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In the present study, we tested whether bioaccumulation in specific tissues of Daphnia magna could explain silver nanoparticle (AgNP) toxicity. Daphnids were exposed to different concentrations of well-suspended AgNPs and AgNO3 . The accumulations of Ag in the whole body, gut, and nongut tissues, as well as the mortality of daphnids, were recorded over a period of 7 d. Regression analysis showed a higher degree of positive correlation between the concentration of Ag in the nongut tissues than gut tissues and the mortality of daphnids. The results strongly suggested that the toxicity of AgNPs could be better explained in terms of bioaccumulation of AgNPs in the nongut tissues. We further tested the maternal transfer of AgNPs in daphnids into the next generation using radioactive tracers, which were able to detect as low as 1.0 to 3.2% of total accumulated Ag transferred to the neonates. The AgNPs significantly affected the reproduction process during the first 2 broods after exposure, whereas AgNO3 only had significant effects on the first brood. It is possible that AgNPs have longer adverse effects than AgNO3 on the reproduction of Daphnia. The present study identified the sensitive site of AgNP toxic action in daphnids and documented the extent of maternal transfer and the significant influence of AgNPs on the reproduction of daphnids. Environ Toxicol Chem 2017;36:3359-3366. © 2017 SETAC.

Traditional metal toxicity tests on organisms have mainly focused on continuous exposure at a fixed concentration. However, organisms are more likely exposed to pollutants intermittently in estuarine environments that are significantly impacted by anthropogenic activity. The present study examined whether different copper (Cu) exposure regimes at an equivalent dose can induce different metabolomics effects on the oysters. An estuarine oyster Crassostrea hongkongensis was exposed to Cu continuously or intermittently at an equal dose (time × concentration) for 6 wk. Continuous exposure regimes included 2 doses of 3.3 μg/L for 24 h and 20 μg/L for 24 h, with corresponding equal doses of 2 intermittent exposure regimes of 20 μg/L for 4 h and 120 μg/L for 4 h, respectively. Time-course measurements suggested that Cu bioaccumulation was comparable at equal low doses between the continuous regime (3.3 μg/L for 24 h) and the intermittent regime (20 μg/L for 4 h), but there was considerable difference for the high dose under different regimes. Nuclear magnetic resonance (NMR)-based metabolomics suggested that continuous and intermittent Cu exposures led to similar metabolite variation pattern in gills at an equal high dose, including decreased amino acids (e.g., aspartate, glycine, isoleucine, leucine, lysine, phenylalanine, threonine, and valine), lower energy-related compounds (e.g., adenosine triphosphate/adenosine diphosphate, acetate, citrate, and glycogen), and altered osmolytes (e.g., homarine and taurine). These biomarkers indicated disturbance of osmotic regulation and energy metabolism induced by Cu exposure regardless of regime. In addition, the 4-h intermittent Cu exposure resulted in slightly fewer adverse effects compared with the corresponding equal-dose continuous exposure. Oysters appeared to recover during the intervals of Cu exposure. The results indicated that metabolomic effects induced by Cu were more dose dependent than the Cu exposure regime. Environ Toxicol Chem 2017;36:2428-2435. © 2017 SETAC.

The Pearl River estuary, southern China, suffers from multiple sources of metal contamination as a result of the rapid industrial development in the region; but the biological impacts of contamination remain unknown. In the present study, a euryhaline oyster, Crassostrea hongkongensis, was collected from different sites of the Pearl River estuary; and various physiological (heart rate, alkaline phosphatase as homeostatic regulation, and glycogen as energy reserve) and cytological (lysosomal membrane stability) biomarkers were quantified to assess this species as a potential bioindicator of metal pollution in contaminated areas. Large variations of metal accumulation levels in the oysters were documented, especially for copper (Cu), zinc (Zn), cadmium (Cd), chromium, and nickel (Ni). Among these metals, the authors demonstrated significant correlations between the digestive gland metal accumulation of Cu, Zn, and Ni and the cellular homeostasis (alkaline phosphatase) and glycogen reserves. Heart rate was positively correlated with Cd but negatively correlated with Cu and Zn concentrations in the gills. Lysosomal membrane stability was significantly inhibited at the most contaminated sites but had no relationship with the accumulated metal concentrations. These measurements indicate that multimetal contamination in the Pearl River estuary impacts the physiological and cytological performance of oysters. Environ Toxicol Chem 2016;35:2577-2586. © 2016 SETAC.

The contaminated oysters discovered in the Pearl River Estuary (Guangdong province, China) contained high levels of metals in their tissues, especially Cu and Zn, indicating that this large and densely urbanized estuary in Southern China suffers from serious metal pollution. The present study aimed to investigate the impacts of multimetal pollution in the Pearl River Estuary on oyster antioxidant and detoxification systems. The responses of various biochemical biomarkers in the ecologically important oyster Crassostrea hongkongensis collected from 7 sites in the Pearl River Estuary were quantified. Significant correlations were demonstrated between the accumulation of Cu and Zn and oxidative stress (lipid peroxidation) and oxidative stress defenses (catalase, glutathione peroxidase) in the oyster gills. Significant correlations between the accumulation of Cd and Cu and detoxification (glutathione and glutathione transferase) in the gills were also documented. Interestingly, metallothionein concentrations were positively correlated with Cd, but negatively correlated with Cu, Ni, and Zn concentrations in the gills. These measurements indicated that Cu in the Pearl River Estuary induced various biochemical responses in the oysters and influenced the susceptibility of oysters to environmental stress. The present study has provided the first evidence of antioxidant and detoxification responses in native contaminated oysters from a field environment seriously contaminated by metals. Coupling biomarkers with tissue metal concentration measurements was a promising approach to identify the metals causing biological impacts in a multimetal-contaminated estuary. Environ Toxicol Chem 2016;35:2798-2805. © 2016 SETAC.

