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To quantify the ecological impacts of organic sediments and environmental dredging, benthic mollusks were chosen as bioindicators of environmental change, measured as sediment organic content and associated parameters. Data on species richness, ecological diversity (which was measured as biodiversity), and abundances were collected alongside sediment and near-bottom water quality data before, during, and after environmental dredging. Organic sediment content was found to have an inverse logarithmic relationship with benthic mollusk biodiversity, species richness, and abundance. Post hoc analyses found that percent dissolved oxygen, which correlates with sediment organic content, was responsible for 29.31–34.12% of the benthic mollusk community variation. Sediments with lower organic content had higher biodiversity (organism densities up to 1 organism m−2), abundance (over 2.0 × 105 organisms m−2), and species richness (organism densities up to 4 organisms m−2). In comparison, sediments with higher organic content had low biodiversity (organism densities 0–1 organisms m−2), abundance (as low as 0 organisms m−2), and species richness (organism densities as low as 0 organisms m−2). [ABSTRACT FROM AUTHOR]

The knowledge of comparative and developmental immunobiology has grown over the years and has been strengthened by the contributions of multi-omics research. High-performance microscopy, flow cytometry, scRNA sequencing, and the increased capacity to handle complex data introduced by machine learning have allowed the uncovering of aspects of great complexity and diversity in invertebrate immunocytes, i.e., immune-related circulating cells, which until a few years ago could only be described in terms of morphology and basic cellular functions, such as phagocytosis or enzymatic activity. Today, invertebrate immunocytes are recognized as sophisticated biological entities, involved in host defense, stress response, wound healing, organ regeneration, but also in numerous functional aspects of organismal life not directly related to host defense, such as embryonic development, metamorphosis, and tissue homeostasis. The multiple functions of immunocytes do not always fit the description of invertebrate organisms as simplified biological systems compared to those represented by vertebrates. However, precisely the increasing complexity revealed by immunocytes makes invertebrate organisms increasingly suitable models for addressing biologically significant and specific questions, while continuing to present the undeniable advantages associated with their ethical and economic sustainability. [ABSTRACT FROM AUTHOR]

Rhodopseudomonas palustris (RP) are known anaerobic bacteria with probiotic properties containing several bioactive compounds and enzymes that benefit aquatic animals. However, studies on the use of RP on aquatic animal species are limited. This study investigated the effects of dietary supplementation with RP formulation on the growth, non-specific immunity, and disease resistance of juvenile ivory shells (Babylonia areolata). The experiment was conducted for 8 weeks, with B. areolata fed a control diet (RP0) and four diets containing four different RP formulations, with doses of 1 (RP1), 5 (RP2), 10 (RP3), and 20 (RP4) g/kg, respectively. Higher levels of R. palustris in the formulation led to increased final weight, weight gain, and specific growth rate in B. areolata. The crude protein content was significantly higher in the RP4 group compared to the RP0 group. However, there was no significant difference in the crude lipid content. Higher levels of R. palustris in the RP4 formulation group increased the trypsin and lipase activities. Dietary supplementation with RP significantly increased the total antioxidant capacity, superoxide dismutase, and catalase activities and decreased the malondialdehyde content in B. areolata. Acid phosphatase and alkaline phosphatase activities were significantly increased in the RP4 group compared to the RP0 group. Dietary RP significantly increased the expression levels of antioxidant-related (superoxide dismutase, Cu/Zn-superoxide dismutase, glutathione S-transferase A-like, ferritin) and immune-related (acid phosphatase, cytochrome c) genes. Higher levels of R. palustris in the formulations RP3 and RP4 increased the survival rate of B. areolata challenged with Vibrio parahaemolyticus. These findings indicate that R. palustris preparation could improve growth performance, muscle composition, and digestive capacity and may act as an immune booster for preventing disease in B. areolata. [ABSTRACT FROM AUTHOR]

Chitons are a group of mollusks (polyplacophores) that differ from the mollusks that are most often found on marine rocky shores (bivalves and gastropods). On the Atlantic coast of North America, chitons have been reported in subtidal habitats from Newfoundland (Canada) to New England (USA). Here, we report the finding of Boreochiton ruber at the mid-to-high intertidal zone in a wave-exposed habitat in Nova Scotia (Canada). After surveying various coastal locations across Nova Scotia for 20 years (2004 to 2024), this was the only intertidal chiton we ever found in such habitats, making this an extremely rare occurrence. Rare species may contribute in unique ways to community functioning, making their reports valuable for biodiversity and ecosystem research. [ABSTRACT FROM AUTHOR]

