Veliger Larvae Research Articles

A new family of freshwater snails with Cretaceous origin from Yunnan, China

Abstract China is a hotspot for species’ discovery nowadays and harbours some of the most threatened environments in the world due to human activities. Here we describe a new remarkable gastropod species from an ancient freshwater lake in Yunnan, south-west China, Squamapex taiji gen. et sp. nov. within a new monospecific family of Truncatelloidea, Squamapicidae fam. nov.. The highly ornamented protoconch of Squamapicidae, with a microstructure distinct from that of the teleoconch, has rarely been found in true freshwater gastropods, being previously mostly known from marine or migratory gastropods with veliger larvae. The most complete family-level and first-dated phylogeny of Truncatelloidea, the largest and most diverse superfamily within Mollusca, based on two mitochondrial and three nuclear markers, shows that Squamapicidae belong to a distinct ancient lineage with Cretaceous origin. Its sister-group, unique protoconch, and distribution suggest a Tethys Ocean origin of this new family.

Evaluation of the teratogenic potency of bulk zinc oxide and its nanoparticles on embryos of the freshwater snail, Helisoma duryi

Bulk zinc oxide (ZnO-BPs) and its nanoparticles (ZnO-NPs) are frequently used in various products for humans. Helisoma duryi embryos can serve as effective model organisms for studying the toxicity of NPs. This study aimed to compare the teratogenic potency of ZnO-BPs and ZnO NPs in the embryonic stages of H. duryi to evaluate the utility of this snail as a bioindicator for ZnO-NPs in the aquatic environment. The mechanisms of teratogenesis were evaluated by determination of the LC50, studying the effect of sub-lethal concentrations of both ZnO forms on the embryos, and studying their enzyme activity, oxidative stress, and biochemical analysis. The SDS-PAGE electrophoresis was undertaken to assess the effect of ZnO-BPs and ZnO NPs on protein synthesis. The results revealed that the veliger stage of H. duryi is the specific stage for bulk and nano ZnO. ZnO-NPs proved to be more toxic to snails’ embryos than ZnO-BPs. Exposure to ZnO influences specific types of defects in development, which in the case of BPs are far less drastic than those caused by NPs. Thus, the toxicity of ZnO-NPs in embryonic development is due to their unique physicochemical properties. The observed malformations include mainly hydropic malformation, exogastrulation, monophthalmia, shell misshapen, and cell lyses. Almost all tested oxidative biomarkers significantly changed, revealing that ZnONPs display more oxidative stress than ZnO-BPs. Also, the low concentration of ZnO induces many disturbances in the organic substances of veliger larvae, such as a decrease in the total protein and total lipid levels and an increase in the glycogen level. The results indicated that ZnO-BPs increase the number of protein bands. Conversely, ZnO-NPs concealed one band from treated egg masses, which was found in the control group. Embryos of snail are an appropriate model to control freshwater snails. This study demonstrates that H. duryi embryos can serve as effective model organisms to study the toxicity of ZnO-NPs.

Oocyte size, early development and hybridisation in two sympatric Chilean species of Fissurella (Mollusca: Vetigastropoda) under laboratory conditions

ABSTRACT Fissurella latimarginata and Fissurella cumingi are two sympatric species found along the southwestern coast of South America. We conducted a comparative analysis of oocyte size and their coats between these species and investigated early development using chemically activated gametes. Additionally, heterologous fertilizations were conducted to assess the presence of a reproductive isolation barrier between the species. The mean diameter of the oocyte and gelatinous coat did not show significant differences between species, but there were differences regarding the vitelline coat. However, the practical utility of this trait as taxonomic character for species discrimination is hindered by variance overlap. Chemical activation of gametes enabled homologous fertilizations, resulting in the production of healthy and viable trochophore larvae. Early development, from fertilisation to hatch of these larvae, takes 28.5–45.5 h in both species. Heterologous fertilizations were also successful, resulting in the hatching of trochophore hybrid larvae, indicating effective interactions between the gametes of both species and viable postzygotic larval development. However, further research should focus on determining if there is asymmetry in gametic compatibility, obtaining veliger larvae, or even more advanced ontogenetic stages.