Low mercury (Hg) concentrations down to several nanograms Hg per gram of wet tissue are documented in certain fish species such as herbivorous fish, and the underlying mechanisms remain speculative. In the present study, bioaccumulation and depuration patterns of inorganic Hg(II) and methylmercury (MeHg) in a herbivorous rabbitfish Siganus canaliculatus were investigated at organ and subcellular levels following waterborne or dietary exposures. The results showed that the efflux rate constants of Hg(II) and MeHg were 0.104 d(-1) and 0.024 d(-1) , respectively, and are probably the highest rate constants recorded in fish thus far. The dietary MeHg assimilation efficiency (68%) was much lower than those in other fish species (∼90%). The predominant distribution of MeHg in fish muscle was attributable to negligible elimination of MeHg from muscle (

With increasing application and commercial production, carbon nanotubes (CNTs) will inevitably be released into aquatic environments and affect the transport and toxicity of toxic metals in ecosystems. The present study examined how CNTs affected the biokinetics and toxicity of a toxic metal, cadmium (Cd), in the freshwater zooplankton Daphnia magna. The authors quantified the dissolved uptake and the 50% lethal concentration (LC50, 48 h and 72 h) of Cd in daphnids in the presence of functionalized multiwalled nanotubes (F-CNTs) with different lengths (10–30 µm vs 0.5–2 µm) and concentrations (4 mg/L and 8 mg/L). Compared with the control treatment without CNTs, both CNTs slowed down the accumulation rate of Cd in D. magna over 8 h of exposure and further reduced the accumulation thereafter. Mechanisms for the reduced Cd uptake were mainly related to the influences of CNTs on the physiological activity of daphnids. The LC50 of D. magna in the presence of Cd and shorter CNTs was almost the same as that of the control group without CNTs. However, the LC50 of the groups with normal CNTs was significantly higher than that of the control group (i.e., F-CNTs decreased Cd toxicity significantly). Meanwhile, CNTs also decreased the tolerance of D. magna to Cd. The present study suggests that different physical properties of CNTs, such as length, need to be considered in the environmental risk assessment of CNTs. Environ Toxicol Chem 2015;34:2824–2832. © 2015 SETAC

Marine mussels have long been used as biomonitors of contamination of trace elements, but little is known about whether variation in tissue trace elements is significantly associated with those of macronutrients and major cations. The authors examined the variability of macronutrients and major cations and their potential relationships with bioaccumulation of trace elements. The authors analyzed the concentrations of macronutrients (C, N, P, S), major cations (Na, Mg, K, Ca), and trace elements (Al, V, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Ba, Pb) in the whole soft tissues of marine mussels Mytilus edulis and Perna viridis collected globally from 21 sites. The results showed that 12% to 84% of the variances in the trace elements was associated with major cations, and the tissue concentration of major cations such as Na and Mg in mussels was a good proxy for ambient seawater concentrations of the major cations. Specifically, bioaccumulation of most of the trace elements was significantly associated with major cations, and the relationships of major cations with trace cations and trace oxyanions were totally opposite. Furthermore, 14% to 69% of the variances in the trace elements were significantly associated with macronutrients. Notably, more than half of the variance in the tissue concentrations of As, Cd, V, Ba, and Pb was explained by the variance in macronutrients in one or both species. Because the tissue macronutrient concentrations were strongly associated with animal growth and reproduction, the observed coupling relationships indicated that these biological processes strongly influenced the bioaccumulation of some trace elements. The present study indicated that simultaneous quantification of macronutrients and major cations with trace elements can improve the interpretation of biomonitoring data.

The accumulation of metals (especially copper) in oysters has led to green-color now being found in Chinese estuaries. In the present study, the authors quantified the depuration of 8 metals (Ag, Cd, Co, Cr, Cu, Ni, Pb, and Zn) in green-colored oysters (Crassostrea sikamea) collected from an estuary that is heavily contaminated by metals as a result of industrial effluent releases. The oysters were depurated under laboratory conditions for 4 mo; the accumulated concentrations and the subcellular distribution of metals were measured at different time intervals. Results showed that the green color of oysters faded to light yellow (nearly normal) after 4 mo of depuration. Depuration of metals could be described by a first-order kinetic process. The calculated overall depuration rate constants of metals were in the range of 0.008 d−1 to 0.024 d−1, with a biological retention half-life of 30 d to 70 d. The depuration rates of green-colored contaminated oysters were significantly higher for Cd, Cu, Cr, and Ni than the rates of oysters from a less contaminated site, whereas the depuration rates of Ag, Co, Pb, and Zn were comparable between the 2 populations. When corrected for the change of oyster tissue weight, the actual efflux rate constants of the metals (0.0708–0.1014 d−1) were much higher than the overall depuration rate constants. Cellular debris and metallothionein-like proteins were the important fractions binding with the metals in the oysters. Significant changes in metal subcellular distribution were observed during the 4-mo depuration for Ag, Cd, Cu, and Zn. Metallothionein-like protein became more important in sequestering the metals during the depuration period, with a concomitant decrease in metals associated with the cellular debris fraction. Environ Toxicol Chem 2014;33:2379–2385. © 2014 SETAC