Trematodes of the family Cyclocoelidae are parasites mainly of the respiratory system of birds and present a cosmopolitan distribution. Although infection with these flukes can result in pathological changes and even bird death, information on their life cycles is scarce and almost entirely based on experimental infection data. Thus, the generation of knowledge on the mollusks that act as natural intermediate hosts of cyclocoelids is necessary and can aid control measures against these air sac trematodes. In the present study, gastropod mollusks collected in an urban stream from Belo Horizonte, Minas Gerais, Brazil, were subjected to the compression technique for the detection of non-emerging larval trematodes. Tailless cercariae with confluent ceca were found in 8/30 (26.7%) specimens of Biomphalaria glabrata and 3/33 (9.1%) specimen of Physella acuta. Samples of the cercariae were subjected to morphological characterization and genetic study (28S, Cox-1, and Nad-1). For comparative purposes, adult trematodes previously collected in the air sac of a common gallinule (Gallinula galeata) found dead in another waterbody from the same region were also characterized. The molecular sequences obtained revealed a high degree of similarity (100% in 28S, 99.2% in Cox-1, and 99.5% in Nad-1) between larval stages found in mollusks and adult parasites found in G. galeata and morphologically identified as Cyclocoelum mutabile. The conspecificity with this widely distributed cyclocoelid was also corroborated by phylogenetic analysis and comparison with isolates of this species previously characterized in Peru and the Czech Republic (99.4–100% and 96.7–97.0% of similarity in Nad-1, respectively). Thus, the integrative analysis carried out in the present work enabled us to identify C. mutabile in mollusks in South America for the first time. The finding of B. glabrata and P. acuta as new intermediate hosts corroborates the importance of freshwater gastropods in the transmission of C. mutabile, as well as the low specificity to the mollusk group, as previously characterized through experimental studies. [ABSTRACT FROM AUTHOR]

Invertebrates in tropical ecosystems are generally considered to have little or no need for a dormant phase due to the stability of the habitat. However, resting stages of aquatic organisms are occasionally found here as well. This fact increases the possibility of transport of tropical organisms by ships' ballast water, which is the main vector for the spread of alien aquatic organisms between continents. During a study of resting stages in the bottom sediments of the island of Taiwan in 2006–2007, nine species of invertebrates were found, invasive or new to the fauna of the island, with some of them forming large banks of resting stages in sediments. [ABSTRACT FROM AUTHOR]

Emerging evidence of rhodolith bed complexity and heterogeneity poses a challenge to monitoring strategies and questions about the role of abiotic factors in controlling the observed morphostructural diversity. Mollusk thanatocoenoses quickly respond to environmental conditions, expressing fidelity to biocoenosis and representing, thus, a useful ecological/paleoecological tool to unravel this heterogeneity. In this research, we studied three distinct rhodolith beds from the Tyrrhenian Sea (Italy), in a range between 40 and 100 m of water depth, together with their mollusk thanatocoenoses, sediment size, and oceanographic conditions. The beds are all heterogeneous and rarely correspond to a specific rhodolith morphotype and shape. On the contrary, the study of the associated thanatocoenoses led to distinguish at least five different assemblages within the known variability of the Coastal detritic (DC) association. DC and deep mud (VP) mollusk species dominate hanatocoenosis A, which is associated with mixed sediment and a low hydrodynamic regime. Thanatocoenosis B corresponds to an assemblage in which DC species mixed with species of adjacent vegetated bottoms in sandy sediment with a medium hydrodynamic. Thanatocoenosis C includes species related to muddy coastal detritic (DE) and offshore detritic (DL) associated with sand and a variable proportion of praline and branch morphotypes. Thanatocoenoses D and E include a wide range of detritic species (DE DC, DL) together with VP and coralligenous (C), associated with exclusive praline and gravelly sand (Thanat. D), or a mixed proportion of branch and praline and mixed sediment (Thanat. E). Our results demonstrate that the study of mollusk thanatocoenoses provides insights into the diversity within and among heterogeneous rhodolith beds. Moreover, rhodoliths, as part of the sediment, create microhabitats suitable for a wide range of mollusk species that contribute to the formation of complex thanatocoenoses. [ABSTRACT FROM AUTHOR]

The effects of temperature manipulation, addition of sperm solution, and exposure to alkalized pH and/or hydrogen peroxide (H2O2) as possible spawning inducers in laboratory-conditioned and unconditioned adults of the yellow clam (Amarilladesma mactroides) were evaluated. In three trials, clams were laboratory-conditioned for 14 days and exposed to thermal shocks (from 20 °C to 23–29 °C), while clams from three additional trials were not conditioned but acclimatized for 45 min before spawning induction. Although conditioning advanced gonad maturity and increased the condition factor, none of the thermal treatments triggered spawning in these first trials. Histological analysis indicated that the gonads of conditioned clams were not mature. Alternatively, unfertilized and fertilized eggs, and larvae were observed after unconditioned clams were induced to spawn. The gonads of unconditioned clams were in an advanced stage of maturity. Exposure to temperature shocks, alone or in combination with the addition of sperm solution and with H2O2, resulted in spawning. Clams exposed to H2O2 kept their valves closed and had a high mortality rate. Temperature manipulation is the most promising stimulus to induce spawning in A. mactroides. As the success of conditioning depends on the stage of gonadal development at the time of capture in the wild, a period longer than 14 days may be necessary. [ABSTRACT FROM AUTHOR]

Spirolides are cyclic imines whose risks to human health have not been sufficiently evaluated. To determine the possible impact of these compounds in Galicia (NW Spain), their presence and concentration in bivalve mollusk were studied from 2014 to 2021. Only 13-desmethyl spirolide C (13desmSPXC) and an isomer have been detected, and always at low concentrations. Mussel, Mytilus galloprovincialis, was the species which accumulated more spirolides, but the presence of its isomer was nearly restricted to cockle, Cerastoderma edule, and two clam species, Venerupis corrugata and Polititapes rhomboides. On average, the highest 13desmSPXC levels were found in autumn-winter, while those of its isomer were recorded in spring-summer. Both compounds showed decreasing trends during the study period. Geographically, the concentration tends to decrease from the southern to the north-eastern locations, but temporal variability predominates over spatial variability. [ABSTRACT FROM AUTHOR]