Cryopreservation of European flat oyster (Ostrea edulis) larvae using a controlled-rate freezer

Larvae of the European flat oyster (Ostrea edulis) were cryopreserved using two protocols originally developed for other bivalvian species with several modifications. Culture of the European flat oyster went through several setbacks due to parasitic diseases, thus, assisted reproductive technologies such as larval cryopreservation can aid in prompt restoration of lost populations and thus facilitate farming of the species. Veliger larvae were diluted in two basic extenders (0.4 M trehalose or 0.4 M sucrose) supplemented with ethylene glycol (EG), dimethyl-sulfoxide (DMSO) or propylene glycol (PG) in concentrations of 10–20% with or without the addition of polyvinylpyrrolidone (average MW-40,000; PVP-40). Larvae were loaded into 0.25-ml or 0.5-ml straws and cryopreserved in a controlled-rate freezer using two slow cooling profiles. Assessment of larval survival 20 min or 24 h post-thaw revealed that the cooling profiles and the type of extender used did not have a significant effect on survival rates. On the other hand, the type of cryoprotectant, straw volume and the time of assessment had a significant main effect on larval survival percentages (p < 0.001 in all three cases). Survival percentages of 70–80% were observed 20 min post-thaw which decreased to 40–60% at 24 h. Generally, EG and DMSO resulted in higher post-thaw larval survival than did PG when larvae were cryopreserved in 0.25-ml straws, whereas, these differences were not detected when 0.5-ml straws were employed. Results of these experiments will allow later studies on long-term survival of cryopreserved European flat oyster larvae.

High‐volume plankton tow net sampling improves eDNA detection of invasive zebra mussels (Dreissena polymorpha) in recently infested lakes

AbstractAquatic invasive species are a serious and growing threat to biodiversity. Zebra and quagga mussels (Dreissena polymorpha and D. rostriformis bugensis) are freshwater invaders causing substantial ecological and economic damage across Europe and North America. Early detection of invasive mussels and other non‐indigenous species is increasingly needed to prevent their establishment and spread. Environmental DNA (eDNA) techniques potentially offer higher sensitivity monitoring tools to complement more conventional methods for surveying adult and juvenile mussels and veliger larvae. eDNA assays are typically performed on small‐volume (0.5–5 L) water samples that are concentrated by filtration prior to extraction and downstream processing. Sampling using a towed plankton net of a larger (64 μm) pore size can process orders of magnitude larger water volumes with the potential for increasing eDNA detection sensitivity. We compared the sensitivities of high‐volume plankton tow net water sampling to filter sampling in three recently infested lakes in Canada and Minnesota, USA. Paired filtration and tow net samples were analyzed for Dreissena DNA using an established quantitative polymerase chain reaction assay for the genus. Higher yields of Dreissena eDNA (more DNA copies) were recovered from plankton tow than from filtered samples in all 33 paired comparisons. In some cases, plankton tow samples were positive for Dreissena eDNA while lower‐volume filtering produced a false negative detection. Our results demonstrate the effectiveness of plankton tow net sampling for eDNA early detection of invasive mussels, a method that can be used exclusively or as a supplement to filter sampling. Our results further suggest that eDNA testing could be incorporated into monitoring programs that routinely use plankton tows for visual detection of invasive mollusk larvae, as well as other aquatic invasive and non‐invasive species ranging from plankton to metazoans, including many fish.

