Gills Of Mussels Research Articles

Integrated Biomarker Response of Oxidative Stress Parameters in the Digestive Glands and Gills of Autochthonous and Invasive Freshwater Mussels from the Sava River, Serbia.

In this study, the activity of oxidative stress parameters superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), and glutathione S-transferase (GST), as well as the concentrations of vitamin E (Vit E) and SH groups in the digestive glands and gills of freshwater mussels Unio pictorum and Sinanodonta woodiana from the Sava River in Serbia were investigated. These parameters were determined in native and invasive mussels under the same environmental conditions. The activities of GSH-Px and GR and the concentration of Vit E were significantly higher in the digestive glands of the autochthonous species U. pictorum than in the invasive species S. woodiana, while the CAT activity and the concentration of SH groups were lower. In the gills of U. pictorum, GSH-Px activity and Vit E concentration were significantly higher, while CAT, GST, and SH groups were lower. Principal component analysis (PCA) showed that oxidative stress parameters were strictly tissue- and species-specific. In addition, integrated biomarker response (IBR) showed a combined response of enzymatic and non-enzymatic oxidative stress parameters depending on the tissue or species studied, indicating different metabolic activities and behaviors of an autochthonous versus an introduced bivalve species.

Comparative transcriptome analysis revealed host defense responses in endosymbiotic gill of Bathymodiolus mussels inhabiting cold seeps and hydrothermal vents

Comparative transcriptome analysis revealed host defense responses in endosymbiotic gill of Bathymodiolus mussels inhabiting cold seeps and hydrothermal vents

Reproductive Strategies and Embryonic Development of Autumn-Spawning Bitterling (Acheilognathus rhombeus) within the Mussel Host.

We investigated the reproductive strategies and embryonic development of Acheilognathus rhombeus (a bitterling species that spawns in autumn) within its freshwater mussel host in the Bongseo Stream, South Korea. By focusing on survival mechanisms during critical stages of embryonic development, the selective use of mussel gill demibranchs by the bitterlings and associated adaptive traits were observed over 1 year. A significant diapause phase occurs at developmental stage D, which lasts for approximately 7 months, allowing embryos to survive winter. Development resumes when the temperature exceeds 10 °C. Minute tubercles on the embryos (crucial for anchoring within the host gill demibranchs and preventing premature ejection) exhibit the largest height during diapause, and the height decreases when developmental stage E is reached, when growth resumes. Acheilognathus rhombeus embryos were observed in 30.5% of the mussels, mostly within the inner gills, thereby maximizing spatial use and oxygen access to enhance survival. These results highlight the intricate relationship between A. rhombeus and its mussel hosts, demonstrating the evolutionary adaptations that enhance reproductive success and survival. This study provides valuable insights into the ecological dynamics and conservation requirements of such symbiotic relationships.

The reproductive costs of bitterling fish and zebra mussel parasitism to a unionid mussel

Unionid mussels are one of the most vulnerable groups of animals, with multitude of factors responsible for the decline of their populations in the freshwater ecosystems. Many of those causes stem from interspecific relationships. Direct data on the reproductive costs incurred to unionids through parasitism by fish and other mussel species are incomplete. We used field experiments to test whether reproduction of the swollen river mussel (Unio tumidus) is affected by parasitism by the European bitterling fish (Rhodeus amarus) and zebra mussel (Dreissena polymorpha). We documented that bitterling brood can develop in the gills of female, male and hermaphrodite mussels. Our research showed that male mussels carried more fish offspring than female U. tumidus and bitterling embryo survival appeared higher in the outer gill demibranchs. Parasitised female U. tumidus suffered reduced fecundity in terms of a reduced number of mature ovulated oocytes and increased oocyte degeneration. Our data lend clear support to the hypothesis that bitterling fish embryos parasitizing mussel gills and epibiont zebra mussel fouling unionid shells represent a major cost to the reproductive success of their host mussels. Extended gonadal activity (multiple spawning) of U. tumidus spreads their spawning time over a longer period, which may mitigate the costs of parasitism.

