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Paralytic shellfish poisoning is a global issue that would benefit from additional screening methods and rapid testing capacities. In this study, we applied 1 H NMR spectroscopy-based metabolomics to identify biomarkers of Paralytic Shellfish Toxin (PST) exposure. We characterized the metabolic phenotypes of field-collected Alaskan mussels Mytilus trossulus across a wide range of bioaccumulated PST levels, from 0 to 1590 μg/100 g. A between-level grouping emerged for high (740-1590 μg/100 g) compared to low/non-detect (0-3.91 μg/100 g) PST levels. High levels of PST contamination in mussels were consistent with alterations to energy and amino acid metabolism, and disturbances in osmoregulation. This research demonstrates the effectiveness of 1 H NMR-based metabolomics in elucidating the biological effects of paralytic shellfish toxin on the health of wild mussel populations, spatial variation, and identifies a metabolic signature indicative of PST contamination in Mytilus trossulus for potential use in a PSP biomarker panel., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Organisms rely on the integrity of the structural materials they produce to maintain a broad range of processes, such as acquiring food, resisting predators or withstanding extreme environmental forces. The production and maintenance of these biomaterials, which are often modulated by environmental conditions, can therefore have important consequences for fitness in changing climates. One well-known example of such a biomaterial is mussel byssus, an array of collagen-like fibers (byssal threads) that tethers a bivalve mollusk securely to benthic marine substrates. Byssus strength directly influences mortality from dislodgement, predation or competition and depends on the quantity and quality of byssal threads produced. We compared the temperature sensitivity of byssal attachment strength of two mussel species common to the west coast of North America, Mytilus trossulus and M. galloprovincialis, when exposed to seawater temperatures ranging from 10 to 24˚C in the laboratory. We found the two species attached equally strong in seawater ≤ 18˚C, but higher temperatures caused byssal thread production rate and quality (break force and extensibility) to be greatly reduced in M. trossulus and increased in M. galloprovincialis, leading to a 2 to 10-fold difference in overall byssus strength between the two species. Using this threshold value (18˚C), we mapped habitat for each species along the west coast of North America based on annual patterns in sea surface temperature. Estimated ranges are consistent with the current distribution of the two species and suggest a potential mechanism by which ocean warming could facilitate the northern expansion of M. galloprovincialis and displacement of native M. trossulus populations., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

Understanding and predicting responses of ectothermic animals to temperature are essential for decision-making and management. The thermal performance curve (TPC), which quantifies the thermal sensitivity of traits such as metabolism, growth and feeding rates in laboratory conditions, is often used to predict responses of wild populations. However, central assumptions of this approach are that TPCs are relatively static between populations and that curves measured under stable temperature conditions can predict performance under variable conditions. We test these assumptions using two latitudinally matched populations of the ecosystem engineer Mytilus trossulus that differ in their experienced temperature variability regime. We acclimated each population in a range of constant or fluctuating temperatures for six weeks and measured a series of both short term (feeding rate, byssal thread production) and long-term (growth, survival) metrics to test the hypothesis that performance in fluctuating temperatures can be predicted from constant temperatures. We find that this was not true for any metric, and that there were important interactions with the population of origin. Our results emphasize that responses to fluctuating conditions are still poorly understood and suggest caution must be taken in the use of TPCs generated under constant temperature conditions for the prediction of wild population responses.

After World War II, thousands of tons of highly toxic chemical warfare agents (CWA) were deposited in the Baltic Sea, the main dumping site locating in the Bornholm Basin. In the present study, Baltic mussels (Mytilus trossulus) were transplanted in the area in cages at two hotspot sites and a reference site at the depths of 35 and 65 m for 2.5 months to study bioaccumulation and biological effects of CWA possibly leaking from the corroding warfare materials. No traces of degradation products of the measured phenylarsenic CWA could be detected in the tissues of mussels. Nevertheless, several biochemical and histochemical biomarkers, geno- and cytotoxicity indicators, and bioenergetic parameters showed significant responses. The Integrated Biomarker Index calculated from the single biomarkers also showed a higher total response at the two hotspot areas compared to the reference site. Although no direct evidence could be obtained confirming the responses being caused specifically by exposure to CWA, the field exposure experiment showed unambiguously that organisms in this sea area are confronting environmental stress affecting negatively their health and this is likely related to chemical contamination, which is possibly connected to the sea-dumped CWA., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Bay mussels (Mytilus trossulus) were transplanted to 18 locations representing a range of potential exposures throughout Puget Sound, WA. Tissues were analyzed for over 200 organic contaminants. Results indicated the widespread exposure of marine organisms to trace levels of organic contaminants including the synthetic opioid oxycodone, present at three urban sites, and the chemotherapy drug melphalan, present at nine locations, at levels that may be of biological concern. Land-use and wastewater outfalls were evaluated as potential sources of CECs to the nearshore. Exposure to alkylphenol ethoxylates was associated with increased impervious surfaces in upland watersheds. A hydrodynamic simulation was performed using the Salish Sea Model to integrate inputs from 99 wastewater sources to Puget Sound. Predictions were consistent with concentrations of several wastewater-associated contaminants and δ 15 N enrichment. These results support the notion that Puget Sound nearshore biota suffer chronic exposures to a suite of contaminants from multiple sources and provide critical to focus future monitoring and management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019. Published by Elsevier B.V.)

