Toxicokinetics and Mussel Watch: Addressing Interspecies Differences for Coastal Cadmium Contamination Assessment.

Bivalves are often employed for biomonitoring contaminants in marine environments; however, in these large-scale programs, unavoidably, using multiple species presents a significant challenge. Interspecies differences in contaminant bioaccumulation can complicate data interpretation, and direct comparisons among species may result in misleading conclusions. Here, we propose a robust framework based on toxicokinetic measurements that accounts for interspecies differences in bioaccumulation. Specifically, via a recently developed double stable isotope tracer technique, we determined the toxicokinetics of cadmium (Cd)─a metal known for its high concentrations in bivalves and significant interspecies bioaccumulation variability─in six widespread bivalve species including mussels (Perna viridis, Mytilus unguiculatus, Mytilus galloprovincialis) and oysters (Magallana gigas, Magallana hongkongensis, Magallana angulata). Results show that oysters generally have higher Cd uptake rate constants (ku: 1.18-3.09 L g-1 d-1) and lower elimination rate constants (ke: 0.008-0.017 d-1) than mussels (ku: 0.21-0.64 L g-1 d-1; ke: 0.018-0.037 d-1). The interspecies differences in tissue Cd concentrations are predominantly due to Cd uptake rather than elimination. Utilizing toxicokinetic parameters to back-calculate Cd concentrations in seawater, we found that the ranking of Cd contamination levels at the six sites markedly differs from those based on tissue Cd concentrations. We propose that this approach will be useful for interpreting data from past and future biomonitoring programs.

Turbid Waters and Clearer Standards: Refining Water Quality Criteria for Coastal Environments by Encompassing Metal Bioavailability from Suspended Particles.

Suspended particulate matter (SPM) carries a major fraction of metals in turbid coastal waters, markedly influencing metal bioaccumulation and posing risks to marine life. However, its effects are often overlooked in current water quality criteria for metals, primarily due to challenges in quantifying SPM's contribution. This contribution depends on the SPM concentration, metal distribution coefficients (Kd), and the bioavailability of SPM-bound metals (assimilation efficiency, AE), which can collectively be integrated as a modifying factor (MF). Accordingly, we developed a new stable isotope method to measure metal AE by individual organisms from SPM, employing the widely distributed filter-feeding clam Ruditapes philippinarum as a representative species. Assessing SPM from 23 coastal sites in China, we found average AEs of 42% for Zn, 26% for Cd, 20% for Cu, 8% for Ni, and 6% for Pb. Moreover, using stable isotope methods, we determined metal Kd of SPM from these sites, which can be well predicted by the total organic carbon and iron content (R2 = 0.977). We calculated MFs using a Monte Carlo method. The calculated MFs are in the range 9.9-43 for Pb, 8.5-37 for Zn, 2.9-9.7 for Cu, 1.4-2.7 for Ni, and 1.1-1.6 for Cd, suggesting that dissolved-metal-based criteria values should be divided by MFs to provide adequate protection to aquatic life. This study provides foundational guidelines to refine water quality criteria in turbid waters and protect coastal ecosystems.

Silica Nanoparticle Size Determines the Mechanisms Underlying the Inhibition of Iron Oxide Nanoparticle Uptake by Daphnia magna.

Aquatic environments are complicated systems that contain different types of nanoparticles (NPs). Nevertheless, recent studies of NP toxicity, and especially those that have focused on bioaccumulation have mostly investigated only a single type of NPs. Assessments of the environmental risks of NPs that do not consider co-exposure regimes may lead to inaccurate conclusions and ineffective environmental regulation. Thus, the present study examined the effects of differently sized silica NPs (SiO2 NPs) on the uptake of iron oxide NPs (Fe2O3 NPs) by the zooplankton Daphnia magna. Both SiO2 NPs and Fe2O3 NPs were well dispersed in the experimental medium without significant heteroaggregation. Although all three sizes of SiO2 NPs inhibited the uptake of Fe2O3 NPs, the underlying mechanisms differed. SiO2 NPs smaller than the average mesh size (∼200 nm) of the filtering apparatus of D. magna reduced the accumulation of Fe2O3 NPs through uptake competition, whereas larger SiO2 NPs inhibited the uptake of Fe2O3 NPs mainly by reducing the water filtration rate of the daphnids. Overall, in evaluations of the risks of NPs in the natural environment, the different mechanisms underlying the effects of NPs of different sizes on the uptake of dissimilar NPs should be considered.