Energy reserves and gut microbiota of marine mussels under combined exposure to pathogens and predation risk

In recent years, environmental issues such as eutrophication have led to proliferation of pathogenic bacteria and deterioration of aquaculture environmental water quality. However, the impact of pathogenic bacteria on the mussel-predator food chain in the marine environment is not yet fully understood. In this experiment, mussels were cultured with crabs (Charybdis japonica) for 14 days. Then crabs were retained or removed and pathogenic bacteria (Vibrio alginolyticus) was added and mussels were cultured for 3 days. C. japonica and V. alginolyticus were used as two stressors to explore their effects on energy content and gut microbiota of Mytilus coruscus. The metabolic parameters (protein, carbohydrate and lipid contents) of the gills and digestive glands, and the gut microbiota of the mussels were measured. The study found that presence of predation risk significantly increased the content of protein but decreased carbohydrate and lipid contents in the gills at 14th day. After 17 days of exposure, the presence of predation risk significantly reduced protein content but significantly increased lipid content in gills of mussel. The protein content in digestive gland was significantly increased, but the lipid content was significantly decreased. The presence of predation risk and pathogens had interactive effect on the protein content in the digestive glands of mussels. The presence of V. alginolyticus and predation risk affected the community structure, species richness, and diversity of gut microbiota. The most abundant phyla noted were Bacteroidota, Proteobacteria, and Firmicutes, and the most abundant genus noted were Prevotella, Klebsiella and Dialister. Alpha diversity analysis and beta diversity analysis revealed that the presence of V. alginolyticus and predation risk altered the microbial community structure. V. alginolyticus plays a positive role in the harmful microbiota of gut microbiota. The proportion of Bacteroidota and Proteobacteria in intestinal bacteria increased significantly. Linear discriminant analysis effect size (LEfSe) showed that the biomarkers in each group are significantly different at the genus level. Significant correlations between metabolic parameters (protein content and carbohydrate content) and the level of KEGG function prediction (amino acid metabolism and carbohydrate metabolism) were observed in gills and digestive glands. This study highlights the priority of energy responses in mussels under dual predator-pathogen pressure, and also highlights the complexity of predator-pathogen interactions. A better understanding of their impact on the gut microbiota and health of marine mussels will help support the sustainability of mussel aquaculture production.

Analysis of Amphora ostrearia var. minor Grunow, a blooming marine diatom (Bacillariophyceae) in French Atlantic coastal waters and its transfer to the genus Tetramphora

A marine amphoroid diatom, responsible for blooms in 2011 and 2019 in French Atlantic waters was studied using morphology and molecular phylogeny. The highest cell concentrations were reached in the region of Marennes Oléron on 19 August 2011 and were related to a greyish discolouration of gills of oysters and mussels. It was identified as Amphora ostrearia var. minor Grunow, based on morphological characteristics. To ensure the correct identification, type material was re-investigated, using the type slide n° 142 (Cleve & Möller 1878. Diatoms. Part II. Esatas Edquists Boktryckeri, Upsala: 49–108) that enabled us to establish the identity of the taxon. Plastid morphology corresponded to Mereschkowsky’s type 8 amphoroid chloroplast, a feature typical for the genus Tetramphora Mereschkowsky. Results of a phylogenetic analysis inferred from a concatenated alignment including the nuclear marker SSU and the chloroplast rbcL indicated that A. ostrearia var. minor was positioned inside the monophyletic clade Tetramphora (Mereschkowsky) Stepanek & Kociolek. This, together with the morphological data, necessitated a transfer of A. ostrearia var. minor Grunow to the genus Tetramphora (Mereschkowsky) Stepanek & Kociolek. In addition, the genetic distances (p-values) from SSU–rbcL sequences were estimated among the species of Tetramphora including T. ostrearia, the generitype. These revealed that the divergence between Tetramphora ostrearia (Brébisson) Mereschkowsky and its variety minor was higher than the mean value within species of the genus. As a consequence, the studied taxon is raised to the species level as Tetramphora minor (Grunow) Nézan, comb. et stat. nov. Toxin analysis of four strains of T. minor revealed the absence of domoic acid production. Pigment analysis of T. ostrearia revealed a high fucoxanthin production, similar to other amphoroid species.