A GalNAc/Gal-specific lectins named CGL and MTL were isolated and characterized from the edible mussels Crenomytilus grayanus and Mytilus trossulus . Amino acid sequence analysis of these lectins showed that they, together with another lectin MytiLec-1, formed a novel lectin family, adopting β-trefoil fold. In this mini review we discuss the structure, oligomerization, and carbohydrate-binding properties of a novel lectin family. We describe also the antibacterial, antifungal, and antiproliferative activities of these lectins and report about dependence of activities on molecular properties. Summarizing, CGL, MTL, and MytiLec-1 could be involved in the immunity in mollusks and may become a basis for the elaboration of new diagnostic tools or treatments for a variety of cancers.

Coastal regions worldwide face increasing management concerns due to natural and anthropogenic forces that have the potential to significantly degrade nearshore marine resources. The goal of our study was to develop and test a monitoring strategy for nearshore marine ecosystems in remote areas that are not readily accessible for sampling. Mussel species have been used extensively to assess ecosystem vulnerability to multiple, interacting stressors. We sampled bay mussels ( Mytilus trossulus ) in 2015 and 2016 from six intertidal sites in Lake Clark and Katmai National Parks and Preserves, in south-central Alaska. Reference ranges for physiological assays and gene transcription were determined for use in future assessment efforts. Both techniques identified differences among sites, suggesting influences of both large-scale and local environmental factors and underscoring the value of this combined approach to ecosystem health monitoring., Competing Interests: Katrina Counihan is employed by the Alaska SeaLife Center., (©2019 Counihan et al.)

This study focused on the alterations that occur in larval molluscan cells after administration of apoptotic inducers and inhibitors used in mammalian cells in response to cold stress. This is the first report on apoptosis modulation in molluscan cells assessed by flow cytometry. Mitochondrial activity, general caspase activation, and membrane integrity of control molluscan cells were compared to those processes in frozen-thawed molluscan cells, primary mouse embryonic fibroblasts, and human colon tumor cells prior to treatment and after incubation with apoptotic inducers or inhibitors. We tested three apoptotic inducers (staurosporine, camptothecin, and mitomycin C, routinely used for the chemical induction of apoptosis in different mammalian cells) and found that only staurosporine resulted in an evident apoptotic increase in molluscan cell cultures: 9.06% early apoptotic cells in comparison with 5.63% in control frozen-thawed cells and 20.6% late apoptotic cells in comparison with 10.68% in controls. Camptothecin did not significantly induce molluscan cell apoptosis but did cause a slight increase in the number of active cells after thawing. Mitomycin C produced similar results, but its effect was less pronounced. In addition, we hypothesize that the use of the apoptotic inhibitors could reduce apoptosis, which is significant after cryopreservation in molluscan cells; however, our attempts failed. Development in this direction is important for understanding the mechanisms of marine organisms' cold susceptibility.

