Fate and impact at molecular level of diatrizoic acid and iohexol contrast agents in Dreissena polymorpha mollusks.

Iodinated contrast media (ICMs) used in X-ray imaging for medical diagnostics are released into wastewater and then encountered in river water at concentrations ranging from several dozen to hundreds of µg/L, and even thousands of µg/L in hospital effluents. ICMs are considered as emerging pollutants as their occurrence and impact on ecosystems and the environment are poorly documented. Even if they are considered inert for humans, aquatic organisms are continuously exposed to ICMs, and their potential deleterious effects are therefore questioned as we have recently demonstrated that they enter into organisms such as the zebra mussels. To answer this question, Dreissena polymorpha were exposed to two ICMs of different osmolality, diatrizoic acid and iohexol, at an environmental concentration (100 µg/L) for 21 days before a depuration phase of 4 days. The occurrence, fate, and impact of both ICMs in these organisms were studied using a metallomic approach. Thus, iodine as well as endogenous copper and zinc were quantified and analyzed in cytosolic extracts of digestive glands, gills, and gonads of mussels by size exclusion chromatography coupled to ICP MS. This work shows that a subcellular fractionation is necessary to distinguish variations in total element content. The cytosolic iodoprotein chromatographic pattern was consistent for the three organs and confirmed the presence of ICMs in cytosols. Additionally, this exploratory work tends to show a weak biological effect of ICMs with a substantial variation of the profile of Cu-binding proteins mostly in the gill cytosol and to a lesser extent, in the digestive gland cytosol.

Density of Limnoperna fortunei larval stages in a cascade of subtropical reservoirs: spatiotemporal variation and environmental influences.

The golden mussel (Limnoperna fortunei) is an invasive bivalve that has established itself in several South American river systems, impacting ecosystem functioning. Reservoir cascades provide their larvae with the means of rapid dispersal, but the relationship between environmental variables and larval stage structure remains unclear. In this study, the density of three L. fortunei larval stages and quantitative detection using DNA are analyzed in a cascade of five reservoirs in the upper Uruguay River Basin and associated with spatiotemporal variation in environmental parameters. The analysis of L. fortunei eDNA presence and absence in freshwater systems appears to be a valuable mapping tool; however, no significant link was found between the eDNA magnitude and the overall larval density. The increase in larval density was related to the fluctuation of environmental parameters over a year, with the highest average larval densities observed in the CN and ITA reservoirs, though no significant difference was observed between the five reservoirs, where D-shaped larvae predominated. During winter, larval density decreased significantly, however, other variables also contribute to species activity and development in the upper Uruguay River Basin reservoirs and may be considered limiting factors. The relationships between environmental parameters were evaluated using a multivariate model. The interaction between reservoir area and precipitation, water temperature, electrical conductivity, and dissolved oxygen had a significant effect on larval density but showed specific influences on each larval stage. Any increase in density was regulated by dissolved oxygen and electrical conductivity content at all larval stages. Furthermore, total phosphorus affected the density of F1 and F3 larvae. The interaction between reservoir area and precipitation, nitrate content, phosphate concentrations, and water temperature had the most influence on the density of F2 and F3 larval stages; the F1 stage was mainly affected by calcium concentrations. The isolated effect of precipitation also contributed to the density of F2 and F3 larvae. Our findings shed light on the interaction between different phases of golden mussel larvae and the main nutrients found in reservoirs, which may be a determining factor in the rise in density of the non-native species in these systems.

Preparation strategies of mussel-inspired chitosan-based biomaterials for hemostasis.

Chitosan (CS) has been extensively studied in wound care for its intrinsic hemostatic and antibacterial properties. However, CS has limiting hemostasis applications on account of its drawbacks such as poor adhesion in humid environments and water solubility at neutral pH. CS-based biomaterials, inspired by mussel-adhesive proteins, serve as a suggested platform by biomedical science. The reports show that the mussel-inspired CS-based hemostatic structure has negligible toxicity and excellent adhesiveness. Biomedicine has witnessed significant progress in the development of these hemostatic materials. This review summarizes the methods for the modification of CS by mussel-inspired chemistry. Moreover, the general method for preparation of mussel-inspired CS-based biomaterials is briefly discussed in this review. This work is expected to give a better understanding of opportunities and challenges of the mussel-inspired strategy for the functionalization of CS-based biomaterials in hemostasis and wound healing. This review is hoped to provide an important perspective on the preparation of mussel-inspired CS-based hemostatic materials.