Responses of zebra and quagga mussels to copper and tribytiltin exposure: Bioconcentration, metabolic and cardiac biomarkers

One of the top ecological priorities is to find sensitive indicators for pollution monitoring. This study focuses on the bioconcentration and responses (condition index, survival, oxygen consumption, heart rates, and oxidative stress and neurotoxic effect biomarkers) of mussels from the Volga River basin, Dreissena polymorpha and Dreissena bugensis, to long-term exposure to toxic chemicals such as tributyltin (TBT, 25 and 100 ng/L) and copper (Cu, 100 and 1000 μg/L). We found that TBT was present in the tissues of zebra and quagga mussels in comparable amounts, whereas the bioconcentration factor of Cu varied depending on its concentration in water. Differences in responses between the two species were revealed. When exposed to high Cu concentrations or a Cu-TBT mixture, quagga mussels had a lower survival rate and a longer heart rate recovery time than zebra mussels. TBT treatment caused neurotoxicity (decreased acetylcholinesterase activity) and oxidative stress (increased levels of thiobarbituric acid reactive substances) in both species. TBT and Cu levels in mussel tissues correlated positively with the condition index, but correlated with the level of acetylcholinesterase in the mussel gills. The principal component analysis revealed three main components: the first consists of linear combinations of 14 variables reflecting TBT water pollution, TBT and Cu levels in mussel tissues, and biochemical indicators; the second includes Cu water concentration, cardiac tolerance, and mussel size; and the third combines weight, metabolic rate, and heart rates. Quagga mussels are less tolerable to contaminants than zebra mussels, so they may be used as a sensitive indicator.

Acetylcholinesterase and dopamine inhibition suppress the filtration rate, burrowing behaviours, and immunological responses induced by bisphenol A in the hemocytes and gills of date mussels, Lithophaga lithophaga

Bisphenol A (BPA), a common industrial chemical with estrogenic activity, has recently gained attention due to its well-documented negative effects on humans and other organisms in the environment. The potential immunotoxicity and neurotoxicity of BPA remain poorly understood in marine invertebrate species. Therefore, the impacts of exposure to BPA on a series of behaviours, immune responses, oxidative stress, neural biomarkers, histology, and the ultrastructure of gills were investigated in the date mussel, Lithophaga lithophaga. After 28 days of exposure to 0.25, 1, 2, and 5 µg/L BPA, hemolymphs from controls and exposed date mussels were collected, and the effects of BPA on immunological parameters were evaluated. Moreover, oxidative stress and neurochemical levels were measured in the gills of L. lithophaga. BPA reduced filtration rates and burrowing behaviour, whereas a 2 µg/L BPA resulted in an insignificant increase after 24 h. The exposure of date mussels to BPA significantly increased total hemocyte counts, a significant reduction in the diameter and phagocytosis of hemocytes, as well as gill lysozyme level. BPA increased lipid peroxidation levels and SOD activity in gills exposed to 2 and 5 µg/L BPA, but decreased GSH levels and SOD activity in 0.25 and 1 µg/L BPA-treated date mussels. Dose-dependent dynamics were observed in the inhibition of acetylcholinesterase activity and dopamine levels. Histological and scanning electron microscope examination revealed cilia erosion, necrosis, inflammation, and hyperplasia formation in the gills. Overall, our findings suggest a relationship between BPA exposure and changes in the measured immune parameters, oxidative stress, and neurochemical disturbances, which may be factored into the mechanisms underlying BPA toxicity in marine molluscs, providing a scientific foundation for marine BPA risk assessment and indicating immunosuppression in BPA-exposed date mussels.

Toxicology assessment of deep-sea mining impacts on Gigantidas platifrons: A comparative in situ and laboratory metal exposure study

Deep-sea toxicology is essential for deep-sea environmental impact assessment. Yet most toxicology experiments are conducted solely in laboratory settings, overlooking the complexities of the deep-sea environment. Here we carried out metal exposure experiments in both the laboratory and in situ, to compare and evaluate the response patterns of Gigantidas platifrons to metal exposure (copper [Cu] or cadmium [Cd] at 100 μg/L for 48 h). Metal concentrations, traditional biochemical parameters, and fatty acid composition were assessed in deep-sea mussel gills. The results revealed significant metal accumulation in deep-sea mussel gills in both laboratory and in situ experiments. Metal exposure could induce oxidative stress, neurotoxicity, an immune response, altered energy metabolism, and changes to fatty acid composition in mussel gills. Interestingly, the metal accumulating capability, biochemical response patterns, and fatty acid composition each varied under differing experimental systems. In the laboratory setting, Cd-exposed mussels exhibited a higher value for integrated biomarker response (IBR) while in situ the Cu-exposed mussels instead displayed a higher IBR value. This study emphasizes the importance of performing deep-sea toxicology experiments in situ and contributes valuable data to a standardized workflow for deep-sea toxicology assessment.