Stimulated Raman Scattering Microscopy Reveals Bioaccumulation of Small Microplastics in Protozoa from Natural Waters.

Microplastics (MPs) are pollutants of global concern, and bioaccumulation determines their biological effects. Although microorganisms form a large fraction of our ecosystem's biomass and are important in biogeochemical cycling, their accumulation of MPs has never been confirmed in natural waters because current tools for field biological samples can detect only MPs > 10 µm. Here, we show that stimulated Raman scattering microscopy (SRS) can image and quantify the bioaccumulation of small MPs (<10 µm) in protozoa. Our label-free method, which differentiates MPs by their SRS spectra, detects individual and mixtures of different MPs (e.g., polyethylene, polypropylene, polyvinyl chloride, polyethylene terephthalate, polystyrene, and poly(methyl methacrylate)) in protozoa. The ability of SRS to quantify cellular MP accumulation is similar to that of flow cytometry, a fluorescence-based method commonly used to determine cellular MP accumulation. Moreover, we discovered that protozoa in water samples from Yangtze River, Xianlin Wastewater Treatment Plant, Lake Taihu and the Pearl River Estuary accumulated MPs < 10 µm, but the proportion of MP-containing cells was low (∼2-5%). Our findings suggest that small MPs could potentially enter the food chain and transfer to organisms at higher trophic levels, posing environmental and health risks that deserve closer scrutiny.

Toward Citizen Science-Based Ocean Acidification Observations Using Smartphone Devices.

pH is a key parameter in many chemical, biological, and biogeochemical processes, making it a fundamental aspect of environmental monitoring. Rapid and accurate seawater pH measurements are essential for effective ocean observation and acidification investigations, resulting in the need for novel solutions that allow robust, precise, and affordable pH monitoring. In this study, a versatile smartphone-based environmental analyzer (vSEA) was used for the rapid measurement of seawater pH in a field study. The feasibility of the use of the vSEA algorithm for pH quantification was explored and verified. When used in conjunction with a three-dimensional (3D)-printed light-proof shell, the quality of captured images is guaranteed. The quantitative accuracy of vSEA pH measurements reached 0.018 units with an uncertainty of <0.01, meeting the requirements of the Global Ocean Acidification Observing Network (GOA-ON) for "weather" goals (permitting a maximum pH uncertainty of 0.02). The vSEA-pH system was successfully applied for on-site pH measurements in coastal seawater and coral systems. The performance of the vSEA-pH system was validated using different real-world samples, and t-test results showed that the vSEA-pH system was consistent with pH measurements obtained using a state-of-the-art benchtop spectrophotometer (t = 1.986, p = 0.7949). The vSEA-pH system is applicable to different types of smartphone devices, making it possible for vSEA-pH to be widely promoted for public citizen use. The vSEA-pH system offers a simple, accurate, and applicable method for the on-site measurement of seawater pH, assisting the large-scale monitoring of ocean acidification by allowing the contribution of citizen science-based data collection.

Label-Free Imaging of Humic Substance Bioaccumulation by Pump-Probe Microscopy.

Humic substances (HS) are the most abundant forms of natural organic matter on the earth surface. Comprised of decomposed plant and animal materials rich in carbon, oxygen, hydrogen, nitrogen, and sulfur complexes, HS facilitate global carbon and nitrogen cycling and the transport of anthropogenic contaminants. While it is known that HS also interact with organisms at different trophic levels to produce beneficial and harmful effects whether HS exert these biological effects through accumulation remains unknown. Current radiolabeling techniques, which only detect the amount of accumulated radiolabels, cannot visualize the transport and accumulation behavior of HS. Here, using a label-free method based on pump-probe microscopy, we show HS entered the protozoan Tetrahymena thermophila, zebrafish embryos, and human cells and exerted direct effects on these organisms. HS accumulated in the nucleus of T. thermophila, chorion pore canals of zebrafish embryos, and nucleus of intestinal and lung cells in a concentration- and time-dependent way. Epigenetic and transcriptomics assays show HS altered chromatin accessibility and gene transcription in T. thermophila. In zebrafish larvae, HS induced neurotoxicity, altering spontaneous muscle contraction and locomotor activity. Detailed images showing HS accumulation in our study reveal new insights on the ecological and environmental behavior of HS.

