Mussel Tissue Research Articles

Characterization of a novel toll-like receptor and activation NF-κB signal pathway in triangle sail mussel Hyriopsis cumingii.

Toll-like receptor (TLR) family plays an important role in innate immunity for detection of and defense against microbial pathogens. In this study, a novel toll-like receptor (HcTLRn) was characterized from freshwater pearl mussel H. cumingii. The complete sequence of HcTLRn was 3725 bp, and the open reading frame (ORF) encoded 718 amino acid residues. Predicted HcTLRn protein possessed seven atypical leucine-rich repeat (LRR) domains, two typical LRR subfamily domains, a C-terminal domain LRR, a transmembrane domain and an intracellular Toll/interleukin-1 (IL-1) receptor domain. Transcripts of HcTLRn were constitutive expressed in the tissues of healthy mussels and were markedly induced in hepatopancreas and gills after lipopolysaccharide (LPS), peptidoglycan (PGN) and polyinosinic polycytidylic acid (ploy I: C) stimulation. Knockdown of HcTLRn in vivo significantly decreased the mRNA levels of TLR pathway transcription factors p65 and p105 as well as antimicrobial peptides (AMPs) including lysozyme (HcLys), theromacin (HcTher), whey acidic protein (HcWAP), LPS-binding protein/bactericidal permeability increasing protein (HcLBP/BPI) 1 and 2 after mussels challenged by LPS. In situ hybridization results showed that HcTLRn mRNA was significantly increased in hemocytes after LPS, PGN and poly I:C stimulation. HcTLRn protein was mainly expressed in hepatopancreas and gills and was significantly increased after LPS stimulation. Moreover, recombinant extracellular domain of HcTLRn (HcTLRn-ECD) proteins could bind to a variety of bacterial and pathogen-associated molecular patterns such as LPS, PGN, and poly I:C in vitro. Subcellular localization results showed that HcTLRn was mainly distributed near the cell membrane and in cytoplasm. Over-expression of HcTLRn activated the NF-κB luciferase reporter in HEK293T cells. Collectively, these results suggested that HcTLRn was a TLR family member that might play an important role in activation of NF-κB signal pathway in Mollusca.

Effects of the toxic dinoflagellate Alexandrium catenella on the behaviour and physiology of the blue mussel Mytilus edulis

The effects of harmful algae on bivalve physiology are complex and involve both physiological and behavioural responses. Studying those responses is essential to better describe and predict their impact on shellfish aquaculture and health risk for humans. In this study we recorded for two months the physiological response of the blue mussel Mytilus edulis from Eastern Canada to a one-week exposure to a paralytic shellfish poisoning producing dinoflagellate strain of Alexandrium catenella, isolated from the St Lawrence estuary, Canada. Mussels in a ‘control’ treatment were fed continuously with a non-toxic diet, while mussels in a ‘starvation’ treatment were fed the same non-toxic diet the first week and subsequently starved for seven weeks. Mussels in a ‘toxic’ treatment received A. catenella for one week before being starved until the end of the experiment. Over a two-month experiment we monitored shell and tissue growth, filtration capacity, respiration rate, byssal attachment strength, valve opening behaviour, and toxin content in tissues. Mussels fed normally on the toxic dinoflagellate and accumulated an average of 51.6 µg STXeq 100 g–1 after one week of exposure. After seven weeks of depuration, about half of the specimen showed levels around 18 µg STXeq 100 g–1. The condition index of exposed mussels (‘toxic’ treatment) decreased rapidly from the start as compared to mussels that received a one-week non-toxic diet (‘starvation’ treatment). Oxygen consumption rates increased in the ‘toxic’ treatment before leveling out with that of mussels from the ‘starvation’ treatment. Valve opening amplitude was lower in the ‘toxic’ treatment during and following the exposure. Average valve closure duration was higher right after the exposure, during the peak of mussel tissue intoxication. No significant change in byssal thread strength was observed through time in each treatment but less force was required to detach mussels from the ‘toxic’ and ‘starvation’ treatments. The number of byssus threads produced by mussels exposed to the toxic dinoflagellate was also lower than in the control group. These results represent advancements in our understanding of the impacts of harmful algae on bivalves and contribute to the development of mitigation measures necessary to both the safety of consumers and the sustainability of aquaculture operations.