Insights into phage-bacteria interaction in cold seep Gigantidas platifrons through metagenomics and transcriptome analyses

Viruses are crucial for regulating deep-sea microbial communities and biogeochemical cycles. However, their roles are still less characterized in deep-sea holobionts. Bathymodioline mussels are endemic species inhabiting cold seeps and harboring endosymbionts in gill epithelial cells for nutrition. This study unveiled a diverse array of viruses in the gill tissues of Gigantidas platifrons mussels and analyzed the viral metagenome and transcriptome from the gill tissues of Gigantidas platifrons mussels collected from a cold seep in the South Sea. The mussel gills contained various viruses including Baculoviridae, Rountreeviridae, Myoviridae and Siphovirdae, but the active viromes were Myoviridae, Siphoviridae, and Podoviridae belonging to the order Caudovirales. The overall viral community structure showed significant variation among environments with different methane concentrations. Transcriptome analysis indicated high expression of viral structural genes, integrase, and restriction endonuclease genes in a high methane concentration environment, suggesting frequent virus infection and replication. Furthermore, two viruses (GP-phage-contig14 and GP-phage-contig72) interacted with Gigantidas platifrons methanotrophic gill symbionts (bathymodiolin mussels host intracellular methanotrophic Gammaproteobacteria in their gills), showing high expression levels, and have huge different expression in different methane concentrations. Additionally, single-stranded DNA viruses may play a potential auxiliary role in the virus–host interaction using indirect bioinformatics methods. Moreover, the Cro and DNA methylase genes had phylogenetic similarity between the virus and Gigantidas platifrons methanotrophic gill symbionts. This study also explored a variety of viruses in the gill tissues of Gigantidas platifrons and revealed that bacteria interacted with the viruses during the symbiosis with Gigantidas platifrons. This study provides fundamental insights into the interplay of microorganisms within Gigantidas platifrons mussels in deep sea.

Genotoxicand biomarker response of mussel gill cells, haemocyte and A549 human lung cells to radiation from X-rays

A comparative study on the genotoxic effect of radiation from X-rays of different levels on Mussel gill cells, haemocyte and human lung cell A549 using the comet assay was carried out. The results revealed an increasing DNA damage with increasing doses of X-ray, single strand breaks and alkali labile sites in the haemocyte of the control had a mean value of 5.92 ± 1.27 while the highest treatment group 15Gray (Gy) had a mean value of 78.65 ± 4.36,withsignificant differences (P <0.05), A mean value of 21.10 ± 13.10 was recorded for Mussel gill cells in the control group while the highest treatment group 15 Gray (Gy) had 85.55 ± 1.43. Statistical analysis revealed significant differences (P <0.05) for univariate tests of the control group and the treatment groups. The A549 human lung cells had a minimal damage in the control group 2.30 ± 0.30 while the highest exposed group had a mean value from the median % tail intensity of 43.10 ± 2.81. Pair wise comparison of gills cells, haemocytes from mussels and A549 human lung cells exposed to 15 Gy of X-rays shows that the highest damaging effect was observed on the gill cells followed by the haemocytes with the least damaging effect on the A549 human lung cells exposed to the same dose rate of X-rays with significant differences (P <0.05) between them.This study revealed that A549 human lung cells were more resistant to DNA damage induced by the X-rays when compared to mussel gill and haemocyte cells and also indicates that mussel gill cells and haemocytes can serve as biomarkers of genotoxicity of radiation exposure using the comet assay.

Host size matters for reproduction: Evolution of spawning preference and female reproductive phenotypes in mussel-symbiotic freshwater bitterling fishes.

Bitterling fishes evolve an idiosyncratic symbiosis with freshwater mussels, in which they are obligated to spawn in the gills of mussels for reproduction. In recent years, freshwater mussel populations have been drastically diminishing, due to accelerating anthropogenic impacts, which can be large threats to the risk of bitterling's extinction cascade (i.e. 'coextinction'). The host mussel size may be an important factor driving the adaptation and evolution of bitterling's reproductive phenotypes. Here we examined the host size preference and morphological adaptation of female bitterling to the host size from 17 localities at the Han River in Korea. Using our developed molecular-based species identification for bitterling's eggs/larvae inside the mussels, we further determined the spawning patterns of seven bitterling species. Mean length of spawned mussels (N = 453) was significantly larger than that of unspawned mussels (N = 1814), suggesting that bitterling prefers to use larger hosts as a spawning ground. Spawning probability was clearly greater as mussel size increases. Results of our reciprocal transplant experiments do provide some evidence supporting the 'bitterling's larger host preference' hypothesis. Interspecific competition appeared to be intense as two fish species often spawned eggs in the same mussel individuals simultaneously. Longer ovipositor and more elongated egg may evolve in females of Tanakia signifer in response to larger host environments. The observed bitterling's spawning preference for large-sized mussels may evolve perhaps because of the fitness advantage in relation to the offspring survival. Our findings further inform on the development of effective conservation and management strategy for the endangered bitterling fishes.