Design of self-healing nanocomposite hydrogels and the application to the detection of human exercise and ascorbic acid in sweat.

Hydrogel sensors have broad application prospects in human motion monitoring and sweat composition detection. However, hydrogel-based sensors are faced with challenges such as low accuracy and poor mechanical properties of analytes detection. Based on mussel-inspired chemistry, we synthesized mesoporous silica@polydopamine-Au (MPS@PDA-Au) nanomaterials and designed a self-healing nanocomposite hydrogel to monitor human movement and ascorbic acid detection in sweat. Mesoporous silica (MPS) possess orderly mesoporous structure. Dopamine (DA) polymerized on the surface of MPS to generate polydopamine (PDA), forming the composite material MPS@PDA-Au. This composite was then embedded into polyvinyl alcohol (PVA) hydrogels through a simple freeze-thaw cycle process. The hydrogels have achieved excellent deformable ability (508.6%), self-healing property (90.5%) and mechanical strength (2.9 MPa). The PVA/MPS@PDA-Au hydrogel sensors had the characteristics of fast response time (123.2 ms), wide strain sensing range (0-500%), excellent fatigue resistance and stability in human detection. The detection range of ascorbic acid (AA) in sweat was wide (8.0 µmol/L-100.0 µmol/L) and the detection limit was low (3.3 µmol/L). Therefore, these hydrogel sensors have outstanding application prospects in human motion monitoring and sweat composition detection.

Pathological and oxidative stress responses of Mytilus galloprovincialis to Vibrio mediterranei infection: An in vivo challenge.

Since the identification of Vibrio mediterranei as a causative agent in mass mortalities of pen shells across the Mediterranean, elucidating its pathogenicity, virulence, and interactions with other bivalves has gained importance. While the cellular and immune responses of bivalves to various Vibrio species have been extensively studied, the infectious characteristics of this Vibrio species, particularly in the context of pen shell outbreaks, remain unclear for other bivalves. Therefore, to evaluate its pathogenicity, we investigated the histological and oxidative effects on the Mediterranean mussel (Mytilus galloprovincialis), a key species in aquaculture. Two distinct infection setups were established: one involving the inoculation of seawater with the bacterial isolate and another involving direct injection of the bacteria into the mussels. After a 24-h exposure period, histological evaluations were conducted on the mantle, gill, and digestive gland tissues of the mussels. Additionally, measurements of superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), and lipid peroxidation levels were performed in the gill and digestive gland tissues. Oxidative responses were significantly elevated in both infection setups compared to the control group, with the directly injected samples exhibiting the highest oxidative responses (p < 0.05). Histological findings indicated that tissue-specific responses to host-pathogen interactions were consistent under both infection conditions. Notable observations included intense hemocytic infiltration in tissues, epithelial hyperplasia, and vacuolization in the gills, as well as focal necrotic areas in the digestive gland. The findings of this study indicate that V. mediterranei, a relatively novel pathogen, can provoke significant acute immune responses and tissue-level reactions in M. galloprovincialis, a species that is both widely distributed and vital to the food chain. These insights into the potential susceptibility of mussels underscore the need for further comprehensive research and inform the development of effective management strategies.

Combined copper isotope and elemental signatures in bivalves and sediments from the Korean coast: Applicability for monitoring anthropogenic contamination.