Acute copper accumulation in Mediterranean mussel and sea snail

In this study, the rate of copper accumulation in mussel (Mytilus galloprovincialis) and sea snail (Rapana venosa) organisms exposed to acutely different non-lethal copper concentrations was investigated for 5 days under laboratory conditions. All soft tissue concentrations were analyzed using an Inductively Coupled Plasma Optical Emission Spectrophotometer. At the end of accumulation, the rate of increase of copper level in mussel and snail soft tissues in different aquariums was found to be 19.19–36.93 and 2.37–7.41 times higher for mussel and snail soft tissues than before exposure, respectively. Differences in metal accumulation at the end of the experiment between species were not statistically significant (p > 0.05). In addition, bio-concentration factor and rates of accumulation values were calculated for both organisms. The corresponding bio-concentration factor values for mussels after exposure to 0.12, 0.36, and 0.60 mg L−1 copper were 406.00, 168.41, and 165.16, respectively, while the corresponding bio-concentration factor values for snails after exposure to 0.12, 0.36, and 0.60 mg L−1 copper were 276.29, 272.12, and 265.73, respectively. While accumulation rate values were 10.15, 12.26, and 20.12 for mussels exposed to 0.12, 0.36, and 0.60 mg L−1 copper, respectively, accumulation rate values were 6.85, 19.76, and 31.69 for snails exposed to 0.12, 0.36, and 0.60 mg L−1 copper, respectively.

Biomarker-based assessment of sublethal toxicity of organic UV filters (ensulizole and octocrylene) in a sentinel marine bivalve Mytilus edulis

The global occurrence of organic UV filters in the marine environment is of increasing ecotoxicological concern. Here we assessed the toxicity of UV filters ensulizole and octocrylene in the blue mussels Mytilus edulis exposed to 10 or 100 μg l−1 of octocrylene and ensulizole for two weeks. An integrated battery of biochemical and molecular biomarkers related to xenobiotics metabolism and cellular toxicity (including oxidative stress, DNA damage, apoptosis, autophagy and inflammation) was used to assess the toxicity of these UV filters in the mussels. Octocrylene (but not ensulizole) accumulated in the mussel tissues during the waterborne exposures. Both studied UV filters induced sublethal toxic effects in M. edulis at the investigated concentrations. These effects involved induction of oxidative stress, genotoxicity (indicated by upregulation of DNA damage sensing and repair markers), upregulation of apoptosis and inflammation, and dysregulation of the xenobiotic biotransformation system. Octocrylene induced cellular stress in a concentration-dependent manner, whereas ensulizole appeared to be more toxic at the lower (10 μg l−1) studied concentration than at 100 μg l−1. The different concentration-dependence of sublethal effects and distinct toxicological profiles of ensulizole and octocrylene show that the environmental toxicity is not directly related to lipophilicity and bioaccumulation potential of these UV filters and demonstrate the importance of using bioassays for toxicity assessment of emerging pollutants in coastal marine ecosystems.

From urban dust and marine sediment to Ginkgo biloba and human serum-a top ten list of Standard Reference Materials (SRMs).

During the past 40years, the National Institute of Standards and Technology (NIST) has developed over 180 natural matrix Standard Reference Materials® (SRMs) for the determination of trace organic constituents in environmental, clinical, food, and dietary supplement matrices. A list of the Top Ten SRMs intended for organic analysis was identified based on selection criteria including analytical challenge to assign certified values, challenges in material preparation, novel matrices, longevity, widespread use, and unique design concept or intended use. The environmental matrix SRMs include air particulate matter, marine sediment, mussel tissue, and human serum with the focus on contaminants such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), chlorinated pesticides, and polybrominated diphenyl ethers (PBDEs). Human serum and plasma SRMs for clinical diagnostic markers including vitamin D metabolites represent clinical analysis, whereas infant formula, multivitamin/multielement tablets, and Ginkgo biloba constitute the food and dietary supplement matrices on the list. Each of the SRMs on the Top Ten list is discussed relative to the selection criteria and significance of the material, and several overall lessons learned are summarized.

An Electrochemical Ti3C2Tx Aptasensor for Sensitive and Label-Free Detection of Marine Biological Toxins.