In aquatic environments, transformation of pollutants by association with functionalized carbon-based nanoparticles can dramatically change their cycling pathways. The present study quantified the uptake and depuration behavior of cadmium and zinc bound with functionalized fullerene nanoparticles (f-nC60) in a freshwater cladoceran, Daphnia magna, in a well-dispersed medium. Metal uptake proceeded with a linear pattern during the 8-h exposure period, and the uptake rate constants (ku) were 1.3-fold to 1.4-fold higher for Cd or comparable for Zn bound with f-nC60 than those of the free ones. The assimilation efficiencies of Cd and Zn bound with f-nC60 were significantly enhanced when compared with those metals bound with algal food. Furthermore, the depuration of metals bound with f-nC60 was relatively slower compared to the depuration of metals bound with carbon nanotubes. A longer exposure to f-nC60 resulted in an even slower depuration of metals. The authors conclude that metal binding with f-nC60 as modified nanoparticles could serve as a new pathway for the elevated metal accumulation in Daphnia. Environ Toxicol Chem 2014;33:1122–1128. © 2014 SETAC

With increasing use of silver nanoparticles (AgNPs), concerns about their potential deleterious effects on aquatic ecosystems have increased. Most previous studies have focused on the toxicity of AgNPs while their bioavailability has been seldom investigated. The present study examined the effects of salinity on the aggregation kinetics as well as the bioavailability of commercial 80-nm citrate-coated AgNPs (c-AgNPs) in the presence or absence of a nonionic surfactant (Tween 20) to marine medaka (Oryzias melastigma). In addition, the uptake of soluble Ag was quantified for comparison and for deducting the uptake of soluble Ag during AgNP exposure by applying a biokinetic model. The authors found that the addition of Tween 20 immediately slowed down the process of aggregation of AgNPs, and an elevated amount of Tween 20 (20 µM) kept AgNPs well dispersed, even in the 30-psu salinity medium. Uptake rate constants (ku ) of AgNPs were less than half the soluble Ag at low salinities (1 psu and 5 psu), while limited bioavailability of c-AgNPs was observed at high salinities (15 psu and 30 psu). However, the Tween 20-stabilized AgNPs (t-AgNPs) were accumulated by medaka at comparable rates as the soluble Ag, indicating the importance of dispersion for bioavailability of AgNPs in a highly ionic environment. The present study provided the first insight of the bioavailability of AgNPs to fish in a high-ionic environment. More studies are needed to gain a full understanding of bioavailability of AgNPs in marine environments.

Traditional metal toxicity tests on organisms have mainly focused on continuous exposure at a fixed concentration. However, organisms are more likely exposed to pollutants intermittently in estuarine environments that are significantly impacted by anthropogenic activity. The present study examined whether different copper (Cu) exposure regimes at an equivalent dose can induce different metabolomics effects on the oysters. An estuarine oyster Crassostrea hongkongensis was exposed to Cu continuously or intermittently at an equal dose (time × concentration) for 6 wk. Continuous exposure regimes included 2 doses of 3.3 μg/L for 24 h and 20 μg/L for 24 h, with corresponding equal doses of 2 intermittent exposure regimes of 20 μg/L for 4 h and 120 μg/L for 4 h, respectively. Time-course measurements suggested that Cu bioaccumulation was comparable at equal low doses between the continuous regime (3.3 μg/L for 24 h) and the intermittent regime (20 μg/L for 4 h), but there was considerable difference for the high dose under different regimes. Nuclear magnetic resonance (NMR)-based metabolomics suggested that continuous and intermittent Cu exposures led to similar metabolite variation pattern in gills at an equal high dose, including decreased amino acids (e.g., aspartate, glycine, isoleucine, leucine, lysine, phenylalanine, threonine, and valine), lower energy-related compounds (e.g., adenosine triphosphate/adenosine diphosphate, acetate, citrate, and glycogen), and altered osmolytes (e.g., homarine and taurine). These biomarkers indicated disturbance of osmotic regulation and energy metabolism induced by Cu exposure regardless of regime. In addition, the 4-h intermittent Cu exposure resulted in slightly fewer adverse effects compared with the corresponding equal-dose continuous exposure. Oysters appeared to recover during the intervals of Cu exposure. The results indicated that metabolomic effects induced by Cu were more dose dependent than the Cu exposure regime. Environ Toxicol Chem 2017;36:2428-2435. © 2017 SETAC.

