parasite S. japonicum. Here, we determined the crystal structure of OhMIF and revealed that there were distinct structural differences between the mammalian and O. hupensis MIFs. Noticeably, there was a projecting and structured C-terminus in OhMIF, which not only regulated the MIF's thermostability but was also critical in the activation of its tautomerase activity. Comparative studies between OhMIF and human MIF (hMIF) by analyzing the tautomerase activity, oxidoreductase activity, thermostability, interaction with the receptor CD74 and activation of the ERK signaling pathway demonstrated the functional differences between hMIF and OhMIF. Our data shed a species-specific light on structural, functional, and immunological characteristics of OhMIF and enrich the knowledge on the MIF family.

Physiological alterations in Pseudosuccinea columella (Mollusca: Gastropoda) after infection by Heterorhabditis baujardi LPP7 (Rhabditida: Heterorhabditidae).

The snail Pseudosuccinea columella participates in the distribution of Fasciola hepatica in the environment by acting as its intermediate host. Therefore, the control of this lymnaeid is one of the ways to prevent hepatic fascioliasis. The objective of this study was to evaluate the susceptibility of P. columella to infective juveniles (IJs) of the entomopathogenic nematode (EPN) Heterorhabditis baujardi in laboratory conditions, as well as to investigate aspects related to the biochemistry and histopathology of snails exposed or not to the EPNs during three weeks. The EPN exposure induced significant reductions in the concentrations of glucose, total proteins and glycogen (gonad-digestive gland complex) in the snails during the onset of the infection, with the levels being restored as the infection progresses. These alterations were accompanied by increased hemolymph activities of aminotransferases and lactate dehydrogenase, as well as the concentrations of uric acid after the first and second weeks of the experiment. The histopathological analyses of the exposed snails revealed cell necrosis at the end of the first week, tissue inflammatory reactions one and two weeks after exposure, and degeneration three weeks afterward in comparison with the unexposed snails. Finally, scanning electronic microscopy revealed proliferation of fibrous connective tissue three weeks after exposure. The results indicate that P. columella is susceptible to H. baujardi. The exposure favored the establishment of a negative energy balance, increased the activity of enzymes related to tissue damages and promoted accumulation of nitrogen compounds in the host snails. Additionally, was observed in P. columella exposed to the EPNs, significant tissue lesions, and demonstrated the strong pathogenic potential of H. baujardi, indicating its possible application for biological control of this snail.

Morphology and activities of cell populations of haemocytes in Oncomelania hupensis following Schistosoma japonicum infection.

Oncomelania hupensis is the only obligatory intermediate host of Schistosoma japonicum, the pathogen of zoonosis schistosomiasis. Haemocytes play a critical role in the cellular immune defence of O. hupensis against S. japonicum challenge. Here, the morphology and classification of haemocytes of O. hupensis were investigated by Giemsa staining and light microscopy, combining with the scanning and transmission electron microscopy and flow cytometry. Granulocytes and hyalinocytes were confirmed as two main types of haemocytes, account for ~ 10% and ~ 90% of all haemocytes, with size varying in 4.3-10.9 µm and 0.4-30.8 µm, respectively. Subpopulations can be identified further by granule feature, shape, size, and surface and inner structure of cells. The heterogeneity in morphology implied varied developmental process and function of haemocyte subpopulations. After the S. japonicum challenge, haemocytes of O. hupensis respond to S. japonicum invasion immediately. The dynamic change of haemocyte subpopulations indicates that the small hyalinocyte could differentiate into a larger one or granulocyte after S. japonicum challenge, and the granulocytes and larger hyalinocytes play leading roles in early defence reaction, but in different ways. Phagocytosis and apoptosis of haemocytes in O. hupensis were proved to be related to immune defence against S. japonicum, with the combined effect of granulocytes and larger hyalinocytes. However, the main pathway of each subpopulation to take effect in different periods need further investigation.

Waves cue distinct behaviors and differentiate transport of congeneric snail larvae from sheltered versus wavy habitats.

Marine population dynamics often depend on dispersal of larvae with infinitesimal odds of survival, creating selective pressure for larval behaviors that enhance transport to suitable habitats. One intriguing possibility is that larvae navigate using physical signals dominating their natal environments. We tested whether flow-induced larval behaviors vary with adults' physical environments, using congeneric snail larvae from the wavy continental shelf (Tritia trivittata) and from turbulent inlets (Tritia obsoleta). Turbulence and flow rotation (vorticity) induced both species to swim more energetically and descend more frequently. Accelerations, the strongest signal from waves, induced a dramatic response in T. trivittata but almost no response in competent T. obsoleta Early stage T. obsoleta did react to accelerations, ruling out differences in sensory capacities. Larvae likely distinguished turbulent vortices from wave oscillations using statocysts. Statocysts' ability to sense acceleration would also enable detection of low-frequency sound from wind and waves. T. trivittata potentially hear and react to waves that provide a clear signal over the continental shelf, whereas T. obsoleta effectively "go deaf" to wave motions that are weak in inlets. Their contrasting responses to waves would cause these larvae to move in opposite directions in the water columns of their respective adult habitats. Simulations showed that the congeners' transport patterns would diverge over the shelf, potentially reinforcing the separate biogeographic ranges of these otherwise similar species. Responses to turbulence could enhance settlement but are unlikely to aid large-scale navigation, whereas shelf species' responses to waves may aid retention over the shelf via Stokes drift.