Ocean acidification impact on the uptake of trace elements by mussels and their biochemical effects

This study delves into the intricate interplay between ocean acidification (OA), metal bioaccumulation, and cellular responses using mussels (Mytilus galloprovincialis) as bioindicators. For this purpose, environmentally realistic concentrations of isotopically labelled metals (Cd, Cu, Ag, Ce) were added to investigate whether the OA increase would modify metal bioaccumulation and induce adverse effects at the cellular level. The study reveals that while certain elements like Cd and Ag might remain unaffected by OA, the bioavailability of Cu and Ce could potentially escalate, leading to amplified accumulation in marine organisms. The present findings highlight a significant rise in Ce concentrations within different mussel organs under elevated pCO2 conditions, accompanied by an increased isotopic fractionation of Ce (140/142Ce), suggesting a heightened potential for metal accumulation under OA. The results suggested that OA influenced metal accumulation in the gills of mussels. Conversely, metal accumulation in the digestive gland was unaffected by OA. The exposure to both trace metals and OA affects the biochemical responses of M. galloprovincialis, leading to increased metabolic capacity, changes in energy reserves, and alterations in oxidative stress markers, but the specific effects on other biomarkers (e.g., lipid peroxidation, some enzymatic responses or acetylcholinesterase activity) were not uniform, suggesting complex interactions between the stressors and the biochemical pathways in the mussels.

Parasitic fish embryos do a “front-flip” on the yolk to resist expulsion from the host

Embryonic development is often considered shielded from the effects of natural selection, being selected primarily for reliable development. However, embryos sometimes represent virulent parasites, triggering a coevolutionary "arms race" with their host. We have examined embryonic adaptations to a parasitic lifestyle in the bitterling fish. Bitterlings are brood parasites that lay their eggs in the gill chamber of host mussels. Bitterling eggs and embryos have adaptations to resist being flushed out by the mussel. These include a pair of projections from the yolk sac that act as an anchor. Furthermore, bitterling eggs all adopt a head-down position in the mussel gills which further increases their chances of survival. To examine these adaptations in detail, we have studied development in the rosy bitterling (Rhodeus ocellatus) using molecular markers, X-ray tomography, and time-lapse imaging. We describe a suite of developmental adaptations to brood parasitism in this species. We show that the mechanism underlying these adaptions is a modified pattern of blastokinesis-a process unique, among fish, to bitterlings. Tissue movements during blastokinesis cause the embryo to do an extraordinary "front-flip" on the yolk. We suggest that this movement determines the spatial orientation of the other developmental adaptations to parasitism, ensuring that they are optimally positioned to help resist the ejection of the embryo from the mussel. Our study supports the notion that natural selection can drive the evolution of a suite of adaptations, both embryonic and extra-embryonic, via modifications in early development.

Impact of face masks weathering on the mussels Mytilus galloprovincialis

The COVID-19 pandemic has triggered an unprecedented need for single-use face masks, leading to an alarming increase in plastic waste globally. Consequently, the improper disposal of face masks has added to the existing burden of plastic pollution in the oceans. However, the complete environmental and marine ecotoxicological impact remains unclear. This study aims to investigate the ecotoxicological impact caused by the weathering of disposable face masks (DFMs) in the marine environment on mussels Mytilus galloprovincialis (M. galloprovincialis ) by assessing biochemical, cytotoxic, and genotoxic effects. The mask leachate was analysed for the presence of nano and microplastics. Furthermore, the leachate was used in in vivo and in vitro toxicity bioassays to assess its impacts on M. galloprovincialis . The in vivo exposure of M. galloprovincialis to face mask leachate for 14 days induced a significant increase in catalase (CAT) activity in mussel gills, although not enough to prevent oxidative damage to cell membranes. DNA damage was also registered in mussel haemocytes after in vivo exposure to mask leachate. The in vitro Neutral Red (NR) cytotoxicity assay indicated that leachate concentrations ≤ 0.5 g/L-1 pose a significant risk to the health of mussel haemocytes, which seems a reliable tool for the cytotoxicity impact assessment of face masks in the marine environment. Therefore, the leachate obtained from face masks in seawater causes oxidative stress, oxidative damage, cytotoxicity, and genotoxicity in M. galloprovincialis , indicating that the plastic burden generated by DFMs in the ocean and its subsequent weathering represents a ubiquitous and invisible threat to the marine biota.