General Research Fund from the Hong Kong Research Grants Council [662610]; Key Project from the Natural Science Foundation of China [21237004]

The toxicity of manufactured silver nanoparticles (AgNPs) has been widely studied, but the influence of AgNPs on the major ions (such as sodium [Na] and calcium [Ca]) regulations are unknown. In the present study, a freshwater cladoceran Daphnia magna was exposed to commercial AgNPs coated with polyvinylpyrrolidone. After 48 h, the Na body content was significantly reduced by AgNO3 exposure, but the Ca body content was significantly increased under AgNO3 and AgNP exposures, respectively. No effect was observed on the body concentrations of Na and Ca at 50 to 500 µg/L AgNPs with 1-µM cysteine addition. Exposure of AgNO3 and AgNPs inhibited the Na influx and elevated the Na efflux. In contrast, their exposure increased the Ca influx, but did not affect the Ca efflux. The results of the present study demonstrated the significant influences of AgNO3 and AgNPs (without cysteine) on Na and Ca regulations. Such effect of AgNPs on Na and Ca regulation disappeared after cysteine addition, indicating that the soluble Ag released from AgNPs played a major role in the ionoregulatory dysfunction. Environ. Toxicol. Chem. 2013;32:913–919. © 2013 SETAC

The Pearl River estuary, southern China, suffers from multiple sources of metal contamination as a result of the rapid industrial development in the region; but the biological impacts of contamination remain unknown. In the present study, a euryhaline oyster, Crassostrea hongkongensis, was collected from different sites of the Pearl River estuary; and various physiological (heart rate, alkaline phosphatase as homeostatic regulation, and glycogen as energy reserve) and cytological (lysosomal membrane stability) biomarkers were quantified to assess this species as a potential bioindicator of metal pollution in contaminated areas. Large variations of metal accumulation levels in the oysters were documented, especially for copper (Cu), zinc (Zn), cadmium (Cd), chromium, and nickel (Ni). Among these metals, the authors demonstrated significant correlations between the digestive gland metal accumulation of Cu, Zn, and Ni and the cellular homeostasis (alkaline phosphatase) and glycogen reserves. Heart rate was positively correlated with Cd but negatively correlated with Cu and Zn concentrations in the gills. Lysosomal membrane stability was significantly inhibited at the most contaminated sites but had no relationship with the accumulated metal concentrations. These measurements indicate that multimetal contamination in the Pearl River estuary impacts the physiological and cytological performance of oysters. Environ Toxicol Chem 2016;35:2577-2586. © 2016 SETAC.

The contaminated oysters discovered in the Pearl River Estuary (Guangdong province, China) contained high levels of metals in their tissues, especially Cu and Zn, indicating that this large and densely urbanized estuary in Southern China suffers from serious metal pollution. The present study aimed to investigate the impacts of multimetal pollution in the Pearl River Estuary on oyster antioxidant and detoxification systems. The responses of various biochemical biomarkers in the ecologically important oyster Crassostrea hongkongensis collected from 7 sites in the Pearl River Estuary were quantified. Significant correlations were demonstrated between the accumulation of Cu and Zn and oxidative stress (lipid peroxidation) and oxidative stress defenses (catalase, glutathione peroxidase) in the oyster gills. Significant correlations between the accumulation of Cd and Cu and detoxification (glutathione and glutathione transferase) in the gills were also documented. Interestingly, metallothionein concentrations were positively correlated with Cd, but negatively correlated with Cu, Ni, and Zn concentrations in the gills. These measurements indicated that Cu in the Pearl River Estuary induced various biochemical responses in the oysters and influenced the susceptibility of oysters to environmental stress. The present study has provided the first evidence of antioxidant and detoxification responses in native contaminated oysters from a field environment seriously contaminated by metals. Coupling biomarkers with tissue metal concentration measurements was a promising approach to identify the metals causing biological impacts in a multimetal-contaminated estuary. Environ Toxicol Chem 2016;35:2798-2805. © 2016 SETAC.

Bivalves are employed widely as biomonitors of metal pollution and proteomics has increasingly been applied to solve ecotoxicological issues. This study aimed to investigate the effects of Cd exposure on the bioaccumulation of other trace elements and reveal the molecular mechanisms using proteomics technologies. The results showed that Cd exposure resulted in remarkable changes in body concentrations of Zn, Cu, Ag, Co, Ni, Pb, and Se in four marine bivalves (scallop Chlamys nobilis, clam Ruditapes philippinarum, mussel Perna viridis, and oyster Saccostrea cucullata). Generally, the bivalves exposed to higher Cd concentration accumulated higher concentrations of Zn, Cu, and Se, but a lower concentration of Co. The accumulation of Ag, Ni, and Pb was specific for different species. The data strongly suggest that the influences of one metal exposure on the bioaccumulation of other metals/metalloids need to be considered in interpreting body concentrations of the elements in the biomonitors. Cd exposure had little effect on bivalve proteomes, and the identified proteins were insufficient to explain the observed disruption of trace element metabolism. However, protein expression signatures composed of the altered proteins could distinguish the clams and the mussels with different body Cd levels. The strong up-regulation of galectin in Cd-exposed oysters indicated the protein as a novel biomarker in environmental monitoring.