Cloning, characterization, and spatio-temporal expression patterns of HdhSPARC and its responses to multiple stressors

SPARC is an extracellular Ca2+-binding, secreted glycoprotein that plays a dynamic role in the growth and development of organisms. This study aimed to describe the isolation, characterization, and expression analysis of HdhSPARC in Pacific abalone (Haliotis discus hannai) to infer its potential functional role. The isolated HdhSPARC was 1633 bp long, encoding a polypeptide of 284 amino acid residues. Structurally, the SPARC protein in abalone is comprised of three biological domains. However, the structure of this protein varied between vertebrates and invertebrates, as suggested by their distinct clustering patterns in phylogenetic analysis. In early development, HdhSPARC was variably expressed, and higher expression was found in veliger larvae. Moreover, HdhSPARC was highly expressed in juvenile abalone with rapid growth compared to their slower-growing counterparts. Among the testicular development stages, the growth stage exhibited higher HdhSPARC expression. HdhSPARC was also upregulated during muscle remodeling and shell biomineralization, as well as in response to different stressors such as heat shock, LPS, and H2O2 exposure. However, this gene was downregulated in Cd-exposed abalone. The present study first comprehensively characterized the HdhSPARC gene, and its spatio-temporal expressions were analyzed along with its responses to various stressors.

Effects of the toxic dinoflagellate Protoceratium reticulatum and its yessotoxins on the survival and feed ingestion of Argopecten purpuratus veliger larvae

The effects of yessotoxins (YTXs) produced by the dinoflagellate Protoceratium reticulatum in the early stages of bivalves have not been studied in detail. The present study evaluates the effects of P. reticulatum and YTXs on the survival and feed ingestion of veliger larvae of Argopecten purpuratus. Larvae were 96 h-exposed to 500, 1000 and 2000 P. reticulatum cells mL−1, and their equivalent YTX extract was prepared in methanol. Results show a survival mean of 82 % at the highest density of dinoflagellate, and 38 % for larvae with the highest amount of YTX extract. Feed ingestion is reduced in the dinoflagellate exposure treatments as a function of cell density. Therefore, the effect of YTXs on A. purpuratus represents a new and important area of study for investigations into the deleterious effects of these toxins in the early stages of the life cycle of this and, potentially, other bivalves.

Laboratory and Field Culture of Larvae of The Slipper Limpet, Crepidula fornicata.

The calyptraeid gastropod mollusk, Crepidula fornicata, has been widely used for studies of larval developmental biology, physiology, and ecology. Brooded veliger larvae of this species were collected by siphoning onto a sieve after natural release by adults, distributed into the culture at a density of 200/L, and fed with Isochrysis galbana (strain T-ISO) at 1 x 105 cells/mL. Shell growth and acquisition of competence for metamorphosis were documented for sibling larvae reared in ventilated 800 mL cultures designed for equilibration to ambient air or to defined atmospheric gas mixtures. Contrasting with these laboratory culture conditions; growth and competence data were also collected for larvae reared in a 15 L flow-through ambient seawater mesocosm located in a field population of reproductive adults. Growth rates and timing of metamorphic competence in the laboratory cultures were similar to those reported in previously published studies. Larvae reared in the field mesocosm grew much faster and metamorphosed sooner than reported for any laboratory studies. Together, these methods are suited for exploring larval development under predetermined controlled conditions in the laboratory as well as under naturally occurring conditions in the field.

Scallop larvae resistant to a pathogenic Vibrio harbor host-associated bacteria with probiotic potential