Saxitoxin (STX) belongs to the family of marine biological toxins, which are major contaminants in seafood. The reference methods for STX detection are mouse bioassay and chromatographic analysis, which are time-consuming, high costs, and requirement of sophisticated operation. Therefore, the development of alternative methods for STX analysis is urgent. Electrochemical analysis is a fast, low-cost, and sensitive method for biomolecules analysis. Thus, in this study, an electrolyte-insulator-semiconductor (EIS) sensor based on aptamer-modified two-dimensional layered Ti3C2Tx nanosheets was developed for STX detection. The high surface area and rich functional groups of MXene benefited the modification of aptamer, which had specific interactions with STX. Capacitance-voltage (C-V) and constant-capacitance (ConCap) measurement results indicated that the aptasensor was able to detect STX with high sensitivity and good specificity. The detection range was 1.0 nM to 200 nM and detection limit was as low as 0.03 nM. Moreover, the aptasensor was found to have a good selectivity and two-week stability. The mussel tissue extraction test suggested the potential application of this biosensor in detecting STX in real samples. This method provides a convenient approach for low-cost, rapid, and label-free detection of marine biological toxins.

In vivo exposure of marine mussels to venlafaxine: bioconcentration and metabolization.

Pharmaceuticals are present in natural waters, thus contributing to the general exposure of aquatic organisms, but few data are available on the accumulation of these substances in marine organisms. The present study evaluated the in vivo bioconcentration of an antidepressant-venlafaxine (VLF)-in marine mussels (Mytilus galloprovincialis) during 7 days of exposure at nominal 10 μg/L concentration, followed by a 7-day depuration period. The bioconcentration factor (BCF) was 265 mL/g dry weight (dw). VLF accumulation reached an average tissue concentration of 2146 ± 156 ng/g dw within 7 days, showing a first-order kinetics process. N-desmethylvenlafaxine (N-VLF) and O-desmethylvenlafaxine (O-VLF) metabolites were quantified in mussel tissues, whereas N,N-didesmethylvenlafaxine (NN-VLF) was only recorded as being detected. These three metabolites were also quantified in water, indicating an active metabolism and VLF excretion in Mediterranean mussels. Complementary experiments conducted at nominal concentrations of 1, 10, and 100 μg/L for 7 days confirmed the VLF bioconcentration and metabolism and allowed us to quantify a supplementary metabolite, i.e., N,O-didesmethylvenlafaxine (NO-VLF), in mussel tissues. These results encourage further research on a more complete characterization of metabolism and on any disturbances linked to bioconcentration of VLF on bivalves.

Evaluation of real-time qPCR-based methods to detect the DNA of the three protozoan parasites Cryptosporidium parvum, Giardia duodenalis and Toxoplasma gondii in the tissue and hemolymph of blue mussels (M. edulis)

The protozoan parasites Cryptosporidium spp., Giardia duodenalis and Toxoplasma gondii can be transmitted to humans through shellfish consumption. No standardized methods are available for their detection in these foods, and the performance of the applied methods are rarely described in occurrence studies. Through spiking experiments, we characterized different performance criteria (e.g. sensitivity, estimated limit of detection (eLD95METH), parasite DNA recovery rates (DNA-RR)) of real-time qPCR based-methods for the detection of the three protozoa in mussel's tissues and hemolymph. Digestion of mussels tissues by trypsin instead of pepsin and the use of large buffer volumes was the most efficient for processing 50g-sample. Trypsin digestion followed by lipids removal and DNA extraction by thermal shocks and a BOOM-based technique performed poorly (e.g. eLD95METH from 30 to >3000 parasites/g). But trypsin digestion and direct DNA extraction by bead-beating and FastPrep homogenizer achieved higher performance (e.g. eLD95METH: 4–400 parasites/g, DNA-RR: 19–80%). Direct DNA recovery from concentrated hemolymph, by thermal shocks and cell lysis products removal was not efficient to sensitively detect the protozoa (e.g. eLD95METH: 10–1000 parasites/ml, DNA-RR ≤ 24%). The bead-beating DNA extraction based method is a rapid and simple approach to sensitively detect the three protozoa in mussels using tissues, that can be standardized to different food matrices. However, quantification in mussels remains an issue.

Impacts of nanoparticles and phosphonates in the behavior and oxidative status of the mediterranean mussels (Mytilus galloprovincialis).