Simple Summary: The current understanding of the origin and rapid diversification of biomineralized skeletal animals and their environmental interactions during the Cambrian Radiation is largely dependent on the phosphatized skeletal remains of early animals. However, many questions remain unresolved about this early Cambrian animal skeletonization bio-event, as identification of primary mineralogy and microstructural organization of these earliest skeletons is often difficult. The comparison of previously poorly understood fibrous microstructures (represented in this study by lamello-fibrillar nacre in early Cambrian mollusk and hyolith shells) with their modern counterparts (coleoid cuttlebones and serpulid tubes) shows key differences in shell microstructures of these animal groups. For example, the aragonitic lamello-fibrillar nacre in some Cambrian shells is herein demonstrated to represent a primitive calcitic prismatic microstructure. This also demonstrates the prevalence of calcitic shells in the Terreneuvian, a period of time when seawater chemistry was thought to predominantly facilitate the precipitation of aragonite skeletons, suggesting that biomineralization of prismatic calcite is strongly biologically controlled. The Precambrian–Cambrian interval saw the first appearance of disparate modern metazoan phyla equipped with a wide array of mineralized exo- and endo-skeletons. However, the current knowledge of this remarkable metazoan skeletonization bio-event and its environmental interactions is limited because uncertainties have persisted in determining the mineralogy, microstructure, and hierarchical complexity of these earliest animal skeletons. This study characterizes in detail a previously poorly understood fibrous microstructure—the lamello-fibrillar (LF) nacre—in early Cambrian mollusk and hyolith shells and compares it with shell microstructures in modern counterparts (coleoid cuttlebones and serpulid tubes). This comparative study highlights key differences in the LF nacre amongst different lophotrochozoan groups in terms of mineralogical compositions and architectural organization of crystals. The results demonstrate that the LF nacre is a microstructural motif confined to the Mollusca. This study demonstrates that similar fibrous microstructure in Cambrian mollusks and hyoliths actually represent a primitive type of prismatic microstructure constituted of calcitic prisms. Revision of these fibrous microstructures in Cambrian fossils demonstrates that calcitic shells are prevalent in the so-called aragonite sea of the earliest Cambrian. This has important implications for understanding the relationship between seawater chemistry and skeletal mineralogy at the time when skeletons were first acquired by early lophotrochozoan biomineralizers. [ABSTRACT FROM AUTHOR]

Big defensins are two-domain antimicrobial peptides (AMPs) that have highly diversified in mollusks. Cg-BigDefs are expressed by immune cells in the oyster Crassostrea gigas, and their expression is dampened during the Pacific Oyster Mortality Syndrome (POMS), which evolves toward fatal bacteremia. We evaluated whether Cg-BigDefs contribute to the control of oyster-associated microbial communities. Two Cg-BigDefs that are representative of molecular diversity within the peptide family, namely Cg-BigDef1 and Cg-BigDef5, were characterized by gene cloning and synthesized by solid-phase peptide synthesis and native chemical ligation. Synthetic peptides were tested for antibacterial activity against a collection of culturable bacteria belonging to the oyster microbiota, characterized by 16S sequencing and MALDI Biotyping. We first tested the potential of Cg-BigDefs to control the oyster microbiota by injecting synthetic Cg-BigDef1 into oyster tissues and analyzing microbiota dynamics over 24 h by 16S metabarcoding. Cg-BigDef1 induced a significant shift in oyster microbiota β-diversity after 6 h and 24 h, prompting us to investigate antimicrobial activities in vitro against members of the oyster microbiota. Both Cg-BigDef1 and Cg-BigDef5 were active at a high salt concentration (400 mM NaCl) and showed broad spectra of activity against bacteria associated with C. gigas pathologies. Antimicrobial specificity was observed for both molecules at an intra- and inter-genera level. Remarkably, antimicrobial spectra of Cg-BigDef1 and Cg-BigDef5 were complementary, and peptides acted synergistically. Overall, we found that primary sequence diversification of Cg-BigDefs has generated specificity and synergy and extended the spectrum of activity of this peptide family. [ABSTRACT FROM AUTHOR]

Mussels have been used as animal models for studying ecotoxicology, biomineralization, and bio-adhesion for many years. Despite a wealth of studies on their shell matrix and byssus proteins, few studies have focused on the extracellular matrix molecules in mussel soft tissues. Extracellular matrix molecules may play important roles in biomineralization, immune reaction, and tissue homeostasis. In the present study, extracellular matrix and mineralization-related molecules in zebra mussel soft tissue were immunolocalized using well-characterized monoclonal antibodies. Our results demonstrate specific immunolocalization for collagen IV, fibronectin, and keratan sulfate in hemocytes; collagen IV in peripheral nerves; and aggrecan, link protein, and collagen XVIII in foot tissue. Laminin, decorin, and osteonectin were also broadly immunolocalized in mussel soft tissues. The distributions of these extracellular matrix molecules in mussel tissues are in line with the cell-mediated shell mineralization hypothesis, providing evidence for the molecules involved in the peripheral nervous system and byssus formation, and explaining the conservation of extracellular matrix molecules during evolution. These results further contribute to establishing zebra mussels as an attractive animal model in biomedical research. [ABSTRACT FROM AUTHOR]