The breeding of the scallop Argopecten purpuratus is threatened by mass mortality events mostly caused by bacterial pathogens of the genus Vibrio affecting larvae and spat cultivated in hatcheries. A strong genetic basis for the resistance of A. purpuratus larvae to vibriosis has been demonstrated, which is associated with an efficient energy metabolism and immunocompetence. However, various factors, including the microbiota composition, have a notable impact on the outcome of an infection in numerous animal models, affecting their resistance. In this context, we aimed to explore the possible contribution of host-associated bacteria to the resistance of scallop larvae to Vibrio bivalvicida VPAP30. To achieve this, we isolated culturable bacteria from four scallop families displaying contrasted resistance to VPAP30 infection. We taxonomically assigned 75 bacterial isolates by 16S rDNA sequencing, and we found that 34 isolates were exclusively associated with resistant larvae. Then, 8 isolates were pre-selected as potential probiotics based on their exclusivity to the resistant families and bibliographic records of their use. Results highlighted three isolates displaying potential, belonging to the genera Psychrobacter, Hydrogenophaga, and Shewanella. The three isolates displayed in vitro antagonistic activity against VPAP30 and a protective effect on scallop veliger larvae against infection. We observed three main aspects of this protective effect: scallop larvae pretreated with each of the three probiotic candidates showed (i) no mortality after 24 h infection, (ii) an earlier induction of immune pattern recognition receptors, and (iii) no increase in total Vibrio spp. counts after infection with VPAP30. The findings demonstrated that scallop larvae resistant to vibriosis carry distinct bacteria possessing probiotic properties suitable for the rearing of scallop larvae. This study reinforces the significance of host resistance-related microorganisms as a potential source for discovering novel endogenous probiotics with biotechnological applications.

Identification and functional analysis of myeloid differentiation factor 88 (MyD88) in early development of Haliotis diversicolor

Myeloid differentiation factor 88 (MyD88) is a universal adaptor protein and plays an important role in the signal transduction of Toll like receptors (TLR) family. In this study, the MyD88 gene from the Haliotis diversicolor (hdMyD88) was identified. The full-length cDNA of hdMyD88 has a 1927 base pairs (bp), with an open reading frame of 1314 bp encoding 437 amino acids including a death domain (DD) at the N-terminus and TIR domain at the C-terminus which are typical features of MyD88 family proteins. Three conserved boxes are also found in the hdMyD88, which are similar to MyD88 in vertebrates. The expression levels of hdMyD88 mRNA at different early embryonic developmental stages of abalone were measured by qPCR revealed that their constitutive expression at all developmental stages analyzed with the considerably highest values at 8 cell stage and the lowest level at the trochosphere stage. Additionally, the mRNA expression of hdMyD88 decreased significantly (P < 0.05) after MyD88-dsRNA soak in the stage of trochosphere and veliger than EGFP-dsRNA group and blank control group. Whole embryo in situ hybridization showed that the positive signals of hdMyD88 were in visceral mass of trochophore larvae and veliger larvae. These results indicate hdMyD88 may could respond to pathogenic infection and may play an important role in early innate immunity in the process of abalone larval development.

Effects of temperature, salinity and diet on embryonic and early larval development in Littorina littorea (Gastropoda: Littorinimorpha)

IntroductionThe common periwinkle Littorina littorea is a delicacy in southern Europe. Enabling aquaculture production would be measure to avoid future over-harvesting and ensure a constant market supply.MethodsHowever, knowledge gaps in reproductive biology needs to be filled to allow setting up rearing protocols.ResultsWe investigated effects of temperature and salinity on embryonic development by incubating eggs at ten different temperatures in steps every 2°C, form 5 to 23°C) and four salinities (20 – 40). Embryonic development was clearly temperature dependent; eggs hatched after three days at the highest temperature while it took 16 days to hatch at the lowest temperature. Calculating effective day-degrees (DDeff) allowed expressing the thermal history of the embryogenesis and concluding that this species hatches on average at 61 (SD 3.7) DDeff. Larval growth- and developmental rates showed a different response to the temperature extremes as snails with the smallest shell lengths at hatch were found for the highest and lowest temperatures (204.5, SD = 11.6, and 202.9µm, SD = 13.7 respectively). Viable larvae only hatched at temperatures above 9°C. Larvae hatched at lower temperatures did not develop a shell or swimming lobes. Salinity and temperature significantly influenced time-to-hatch and hatching success; the lowest salinity (20PSU) lead to later hatching (80-100DDeff) and lower hatching success compared to higher salinities (25, 30, and 40, hatch around 40-50 DDeff). Neither hatch rate nor time-to-hatch showed differences among the higher salinities, suggesting a salinity threshold below 25. There was no significant interaction between salinity and temperature on time-to-hatch or hatching success. Larval diet had a significant effect on growth rates during the planktonic stage. Veliger larvae fed on two different live microalgae mono-diets (Dunaliella tertiolecta and Rhodomonas baltica) successfully passed their pelagic phase at the same time with no differences in growth rates, while veliger larvae fed on either of two preserved microalgae products (Nannochloropis sp. and Tetrasemis sp. frozen pastes) were not growing.DiscussionWe conclude that embryonal development can be successfully completed at temperatures between 7 and 20°C, while lower or higher temperatures should be avoided. Successfully hatched larvae can be fed on mono-microalgae diets until settlement.