The current study investigated the exposure of the Mediterranean mussel (Mytilus galloprovincialis) to gold nanoparticles decorated zinc oxide (Au-ZnO NPs) and phosphonate [Diethyl (3-cyano-1-hydroxy-1-phenyl-2-methylpropyl)] phosphate (PC). The mussels were exposed to concentrations of 50 and 100 µg L-1 of both compounds alone, as well as to a mixture of both pollutants (i.e. Mix). The singular and the combined effect of each pollutant was investigated by measuring the concentration of various metals (i.e., Cu, Fe, Mn, Zn and Au) in the the digestive glands and gills of mussels, their filtration capacity (FC), respiration rate (RR) and the response of oxidative biomarkers, respectively, following 14 days of exposure. The concentrations of Cu, Fe, Mn, Zn and Au increased directly with Au-ZnO NPs in mussel tissues, but significantly only for Zn. In contrast, the mixture of Au-ZnO100 NPs and PC100 did not induce any significant increase in the content of metals in digetsve glands and gills, suggesting antagonistic interactions between contaminants. In addition, FC and RR levels decreased following exposure to Au-ZnO100 NPs and PC100 treatments and no significant alterations were observed after the exposure to 50 µg.L-1 of both contaminants and Mix. Hydrogen peroxide (H2O2) level, GSH/GSSG ratio, superoxide dismutase (SOD), catalase (CAT) and acetylcholinesterase (AChE) activities showed significant changes following the exposure to both Au-ZnO NPs and PC, in the gills and the digestive glands of the mussel. However, no significant modifications were observed in both organs following the exposure to Mix. The current study advances the understanding of the toxicity of NPs and phosphonates on M. galloprovincialis and sets the path for future ecotoxicological studies regarding the synergic effects of these substances on marine species. Moreover, the current experiment suggests that the oxidative stress and the neurotoxic pathways are responsive following the exposure of marine invertebrates to both nanoparticles and phosphonates, with potential antagonist interactions of these substances on the physiology of targeted species.

The mussel caging approach in the assessment of trace metal contamination in southern Mediterranean coastal waters: a multi-biomarker study.

In the framework of the biomonitoring programme of the Gulf of Annaba (north-eastern Algeria), this study aims to assess the health status of the Gulf by transplanting the brown mussel Perna perna (Linnaeus 1758) for 12 weeks (June 2017-August 2017) at three sites. The concentrations of copper (Cu), zinc (Zn), lead (Pb) and cadmium (Cd) and as well as a battery of enzymatic and non-enzymatic biomarkers were measured in mussels before and after the transplantation period. Furthermore, analysis of trace metals was performed on the surface layer of the sediment of all the sites. A significant increase in the Cu and Pb concentrations in the mussel dry tissues was observed after the transplantation period and followed the order of metal contamination of the surface layer sediments, indicating a relationship between the bioaccumulation of metals and their bioavailability at each site. Unlike those of Cu and Pb, Cd and Zn concentrations did not reach significant levels of bioaccumulation at any of the three study sites. The biomarker response results were complementary to the measured metal concentrations in the mussel tissues and were associated with the metal accumulation index. Metal bioaccumulation in mussels and supporting biomarker response results identified the most important pollution point sources in the area.

Bioaccumulation of lead (Pb) and mercury (Hg) in green mussel Perna viridis (Linnaeus, 1758) in Cengkok Coastal Waters, Banten Bay, Indonesia

Banten Bay potentially receives pollutants such as heavy metals from various anthropogenic activities operating around the bay. Cengkok Coastal Waters, as part of Banten Bay, has a variety of fishery commodities, including green mussels (Perna viridis). This paper identifies the risks of heavy metal contamination by quantifying the bioaccumulation of Pb and Hg in the green mussel tissue. The study was conducted from March to August 2019. Analytical results indicate concentrations of Pb and Hg in the range of 0.005-0.044 mg/kg and 0.012-0.036 mg/kg, respectively. These results were still below the established national standards by SNI (2009) for fresh food and BPOM (2018) for processed food. These concentrations suggest that green mussel found in Cengkok Coastal Waters and surrounding areas are still suitable for consumption by the local community as long as they do not exceed the permissible limits, namely, 0.7 kg meat/week (for children) and 2.4 kg meat/week (for adults).