Higher Risks of Copper Toxicity in Turbid Waters: Quantifying the Bioavailability of Particle-Bound Metals to Set Site-Specific Water Quality Criteria.

In coastal waters, particulate metals constitute a substantial fraction of the total metals; however, the prevalent water quality criteria are primarily based on dissolved metals, seemingly neglecting the contribution of particulate metals. Here we developed a method to quantify the toxicity risk of particulate metals, and proposed a way to calculate modifying factors (MFs) for setting site-specific criteria in turbid waters. Specifically, we used a side-by-side experimental design to study copper (Cu) bioaccumulation and toxicity in an estuarine clam, Potamocorbula laevis, under the exposure to "dissolved only" and "dissolved + particulate" 65Cu. A toxicokinetic-toxicodynamic model (TK-TD) was used to quantify the processes of Cu uptake, ingestion, assimilation, egestion, and elimination, and to relate mortality risk to tissue Cu. We find that particulate Cu contributes 40-67% of the Cu bioaccumulation when the suspended particulate matter (SPM) ranges from 12 to 229 mg L-1. The Cu-bearing SPM also increases the sensitivity of organisms to internalized Cu by decreasing the internal threshold concentration (CIT) from 141 to 76.8 µg g-1. MFs were derived based on the TK-TD model to consider the contribution of particulate Cu (in the studied SPM range) for increasing Cu bioaccumulation (MF = 1.3-2.2) and toxicity (MF = 2.3-3.9). Water quality criteria derived from dissolved metal exposure need to be lowered by dividing by an MF to provide adequate protection. Overall, the method we developed provides a scientifically sound framework to manage the risks of metals in turbid waters.

Enhancing Sediment Bioaccumulation Predictions: Isotopically Modified Bioassay and Biodynamic Modeling for Nickel Assessment.

Quantifying metal bioaccumulation in a sedimentary environment is a valuable line of evidence when evaluating the ecological risks associated with metal-contaminated sediments. However, the precision of bioaccumulation predictions has been hindered by the challenges in accurately modeling metal influx processes. This study focuses on nickel bioaccumulation from sediment and introduces an innovative approach using the isotopically modified bioassay to directly measure nickel assimilation rates in sediment. Tested in sediments spiked with two distinct nickel concentrations, the measured Ni assimilation rates ranged from 35 to 78 ng g-1 h-1 in the Low-Ni treatment and from 96 to 320 ng g-1 h-1 in the High-Ni treatment. Integrating these rates into a biodynamic model yielded predictions of nickel bioaccumulation closely matching the measured results, demonstrating high accuracy with predictions within a factor of 3 for the Low-Ni treatment and within a factor of 1 for the High-Ni treatment. By eliminating the need to model metal uptake from various sources, this streamlined approach provides a reliable method for predicting nickel bioaccumulation in contaminated sediments. This advancement holds promise for linking bioaccumulation with metal toxicity risks in sedimentary environments, enhancing our understanding of metal-contaminated sediment risks and providing valuable insights to support informed decision-making in ecological risk assessment and management.

Isotopically Modified Bioassay Bridges the Bioavailability and Toxicity Risk Assessment of Metals in Bedded Sediments.