This study investigates the applicability of elemental and Cu isotope compositions in sediments and bivalves from the Korean coast to monitor anthropogenic Cu contamination. Sediments with high Cu (>64.4 mg/kg) and/or moderate enrichment levels (EFCu) exhibit homogenous δ65CuAE647 values (-0.12 to +0.16 ‰), suggesting similar anthropogenic Cu fingerprints along the Korean coast. Sediments with Cu concentrations near natural background levels (< 20.6 mg/kg) display large isotopic variability (Δ65Cumax-mim: ~0.8 ‰), encompassing those from sediments under anthropic influences. We hypothesize that Cu isotopic compositions of Korean geology are heterogeneous, therefore, natural end-members of source mixing models should be established locally at small scales. Cu concentrations in Oysters correlate with sediments, and their isotopic compositions are more suitable for monitoring Cu contamination, while mussel's regulatory mechanisms seem to affect source records. The current Cu isotope data will help to detect shifts attributable to anthropic contamination in future biomonitoring.

Freshwater Mussel Viromes Increase Rapidly in Diversity and Abundance When Hosts Are Released from Captivity into the Wild.

Freshwater mussels (order: Unionida) are highly imperiled globally and are increasingly the focus of captive propagation efforts to protect and restore wild populations. The Upper Tennessee River Basin (UTRB) in Virginia is a freshwater biodiversity hotspot hosting at least 45 of North America's ~300 species of freshwater mussels, including 21 threatened and endangered species listed under the U.S. Endangered Species Act. Recent studies have documented that viruses and other microbes have contributed to freshwater mussel population declines in the UTRB. We conducted a multi-year longitudinal study of captive-reared hatchery mussels released to restoration sites throughout the UTRB to evaluate their viromes and compare them to captive hatchery environments. We documented 681 viruses from 27 families. The hatchery mussels had significantly less viruses than those deployed to wild sites, with only 20 viruses unique to the hatchery mussels. After the hatchery mussels were released into the wild, their number of viruses initially spiked and then increased steadily over time, with 451 viruses in total unique to the mussels in the wild. We found Clinch densovirus 1 (CDNV-1), a virus previously associated with mass mortality events in the Clinch River, in all samples, but the wild site mussels consistently had significantly higher CDNV-1 levels than those held in the hatchery. Our data document substantial differences between the viruses in the mussels in the hatchery and wild environments and rapid virome shifts after the mussels are released to the wild sites. These findings indicate that mussel release programs might benefit from acclimatization periods or other measures to mitigate the potential negative effects of rapid exposure to infectious agents found in natural environments.

In vivo bioaccumulation and responses of hemocytes of mussels Perna viridis to microplastics and nanoplastics exposure.

Growing micro- and nano-plastic (MNPs) pollution in the environment poses a threat to marine animals. Due to their excellent filtration capacity, bivalves can easily ingest MNPs, which could be translocated to open circulation system with potential risks. In the present study, the accumulation and elimination of MNPs (200 nm and 1 µm) in the mussel hemolymph serum and hemocytes were firstly quantified, and the differential sensitiveresponses of two subpopulations of hemocytes were then explored by in vivo exposure under environmentally relevant concentration of MNPs (200 µg/L). We demonstrated that MNPs were readily translocated into hemolymph serum, but were immediately followed by efficient internalization by hemocytes. Remarkably, concentrations of MNPs in hemolymph were only 0.63 and 0.39 times lower than the ambient exposure concentration. Granulocytes displayed a much higher potential of accumulating MNPs than the agranulocytes. MPs were more readily internalized by granulocytes, with their estimated maximum bioaccumulation factor (BCF) of 0.29 L/g. Due to the primary function of phagocytic encapsulation of MNPs by granulocytes, lysosome features especially the decline of subsequent lysosome membrane potential could be a potential sensitive biomarker in response to MNPs exposure. Our results provided insights on the bioaccumulation of MNPs at the cellular levels in marine bivalves.

"Synergistic anticoagulant and endothelial regeneration strategy" based on mussel-inspired phospholipid copolymer coating and bioactive zeolitic imidazolate frameworks-90 to maintain the patency of CoCr stent.