Microplastics in Invasive Freshwater Mussels (Dreissena sp.): Spatiotemporal Variation and Occurrence With Chemical Contaminants

Invasive zebra and quagga mussels (Dreissena spp.) in the Great Lakes of North America are biomonitors for chemical contaminants, but are also exposed to microplastics (<5 mm). Little research has examined in situ microplastic ingestion by dreissenid mussels, or the relationship between microplastics and chemical contaminants. We measured microplastics and chemical contaminants in mussel tissue from Milwaukee Harbor (Lake Michigan, United States) harvested from reference locations and sites influenced by wastewater effluent and urban river discharge. Mussels were deployed in cages in the summer of 2018, retrieved after 30 and 60 days, sorted by size class, and analyzed for microplastics and body burdens of three classes of contaminants: alkylphenols, polyaromatic hydrocarbons, and petroleum biomarkers. Microplastics in mussels were higher in the largest mussels at the wastewater-adjacent site after 30 days deployment. However, there was no distinction among sites for microplastics in smaller mussels, and no differences among sites after 60 days of deployment. Microplastics and chemical contaminants in mussels were not correlated. Microplastics have a diversity of intrinsic and extrinsic factors which influence their ingestion, retention, and egestion by mussels, and which vary relative to chemicals. While dreissenid mussels may not serve as plastic pollution biomonitors like they can for chemical contaminants, microplastics in dreissenid mussels are widespread, variable, and have unknown effects on physiology, mussel-mediated ecosystem processes, and lake food webs. These data will inform our understanding of the spatial distribution of microplastics in urban freshwaters, the role of dreissenid mussels in plastic budgets, and models for the fate of plastic pollution.

Effect of dissolved oxygen deficiency on 210Pb content in mussel's tissues at the Johor strait, Malaysia

Aim: To investigate the effect of dissolved oxygen deficiency on the level of 210Pb in mussel's tissues, Perna viridis which has been reported to be highly tolerant to hypoxia. Methodology: Mussels (Perna viridis), seawater and sediment samples were collected from nine stations along the Johor Straits during North-east monsoon (November 2017), inter-monsoon (March 2018), and South-west monsoon (August 2018). The 210Pb contents were extracted and estimated by Gross Alpha-beta Counter. Results: The lowest dissolved oxygen was recorded at Station 4 which decreased from 2.14 mg l-1 to 2.10 mg l-1 and subsequently to 1.63 mg l-1 during first to third samplings. In sediment, 210Pb activities increased with the increase in organic matter. Meanwhile, in mussels, the 210Pb activities showed almost similar trend with sediment where 210Pb activities were much higher adjacent to the causeway structure, especially at the East part of Johor Strait. The level of 210Pb in mussel's tissue and sediment also increased with decreasing level of dissolved oxygen. Interpretation: A causeway structure disrupted the water quality of Johor Straits and caused almost hypoxic conditions nearby. The coverage of oxygen depletion expanded during inter-monsoon. An inverse relationship between 210Pb activities and dissolved oxygen prove that deficiency of dissolved oxygen potentially influenced the bioaccumulation of 210Pb in mussels to correspond to the build-up of concentration factor in their tissues which exceeded the recommended value.

Responses to β-alanine and carnosine supplementation of mussel Mytilus coruscus as revealed by UPLC–MS/MS based untargeted metabolomics

Histidine-containing dipeptides (HCDs) have been demonstrated with important physiological functions in vertebrates. This study was executed to investigate the contents and metabolic profiles of HCDs in an invertebrate, the mussel Mytilus coruscus was selected. The carnosine/anserine analysis indicated that M. coruscus contain anserine with a higher concentration than that of carnosine, and the increases in HCDs concentrations due to β-alanine injection and carnosine injection were observed, confirming that Mytilus belongs to a species with the ability of HCDs synthesis. Further metabolomics analysis was performed based on ultra-performance liquid chromatography-tandem mass spectrometry technology to explore the effects of β-alanine injection and carnosine injection, respectively, on the whole mussel tissue. A total of 1,868 significantly differential metabolites (SDMs) were detected between the β-alanine injected, carnosine injected, and the control M. coruscus groups. The SDMs induced by β-alanine injection were enriched mainly in energy metabolism, lipid metabolism, and amino acid metabolism of mussel, while carnosine injection mainly disrupted arachidonic acid metabolism, steroid hormone biosynthesis, and retinol metabolism in the mussel. The activities of antioxidant enzymes from the whole mussel tissue extracts before and after β-alanine/carnosine injection were measured, and both treatments showed significant elevation of enzymatic activities of superoxide dismutase and catalase. Considering the positive effects of β-alanine injection on mussel metabolism, this study provided a clue for the potential application of β-alanine supplementation in mussel farming.