The application of bioavailability-based risk assessment for the management of contaminated sediments requires new techniques to rapidly and accurately determine metal bioavailability. Here, we designed a multimetal isotopically modified bioassay to directly measure the bioavailability of different metals by tracing the change in their isotopic composition within organisms following sediment exposure. With a 24 h sediment exposure, the bioassay sensed significant bioavailability of nickel and lead within the sediment and determined that cadmium and copper exhibited low bioavailable concentrations and risk profiles. We further tested whether the metal bioavailability sensed by this new bioassay would predict the toxicity risk of metals by examining the relationship between metal bioavailability and metal toxicity to chironomid larvae emergence. A strong dose-toxicity relationship between nickel bioavailability (nickel assimilation rate) and toxicity (22 days emergence ratio) indicated exposure to bioavailable nickel in the sediment induced toxic effects to the chironomids. Overall, our study demonstrated that the isotopically modified bioassay successfully determined metal bioavailability in sediments within a relatively short period of exposure. Because of its speed of measurement, it may be used at the initial screening stage to rapidly diagnose the bioavailable contamination status of a site.

Measuring Metal Uptake and Loss in Individual Organisms: A Novel Double Stable Isotope Method and its Application in Explaining Body Size Effects on Cadmium Concentration in Mussels.

Interindividual variabilities in metal bioaccumulation confound our interpretation of the biomonitoring data. Measuring metal toxicokinetics in organism "individuals" may provide insights into the processes underlying the variabilities. Therefore, we developed a double stable isotope method that can simultaneously measure uptake and elimination of metals in individual organisms and thus the distribution of the toxicokinetic parameters. Specifically, we exposed organisms to both isotopes (113Cd and 114Cd; Cd = cadmium) during the first stage and to only one isotope (114Cd) during the second stage. Metal uptake and elimination rate constants (i.e., ku and ke) were simultaneously estimated from the content of the two isotopes measured in each organism at the end of the second stage. We applied the method to investigate the interindividual variability in Cd concentrations caused by body size in two marine mussel species. Cd concentrations are higher in larger Xenostrobus atratus but lower in smaller Perna viridis. Size-dependent Cd uptake is found to be responsible for size effects on Cd concentrations in the mussels and the interspecies differences in the relationship between Cd concentration and body size. Specifically, Cd ku increases with size in X. atratus (0.057-0.297 L g-1 d-1) but decreases with size in P. viridis (0.155-0.351 L g-1 d-1). In contrast, Cd ke is not influenced by body size (X. atratus: 0.002-0.060 d-1; P. viridis: 0.008-0.060 d-1). Overall, we extended the applicability of the stable isotope methods to measure metal toxicokinetics in "individual" organisms, providing a readily available tool for investigating problems related to metal bioaccumulation.

Application of a Multi-Metal Stable-Isotope-Enriched Bioassay to Assess Changes to Metal Bioavailability in Suspended Sediments.

The direct measurement of particulate contaminant bioavailability is a challenging aspect for the environmental risk assessment of contaminated sites. Here, we demonstrated a multi-metal stable-isotope-enriched bioassay to simultaneously measure the bioavailability of Cd, Cu, and Zn in naturally contaminated sediments following differing periods of resuspension treatment. Freshwater filter-feeding clams were pre-labeled with the isotopes 114Cd, 65Cu, and 68Zn to elevate isotope abundances in their tissues and then exposed to metal-contaminated suspended sediments. The assimilation of sediment-associated metals by clams would decrease the isotope ratios (Cd114/111, Cu65/63, and Zn68/64) in tissues, providing a direct measurement of metal bioavailability. For the sediments tested here, the method revealed bioavailable cadmium and non-bioavailable copper in sediments but was inconclusive for zinc. With a longer resuspension time, the bioavailability of particulate cadmium increased, but that of copper was unaffected. Metal bioavailability predicted using traditional wet-chemical extraction methods was inconsistent with these findings. The study indicated that multi-metal stable-isotope-enriched bioassay provides a new tool for directly assessing metal bioavailability in sediments, and this method is amenable for use in in situ assessments.

Predicting Risks of Cadmium Toxicity in Salinity-Fluctuating Estuarine Waters Using the Toxicokinetic-Toxicodynamic Model.