Given the risks of poor patient compliance and bleeding associated with current dual antiplatelet therapies, it is urgent to develop the next generation of cardiovascular stents with anticoagulation and rapid endothelialization capabilities. Inspired by the prominent bioactivity and bioavailability of zeolitic imidazolate framework-90 (ZIF-90) in driving endothelial cell (EC) morphogenesis, this research proposes a "synergistic anticoagulant and endothelial regeneration strategy" depending on mussel-inspired phospholipid copolymer (MIPC) and ZIF-90. Depending on the copolymerization of the catechol with dopamine (Dopa) monomers, Dopa/MIPC coating was immobilized on the surface of CoCr via a one-pot process for resisting the initial thrombosis induced by platelets and fibrinogen. Meanwhile, ZIF-90 was loaded on the coating via coordination effect, aiming to accelerate the proliferation and migration of ECs. Compared with CoCr, the well-designed CoCr-Dopa/MIPC@ZIF-90 not only reduced fibrinogen adhesion by approximately 40 % and platelet adhesion by almost 55 %, but also promoted the proliferation and migration of ECs significantly in vitro. Furthermore, the blood flow velocity of CoCr-Dopa/MIPC@ZIF-90 stent was similar to natural aorta and ECs coverage on it was greatly strengthened after 30 days in a rat aorta vascular stent implantation model. Collectively, CoCr-Dopa/MIPC@ZIF-90 exhibited obvious superiority in reducing the formation of thrombus and promoting endothelial regeneration, which might meet the high requirement for the next generation of vascular stent.

Advances in Mussel Adhesion Proteins and Mussel-Inspired Material Electrospun Nanofibers for Their Application in Wound Repair.

Mussel refers to a marine organism with strong adhesive properties, and it secretes mussel adhesion protein (MAP). The most vital feature of MAP is the abundance of the 3,4-dihydroxyphenylalanine (DOPA) group and lysine, which have antimicrobial, anti-inflammatory, antioxidant, and cell adhesion-promoting properties and can accelerate wound healing. Polydopamine (PDA) is currently the most widely used mussel-inspired material characterized by good adhesion, biocompatibility, and biodegradability. It can mediate various interactions to form functional coatings on cell-material surfaces. Nanofibers based on MAP and mussel-inspired materials have been exerting a vital role in wound repair, while there is no comprehensive review presenting them. This Review introduces the structure of MAPs and their adhesion mechanisms and mussel-inspired materials. Second, it introduces the functionalized modification of MAPs and their inspired materials in electrospun nanofibers and application in wound repair. Finally, the future development direction and coping strategies of MAP and mussel-inspired materials are discussed. Moreover, this Review can offer novel strategies for the application of nanofibers in wound repair and bring about new breakthroughs and innovations in tissue engineering and regenerative medicine.

Antidepressants as new endocrine disruptors? - transcriptomic profiling in gonads of Mytilus trossulus exposed to norfluoxetine.

In this study an impact of norfluoxetine (NFLU) on Mytilus trossulus gonads was investigated focusing on sex-related differences in hormonal changes, gene expression, and transcriptomic profiling. Sex-specific differences in gonadal serotonin levels were found. NFLU stimulates serotonin synthesis and/or transport in female gonads, potentially accelerating oocyte maturation and gamete release. In males, NFLU decreases serotonin level what likely leads to impeding sperm maturation and thus spawning delay. Transcriptomic analyses highlighted the presence of NFLU-induced changes in gene expression related to gametogenesis and neurotransmission. In females, NFLU upregulated genes associated with oocyte development and downregulated those involved in sperm maturation. NFLU-treated males exhibited mixed effects in their genes in relation to spermatogenesis. Additionally, sex-related differences in the expression of the CYP450 genes responsible for detoxification were found. Overall, norfluoxetine acts as an endocrine-disrupting chemical and impacts gonadal serotonin levels and gene expression, potentially disrupting reproductive success of M. trossulus.

Bivalve shell growth from molecular to sclerochronological scale: Environment and intrinsic factors control increment deposition.

Biomineralisation of bivalve shells raises questions at the level of genes to the final calcified product. For the first time, gene expression has been studied in association with growth increment deposition in the mussel Mytilus galloprovincialis. A short-term experiment highlighted that biomineralisation genes exhibit a rhythm of expression consistent with the observed tidal increment formation. Long-term mark-recapture experiments were conducted in three Mediterranean environments and revealed that the mussel shells harbour complex incrementation regimes, consisting of daily, tidal and a mixed periodicity of ∼1.7 growth increment.d-1 formed. The latter is likely related to the local tidal regime, although the mussels were continuously submerged and exposed to a small tidal range. The pattern of growth increments shifted from mixed to daily in Mediterranean lagoon, and to tidal at sea, probably linked to biological clocks. Based on our results and the literature, a hypothetical model for mussel shell increment formation in various habitats is proposed.