Simple Summary: Arion vulgaris has become a major invasive pest slug in Europe, causing extensive damage to many crops. To control this pest, the use of chemical molluscicides remains the most important. However, despite the proved efficacy, they still have detrimental environmental effects. We performed two double-replicated laboratory studies testing molluscicide pellets with metaldehyde (3% and 5%) and iron phosphate (1%) and found the reluctance of slugs to eat a full lethal dose regardless of whether the poison is stronger or weaker. As a consequence, slugs remain alive and only reduce their herbivory by half; the remaining granules or their parts are the main source of toxic effects of molluscicides in the environment. Moreover, a higher metaldehyde content of the pellets does not lead to lower herbivory. The results showed that a new application of molluscicides could be useful; the application rate should be decreased according to the ability of slugs to eat a certain amount of molluscicide pellets. Arion vulgaris are mostly controlled using chemical molluscicide products, and the detrimental environmental effects of these molluscicides can be reduced by decreasing the number of pellets applied per unit area. The objective of this study was to compare three slug control methods during two double-replicated seven-day laboratory experiments, in which slugs could choose the number of pellets with metaldehyde (3% or 5%) or iron phosphate (1%) and different types of food: leafy plants (lettuce), root vegetables (carrot), a cereal-based diet (oatmeal), or an animal-based diet (dry cat food). Slugs were irrigated and allowed to recover. We found a reluctance of slugs to eat big amounts of pellets and, therefore, to reach a full lethal dose, which resulted in low mortality (the rate was only 2.1%), regardless of whether the poison was stronger or weaker. Herbivory of slugs was in some cases reduced by half, but no treatments resulted in slugs to stop eating. Pellets with 3% metaldehyde were significantly more acceptable than pellets with 5% metaldehyde (uneaten pellets were left). The results showed that the new application of molluscicides could be useful; the application rate should be decreased according to the slugs number and ability of slugs to eat a certain amount of molluscicide pellets. [ABSTRACT FROM AUTHOR]

Marine organisms are prolific resources of guanidine-containing natural products with intriguing structures and promising biological activities. These molecules have therefore attracted the attention of chemists and biologists for their further studies towards potential drug leads. This review focused on the guanidine alkaloids derived from marine sources and discussed the recent progress on their isolation, synthesis and biological activities, covering the literature from the year 2010 to the present. [ABSTRACT FROM AUTHOR]

Over the last eight years newtheories regarding nucleation, crystal growth, and polymorphism have emerged. Many of these theories were developed in response to observations in nature, where classical nucleation theory failed to account for amorphous mineral precursors, phases, and particle assembly processes that are responsible for the formation of invertebrate mineralized skeletal elements, such as the mollusk shell nacre layer (aragonite polymorph) and the sea urchin spicule (calcite polymorph). Here, we summarize these existing nucleation theories and place them within the context of what we know about biomineralization proteins, which are likely participants in the management of mineral precursor formation, stabilization, and assembly into polymorphs. With few exceptions, much of the protein literature confirms that polymorph-specific proteins, such as those from mollusk shell nacre aragonite, can promote polymorph formation. However, past studies fail to provide important mechanistic insights into this process, owing to variations in techniques, methodologies, and the lack of standardization in mineral assay experimentation. We propose that the way forward past this roadblock is for the protein community to adopt standardized nucleation assays and approaches that are compatible with current and emerging nucleation precursor studies. This will allow cross-comparisons, kinetic observations, and hopefully provide the information that will explain how proteins manage polymorph formation and stabilization. [ABSTRACT FROM AUTHOR]

Since the 1990s, a number of terminal alkynyl residue-containing cyclic/acyclic peptides have been identified from marine organisms, especially cyanobacteria and marine mollusks. This review has presented 66 peptides, which covers over 90% marine peptides with terminal alkynyl fatty acyl units. In fact, more than 90% of these peptides described in the literature are of cyanobacterial origin. Interestingly, all the linear peptides featured with terminal alkyne were solely discovered from marine cyanobacteria. The objective of this article is to provide an overview on the types, structural characterization of these unusual terminal alkynyl fatty acyl units, as well as the sources and biological functions of their composed peptides. Many of these peptides have a variety of biological activities, including antitumor, antibacterial, antimalarial, etc. Further, we have also discussed the evident biosynthetic origin responsible for formation of terminal alkynes of natural PKS (polyketide synthase)/NRPS (nonribosome peptide synthetase) hybrids. [ABSTRACT FROM AUTHOR]

The invasive mollusk Crepidula fornicata, occurring in large amounts in bays along the French Northeastern Atlantic coasts, may have huge environmental effects in highly productive ecosystems where shellfish are exploited. The present study aims at determining the potential economic value of this marine species in terms of exploitable substances with high added value. Lipid content and phospholipid (PL) composition of this mollusk collected on the Bourgneuf Bay were studied through four seasons. Winter specimens contained the highest lipid levels (5.3% dry weight), including 69% of PLs. Phosphatidylcholine (PC) was the major PL class all year, accounting for 63.9% to 88.9% of total PLs. Consequently, the winter specimens were then investigated for PL fatty acids (FAs), and free sterols. Dimethylacetals (DMAs) were present (10.7% of PL FA + DMA mixture) revealing the occurrence of plasmalogens. More than forty FAs were identified, including 20:5n-3 (9.4%) and 22:6n-3 (7.3%) acids. Fourteen free sterols were present, including cholesterol at 31.3% of the sterol mixture and about 40% of phytosterols. These data on lipids of C. fornicata demonstrate their positive attributes for human nutrition and health. The PL mixture, rich in PC and polyunsaturated FAs, offers an interesting alternative source of high value-added marine lecithin. [ABSTRACT FROM AUTHOR]