In estuaries, salinity fluctuates rapidly and continuously, greatly affecting the bioavailability and thus toxicity of contaminants, especially metals, causing difficulties in deriving site-specific water quality criteria. We developed a method for predicting the toxicity of the metal cadmium (Cd) in estuarine waters of any salinity fluctuation scenario. Cd bioaccumulation and toxicity were measured in an estuarine clam Potamocorbula laevis under stable salinities (salinity = 5, 15, 25) and fluctuating salinities (5-25), using the toxicokinetic-toxicodynamic (TK-TD) framework. Cd bioaccumulation decreases with increasing salinity; whereas intrinsic Cd sensitivity of organisms reaches the minimum at an intermediate salinity around 20. At each specific Cd level, interpolating TK-TD parameters measured at the stable salinities well predicts the Cd bioaccumulation and toxicity under fluctuating salinities. To extend the model for various Cd levels, the biotic ligand model (BLM) was integrated into the TK-TD framework. The BLM-based TK-TD model was successfully applied to scenarios of simulated and monitored salinity fluctuations in estuarine waters, for which the median lethal concentrations and no-effect concentrations (2.0-3.1 µg L-1) of Cd were derived. Overall, we integrated the BLM and TK-TD models and provided a useful tool for predicting metal risks and deriving criteria values for salinity-fluctuating estuarine waters.

Physiologically Based Pharmacokinetic Model for the Biotransportation of Arsenic in Marine Medaka (Oryzias melastigma).

The toxicity of arsenic (As) targets specific tissues of organisms, while the biotransportation of As among the tissues of fish remains poorly understood. In the present study, radiotracer techniques followed by a physiologically based pharmacokinetic (PBPK) modeling were applied to simulate the biotransportation (absorption, distribution, and elimination) of 73As(V) and biotransformation of As(V) in the marine medaka Oryzias melastigma after waterborne As exposure. Fish were simulated by a six-compartment model by assuming that blood was the intermediate exchange among different compartments (gill, intestine, liver, head, and carcass). Modeling suggested that intestine and gill were the uptake, exchange, as well as elimination sites of waterborne As, while carcass and head were the main storage sites. Intestine played a vital role in the metabolism of As(V) by biotransforming inorganic As into arsenobetaine (AsB), possibly because of the important role of gut microbiota. The correlation between the PBPK model constants and the As speciation (e.g., AsB %, inorganic As %, and methylated As %) indicated that AsB tended to be stored in the tissues rather than being depurated, while inorganic and methylated As were more easily transferred from tissues to the blood and eliminated. Modeling simulation coupling with biotransformation for the first time demonstrated that the fish intestine was the main metabolic site, and synthesis of AsB as mediated by the microbiota in the intestine contributed to the high As bioaccumulation in marine fish.

Unravelling Metal Speciation in the Microenvironment Surrounding Phytoplankton Cells to Improve Predictions of Metal Bioavailability.

A lack of knowledge on metal speciation in the microenvironment surrounding phytoplankton cells (i.e., the phycosphere) represents an impediment to accurately predicting metal bioavailability. Phycosphere pH and O2 concentrations from a diversity of algae species were compiled. For marine algae in the light, the average increases were 0.32 pH units and 0.17 mM O2 in the phycosphere, whereas in the dark the average decreases were 0.10 pH units and 0.03 mM O2, in comparison to bulk seawater. In freshwater algae, the phycosphere pH increased by 1.28 units, whereas O2 increased by 0.38 mM in the light. Equilibrium modeling showed that the pH alteration influenced the chemical species distribution (i.e., free ion, inorganic complexes, and organic complexes) of Al, Cd, Co, Cu, Fe, Hg, Mn, Ni, Pb, Sc, Sm, and Zn in the phycosphere, and the O2 fluctuation increased oxidation rates of Cu(I), Fe(II) and Mn(II) from 2 to 938-fold. The pH/O2-induced changes in phycosphere metal chemistry were larger for freshwater algae than for marine species. Reanalyses of algal metal uptake data in the literature showed that uptake of the trivalent metals (Sc, Sm and Fe), in addition to divalent metals, can be better predicted after considering the phycosphere chemistry.