Climate-driven shifts in freshwater biodiversity will impact mitigation costs for hydropower.

Climate change is forecasted to drastically alter freshwater fish and mussel species distribution. Hydropower dam reservoirs, which modify downstream thermal regimes, may interact with climate change's impact on species distribution. This distribution shift may feedback, affecting hydropower operation costs through environmental compliance. We investigated how freshwater species distribution will shift due to climate change and hydropower reservoirs in the conterminous United States (CONUS), and how this will affect biodiversity mitigation costs for privately-owned hydropower plants. In general, using environmental niche modeling, we found that climate change increased the range of both freshwater fish and mussel species on average. For fish, this was mainly due to the expanded habitat for warm-water and cool-water fish species despite the diminish in habitat for cold-water species. Compared to climate change, thermal stratification of hydropower reservoirs had a small impact on the future range changes of these species in the tailwaters but showed an interaction with the effect of climate change on species range. Geographically, we projected an increase of species richness in the west and a decrease in the central and east of CONUS for fish, while projecting uniform increase for mussels. With this shift in species distribution, we estimated that the Northwest region will face the largest increase in mitigation cost, while the majority of plants in the Southeast will experience a decrease in cost.

Finding food: how generalist predators use contact-chemosensory information to guide prey preferences.

Understanding the processes that guide carnivores in finding and selecting prey is a fundamental, unresolved challenge in sensory biology. To our knowledge, no published work has yet revealed the complete structural identities of compounds that cue preferences by generalist predators for different prey species. With this research imperative in mind, we determined the chemistry driving consumer preferences for live intact prey using two generalist predatory species (sea stars, Pisaster ochraceus; whelks, Acanthinucella spirata), along with two foundation prey species (mussels, Mytilus californianus; barnacles, Balanus glandula), inhabiting rocky, wave-swept shores. Each prey species is known to secrete either a 29.6 kDa (named 'KEYSTONEin') or a 199.6 kDa (named 'MULTIFUNCin') glycoprotein as a contact-chemical cue. Here, experimental manipulations utilized faux prey consisting of cleaned barnacle or mussel shells infused with KEYSTONEin, MULTIFUNCin or seawater (control) gels. Whelks exhibited a strong penchant for MULTIFUNCin over KEYSTONEin, irrespective of shell type. In contrast, sea stars generally preferred KEYSTONEin over MULTIFUNCin, but this preference shifted depending on the experimental context in which they encountered physical (shell) and chemical (glycoprotein) stimuli. This study ultimately demonstrates clear and contrasting chemical preferences between sea stars and whelks. It highlights the importance of experimental setting in determining chemical preferences. Finally, it shows that prey preferences by these predators hinge only on one or two contact-protein cues, without the need for quality coding via fluid-borne compounds, low-molecular-weight substances or mixture blends.

Mussel-inspired bifunctional coating for long-term stability of oral implants.

Peri-implantitis and osseointegration failure present considerable challenges to the prolonged stability of oral implants. To address these issues, there is an escalating demand for a resilient implant surface coating that seamlessly integrates antimicrobial features to combat bacteria-induced peri­implantitis, and osteogenic properties to promote bone formation. In the present study, a bio-inspired poly(amidoamine) dendrimer (DA-PAMAM-NH2) is synthesized by utilizing a mussel protein (DA) known for its strong adherence to various materials. Conjugating DA with PAMAM-NH2, inherently endowed with antibacterial and osteogenic properties, results in a robust and multifunctional coating. Robust adhesion between DA-PAMAM-NH2 and the titanium alloy surface is identified using confocal laser scanning microscopy (CLSM) and attenuated total reflectance-infrared (ATR-IR) spectroscopy. Following a four-week immersion of the coated titanium alloy surface in simulated body fluid (SBF), the antimicrobial activity and superior osteogenesis of the DA-PAMAM-NH2-coated surface remain stable. In contrast, the bifunctional effects of the PAMAM-NH2-coated surface diminish after the same immersion period. In vivo animal experiments validate the enduring antimicrobial and osteogenic properties of DA-PAMAM-NH2-coated titanium alloy implants, significantly enhancing the long-term stability of the implants. This innovative coating holds promise for addressing the multifaceted challenges associated with peri­implantitis and osseointegration failure in titanium-based implants. STATEMENT OF SIGNIFICANCE: Prolonged stability of oral implants remains a clinically-significant challenge. Peri-implantitis and osseointegration failure are two important contributors to the poor stability of oral implants. The present study developed a mussel-bioinspired poly(amidoamine) dendrimer (DA-PAMAM-NH2) for a resilient implant surface coating that seamlessly integrates antimicrobial features to combat bacteria-induced peri­implantitis, and osteogenic properties to promote bone formation to extend the longevity of oral implants.