Quantification of lipid droplets in hatchery reared veliger larvae of the green-lipped mussel, Perna canaliculus

Domestication efforts and selective breeding in bivalve aquaculture would greatly benefit from reliable screening biomarkers in the early life stages of development (e.g., larvae). In this study the lipids remaining at the completion of embryo development in selectively-bred/hatchery reared green-lipped mussel, Perna canaliculus, were targeted and quantified using two approaches: confocal laser scanning microscopy and image analysis (CLSM/IA) on Oil Red-O stained larva (i.e., lipid droplet number and volume distribution), and Iatroscan thin layer chromatography/flame ionization detection (TLC/FID) (i.e., energetic and structural lipid classes content). Four groups of newly formed pre-feeding veliger larvae, produced from two genetically distinct families (FA, FB) that completed embryo development at either low (L, 17 °C) or high (H, 21 °C) temperatures, were evaluated. The FB produced offspring better equipped to deal with higher temperatures during embryonic development, with greater survival, higher normal development, and larger larvae than FA when developing at 21 °C. In terms of lipids content, CLSM/IA showed that each individual larva had overall between 83 and 157 lipid droplets (ranging in volume from 2.5 to 294 μm3). The TLC/FID estimation of mean total lipid content ranged from 4.1 to 6.9 ƞg larva−1 (29–39% structural lipid and 62–69% energetic lipid). Both lipid quantification methods were successful in detecting differences among larval groups with different genetic backgrounds, as well as providing insight into the ability to cope with changes in the growing environment when relying on maternal energy sources (triacylglycerol-TAG identified as the main source of energy for early development). Stronger differences between the two families were identified using CLSM/IA than with TLC/FID (i.e., with FA having overall more and larger lipid droplets). However, TLC/FID was able to distinguish FA larvae reared at high temperatures from the other groups (due to lower levels of TAG, 46% of total lipids, versus 52–56% in the other groups), therefore being more sensitive to the pooled population response compared to the individual lipid droplet quantification in CLSM/IA. The application of these tools, when fully developed, could be extended to controlled commercial operations to diagnose and predict larval performance, enabling more directed early screening and informing production and breeding decisions.

Integration of physiological and gene expression analyses to reveal biomarkers for protein dynamic mechanisms regulating higher growth and survival among larval oyster families (Crassostrea gigas)

The oyster Crassostrea gigas is a major, global aquaculture species. As with any domestically-farmed species, the characterization of breeding lines that yield desired phenotypes is of immense value. An understanding of the fundamental biological bases of such phenotypes is needed to enhance aquaculture production. The aim of our study was to investigate the mechanisms of protein metabolic dynamics and energy allocation in oyster larvae. A series of controlled crosses yielded full-sibling larval families that allowed for measurements of integrative physiological processes during development. Experimentally, phenotypic contrasts between larval families were assayed by measuring: (1) growth and survival, (2) utilization of energy reserves of lipid and protein, (3) rates of protein synthesis and turnover, (4) respiration rates, and (5) transcriptome gene expression. Initially, newly-formed 2-day-old veliger larvae from four different families had similar sizes and physiologies, as measured by respiration, protein synthesis, turnover and content, the amount of energy allocated to protein synthesis, and gene expression pattern. Upon feeding, notable phenotypic contrasts became evident in different families. The larval family with faster growth had higher rates of protein synthesis and allocated a higher percentage of available energy to that single process. Based on family-specific differences, a series of samples was selected for developmental time-course analysis of changes in RNA pools. Principal component analyses of family-specific differential gene expression, combined with measured biochemical and physiological processes, led to the identification of two ribosomal gene biomarkers for protein synthesis. Such biomarkers could be potentially valuable tools for assessing complex traits that regulate physiological state, leading to optimization of breeding programs for oyster aquaculture.