Interest in the presence of pharmaceutically active compounds in the aquatic environment has been growing for over 20 years, yet very few studies deal with the metabolism of pharmaceuticals in marine organisms. In this study, the bioaccumulation under short-term conditions and metabolism of diclofenac were investigated. Mytilus trossulus was used as a representative of the Baltic benthic fauna. The mussels were exposed to diclofenac at a concentration of 133.33 μg/L for five days, following a five-day depuration phase. The highest concentration of diclofenac (7.79 μg/g dw) in tissues was determined on day 3. Subsequently, the concentration of diclofenac in tissues decreased rapidly to 0.86 μg/g dw on day 5. After five days of depuration, the concentration of diclofenac was 0.21 μg/g dw. Hydroxylated diclofenac metabolites were found both in tissues of mussels and water. This study shows that M. trossulus has the potential to accumulate diclofenac and metabolize it to 4-OH and 5-OH diclofenac., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This paper describes changes in the content of free steroid hormones e.g. testosterone (T), estradiol-17β (E2), estrone (E1) and estriol (E3) of Mytilus trossulus from the southern Baltic Sea as a function of season, stage of gametogenesis, sex, tissue (gonadal and somatic) and depth. The highest levels of T, E2, E1 and E3 were found in mussels sampled in spring and summer while the lowest levels were found in winter. This pattern was stable and was seen in both sexes and tissues in mussels from both mussel beds. The spring and summer peaks in steroid levels (SL) coincided with advanced levels of gametogenesis (the highest gonadal index, GI) of our model species. But, the lowest GI (autumn) and the lowest steroids content (winter) did not overlap. Instead, water temperature increase was followed by increase of SL and vice versa. This suggests that steroids may not be actively involved in the early stages of gamete development and does not preclude them from potentially being involved as endogenous modulators in the final stages of reproduction (e.g. spawning). Hence, observed fluctuations in SL in our model species are unlikely to be caused by reproductive cycle but are rather of unknown nature, likely linked with environmental conditions. Sex-related differences in steroid content included estrogen domination in females and androgen domination in males. A trend towards higher level of steroids in gills than in gonads was found, supporting the hypothesis about an exogenous origin of steroids in bivalves. However, based on the present results, we cannot exclude the possibility that these steroids have both an endogenous and exogenous origin., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Marine mussels (Mytilus trossulus) attach to a wide variety of surfaces underwater using a protein adhesive that is cured by the surrounding seawater environment. In this study, the influence of environmental post-processing on adhesion strength was investigated by aging adhesive plaques in a range of seawater pH conditions. Plaques took 8-12 days to achieve full strength at pH 8, nearly doubling in adhesion strength (+94%) and increasing the work required to dislodge (+59%). Holding plaques in low pH conditions prevented strengthening, causing the material to tear more frequently under tension. The timescale of strengthening is consistent with the conversion of DOPA to DOPA-quinone, a pH dependent process that promotes cross-linking between adhesive proteins. The precise arrangement of DOPA containing proteins away from the adhesive-substratum interface emphasizes the role that structural organization can have on function, an insight that could lead to the design of better synthetic adhesives and metal-coordinating hydrogels.

During summer 2014-2017, wild mytilid mussels, highly infested with the pathogenic Coccomyxa-like microalgae, were collected along the Estuary and northwestern part of Gulf of St. Lawrence (Québec, Canada). Molecular identification showed that algae can be assigned to a single taxon, Coccomyxa sp. (KJ372210), whereas hosts are represented by Mytilus edulis, M. trossulus and hybrid between these two species. This is the first record of M. trossulus and hybrid among hosts of this pathogenic alga. Our results are indicative of a possible widespread distribution of Coccomyxa sp. in the Lower St. Lawrence Estuary and along coastal waters of Canadian Maritime provinces., (Copyright © 2018 Elsevier Inc. All rights reserved.)