In the present study, the dissolved uptake, dietary assimilation, and elimination of arsenic (initially added as arsenate) in the freshwater cladoceran Daphnia magna were examined. A biphasic correlation between the arsenic uptake rate and its ambient concentration, as well as a two-saturation-site arsenic uptake competition with phosphate was observed. The calculated uptake rate constant, as influenced by the ambient phosphorus concentration, ranged from 0.035 to 0.35 L/g/d. Food concentration substantially decreased (by 23.2-64.4%) the arsenic assimilation efficiency with the incipient limiting algal food concentration of 3.86 mg/L dry weight. Arsenic assimilation by the daphnids was independent of their own phosphorus status, but was lower when their algal diet was phosphorus-limited and thus contained a higher proportion of arsenite due to the enhanced biotransformation. Arsenic efflux rate constant ranged from 0.34 to 0.44 d � 1 with increased food concentration slightly facilitating its loss. Excretion, accounting for 51.3 to 60.6% of total loss, was the dominant pathway for arsenic elimination with a remarkable contribution from offspring production (24.7- 29.8%), whereas molting (3.64-4.05%) and egestion (7.9-11.9%) had minor roles only. According to the well-established biokinetic model, dietary assimilation was predicted to be the main pathway for arsenic bioaccumulation in the daphnids, and arsenic has a great potential to be biodiminished along the food chain. Environ. Toxicol. Chem. 2012;31:1283-1291. # 2012 SETAC

The harpacticoid copepod Tigriopus japonicus has a wide geographical distribution and is considered as a suitable model species for the assessment of toxicity of marine pollutants. The aim of the present study was to test the impacts of polychlorinated biphenyl (PCB) 126 (3,3',4,4',5-pentachlorobiphenyl) on the growth, development, and reproduction of T. japonicus in two successive generations. We first quantified the 96-h 50% lethal concentration (2.83 mg/L; all reported concentrations are nominal values), the no-observed-effect concentration (0.6 mg/L), and the lowest-observed-effect concentration (LOEC; 1.2 mg/L) of PCB126 in the nauplii. Nauplii were more sensitive than the adults, which still survived at the highest tested PCB126 concentration (8 mg/L). In the chronic toxicity testing, 10 life history traits were quantified for T. japonicus. No obvious effect on any of these traits was observed in the first generation (F0) at tested concentrations (100 µg/L) lower than the LOEC. During the second generational life-cycle exposure (F1), however, PCB126 had an obvious toxic effect on the reproduction (1 µg/L) and growth (0.1 µg/L). Thus, copepods became more sensitive to PCB126 exposure as generations developed. Among the different traits tested, body size was the most sensitive parameter. Reproduction (fecundity, number of clutches, nauplii/clutch) and intrinsic population growth were also significantly impacted by PCB exposure. The survivorship, sex ratio, hatching time, and development were not affected. Environmental risk assessment of contaminants must therefore be based on a long-term multigenerational exposure to provide a realistic measurement of the influences of pollutants on aquatic life.

Metal pollution levels in estuarine and coastal environments have been widely reported, but few documented reports exist of severe contamination in specific environments. Here, we report on a metal-contaminated estuary in Fujian Province, China, in which blue oysters (Crassostrea hongkongensis) and green oysters (Crassostrea angulata) were discovered to be contaminated with Cu and other metals. Extraordinarily high metal concentrations were found in the oysters collected from the estuary. Comparison with historical data suggests that the estuary has recently been contaminated with Cr, Cu, Ni, and Zn. Metal concentrations in blue oysters were as high as 1.4 and 2.4% of whole-body tissue dry wt for Cu and Zn, respectively. Cellular debris was the main subcellular fraction binding the metals, but metal-rich granules were important for Cr, Ni, and Pb. With increasing Cu accumulation, its partitioning into the cytosolic proteins decreased. In contrast, metallothionein-like proteins increased their importance in binding with Zn as tissue concentrations of Zn increased. In the most severely contaminated oysters, only a negligible fraction of their Cu and Zn was bound with the metal-sensitive fraction, which may explain the survival of oysters in such contaminated environments.

Silver nanoparticles (AgNP) are now widely used as antibacterial products, and their potential toxicities in aquatic organisms are a matter of increasing concern. In the present study, we conducted experiments to reveal the acute and chronic toxicities of AgNP and its bioaccumulation from both aqueous and dietary sources in a model freshwater cladoceran, Daphnia magna. No mortality was observed in 48-h acute toxicity testing when the daphnids were exposed up to 500 µg Ag/L as AgNP. The AgNP accumulation reached as high as 22.9 mg Ag/g dry weight at the highest AgNP concentration tested (500 µg/L). In contrast, D. magna was extremely sensitive to free Ag ion (Ag+, added as AgNO3), with a measured 48-h 50% lethal concentration of 2.51 µg/L. Thus, any AgNP potential acute toxicity may be caused by the release of Ag+ into the solution. During the 21-d chronic exposure, dietborne AgNO3 had the most significant influence on reproduction, whereas waterborne AgNP had the most significant inhibition on growth. Significant delay and decrease of reproduction in daphnids exposed to dietborne AgNO3 occurred at a dissolved Ag concentration of 0.1 µg/L added to the algae. Significant inhibitions of growth and reproduction were also found for the AgNP exposure, with the lowest observed effective concentration of 5 µg/L and 50 µg/L, respectively. Chronic effects of AgNP were probably caused by the low food quality of algae associated with AgNP and the low depuration of ingested AgNP. Environmental risk assessments of AgNP should therefore include tests on the chronic toxicity to aquatic organisms as well as the direct and indirect effects of AgNP resulting from the release of Ag+ into the environment. Environ. Toxicol. Chem. 2011; 30:885–892. © 2010 SETAC