Modulation of thermal stress response by prostaglandins in gills of the blue mussel Mytilus edulis

Mytilus edulis has experienced massive mortality events associated with thermal stress that have been documented in wild as well as cultivated organisms, since this species is traditionally farmed in intertidal areas that are exposed to temperature variations. The stress responses linked to changes in temperature are modulated by prostaglandins (PG), which induce apoptosis, synthesis of heat shock proteins and antioxidant enzymes, among others. PG are mainly synthesized from arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA), but PG-like compounds can be produced through non-enzymatic oxidation of fatty acids in response to increased reactive oxygen species (ROS). In this study, we applied acute (a single temperature increase of 13 °C, from 12 °C to 25 °C) or chronic (cyclical temperature increases and decreases of 13 °C for eight days) thermal stress to mussels fed a microalga mix of Nannochloropsis oculata and Pavlova lutheri. The response was quantified by measuring PG produced enzymatically by cyclooxygenase (COX) from ARA, EPA, and DHA (PGE2, PGE3, 13-HDHA, respectively), non-enzymatic PG (13,14-dihydro-15-keto-PGE2 PGF3α and 8-isoprostane). PGE2, PGE3, and 13-HDHA levels were higher in the gills of mussels exposed to acute stress compared to chronic stress while 3,14-dihydro-15-keto-PGE2 showed no significant differences as a result of chronic or acute stress. PGF3α had lower levels in mussels exposed to chronic stress compared to those under acute stress. In contrast, 8-isoprostane was significantly higher in mussels exposed to chronic stress compared to acute stress. Mussels exposed to acute stress showed an increase in hemocyte infiltration in the gills, while those exposed to chronic stress showed a decrease in gill filament width compared to mussels submitted to acute stress; both effects are associated with PG modulation. These findings suggest that acute and chronic stressors have distinct effects on lipid composition, prostaglandin levels, and gill filament morphology, highlighting potential adaptations or responses to different stress conditions. Understanding the mechanisms by which PG and PG-like compounds modulate the thermal stress response in mussels will contribute to the understanding of climate change impacts on bivalves and the possible adaptation processes involved.

Responses of Mytilus galloprovincialis in a Multi-Stressor Scenario: Effects of an Invasive Seaweed Exudate and Microplastic Pollution under Ocean Warming.

Microplastic pollution, global warming, and invasive species are known threats to marine biota, but the impact of their simultaneous exposure is still not well understood. This study investigated whether the toxic effects posed by the invasive red seaweed Asparagopsis armata exudate (2%) to the mussel Mytilus galloprovincialis are amplified by a 96 h exposure to increased temperature (24 °C) and polyethylene microplastics (PE-MPs, 1 mg/L). Biochemical (neurotoxicity, energy metabolism, oxidative stress, and damage) and physiological (byssal thread production) responses were evaluated. The number of produced byssus greatly decreased under concomitant exposure to all stressors. The antioxidant defences were depleted in the gills of mussels exposed to temperature rises and PE-MPs, regardless of exudate exposure, preventing oxidative damage. Moreover, the heat shock protein content tended to decrease in all treatments relative to the control. The increased total glutathione in the mussels' digestive gland exposed to 24 °C, exudate, and PE-MPs avoided oxidative damage. Neurotoxicity was observed in the same treatment. In contrast, the energy metabolism remained unaltered. In conclusion, depending on the endpoint, simultaneous exposure to A. armata exudate, PE-MPs, and warming does not necessarily mean an amplification of their single effects. Studies focusing on the impact of multiple stressors are imperative to better understand the underlying mechanisms of this chronic exposure.

An Exploration of Novel Bioactives from the Venomous Marine Annelid Glycera alba.