Cooperative effects of magnesium ions and acidic polypeptides originating from a family of proteins known as Asprich (mollusk Atrina rigida) were studied. In our previous studies, these two acidic polypeptides were found to be effective in controlling the morphology of the calcium carbonate mineral, the main inorganic constituent of prismatic layer of the mollusk shell. Since these Asprich sequences are believed to contain a putative magnesium binding domain, the morphology-controlling effects were further investigated with the addition of magnesium ions. The mineral morphology was dramatically changed by the combined influence of each polypeptides and the magnesium ions, substantiating the recognized importance of magnesium in the formation of calcium carbonate-based biominerals. [ABSTRACT FROM AUTHOR]

The Eastern oyster (Crassostrea virginica) is a useful, robust model marine organism for tissue metabolism studies. Its relatively few organs are easily delineated and there is sufficient understanding of their functions based on classical assays to support interpretation of advanced spectroscopic approaches. Here we apply high-resolution proton nuclear magnetic resonance (1H NMR)-based metabolomic analysis to C. virginica to investigate the differences in the metabolic profile of different organ groups, and magnetic resonance imaging (MRI) to non-invasively identify the well separated organs. Metabolites were identified in perchloric acid extracts of three portions of the oyster containing: (1) adductor muscle, (2) stomach and digestive gland, and (3) mantle and gills. Osmolytes dominated the metabolome in all three organ blocks with decreasing concentration as follows: betaine > taurine > proline > glycine > β-alanine > hypotaurine. Mitochondrial metabolism appeared most pronounced in the adductor muscle with elevated levels of carnitine facilitating β-oxidation, and ATP, and phosphoarginine synthesis, while glycogen was elevated in the mantle/gills and stomach/digestive gland. A biochemical schematic is presented that relates metabolites to biochemical pathways correlated with physiological organ functions. This study identifies metabolites and corresponding 1H NMR peak assignments for future NMR-based metabolomic studies in oysters. [ABSTRACT FROM AUTHOR]

In humans, injuries and diseases can result in irreversible tissue or organ loss. This well-known fact has prompted several basic studies on organisms capable of adult regeneration, such as amphibians, bony fish, and invertebrates. These studies have provided important biological information and helped to develop regenerative medicine therapies, but important gaps concerning the regulation of tissue and organ regeneration remain to be elucidated. To this aim, new models for studying regenerative biology could prove helpful. Here, the description of the cephalic tentacle regeneration in the adult of the freshwater snail Pomacea canaliculata is presented. In this invasive mollusk, the whole tentacle is reconstructed within 3 months. Regenerating epithelial, connective, muscular and neural components are already recognizable 72 h post-amputation (hpa). Only in the early phases of regeneration, several hemocytes are retrieved in the forming blastema. In view of quantifying the hemocytes retrieved in regenerating organs, granular hemocytes present in the tentacle blastema at 12 hpa were counted, with a new and specific computer-assisted image analysis protocol. Since it can be applied in absence of specific cell markers and after a common hematoxylin-eosin staining, this protocol could prove helpful to evidence and count the hemocytes interspersed among regenerating tissues, helping to unveil the role of immune-related cells in sensory organ regeneration. [ABSTRACT FROM AUTHOR]

River water that receives treated wastewater can be contaminated by pathogens including enteric viruses due to fecal pollution, which may represent an important public health hazard. There is a great diversity of enteric viruses and fecal bacteriophages, especially F-specific RNA bacteriophages (FRNAPHs), are commonly proposed as indicators of viral pollution due to a variety of characteristics such as their structural similarities to the main enteric viruses, their high concentrations in raw wastewater and their environmental survival rate, which is better than other cultivable enteric viruses. However, evaluating the viral contamination of water on the basis of FRNAPH concentration levels continues to present a challenge. This is because the quality of detection is strongly dependent on the quantity of viral particles, high spatio-temporal variabilities and the physico-chemical conditions of the water during sampling. To overcome these limitations, the present study aims to evaluate whether the bivalve mollusk Dreissena polymorpha (zebra mussel) could be considered a suitable experimental model for assessing the viral contamination of rivers. In order to determine this, the capacity of D. polymorpha to accumulate FRNAPHs and assimilate them into their soft tissue was studied. This provided a proof of concept for the use of D. polymorpha to evaluate the viral contamination of surface water. Two experiments were conducted: (1) an in situ experiment to confirm that zebra mussels naturally accumulated FRNAPHs and (2) a laboratory experiment to determine the accumulation and depuration kinetics of FRNAPHs in D. polymorpha tissue. The study highlights the capacity of the mussels to accumulate infectious FRNAPHs both on a laboratory scale under controlled conditions as well as in situ at different sites that are representative of different bodies of water. An analysis of the mussels' soft tissue showed that they were capable of reflecting the water's contamination level very quickly (within less than 24 h). Moreover, the soft tissue retained the viral load much longer than the water due to a low depuration rate. The analysis of FRNAPH concentrations in mussels exposed in situ suggested that there were differences in contamination levels between sites. These preliminary results underline the potential utility of zebra mussels in assessing viral contamination by measuring the accumulation of FRNAPHs in their tissue. This may ultimately enable stakeholders to use zebra mussels as a means of monitoring viral pollution in surface water. [ABSTRACT FROM AUTHOR]