Metallothionein-like proteins (MTLPs) are important inducible ligands for Cd detoxification, but whether MTLPs are involved in Cd bioaccumulation remains unclear. The present study investigated the potential interrelationships between MTLP turnover and Cd biokinetics in the scallop Chlamys nobilis with or without dissolved Cd pre-exposure. After Cd pre-exposure, increased synthesis rate and standing stock concentration of MTLPs as well as increased dissolved Cd uptake rate, were found in C. nobilis. However, the level of newly sequestrated Cd by MTLPs in the pre-exposed scallops was similar to that of the controls, indicating that MTLPs had no effect on Cd uptake from the dissolved phase. Instead, Cd uptake rate was positively related to the Cd binding rate of cellular debris and organelles. In addition, no significant difference was observed in the Cd dietary assimilation efficiency of the scallops, showing that MTLPs played a minimal role in Cd uptake from the dietary phase. Conversely, the Cd efflux rate constant decreased by 68 to 75% in the pre-exposed scallops in comparison with that in the control. Correspondingly, progressive elevations of MTLP-bound Cd and standing stock MTLP concentration were observed in the pre-exposed individuals during the Cd elimination period. In the pre-exposed scallops, the faster de novo MTLP synthesis could remobilize and recycle more Cd from other intracellular pools, effectively reducing the Cd elimination rate. The present study thus showed the differential roles of MTLPs in Cd uptake and elimination in the bivalves. Environ. Toxicol. Chem. 2011; 30:738–746. © 2011 SETAC

Microcystis aeruginosa is a dominant cyanobacterium commonly found in Chinese freshwater ecosystems during phytoplankton blooms, and copper sulfate is one of the most frequently used algicides for controlling the nuisance blooms. In this study, we examined the effects of varied water chemistry (dissolved organic matter, pH, and hardness) on copper (Cu) toxicity to M. aeruginosa, as well as the Cu short-term uptake kinetics, using 67Cu as a radioactive tracer. Elevated dissolved organic matter concentrations resulted in a reduction of Cu toxicity, and increasing pH in the range 6.7 to 8.5 led to an increase of Cu toxicity based on the ambient dissolved Cu concentration. The variation of growth inhibition was much more dependent on the intracellular Cu concentration than on the total ambient Cu or calculated free Cu2+ concentration; thus, the biotic ligand model could be used to explain the Cu toxicity to M. aeruginosa under different water chemistry conditions. We demonstrated that M. aeruginosa were extremely sensitive to Cu toxicity compared with other freshwater phytoplankton species based on the intracellular Cu concentration. The median inhibition concentration of intracellular Cu was as low as 3.3 to 13.1 × 10−18 mol Cu/cell for all treatments. The Cu short-term uptake kinetics in M. aeruginosa could be explained by the free-ion activity model. The uptake rate, however, could not explain the discrepancy in Cu sensitivity between M. aeruginosa and other phytoplankton species. Other mechanisms might explain the extreme sensitivity of M. aeruginosa to Cu toxicity. Environ. Toxicol. Chem. 2010;29:2260–2268. © 2010 SETAC

Subcellular metal distribution may play an important role in the bioaccumulation and trophic transfer of metals in marine food chains. In the present study, we preexposed the green mussel Perna viridis to Ag and Cd and quantified their trophic transfer efficiencies to two predators (whelks [Thais clavigera] and fish [Terapon jarbua]). For the mussels, more Ag was distributed in the metal-rich granule (MRG) fraction following Ag exposure, and more Cd was distributed in the metallothionein-like protein following Cd exposure. In addition, Cd was mainly bound with the proteins having a molecular size of approximately 20 kDa. After being fed with metal-exposed mussels, the assimilation efficiencies of Ag decreased significantly (from 77 to 29-60% in whelks and from 9 to 2% in fish) with an increasing percentage of Ag deposited in the MRG fraction of the prey. In contrast, the assimilation efficiencies of Cd remained comparable (81-85% in whelks and 6-8% in fish), because its partitioning in the soluble fraction of different treatments of the prey was similar. The efflux of Ag and Cd in the two predators was comparable after feeding on preexposed mussel prey. Our results imply that the subcellular distribution of metals in prey may affect the dietary assimilation of metals in predators, but such influence is clearly metal-specific. The present study may lead to a better understanding of metal trophic transfer in different marine food chains.