Stability and expression patterns of housekeeping genes in Mediterranean mussels (Mytilus galloprovincialis) under field investigations.

The use of marine mussels as biological models encompasses a broad range of research fields, in which the application of RNA analyses disclosed novel biomarkers of environmental stress and investigated biochemical mechanisms of action. Quantitative real-time PCR (qPCR) is the gold standard for these studies, and despite its wide use and available protocols, it may be affected by technical flaws requiring reference gene data normalization. In this study, stability of housekeeping genes commonly employed as reference genes in qPCR analyses with Mytilus galloprovincialis was explored under field conditions. Mussels were collected from farms in the Northwestern Adriatic Sea. The sampling strategy considered latitudinal gradients of environmental parameters (proxied by location), gender, and their interactions with seasonality. Analyses of gene stability were performed using different algorithms. BestKeeper and geNorm agreed that combination of the ribosomal genes 18S ribosomal RNA (18S) and 28S ribosomal RNA (28S) was the best normalization strategy in the conditions tested, which agrees with available evidence. NormFinder provided different normalization strategies, involving combinations of tubulin (TUB)/28S (Gender/Season effect) or TUB/helicase (HEL) (Location/Season effect). Since NormFinder considers data grouping and computes both intra- and inter-group stability variations, it should work better with complex experimental designs and dataset structuring. Under the selected normalization strategies, expressions of the variable housekeeping genes actin (ACT) and elongation factor-1α (EF1) correlated with seasonal and latitudinal changes of abiotic environmental factors and mussel physiological status. Results point to consider ACT and EF1 expressions as molecular biomarkers of mussel general physiological status in field studies.

Interactive impact of residual pyrethroid compounds used in the Chilean salmon farming industry and coastal acidification conditions on the feeding performance of farmed mussels in northern Patagonia.

The use of pyrethroids in aquaculture has been an important component of achieving a thriving salmon farming industry in Chile. While the residual presence of such substances is known to depend on environmental conditions, most ecotoxicological studies to date have not considered environmental context. Here, we conducted oceanographic monitoring combined with experiments aiming to estimate the effects of two pyrethroids on the feeding rates of larvae of farmed mussels, Mytilus chilensis. In additional experiments, mussel spats were exposed to both pyrethroids, but under contrasting temperature/pH so as to mimic winter and summer conditions. Experiments mimicking spring conditions revealed that both pyrethroid substances affected the feeding of mussel larvae as a function of concentration. Conversely, significant impact of pyrethroids on adults were not observed with regard to temperature and pH, but a significant impact of low temperature/low pH condition on ingestion rates was confirmed. Given the current status of increasing ocean acidification, the results of this study are expected to provide useful information with regard to achieving sustainable mussel aquaculture, especially considering both activities occur in similar geographic areas, and the expansion of salmon farming areas is ongoing in Chile.

Heat Hardening Ameliorates Apoptotic and Inflammatory Effects Through Increased Autophagy in Mussels.