Simple Summary: Sustainable solutions to the spreading of invasive species are difficult to find due to the absence of biological information about basic immune mechanisms of the target pests. Here, we present evidence of the effects of a commercially available roundworm, Phasmarhabditis hermaphrodita, against the invasive apple snail Pomacea canaliculata. The effects are principally evaluated in terms of snail survival and immune activation. Via molecular and microscopy-based approaches, we demonstrate that dosage and temperature are critical in determining the effects of the roundworm, and that the apple snail response to this immune challenge involves different organs. To our knowledge, these findings are the first demonstration that a P. hermaphrodita-based molluscicide can effectively kill P. canaliculata and that the snail can mount a multi-organ response against this pathogenic roundworm. The spreading of alien and invasive species poses new challenges for the ecosystem services, the sustainable production of food, and human well-being. Unveiling and targeting the immune system of invasive species can prove helpful for basic and applied research. Here, we present evidence that a nematode (Phasmarhabditis hermaphrodita)-based molluscicide exerts dose-dependent lethal effects on the golden apple snail, Pomacea canaliculata. When used at 1.7 g/L, this biopesticide kills about 30% of snails within one week and promotes a change in the expression of Pc-bpi, an orthologue of mammalian bactericidal/permeability increasing protein (BPI). Changes in Pc-bpi expression, as monitored by quantitative PCR (qPCR), occurred in two immune-related organs, namely the anterior kidney and the gills, after exposure at 18 and 25 °C, respectively. Histological analyses revealed the presence of the nematode in the snail anterior kidney and the gills at both 18 and 25 °C. The mantle and the central nervous system had a stable Pc-bpi expression and seemed not affected by the nematodes. Fluorescence in situ hybridization (FISH) experiments demonstrated the expression of Pc-bpi in circulating hemocytes, nurturing the possibility that increased Pc-bpi expression in the anterior kidney and gills may be due to the hemocytes patrolling the organs. While suggesting that P. hermaphrodita-based biopesticides enable the sustainable control of P. canaliculata spread, our experiments also unveiled an organ-specific and temperature-dependent response in the snails exposed to the nematodes. Overall, our data indicate that, after exposure to a pathogen, the snail P. canaliculata can mount a complex, multi-organ innate immune response. [ABSTRACT FROM AUTHOR]

Tetrodotoxin (TTX) is a potent natural toxin causative of human food intoxications that shares its mechanism of action with the paralytic shellfish toxin saxitoxin (STX). Both toxins act as potent blockers of voltage-gated sodium channels. Although human intoxications by TTX were initially described in Japan, nowadays increasing concern about the regulation of this toxin in Europe has emerged due to its detection in fish and mollusks captured in European waters. Currently, TTX is only regularly monitored in Dutch fishery products. However, the European Food Safety Authority (EFSA) has established a safety level of 44 µg/kg TTX as the amount of toxin that did not cause adverse effects in humans. This level was extrapolated considering initial data on its acute oral toxicity and EFSA remarked the need for chronic toxicity studies to further reduce the uncertainty of future toxin regulations. Thus, in this work, we evaluated the oral chronic toxicity of TTX using the safety levels initially recommended by EFSA in order to exclude potential human health risks associated with the worldwide expanding presence of TTX. Using internationally recommended guidelines for the assessment of oral chronic toxicity, the data provided here support the proposed safety level for TTX as low enough to prevent human adverse effects of TTX even after chronic daily exposure to the toxin. However, the combination of TTX with STX at doses above the maximal exposure level of 5.3 µg/kg body weight derived by EFSA increased the lethality of TTX, thus confirming that both TTX and paralytic shellfish toxins should be taken into account to assess human health risks. [ABSTRACT FROM AUTHOR]

Heavy metal pollution has been on the rise with serious implications for the wellbeing of aquatic ecosystems. Benthic sediments and freshwater mollusk (snail): Mellanoides tuberculata were sampled from five stations for determination of heavy metals concentrations and measurement of antioxidant enzyme activities. The spatial variation was studied using an enrichment factor, potential ecological risk index, and mean probable effect limit quotient (mPELq). From the results, Cu, Zn, Cd, Cr, Pb, Ni, and Co contamination levels were high at stations S3, S4, and S5 with an mPEL quotient of 94.40%. The variation of metal concentration and Enrichment factor were in the order S5 > S3 > S4 > S2 > S1, which was attributed to anthropogenic influences at the catchment due to industrial activities and atmospheric deposition of metals. Station five in this study is downstream and requires the most monitoring and management to prevent several ecological risks of metal pollutants in River Kaduna. [ABSTRACT FROM AUTHOR]