This study described the occurrence of abnormalities in bivalve larvae from the Puck Bay. Analyses of plankton samples collected in 2012-2013 showed that larval Mytilus trossulus, Mya arenaria, and Cerastoderma glaucum exhibited abnormalities that could indicate adverse environmental impacts. The deformities were mainly in shells, but missing soft tissue fragments and protruding vela were also noted. In addition to larval studies, we analyzed benthic postlarvae of Mytilus trossulus. Interestingly, grooves and notches at different locations of the prodissoconch, dissoconch, and shell margin were observed. Some of these deformations were reminiscent of the indentations found on the shell edge of larvae. Comparing the proportion of abnormal postlarvae to larvae with shell abnormalities suggested that the survival of larvae with shell abnormalities was low. Overall, our results suggested that the ratio of abnormal bivalve larvae could be used as an indicator of the biological effects of hazardous substances in the pelagic environment., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In the present work we compared the type and frequency of gonadal abnormalities among Mytilus trossulus populations from the Gulf of Gdańsk, Baltic Sea. Observed histopathologies were grouped as regressive changes (RC, gonadal atresia (GA) and regression (GR)), progressive changes (PC, gonadal tumors) and intersex. Sex-based and spatial differences in frequency of RC were found, with the highest frequency of RC and PC found in mussels from polluted station B followed by mussels from station A located near a purification plant outlet. Bivalves from the reference area had the lowest frequency of RC. In order to confirm biomarker applicability of RC, an exposure experiment with model xenoestrogen 17α-ethinylestradiol (EE2) was performed. The exposure of M. trossulus to 50 and 500ngdL -1 of EE2 resulted in an increased frequency of gonadal regression and atresia, including melanized hemocytes infiltration in seminiferous tubules. We thus suggest that these changes can serve as biomarkers of endocrine disrupting compounds in biomonitoring studies., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The aim of the study was to investigate abiotic and biotic factors influencing the accumulation of endocrine disrupting compounds (EDCs) such as bisphenol A (BPA), 4-tert-octylphenol (OP) and 4-nonylphenol (NP) in mussels Mytilus trossulus from the Gulf of Gdansk (Southern Baltic). The key abiotic factor influencing BPA, OP and NP accumulation in mussels is their hydrophilicity/lipophilicity, which affects their main assimilation routes - by digestive tract for the more lipophilic OP and NP, and additionally by the gills for the less lipophilic BPA. As a result, high condition index (i.e. higher soft tissue weight) is more often correlated with high concentrations of OP and NP in mussels than with BPA. Furthermore, alkylphenols have 6-8 times greater accumulative potential than BPA. Concentration of the studied compounds was lower in females than in males following spawning, and the effect lasted longer for BPA than for alkylphenols. The influence of season and hydrological conditions on BPA, OP, NP in the mussel was more pronounced than the proximity of external sources of these compounds. An increase in water temperature in summer probably stimulated the solubility of BPA, the least lipophilic of the studied compounds, and led to increased assimilation of this compound from water (through gills). On the other hand, high OP and NP concentrations in mussels occurred in spring, which was caused by increased surface run-off and sediments resuspension., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The occurrence of freshwater harmful algal bloom toxins impacting the coastal ocean is an emerging threat, and the potential for invertebrate prey items to concentrate toxin and cause harm to human and wildlife consumers is not yet fully recognized. We examined toxin uptake and release in marine mussels for both particulate and dissolved phases of the hepatotoxin microcystin, produced by the freshwater cyanobacterial genus Microcystis. We also extended our experimental investigation of particulate toxin to include oysters (Crassostrea sp.) grown commercially for aquaculture. California mussels (Mytilus californianus) and oysters were exposed to Microcystis and microcystin toxin for 24h at varying concentrations, and then were placed in constantly flowing seawater and sampled through time simulating riverine flushing events to the coastal ocean. Mussels exposed to particulate microcystin purged the toxin slowly, with toxin detectable for at least 8 weeks post-exposure and maximum toxin of 39.11ng/g after exposure to 26.65μg/L microcystins. Dissolved toxin was also taken up by California mussels, with maximum concentrations of 20.74ng/g after exposure to 7.74μg/L microcystin, but was purged more rapidly. Oysters also took up particulate toxin but purged it more quickly than mussels. Additionally, naturally occurring marine mussels collected from San Francisco Bay tested positive for high levels of microcystin toxin. These results suggest that ephemeral discharge of Microcystis or microcystin to estuaries and the coastal ocean accumulate in higher trophic levels for weeks to months following exposure., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