Making Acute Tests More Ecologically Relevant: Cadmium Bioaccumulation and Toxicity in an Estuarine Clam under Various Salinities Modeled in a Toxicokinetic-Toxicodynamic Framework.

Salinity has considerable effects on the toxicity of metals in estuarine waters. The effects of salinity are manifold, making it difficult to summarize for risk assessments. In this study, we separated and quantified the multiple effects of salinity on cadmium (Cd) in a toxicokinetic-toxicodynamic framework. The estuarine clam, Potamocorbula laevis, was used as a model organism. Cd bioaccumulation was measured using a stable-isotope-tracer technique; in parallel, toxicity tests were conducted. With the increase of salinity from 5 to 30, Cd uptake decreased monotonically. In contrast, the intrinsic sensitivity of organisms, measured by the toxicodynamic parameters, reached its minimum at intermediate salinities (i.e., 10 to 20). The overall salinity effects were dominated by the effects on Cd bioaccumulation; therefore, Cd toxicity decreased monotonically with the increases of salinity. The model developed in this study could provide predictions of no-effect concentration (1.7 to 34.9 µg L-1, end point mortality) and the median lethal concentration (LC50) of Cd at different salinities. In conclusion, we developed a framework for quantifying the multiple effects of salinity and a method for estimating no-effect concentration from acute toxicity tests, which can be used for better assessments of metal risks in estuarine waters.

Why Does Cysteine Enhance Metal Uptake by Phytoplankton in Seawater but Not in Freshwater?

Low-molecular-weight weak ligands such as cysteine have been shown to enhance metal uptake by marine phytoplankton in the presence of strong ligands, but the effect is not observed in freshwater. We hypothesized that these contrasting results might be caused by local cysteine degradation and a Ca effect on metal-ligand exchange kinetics in the boundary layer surrounding the algal cells; newly liberated free metal ions cannot be immediately complexed in seawater by Ca-bound strong ligands but can be rapidly complexed by free ligands at low-Ca levels. The present results consistently support this hypothesis. At constant bulk Cd2+ concentrations, buffered by strong ligands: (1) at 50 mM Ca, cysteine addition significantly enhanced Cd uptake in high-Ca preacclimated euryhaline Chlamydomonas reinhardtii (cultured with cysteine as a nitrogen source to enhance local Cd2+ liberation via cysteine degradation); (2) at 0.07 mM Ca, this enhancement was not observed in the algae; (3) at 50 mM Ca, the enhancement disappeared when C. reinhardtii were cultured with ammonium (to inhibit cysteine degradation and local Cd2+ liberation); (4) cysteine addition did not enhance Cd uptake by cysteine-cultured marine Thalassiosira weissflogii when the concentration of immediately reacting strong ligands was sufficient to complex local Cd2+ liberation.

Modeling the Toxicokinetics of Multiple Metals in the Oyster Crassostrea hongkongensis in a Dynamic Estuarine Environment.

Metal contamination is a major problem in many estuaries. Toxicokinetic models are useful tools for predicting metal accumulation in estuarine organisms and managing the associated ecological risks. However, obtaining toxicokinetic parameter values with sufficient predictive power is challenging for dynamic estuarine waters. In this study, we determined the toxicokinetics of multiple metals in the oyster Crassostrea hongkongensis in a dynamic estuary polluted by metals using a 48 day transplant experiment. During the experiment, metal concentrations in oysters, water, and suspended particles were intensively monitored at 3 day intervals. The toxicokinetic parameters were then estimated using the Markov chain Monte Carlo (MCMC) method. The calibrated model was capable of successfully simulating the time-course of metal bioaccumulation in oysters and was further validated by predicting the bioaccumulation at another site in the estuary. Furthermore, the model was used to assess the relative importance of different pathways in metal bioaccumulation. With the MCMC method, distributions instead of single values were assigned to model parameters. This method makes the model predictions probabilistic with clearly defined uncertainties, and they are thus particularly useful for the risk assessment of metals in aquatic systems.

Two-Compartment Kinetic Modeling of Radiocesium Accumulation in Marine Bivalves under Hypothetical Exposure Regimes.