Histone methylation patterns are important epigenetic regulators of mammalian development, notably through stem cell identity maintenance by chromatin remodeling and transcriptional control of pluripotency genes. But, the implications of histone marks are poorly understood in distant groups outside vertebrates and ecdysozoan models. However, the development of the Pacific oyster Crassostrea gigas is under the strong epigenetic influence of DNA methylation, and Jumonji histone-demethylase orthologues are highly expressed during C. gigas early life. This suggests a physiological relevance of histone methylation regulation in oyster development, raising the question of functional conservation of this epigenetic pathway in lophotrochozoan. Quantification of histone methylation using fluorescent ELISAs during oyster early life indicated significant variations in monomethyl histone H3 lysine 4 (H3K4me), an overall decrease in H3K9 mono- and tri-methylations, and in H3K36 methylations, respectively, whereas no significant modification could be detected in H3K27 methylation. Early in vivo treatment with the JmjC-specific inhibitor Methylstat induced hypermethylation of all the examined histone H3 lysines and developmental alterations as revealed by scanning electronic microscopy. Using microarrays, we identified 376 genes that were differentially expressed under methylstat treatment, which expression patterns could discriminate between samples as indicated by principal component analysis. Furthermore, Gene Ontology revealed that these genes were related to processes potentially important for embryonic stages such as binding, cell differentiation and development. These results suggest an important physiological significance of histone methylation in the oyster embryonic and larval life, providing, to our knowledge, the first insights into epigenetic regulation by histone methylation in lophotrochozoan development. [ABSTRACT FROM AUTHOR]

We evaluated the prevalence of influenza A virus (IAV) in different species of bivalves inhabiting natural water bodies in waterfowl habitat along the Delmarva Peninsula and Chesapeake Bay in eastern Maryland. Bivalve tissue from clam and mussel specimens (Macoma balthica, Macoma phenax, Mulinia sp., Rangiacuneata, Myaarenaria, Guekensia demissa, and an undetermined mussel species) from five collection sites was analyzed for the presence of type A influenza virus by qPCR targeting the matrix gene. Of the 300 tissue samples analyzed, 13 samples (4.3%) tested positive for presence of influenza virus A matrix gene. To our knowledge, this is the first report of detection of IAV in the tissue of any bivalve mollusk from a natural water body. [ABSTRACT FROM AUTHOR]

Shellfish purple, also known as Tyrian purple and royal purple, has a long history, which has been revealed and documented in recent years through valid physicochemical studies using sophisticated techniques. The aim of the work was to summarize the conclusions of these studies and to describe the results of two unpublished investigations regarding the (i) identification of shellfish purple in a textile (4th century BCE) from ancient Macedonia and (ii) dramatic effect of the dyeing conditions on the composition of the purple dye. Moreover, a critical discussion is included about the discovery of the shellfish pigment and dye based on the available scientific evidence. Previously published reports describing the identification of the shellfish colorant in objects of the cultural heritage were carefully summarized. Shellfish purple was not used only as colorant, but it served other purposes as emphasized in this review. In particular, examples for the use of shellfish purple in medicine, grave goods and fillers and plasters in walls, were described. Examples of materials and methods that were used in the past to produce "fake" purple, imitating the aesthetic result of the valuable royal marine material were summarized. Finally, the solubility of indigoids was discussed using modern approaches of physical chemistry. [ABSTRACT FROM AUTHOR]

Simple Summary: Bivalve sex is thought to fluctuate depending on environmental conditions. So far, there has been no investigation on the phenotypic stability of sex in the commercially important Yesso scallop Mizuhopecten yessoensis. The present study revealed that the sex of the Yesso scallop is stable after initial sex differentiation and that this species maintains a sex-stable maturation system throughout its life. In addition, gonad differentiation for each sex was precisely characterized by using molecular markers throughout the maturational cycle. The objective of the present study was to analyze the phenotypic stability of sex after sex differentiation in the Yesso scallop, which is a gonochoristic species that has been described as protandrous. So far, no study has investigated in detail the sexual fate of the scallop after completion of sex differentiation, although bivalve species often show annual sex change. In the present study, we performed a tracking experiment to analyze the phenotypic stability of sex in scallops between one and two years of age. We also conducted molecular marker analyses to describe sex differentiation and gonad development. The results of the tracking experiment revealed that all scallops maintained their initial sex phenotype, as identified in the last reproductive period. Using molecular analyses, we characterized my-dmrt2 and my-foxl2 as sex identification markers for the testis and ovary, respectively. We conclude by proposing that the Yesso scallop is a sex-stable bivalve after its initial sex differentiation and that it maintains a sex-stable maturation system throughout its life. The sex-specific molecular markers identified in this study are useful tools to assess the reproductive status of the Yesso scallop. [ABSTRACT FROM AUTHOR]

Various environmental and ecotoxicological aspects related to applications of neonicotinoid insecticides are assessed. Dosages of neonicotinoids applied in seed coating materials were determined and are compared to other applications (spray and granule). Environmental levels in soils and affecting factors in translocation are discussed. Excretion of neonicotinoids via guttation from coated maize seeds up to two months upon emergence, as well as cross-contamination of plants emerged from non-coated seeds or weeds nearby have been demonstrated. Contamination of surface waters is discussed in scope of a worldwide review and the environmental fate of the neonicotinoid active ingredients and the formulating surfactant appeared to be mutually affected by each other. Toxicity of neonicotinoid active ingredients and formulations on Daphnia magna completed with some investigations of activity of the detoxifying glutathione S-transferase enzyme demonstrated the modified toxicity due to the formulating agents. Electrophysiological results on identified central neurons of the terrestrial snail Helix pomatia showed acetylcholine antagonist (inhibitory) effects of neonicotinoid insecticide products, but no agonist (ACh-like) effects were recorded. These data also suggested different molecular targets (nicotinergic acetylcholine receptors and acetylcholine esterase enzyme) of neonicotinoids in the snail central nervous system., Competing Interests: The authors declare no conflict of interest.