Exposure to Alexandrium spp. impairs the development of Green-lipped mussel (Perna canaliculus) embryos and larvae

The green-lipped mussel (GLM) Perna canaliculus is an economically, ecologically, and culturally important species in Aotearoa New Zealand. Since 2011, harmful algal blooms (HABs) of Alexandrium spp. have occurred annually in the Marlborough Sounds, the largest GLM aquaculture region in New Zealand. Across a similar timeframe, there has been a severe reduction in wild spat (juvenile mussel) catch. This research investigated the effects of Alexandrium pacificum (which produces paralytic shellfish toxins; PSTs) and A. minutum (a non-producer of PSTs) on the development of four GLM larval life stages (gametes, embryos, D-stage and settlement).Early life stages of GLM were exposed to environmentally relevant concentrations of Alexandrium spp. as whole cell, lysate and filtrate treatments. A 48-h exposure of embryos to whole A. pacificum cells at 500 cells mL−1 caused lysis of embryos, severe abnormalities, and reduced development through to veliger (D-stage) larvae by 85%. GLM growth was impaired at cell concentrations as low as 250 cells mL−1 during a 4-day exposure of D-stage larvae to both Alexandrium spp. Exposure of GLM to both whole and lysed treatments of Alexandrium spp. at 500 cells mL−1 resulted in halved larval growth rates (2.00 µm day−1 vs 4.48 µm day−1 in the control) and growth remained impeded during a 4-day recovery period.Both A. pacificum and A. minutum were found to negatively impact D-larvae. Both whole-cell and lysed-cell treatments of A. pacificum had similar negative effects, suggesting that Alexandrium spp. toxicity to D-larvae is independent of PSTs. Additionally, cell membrane-free treatments of A. pacificum had no negative effects on embryo development, indicating that cell surface-associated bioactive compounds may be responsible for the observed negative effects during this early life stage.Conversely, non-PST-producing A. minutum was toxic to D-stage larvae but not to embryos; larval growth was reduced following a brief 1 h exposure of sperm to cell membrane-free treatments of A. pacificum. No effects were recorded in GLM larvae exposed during settlement, highlighting the potential for differences in susceptibility of early life stages to Alexandrium spp. exposure and the influence of exposure durations.In the wild, blooms of Alexandrium spp. can persist for several months, reaching cell densities higher than those investigated in the present study, and as such may be detrimental to the vulnerable early life stages of GLM.

A Diet Rich in HUFAs Enhances the Energetic and Immune Response Capacities of Larvae of the Scallop Argopecten purpuratus.