Low mercury (Hg) concentrations down to several nanograms Hg per gram of wet tissue are documented in certain fish species such as herbivorous fish, and the underlying mechanisms remain speculative. In the present study, bioaccumulation and depuration patterns of inorganic Hg(II) and methylmercury (MeHg) in a herbivorous rabbitfish Siganus canaliculatus were investigated at organ and subcellular levels following waterborne or dietary exposures. The results showed that the efflux rate constants of Hg(II) and MeHg were 0.104 d(-1) and 0.024 d(-1) , respectively, and are probably the highest rate constants recorded in fish thus far. The dietary MeHg assimilation efficiency (68%) was much lower than those in other fish species (∼90%). The predominant distribution of MeHg in fish muscle was attributable to negligible elimination of MeHg from muscle ( 0) and efficient elimination of MeHg from gills (0.12 d(-1) ), liver (0.17 d(-1) ), and intestine (0.20 d(-1) ), as well as efficient transportation of MeHg from other organs into muscle. In contrast, Hg(II) was much more slowly distributed into muscle but was efficiently eliminated by the intestine (0.13 d(-1) ). Subcellular distribution indicated that some specific membrane proteins in muscle were the primary binding pools for MeHg, and both metallothionein-like proteins and Hg-rich granules were the important components in eliminating both MeHg and Hg(II). Overall, the present study's results suggest that the low tissue Hg concentration in the rabbitfish was partly explained by its unique biokinetics. Environ Toxicol Chem 2016;35:2074-2083. © 2016 SETAC.

With increasing application and commercial production, carbon nanotubes (CNTs) will inevitably be released into aquatic environments and affect the transport and toxicity of toxic metals in ecosystems. The present study examined how CNTs affected the biokinetics and toxicity of a toxic metal, cadmium (Cd), in the freshwater zooplankton Daphnia magna. The authors quantified the dissolved uptake and the 50% lethal concentration (LC50, 48 h and 72 h) of Cd in daphnids in the presence of functionalized multiwalled nanotubes (F-CNTs) with different lengths (10-30 µm vs 0.5-2 µm) and concentrations (4 mg/L and 8 mg/L). Compared with the control treatment without CNTs, both CNTs slowed down the accumulation rate of Cd in D. magna over 8 h of exposure and further reduced the accumulation thereafter. Mechanisms for the reduced Cd uptake were mainly related to the influences of CNTs on the physiological activity of daphnids. The LC50 of D. magna in the presence of Cd and shorter CNTs was almost the same as that of the control group without CNTs. However, the LC50 of the groups with normal CNTs was significantly higher than that of the control group (i.e., F-CNTs decreased Cd toxicity significantly). Meanwhile, CNTs also decreased the tolerance of D. magna to Cd. The present study suggests that different physical properties of CNTs, such as length, need to be considered in the environmental risk assessment of CNTs.

Marine mussels have long been used as biomonitors of contamination of trace elements, but little is known about whether variation in tissue trace elements is significantly associated with those of macronutrients and major cations. The authors examined the variability of macronutrients and major cations and their potential relationships with bioaccumulation of trace elements. The authors analyzed the concentrations of macronutrients (C, N, P, S), major cations (Na, Mg, K, Ca), and trace elements (Al, V, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Ba, Pb) in the whole soft tissues of marine mussels Mytilus edulis and Perna viridis collected globally from 21 sites. The results showed that 12% to 84% of the variances in the trace elements was associated with major cations, and the tissue concentration of major cations such as Na and Mg in mussels was a good proxy for ambient seawater concentrations of the major cations. Specifically, bioaccumulation of most of the trace elements was significantly associated with major cations, and the relationships of major cations with trace cations and trace oxyanions were totally opposite. Furthermore, 14% to 69% of the variances in the trace elements were significantly associated with macronutrients. Notably, more than half of the variance in the tissue concentrations of As, Cd, V, Ba, and Pb was explained by the variance in macronutrients in one or both species. Because the tissue macronutrient concentrations were strongly associated with animal growth and reproduction, the observed coupling relationships indicated that these biological processes strongly influenced the bioaccumulation of some trace elements. The present study indicated that simultaneous quantification of macronutrients and major cations with trace elements can improve the interpretation of biomonitoring data.

The discovery of colored oysters in an estuary in Southern China raised questions of how these oysters could survive in such an unpredictable estuary. In the present study, the authors conducted a transplant experiment using oysters Crassostrea hongkongensis in the estuary to quantify the changes of metal biokinetics (Cd and Zn) and the responses of metallothionein-like proteins. Oysters in the transplantation experiment accumulated extremely high concentrations of Cd, Cu, Cr, Ni, and Zn. The present study provided the field evidence of the time-course coupling relationship between Cd and Zn bioaccumulation. Over the 2-mo transplantation, the uptake rate constants of Cd and Zn varied greatly during the early stage of exposure but subsequently became comparable among the different locations. The dietary Zn assimilation remained relatively constant, whereas the Cd assimilation increased gradually with increasing period of exposure. No notable difference was seen in dietary metal assimilation among the different locations. In contrast, the efflux rates quantified by the end of transplantation were 26% to 42% higher for Cd and 12% to 37% higher for Zn than that in the unexposed oysters. The authors further demonstrated that the incoming Cu and Zn were not stored in metallothionein-like pools and that metallothionein-like proteins synthesis and breakdown were little affected by metal exposure. Overall, the authors' transplantation experiments suggested that oysters displayed a rather weak ability to modify their biokinetics and metallothionein turnover under metal exposure. Environ Toxicol Chem 2015;34:1818–1825. © 2015 SETAC