Interpreting the variable concentrations of (137)Cs in the field biological samples requires mechanistic understanding of both environmental and biological behavior of (137)Cs. In this study, we used a two-compartment model to estimate and compare the (137)Cs biokinetics in three species of subtropical marine bivalves. Significant interspecific difference of (137)Cs biokinetics was observed among oysters, mussels, and scallops. There was considerable (137)Cs assimilation from phytoplankton in the bivalves, but the calculated trophic transfer factors were generally between 0.04 and 0.4. We demonstrated a major efflux of radiocesium in the scallops (with a rate constant of 0.207 d(-1)), whereas the efflux was comparable between oysters and mussels (0.035-0.038 d(-1)). A two-compartment kinetic model was developed to simulate the (137)Cs accumulation in the three bivalves under four hypothetical exposure regimes. We showed that the bivalves respond differently to the exposure regimes in terms of time to reach equilibrium, equilibrium concentration, and maximum concentration. Bivalves suffering more frequent intermittent exposure may have higher maximum concentrations than those receiving less frequent exposure. The interspecific difference of (137)Cs accumulation in bivalves has important implications for biomonitoring and implementing management techniques. This study represents one of the first attempts to combine both dissolved and dietary pathways to give a realistic simulation of (137)Cs accumulation in marine bivalves under dynamic exposure regimes.

Speciation of Cu and Zn in Two Colored Oyster Species Determined by X-ray Absorption Spectroscopy.

In oysters, Cu and Zn concentration can reach extraordinarily high (1-2% of tissue dry weights), leading to intense green or blue colors in oyster tissues. It is thus puzzling how oysters detoxify the excessively accumulated metals. Here, we used X-ray absorption spectroscopy to examine the Cu and Zn speciation in two "colored" oysters (Crassostrea hongkongensis and Crassostrea sikamea) collected from a severely metal-contaminated estuary. In contaminated oysters, we found a much higher proportion of tissue Cu and Zn as oxygen- or nitrogen-bonded species, contrasting to the sulfur-bonded (thiolate) species in normal oysters. Speciation of Cu and Zn in mantle was similar to that in gills, both of which were different from that in the digestive gland. In C. sikamea, the difference of metal speciation between normal and contaminated individuals was of similar pattern but less pronounced than that in C. hongkongensis. In normal oysters, Cu existed mainly as Cu(I) bound to sulfur, whereas in contaminated oysters mainly as Cu(II) bound to oxygen or nitrogen. Our study provided direct and semiquantitative information on the changes of metal speciation in contaminated oysters, indicating that oysters could efficiently detoxify the excessively accumulated Cu and Zn by storing them in oxygen- and nitrogen-bonded complexes in metal-rich environments.

Metabolome analysis revealed the critical role of betaine for arsenobetaine biosynthesis in the marine medaka (Oryzias melastigma).

Arsenobetaine (AsB), a non-toxic arsenic (As) compound found in marine fish, structurally resembles betaine (GB), a common methyl donor in organisms. This study investigates the potential role of GB in AsB synthesis in marine medaka (Oryzias melastigma) using metabolomic analysis. Dietary exposure to arsenate (As(V)) and varying GB concentrations (0.05% and 0.1% in diets) increased total As and AsB bioaccumulation, particularly in marine medaka muscle. Metabolomic analysis revealed that GB played a crucial role in promoting up-regulation in methylthioadenosine (MTA) by modulating the methionine cycle and down-regulation in glutathione (GSH) by modulating the glutathione cycle. Methionine metabolism and GSH, potentially binding again to exogenous GB, could synchronously produce more non-toxic AsB. Combining verification experiments of differential metabolites of Escherichia coli in vitro, GB, GSH, S-adenosylmethionine (SAM), and arsenocholine (AsC) entered methionine and glutathione metabolism pathways to generate more AsB. These findings underscore the GB's crucial regulatory role in modulating the synthesis of AsB. This study provides vital insights into the interplay between the structural analogs GB and AsB, offering specific strategies to enhance the detoxification mechanisms of marine fish in As-contaminated environments.