Massive mortalities in farmed larvae of the scallop Argopecten purpuratus have been associated with pathogenic Vibrio outbreaks. An energetic trade-off between development-associated demands and immune capacity has been observed. Given that highly unsaturated fatty acids (HUFAs) are essential nutrients for larval development, we evaluated the effect of diets based on microalgae low and high in HUFAs (LH and HH, respectively) on the energetic condition and the immune response of scallop larvae. The results showed that the HH diet increased cellular membrane fluidity in veliger larvae. The routine respiration rate was 64% higher in the HH-fed veligers than in the LH-fed veligers. Additionally, the metabolic capacity tended to be higher in the HH-fed veligers than in the LH-fed veligers after the Vibrio challenge. After the challenge, the HH-fed veligers presented higher transcript induction of ApTLR (immune receptor) and ApGlys (immune effector) genes, and the HH-fed pediveligers presented higher induction of ApLBP/BPI1 (antimicrobial immune effector) gene, than the LH-fed larvae. Furthermore, the HH-fed veligers controlled total Vibrio proliferation (maintaining near basal levels) after the bacterial challenge, while the LH-fed veligers were not able to control this proliferation, which increased three-fold. Finally, the HH-fed larvae showed 20-25% higher growth and survival rates than the LH-fed veligers. Overall, the results indicated that the administration of a HH diet increases cell membrane fluidity and energy metabolic capacity, which in turn enhances immunity and the ability to control Vibrio proliferation. The administration of microalgae high in HUFAs would be a promising strategy for improving scallop larval production efficiency.

Molecular Characterization of Tropomyosin and Its Potential Involvement in Muscle Contraction in Pacific Abalone.

Tropomyosin (TPM) is a contractile protein responsible for muscle contraction through its actin-binding activity. The complete sequence of TPM in Haliotis discus hannai (Hdh-TPM) was 2160 bp, encoding 284 amino acids, and contained a TPM signature motif and a TPM domain. Gene ontology (GO) analysis based on the amino acid sequence predicted Hdh-TPM to have an actin-binding function in the cytoskeleton. The 3D analysis predicted the Hdh-TPM to have a coiled-coil α-helical structure. Phylogenetically, Hdh-TPM formed a cluster with other TPM/TPM1 proteins during analysis. The tissue-specific mRNA expression analysis found the higher expression of Hdh-TPM in the heart and muscles; however, during embryonic and larval development (ELD), the higher expression was found in the trochophore larvae and veliger larvae. Hdh-TPM expression was upregulated in fast-growing abalone. Increasing thermal stress over a long period decreased Hdh-TPM expression. Long-term starvation (>1 week) reduced the mRNA expression of Hdh-TPM in muscle; however, the mRNA expression of Hdh-TPM was significantly higher in the mantle, which may indicate overexpression. This study is the first comprehensive study to characterize the Hdh-TPM gene in Pacific abalone and to report the expression of Hdh-TPM in different organs, and during ELD, different growth patterns, thermal stress, seasonal changes, and starvation.

Silver nanoparticle toxicity to the larvae of oyster Crassostrea angulata: Contribution of in vivo dissolution

Understanding the toxic mechanism of silver nanoparticles (AgNPs) is crucial for it risk assessment in marine environment, but the role of Ag+ release in the AgNP toxicity to marine biota is not yet well addressed. This study investigated the toxicity of AgNPs to the veliger larvae of oyster Crassostrea angulata, with a specific focus on the possibility of the involvement of in vivo dissolution of AgNPs in the toxicity via an aggregation-induced emission luminogen (AIEgen)-based imaging technique. AgNO3 exhibited significantly greater toxicity than AgNPs based on the total Ag, as indicated by lower 50 % growth inhibition concentration (EC50). The average concentration of soluble Ag in seawater at the EC50 of AgNPs was far lower than the EC50 of AgNO3, indicating that the AgNP toxicity could not be fully explained by the dissolved Ag in the medium. Despite the comparable soluble Ag concentration in seawater for both treatments, more Ag was accumulated in the larvae exposed to AgNPs, suggesting their ability to directly ingest particulate Ag, which was further confirmed by the presence of AgNPs aggregates in the esophagus and stomach. With the application of AIEgen-based imaging technique, in vivo dissolution of AgNPs in oyster larvae was thoroughly verified by an increase in Ag(I) content in the larvae exposed to AgNPs after depuration. The results collectively implied that apart from the Ag released in the medium, the Ag dissolved from the ingested AgNPs may also greatly contribute to the toxicity of AgNPs toward the oyster larvae. The findings of this work shed new light on the bioavailability and toxicity of AgNPs in marine environment.