The immense biodiversity of marine invertebrates makes them high-value targets for the prospecting of novel bioactives. The present study investigated proteinaceous toxins secreted by the skin and proboscis of Glycera alba (Annelida: Polychaeta), whose congenerics G. tridactyla and G. dibranchiata are known to be venomous. Proteomics and bioinformatics enabled the detection of bioactive proteins that hold potential for biotechnological applications, including toxins like glycerotoxins (GLTx), which can interfere with neuromuscular calcium channels and therefore have value for the development of painkillers, for instance. We also identified proteins involved in the biosynthesis of toxins. Other proteins of interest include venom and toxin-related bioactives like cysteine-rich venom proteins, many of which are known to interfere with the nervous system. Ex vivo toxicity assays with mussel gills exposed to fractionated protein extracts from the skin and proboscis revealed that fractions potentially containing higher-molecular-mass venom proteins can exert negative effects on invertebrate prey. Histopathology, DNA damage and caspase-3 activity suggest significant cytotoxic effects that can be coadjuvated by permeabilizing enzymes such as venom metalloproteinases M12B. Altogether, these encouraging findings show that venomous annelids are important sources of novel bioactives, albeit illustrating the challenges of surveying organisms whose genomes and metabolisms are poorly understood.

Short-term exposure to polystyrene microplastics hampers the cellular function of gills in the Mediterranean mussel Mytilus galloprovincialis

Plastic is undoubtedly the most useful and versatile polymeric material that man has developed in the last two centuries Despite the societal benefits, plastic is now a serious global issue because it is persistent and may bioaccumulate into aquatic biota as microplastics (MPs). This study was designed to evaluate the daily uptake and cellular effects due to a short-term (up to 72 h) exposure to 3 μm red polystyrene MPs (50 beads/mL) in the gills of the Mediterranean mussel Mytilus galloprovincialis, chosen as model species for its ecological and commercial relevance. After measuring the daily uptake of MPs and detecting their presence within the branchial epithelium at all the exposure time-points (T24, T48, T72), some cleaning mechanisms were observed by neutral and acid mucous secretions at mussel gills. The protonic Nuclear Magnetic Resonance (1H NMR)-based metabolomics, combined with chemometrics, allowed to comprehensively explore the time-dependent metabolic disorders triggered by MPs in mussel gills over the short-term trial. Specifically, the clear clustering between MP-treated mussel gills and those from control, together with the grouping for experimental time-points as depicted by the Principal Component Analysis (PCA), were due to changes in the amino acids and energy metabolism, disturbances in the osmoregulatory processes, as well as in the cholinergic neurotransmission. Moreover, as evidenced by enzymatic assays, even the oxidative defense systems and lipid metabolism were hampered by MP exposure. Overall, these findings provides the first insights into the early time-dependent mechanisms of toxicity of polystyrene MPs in marine mussels, and underline the potential environment and human health risk posed by MPs contamination.

Copper-Induced Ionoregulatory Disturbance, Histopathology, and Transcriptome Responses in Freshwater Mussel (Anodonta woodiana) Gills

Copper (Cu) contamination has become a severe problem in freshwater environments worldwide. The freshwater mussel Anodonta woodiana is used as a unique bioindicator to monitor Cu contamination in freshwater environments. However, Cu toxicity and response mechanisms in A. woodiana are still largely unknown. A sublethal acute exposure experiment (2.0 mg/L Cu exposure for 72 h) was conducted to investigate the effects of Cu bioaccumulation on ionoregulatory homeostasis, histological features, and transcriptome responses using A. woodiana gills as indicator tissue. The gill bioaccumulation capacity was up to 474. Cu bioaccumulation decreased Na+ and Mg2+ concentrations (p < 0.05) by 82% and 17%, respectively, and induced cilia loss, epithelial desquamation, and filament atrophy of the gills. Transcriptome analysis identified 3160 differentially expressed genes (DEGs), including 1870 upregulated and 1290 downregulated genes. GO enrichment analysis showed that cellular processes, metabolic processes, biological regulation, and responses to stimuli contained the most DEGs in the biological processes. KEGG pathway analysis showed that apoptosis, arginine and proline metabolism, the toll-like receptor signaling pathway, apoptosis-multiple species, histidine metabolism, beta-alanine metabolism, cytokine–cytokine receptor interaction, and the p53 signaling pathway were significantly enriched. These findings provide comprehensive evidence for exploring Cu toxicity and response mechanisms in freshwater mussels.