In contemporary ecosystems, organisms are increasingly confronted with suboptimal living conditions. We aimed to understand the role of ecosystem engineering species in suboptimal habitats from a population inhabiting the species range margin in naturally stressful conditions. We determined the impact of 2-4 cm sized patches of dwarfed mussels Mytilus trossulus close to its lower salinity limit in the North-Eastern Baltic Sea, on epibenthic community patterns. Mussels affected total macrofaunal abundance and biomass and the taxonomic and functional community structure based on abundances, as well as the species composition of macrofauna. Mussels did not affect ephemeral algae or sediment chlorophyll content, but increased the abundance, biomass, richness, and diversity of grazers, within a radius approximately twelve times the size of mussel patches. We can expect marginal populations of ecosystem engineers in suboptimal habitats to contribute to spatial heterogeneity in biotic patterns and eventual ecosystem stability., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In the present study, a new Gal/GalNAc specific lectin from the mussel Mytilus trossulus (designated as MTL) was identified, and its expression levels, both in tissues and toward pathogen stimulation, were then characterized. The MTL primary structure was determined via cDNA sequencing. Deduced sequence of 150 amino acid residues showed 89% similarity to lectins from the mussels Crenomytilus grayanus and Mytilus galloprovincialis that were the first members of a new family of zoolectins. The results indicated that the MTL might be involved in immune response toward pathogen infection, and it might perform different recognition specificity toward bacteria or fungi., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The aim of this study was to identify and quantify natural steroid hormones (17β-estradiol E2, estriol E3, estrone E1, testosterone T) and xenoestrogen (17α-ethinylestradiol EE2) in gills and gonads of Mytilus trossulus from the Gulf of Gdańsk, Poland, using the LC-MS technique based on the enzymatic digestion of tissues, SPE extraction, and subsequent LC-MS analysis of the eluates. As a result, spatial differences in several steroid hormone concentrations were detected. While the highest concentrations of testosterone and natural estrogens were documented in mussels collected at the reference station (app. 13 ng g(-1) wet weight for T, 9 ng g(-1)w.w. for E2 and 3.5 ng g(-1)w.w. for E1), decreased levels of natural steroids and increased levels of 17α-ethinylestradiol (EE2) were determined in individuals from the sewage treatment plant zone (app. 5 ng g(-1)w.w. for T, 3 ng g(-1)w.w. for E1, 1 ng g(-1)w.w. for E2 and E3, 1 ng g(-1)w.w. for EE2). No statistically significant tissue-related changes in steroids concentrations were found, but a trend towards higher steroids level in gills than in gonad was observed., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Mytilus species are important organisms in marine systems being highly abundant and widely distributed along the coast of Europe and worldwide. They are typically used in biological effects studies and have a suite of biological effects endpoints that are frequently measured and evaluated for stress effects in laboratory experiments and field monitoring programmes. Differences in bioaccumulation and biological responses of the three Mytilus species following exposure to copper (Cu) were investigated. A laboratory controlled exposure study was performed with three genetically confirmed Mytilus species; M. galloprovincialis, M. edulis and M. trossulus. Chemical bioaccumulation and biomarkers were assessed in all three Mytilus species following a 4 day and a 21 day exposure to waterborne copper concentrations (0, 10, 100 and 500μg/L). Differences in copper bioaccumulation were measured after both 4 and 21 days, which suggests some physiological differences between the species. Furthermore, differences in response for some of the biological effects endpoints were also found to occur following exposure. These differences were discussed in relation to either real physiological differences between the species or merely confounding factors relating to the species natural habitat and seasonal cycles. Overall the study demonstrated that differences in chemical bioaccumulation and biomarker responses between the Mytilus spp. occur with potential consequences for mussel exposure studies and biological effects monitoring programmes. Consequently, the study highlights the importance of identifying the correct species when using Mytilus in biological effects studies., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Flow cytometry was investigated as an alternative to visual haemocytology for potentially higher-throughput and less subjective detection of neoplasia in Mytilus trossulus. In contrast to previous studies of ploidy in the Mytilus spp. complex, distinct tetra- and pentaploidal neoplastic cells were rare and a wide range of aneuploidy peaks from 1.4n to 5.5n were detected for late-stage leukemic animals. There was no correlation between aneuploidy and the number of diseased cells for early and intermediate disease stages. Formation of aneuploidy and neoplasia progression might not be simultaneous, and DNA content analysis using flow cytometry was only useful for detecting late stages of the disease., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Blue mussels (Mytilus trossulus) were transplanted in cages for three months in two Swedish coastal areas in the Bothnian Sea (northern Baltic Sea) to investigate the interactions between analysed environmental chemicals and biological responses. A wide array of biological parameters (biomarkers) including antioxidant and biotransformation activity, geno-, cyto- and neurotoxic effects, phagocytosis, bioenergetic status and heart rate were measured to detect the possible effects of contaminants. Integrated Biomarker Response index and Principal Component Analysis performed on the individual biological response data were able to discriminate between the two study areas as well as the contaminated sites from their respective local reference sites. The two contaminated sites outside the cities of Sundsvall (station S1) and Gävle (station G1) were characterised by different biomarker response patterns. Mussels at station S1 showed a low condition index, increased heart rate recovery time and phagocytosis activity coinciding with the highest tissue concentrations of some trace metals, polycyclic aromatic hydrocarbons and organotins. At station G1 the highest organochlorine pesticide concentration was recorded as well as elevations in glutathione S-transferase activity, thiamine content and low lysosomal membrane stability. Significant variability in the geno- and cytotoxic responses and bioenergetic status was also observed at the different caging stations. The results obtained suggest that different chemical mixtures present in the study areas cause variable biological response patterns in organisms., (Copyright © 2013 Elsevier B.V. All rights reserved.)