The severity, frequency, and duration of extreme events, in the context of global warming, have placed many marine ecosystems at high risk. Therefore, the application of methods that can mediate the impacts of global warming on marine organisms seems to be an emerging necessity in the near term. In this context, enhancing the thermal resilience of marine organisms may be crucial for their sustainability. It has been shown that the repeated time-limited exposure of an organism to an environmental stimulus modifies its response mode, thus enhancing resilience and allowing adaptation of the physiological and developmental phenotype to environmental stress. In the present study, we investigated the "stress memory" effect caused by heat hardening on Mytilus galloprovincialis cellular pathways to identify the underlying biochemical mechanisms that enhance mussel thermal tolerance. Heat hardening resulted in increased ETS activity and ATP production and increased autophagic performance at all elevated temperatures (24 °C, 26 °C, and 28 °C). Furthermore, at these increased temperatures, apoptosis and inflammation remain at significantly lower levels in pregnant individuals than in nonhardened individuals. Autophagy, as a negative regulator of apoptosis, may lead to decreased damage to surrounding cells, which in turn alleviates inflammatory effects. In conclusion, the exposure of mussels to heat hardening seems to provide a physiological response that enhances heat tolerance and increases cell survival through increased energy production and reduced cell death and inflammatory responses. The latter can be utilized for the management and conservation of aquatic species of economic value or endangered status.

Benchmarking sample pooling for epigenomics of natural populations.

DNA methylation (DNAm) is a mechanism for rapid acclimation to environmental conditions. In natural systems, small effect sizes relative to noise necessitates large sampling efforts to detect differences. Large numbers of individually sequenced libraries are costly. Pooling DNA prior to library preparation may be an efficient way to reduce costs and increase sample size, yet there are to date no recommendations in ecological epigenetics research. We test whether pooled and individual libraries yield comparable DNAm signals in a natural system exposed to different pollution levels by generating whole-epigenome data from two invasive molluscs (Corbicula fluminea, Dreissena polymorpha) collected from polluted and unpolluted localities (Italy). DNA of the same individuals were used for pooled and individual epigenomic libraries and sequenced with equivalent resources per individual. We found that pooling effectively captures similar genome-wide and global methylation signals as individual libraries, highlighting that pooled libraries are representative of the global population signal. However, pooled libraries yielded orders of magnitude more data than individual libraries, which was a consequence of higher coverage. We would therefore recommend aiming for a high initial coverage of individual libraries (15×) in future studies. Consequently, we detected many more differentially methylated regions (DMRs) with the pooled libraries and a significantly lower statistical power for regions from individual libraries. Computationally pooled data from the individual libraries produced fewer DMRs and the overlap with wet-lab pooled DMRs was relatively low. We discuss possible causes for discrepancies, list benefits and drawbacks of pooling, and provide recommendations for future epigenomic studies.

Blood-responsive mussel-inspired hydrogels for hemostasis, antibacterial action, and wound healing.

Rapid hemostasis, potent antimicrobial activity, and efficient wound management are critical factors in enhancing the survival of trauma patients. Chitosan, as a green and sustainable biomaterial with low cost, degradability and biocompatibility, is widely used in the biomedical field. However, chitosan dissolves in an acidic environment, which is not conducive to wound healing. In this study, chitosan was chemically modified to address this limitation. A mussel-inspired hydrogel composed of caffeic acid-grafted chitosan, gallic acid-grafted chitosan, and oxidized microcrystalline cellulose (CHI-C/CSG/OMCC) was designed. This hydrogel exhibits blood-responsive gelation behavior and offers a synergistic combination of tissue adhesion, antimicrobial properties, and tissue repair capabilities. The carboxyl, hydroxyl, phenolic hydroxyl and aldehyde groups within the hydrogel system endowed the hydrogel with excellent adhesion properties (53.1 kPa adhesion strength to porcine skin-adherent tissues), biocompatibility, and excellent antimicrobial properties. Surprisingly, this hydrogel not only achieved rapid and effective hemostasis, but also effectively promoted wound healing in a mouse skin injury model. In addition, its remarkable efficacy in stopping bleeding within approximately 2 min without rebleeding was demonstrated in a porcine model of acute gastrointestinal hemorrhage in the esophagus, stomach, and intestines. This blood-responsive ternary hydrogel offers a promising alternative to wound management materials due to its excellent overall performance and superior efficacy in all phases of wound healing.