Spatiotemporal change of cesium-137 in the Pacific coast of Tohoku, Japan: The mussel watch approach

We measured radiocesium in mussel tissue collected from the Pacific coast of Tohoku from 2011 to 2015 to investigate the temporal and spatial dynamics of radiocesium in the coastal area. Radioactive 137Cs was detected in all the samples collected in 2011, but it was not found in samples from localities north of Sendai after 2012. In contrast, 137Cs was detected in many sites in the Fukushima area even from 2012 to 2015. The fluctuation of 137Cs concentration in mussel tissue seems to reflect the 137Cs concentration in suspended particles in the seawater, suggesting that there was an influx of soil deposition and resuspension of seabed sediment. These results suggest that the 137Cs concentration in mussel tissue sensitively indicates the 137Cs concentration in the environment, and that the “mussel watch” approach is an effective way to understand the dynamics of radiocesium concentrations in coastal areas.

A Simple and Rapid Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantitation of Pharmaceuticals and Related Compounds in Mussels and Oysters.

A simple, rapid ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method has been developed and optimized for the quantitation of a range of pharmaceuticals, metabolites, and related bioactive compounds in the bivalve mollusc species mussels (Mytilus edulis) and Pacific oysters (Crassostrea gigas). Shellfish tissues were extracted using a simple solvent-based extraction method prior to concentration and purification by pass-through solid-phase extraction and quantified using stable isotope dilution MS/MS. The analytes covered a range of therapeutic classes including antidepressants, anticonvulsants, beta-blockers, and antiplatelets. Of the 34 compounds included in the present study initially, 28 compounds were found to demonstrate acceptable performance. Performance was assessed by examining extraction efficiencies, matrix effects, sensitivity, and within- and between-batch precision. The results show that as indicated by acceptable HorRat and accuracy values, the method is fit for purpose. Application of this method to environmental mussel and oyster samples revealed the presence of 12 compounds at quantifiable concentrations, with the antidepressant sertraline being present at the highest level, reaching a concentration of 6.12 ng/g in mussel tissue. © 2021 Crown copyright. Environmental Toxicology and Chemistry 2021;40:3263-3274. © 2021 SETAC. This article is published with the permission of the Controller of HMSO and the Queen's Printer for Scotland.

Mission impossible: Reach the carrion in a lithium pollution and marine warming scenario

In this study we investigated the independent and synergistic effects of lithium (Li, 0.08 mM) contamination and the rising seawater temperature (21 °C; control- 15 °C) on survival and trophic interactions (foraging behaviour, success, search time, carrion preference, feeding time, and tissue consumption-the dry weight basis) of the opportunistic intertidal scavenger Tritia neritea. Trophic interactions were assessed in a two-choice test using a Y-maze design using the same amount of two carrion species (Solen marginatus and Mytilus galloprovincialis) given to all snails simultaneously. Lithium pollution and synergestic warming have the effect of reducing the survival rate of T. neritea, triggering potential global change scenarios. The foraging behaviour of T. neritea under Li-contaminated conditions was characterised by a decrease in the snail's effectiveness in finding a carrion. Lithium changes the feeding behaviour as well as increasing the time it takes for snails to reach their food. T. neritea did not show preference for any of the carrion species offered in controls, but a shift in feeding behaviour towards more energetic carrion under Li contamination which may indicate a strategy to compensate for the greater energy expenditure necessary to survive. There were no differences in feeding time at the different treatments and regardless of the treatment tested T. neritea consumed more mussels tissue probably due to its greater palatability. Results showing foraging modifications in an intertidal scavenger mollusc in global change scenarios indicate potential changes in complex trophic interactions of marine food webs.

Speciation analysis of organoarsenic species in marine samples: method optimization using fractional factorial design and method validation