The peptide FMRFamide is one of the well-known peptides involved in multiple physiological processes in the phylum Mollusca. In this study, a FMRFamide gene (GenBank accession No. KJ933411) was identified in a cuttlefish species called Sepiella japonica and was designated as SjFMRFamide. The total length of the SjFMRFamide sequence was found to be 1880 bp while the open reading frame contained 996 bp encoding a protein of 331 amino acid residues with a predicted isoelectric point (pI) and molecular weight (MW) of 9.18 and 38.8 kDa along with a 333 bp 5′-untranslated region (UTR) and 551 bp 3′-UTR. The deduced SjFMRFamide precursor protein contains one signal peptide and expresses four kinds FMRFamide-related peptides including a single copy of FLRFamide, ALSGDAFLRFamide, and FIRFamide and multiple copies of FMRFamide. Results of phylogenetic relation analysis strongly indicated that the sequence of this gene shares high identity with the genes of known FMRFamides. Spatial expression analysis indicated the highest mRNA expression of SjFMRFamide in the brain of male and female cuttlefishes among the eight tissues analyzed. An in situ hybridization assay of the brain indicated that SjFMRFamide was transcribed in several functional lobes, which suggests that it might be related to many physiological regulatory mechanisms. This is the first study describing FMRFamide in S. japonica and the results may contribute to future studies of neuropeptide evolution or may prove useful for the development of aquaculture methods for this cuttlefish species. [ABSTRACT FROM AUTHOR]

Lucina pectinata is a clam that lives in sulfide-rich environments and houses intracellular sulfide-oxidizing endosymbionts. To identify new Lucina pectinata proteins, we produced libraries for genome and transcriptome sequencing and assembled them de novo. We searched for histone-like sequences using the Lucina pectinata histone H3 partial nucleotide sequence against our previously described genome assembly to obtain the complete coding region and identify H3 coding sequences from mollusk sequences in Genbank. Solen marginatus histone nucleotide sequences were used as query sequences using the genome and transcriptome assemblies to identify the Lucina pectinata H1, H2A, H2B and H4 genes and mRNAs and obtained the complete coding regions of the five histone genes by RT-PCR combined with automated Sanger DNA sequencing. The amino acid sequence conservation between the Lucina pectinata and Solen marginatus histones was: 77%, 93%, 83%, 96% and 97% for H1, H2A, H2B, H3 and H4, respectively. As expected, the H3 and H4 proteins were the most conserved and the H1 proteins were most similar to H1's from aquatic organisms like Crassostrea gigas, Aplysia californica, Mytilus trossulus and Biomphalaria glabrata . The Lucina pectinata draft genome and transcriptome assemblies, obtained by semiconductor sequencing, were adequate for identification of conserved proteins as evidenced by our results for the histone genes.

Marine organisms are prolific resources of guanidine-containing natural products with intriguing structures and promising biological activities. These molecules have therefore attracted the attention of chemists and biologists for their further studies towards potential drug leads. This review focused on the guanidine alkaloids derived from marine sources and discussed the recent progress on their isolation, synthesis and biological activities, covering the literature from the year 2010 to the present., Competing Interests: The authors declare no conflict of interest.

Since the 1990s, a number of terminal alkynyl residue-containing cyclic/acyclic peptides have been identified from marine organisms, especially cyanobacteria and marine mollusks. This review has presented 66 peptides, which covers over 90% marine peptides with terminal alkynyl fatty acyl units. In fact, more than 90% of these peptides described in the literature are of cyanobacterial origin. Interestingly, all the linear peptides featured with terminal alkyne were solely discovered from marine cyanobacteria. The objective of this article is to provide an overview on the types, structural characterization of these unusual terminal alkynyl fatty acyl units, as well as the sources and biological functions of their composed peptides. Many of these peptides have a variety of biological activities, including antitumor, antibacterial, antimalarial, etc. Further, we have also discussed the evident biosynthetic origin responsible for formation of terminal alkynes of natural PKS (polyketide synthase)/NRPS (nonribosome peptide synthetase) hybrids., Competing Interests: The authors declare no conflict of interest.

The Eastern oyster (Crassostrea virginica) is a useful, robust model marine organism for tissue metabolism studies. Its relatively few organs are easily delineated and there is sufficient understanding of their functions based on classical assays to support interpretation of advanced spectroscopic approaches. Here we apply high-resolution proton nuclear magnetic resonance ((1)H NMR)-based metabolomic analysis to C. virginica to investigate the differences in the metabolic profile of different organ groups, and magnetic resonance imaging (MRI) to non-invasively identify the well separated organs. Metabolites were identified in perchloric acid extracts of three portions of the oyster containing: (1) adductor muscle, (2) stomach and digestive gland, and (3) mantle and gills. Osmolytes dominated the metabolome in all three organ blocks with decreasing concentration as follows: betaine > taurine > proline > glycine > ß-alanine > hypotaurine. Mitochondrial metabolism appeared most pronounced in the adductor muscle with elevated levels of carnitine facilitating ß-oxidation, and ATP, and phosphoarginine synthesis, while glycogen was elevated in the mantle/gills and stomach/digestive gland. A biochemical schematic is presented that relates metabolites to biochemical pathways correlated with physiological organ functions. This study identifies metabolites and corresponding (1)H NMR peak assignments for future NMR-based metabolomic studies in oysters.