Lipid Profiling of Pacific Abalone (Haliotis discus hannai) at Different Developmental Stages Using Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry.

Pacific abalone (Haliotis discus hannai) is a commercially important mollusk; therefore, improvement of its growth performance and quality has been emphasized. During embryonic development, abalones undergo a series of distinct larval stages, including swimming veliger larvae, juveniles, and mature individuals, and their biomolecular composition varies depending on the developmental stage. Therefore, in the present study, we performed untargeted lipid profiling of abalone tissues at different developmental stages as well as the hemolymph of mature female and male abalones using ultrahigh-performance liquid chromatography-tandem mass spectrometry. These profiles can provide meaningful information to understand compositional changes in lipids through abalone metamorphosis and development. A total of 132 lipids belonging to 15 classes were identified from abalone tissues at different developmental stages. Moreover, 21 lipids belonging to 8 classes were identified from the hemolymph of mature abalones. All data were processed following strict criteria to provide accurate information. Triglycerides and phosphatidylcholines were the major lipid components identified in both tissues and hemolymph, accounting for, respectively, 27% and 15% of all lipids in tissues and, respectively, 24% and 38% of all lipids in the hemolymph. Of note, lysophosphatidylcholine was only detected in the tissues of mature abalones, paving the way for further analyses of abalone lipids based on developmental stages. The present findings offer novel insights into the lipidome of abalone tissues and hemolymph at different developmental stages, building a foundation for improving the efficiency and quality of abalone aquaculture.

Ontogenic development of digestive enzymes in veliger larvae of dog conch (Laevistrombus canarium)

Ontogenic development of digestive enzymes in veliger larvae of dog conch (Laevistrombus canarium)

A mosaic of conserved and novel modes of gene expression and morphogenesis in mesoderm and muscle formation of a larval bivalve

The mesoderm gives rise to several key morphological features of bilaterian animals including endoskeletal elements and the musculature. A number of regulatory genes involved in mesoderm and/or muscle formation (e.g., Brachyury (Bra), even-skipped (eve), Mox, myosin II heavy chain (mhc)) have been identified chiefly from chordates and the ecdysozoans Drosophila and Caenorhabditis elegans, but data for non-model protostomes, especially those belonging to the ecdysozoan sister clade, Lophotrochozoa (e.g., flatworms, annelids, mollusks), are only beginning to emerge. Within the lophotrochozoans, Mollusca constitutes the most speciose and diverse phylum. Interestingly, however, information on the morphological and molecular underpinnings of key ontogenetic processes such as mesoderm formation and myogenesis remains scarce even for prominent molluscan sublineages such as the bivalves. Here, we investigated myogenesis and developmental expression of Bra, eve, Mox, and mhc in the quagga mussel Dreissena rostriformis, an invasive freshwater bivalve and an emerging model in invertebrate evodevo. We found that all four genes are expressed during mesoderm formation, but some show additional, individual sites of expression during ontogeny. While Mox and mhc are involved in early myogenesis, eve is also expressed in the embryonic shell field and Bra is additionally present in the foregut. Comparative analysis suggests that Mox has an ancestral role in mesoderm and possibly muscle formation in bilaterians, while Bra and eve are conserved regulators of mesoderm development of nephrozoans (protostomes and deuterostomes). The fully developed Dreissena veliger larva shows a highly complex muscular architecture, supporting a muscular ground pattern of autobranch bivalve larvae that includes at least a velum muscle ring, three or four pairs of velum retractors, one or two pairs of larval retractors, two pairs of foot retractors, a pedal plexus, possibly two pairs of mantle retractors, and the muscles of the pallial line, as well as an anterior and a posterior adductor. As is typical for their molluscan kin, remodelling and loss of prominent larval features such as the velum musculature and various retractor systems appear to be also common in bivalves.