Organoarsenic species in marine matrices have been studied for many years but knowledge gaps still exist. Most literature focuses on monitoring of arsenic (As) species using previously published methods based on anion- and cation-exchange high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS). These studies are often limited to few As species and/or only specific method performance characteristics are described. Most marine certified reference materials (CRMs) are only certified for arsenobetaine (AB) and dimethylarsinate (DMA), making it difficult to evaluate the accuracy of analytical methods for other organoarsenic species. To address these gaps, the main objective of this work was to develop and validate a method for speciation analysis of a broad range of organoarsenic species in marine matrices. Optimum extraction conditions were identified through a 27–3 fractional factorial design using blue mussel as test sample. The effects of sample weight, type and volume of extraction solution, addition of H2O2 to the extraction solution, extraction time and temperature, and use of ultrasonication were investigated. The highest As recoveries were obtained by using 0.2 g as sample weight, 5 mL of aqueous methanol (MeOH:H2O, 50% v/v) as extractant, extraction carried out at 90 °C for 30 min, and without ultrasonication. Anion- and cation-exchange HPLC-ICP-MS settings were subsequently optimized. The method detected a total of 33 known and unknown As species within a run time of 23 and 20 min for cation-exchange and anion-exchange, respectively. A single-laboratory validation was conducted using several marine CRMs: BCR 627 (tuna fish tissue), ERM-CE278k (mussel tissue), DORM-4 (fish protein), DOLT-5 (dogfish liver), SQID-1 (cuttlefish), TORT-3 (lobster hepatopancreas), and CRM 7405-b (hijiki seaweed). Method performance characteristics were evaluated based on selectivity, limits of detection and quantification, linearity, trueness, precision, and measurement uncertainty. This work proposes an extraction procedure which allowed satisfactory quantification of As species with low solvent and energy consumption, supporting “Green Chemistry” principles. The study also presents a new set of As speciation data, including methylated arsenic species and arsenosugars, in recently issued marine CRMs, which will be valuable for future speciation studies on As. This work is the first to report a total of 33 different As species in marine CRMs.Graphical abstract

Mobility, ecotoxicity, bioaccumulation and sources of trace elements in the bottom sediments of the Ro\u017cn\xf3w reservoir

The aim of the study was to use of geochemical, chemical, ecotoxicological and biological indicators for a comprehensive assessment of ecological risks related to the mobility, ecotoxicity and bioavailability of trace elements in the bottom sediment of the Rożnów reservoir. The study found three elements deserving attention in the sediments: cadmium, nickel and chromium. Cadmium proved to be the most mobile and bioavailable, although the total cadmium content and geochemical indicators did not reveal any risk to organisms. Geochemical indicators showed that the sediments are contaminated with nickel and chromium, but both elements had a low bioaccumulation factor. Fractional analysis also revealed relatively low mobility of Cr and Ni and a higher potential risk of bioavailability for nickel. Most of the tested sediment samples had low toxicity in relation to the tested organisms. For H. incongruens, 11% of the samples were non-toxic, 50% of the samples had low toxicity, and 39% of the samples were toxic. For A. fischeri, no toxicity was found in 7% of the samples, low toxicity in 76% of the samples and toxicity in 17% of the sediment samples. The As, Cd, Cu content in the F1 fraction correlated significantly positively with the content of these metals in mussel tissues. Both biotesting and chemical analysis can reveal a potential risk to aquatic organisms. For a real assessment of the ecological risks associated with trace elements, it is necessary to use bioindicators taken from the environment and exposed to trace elements in situ.

Distribution of polycyclic aromatic hydrocarbons (PAHs) in the bottom sediments of a dam reservoir, their interaction with organic matter and risk to benthic fauna

PurposeThe aims of the study were to investigate the interaction between fractions of organic matter and polycyclic aromatic hydrocarbons (PAHs) in bottom sediments and to use mussels as passive biomonitors and consensus-based sediment quality guidelines for ecological risk assessment in sediments.MethodsBottom sediment samples were taken from 46 points located in the Rożnów reservoir (Poland). The sediment organic matter (SOM) characteristics included total carbon (TC), total organic carbon (TOC), humic acid carbon (Cha), fulvic acid carbon (Cfa), non-hydrolysing carbon (Cnh), and dissolved organic carbon (DOC). The extraction procedure was carried out in bottom sediments as well as in freeze-dried mussel tissue samples to directly determine the accumulation potential of PAHs to the living organisms in their natural environment.ResultsThe content of organic matter fractions was in the following order: Cfa (fulvic acid) > Cnh (non-hydrolysing carbon) > Cha (humic acid) > DOC (dissolved organic carbon). The mean ∑16PAHs (μg kg−1) concentration was 1755.2 ± 724 (total) and 256 ± 254 (bioavailable) in sediments and 1740 ± 72.2 in the mussel tissues. A significant positive correlation was found between the concentration of PAHs in the mussel tissues and the total and bioavailable concentration of PAHs in bottom sediments.ConclusionThe PAH concentration in bottom sediments depended on the stabile carbon forms Cnh. Principal component analysis (PCA) suggests that the fine fraction can significantly increase the bioavailability of PAHs and can be an important factor in the distribution of PAHs in the sediments.