Silver (Ag) is an important metal contaminant in many coastal waters and often is accompanied by high nutrient concentrations in the effluent outfall. The biological uptake of Ag by the marine diatom Thalassiosira pseudonana at various levels of nitrate, ammonium, and phosphate was examined under different growth conditions of the cells. The biological uptake of Ag increased significantly with increasing concentrations of nitrate, ammonium, and phosphate, presumably because of the increase in algal growth rate or increasing sulfur-containing ligands available for Ag transport. The calculated Ag uptake rate constants increased by 3 to 16 times with increasing nitrate and ammonium concentration from 5.88 microM to 176 microM. The assimilation efficiency (AE) of Ag by the coastal copepod Acartia spinicauda was quantified under different ecological and chemical conditions of the diatom prey. The Ag AEs were in the range of 3 to 23%, and increased with decreasing diatom food concentrations or when the diatoms were grown at a lower nutrient level. Significant correlations were demonstrated between the AE and the distribution of Ag in the diatom's cytoplasm, the Ag concentration factor in the diatoms, Ag retention in the particles during the feeding period, and the Ag gut passage time in the copepods. Desorption within the copepod's gut appears to play a critical role in Ag assimilation and partially accounts for the variability of Ag AEs under different food and geochemical conditions. Our study highlights that several geochemical and physiological processes all significantly affect Ag trophic transfer in marine copepods. Given the dependence of Ag transfer on its concentration in ingested particles and food concentration, dietary uptake of Ag is probably variable in natural environments. The influence of eutrophication on Ag trophic transfer is dependent on the degree to which each kinetic parameter is affected by nutrient enrichments.

Kinetic uptake of Cd, Se(IV), and Zn by Daphnia magna from the dissolved phase was determined using radiotracer techniques in moderately hard water. The metal influx rate and distribution in the soft tissue and the exoskeleton of the daphnids as influenced by metal concentration, inorganic ligands including pH, Ca2+ and SO42−, and body size were quantified. When the metal concentrations were 50%). For Cd and Se, dietary exposure is dominant when the bioconcentration factors of these metals in phytoplankton are at the high end.

The absorption efficiency (AE) of benzo[a]pyrene (BaP), a representative polycyclic aromatic hydrocarbon compound, in the green mussel Perna viridis was measured under different biological and food conditions (body size, food quantity, and food species). Uptake of BaP from the aqueous phase by different species of phytoplankton and green mussels was also experimentally quantified. A bioenergetic-based kinetic model was then used to separate the exposure pathways of BaP accumulation in the green mussels. Our results demonstrated that the AE of BaP in the green mussels ranged between 17 and 47%. Body size did not significantly affect the AE of BaP in the mussels. The AEs increased with a decrease in food availability, largely as a result of lengthening the gut passage of chemicals. The AEs differed by up to 2.3 times (17-39%) among the different food diets tested (four phytoplankton diets and one sediment diet). The BaP associated with the sediment was assimilated least efficiently. The difference in gut passage also appeared to account partially for the difference in AEs observed among different food types. No correlation between the AE and the distribution of BaP in the cytoplasm of phytoplankton cells was found. The concentration factor of BaP measured in four different marine phytoplankton species was in the range of 10(4)-5 x 10(4) (L/kg). The calculated uptake rate constant of BaP in the green mussels was 6.37 L/g/d, and the efficiency of absorption of BaP from the aqueous phase was 6.6 to 8.8%. The model calculation indicated that under most circumstances, >56% of BaP in mussels can indeed be derived from the uptake from the aqueous phase. However, the relative importance of aqueous and dietary exposure can be affected by the variation of AE, mussel's ingestion rate, and concentration of BaP in ingested food particles.

The accumulation of metals (especially copper) in oysters has led to green-color now being found in Chinese estuaries. In the present study, the authors quantified the depuration of 8 metals (Ag, Cd, Co, Cr, Cu, Ni, Pb, and Zn) in green-colored oysters (Crassostrea sikamea) collected from an estuary that is heavily contaminated by metals as a result of industrial effluent releases. The oysters were depurated under laboratory conditions for 4 mo; the accumulated concentrations and the subcellular distribution of metals were measured at different time intervals. Results showed that the green color of oysters faded to light yellow (nearly normal) after 4 mo of depuration. Depuration of metals could be described by a first-order kinetic process. The calculated overall depuration rate constants of metals were in the range of 0.008 d(-1) to 0.024 d(-1) , with a biological retention half-life of 30 d to 70 d. The depuration rates of green-colored contaminated oysters were significantly higher for Cd, Cu, Cr, and Ni than the rates of oysters from a less contaminated site, whereas the depuration rates of Ag, Co, Pb, and Zn were comparable between the 2 populations. When corrected for the change of oyster tissue weight, the actual efflux rate constants of the metals (0.0708-0.1014 d(-1) ) were much higher than the overall depuration rate constants. Cellular debris and metallothionein-like proteins were the important fractions binding with the metals in the oysters. Significant changes in metal subcellular distribution were observed during the 4-mo depuration for Ag, Cd, Cu, and Zn. Metallothionein-like protein became more important in sequestering the metals during the depuration period, with a concomitant decrease in metals associated with the cellular debris fraction.

Several important sediment geochemical factors (particle size, contact time, artificial sediment composition, acid volatile sulfide, and pure inorganic particles coated or uncoated with humic acid) were examined for their influences on the assimilation of Cd, Cr, and Zn in the marine clam Ruditapes philippinarum. Metal desorption and phase speciation were concurrently measured for these particles. Differences in particle size did not significantly affect metal assimilation. Increasing contact time with sediments decreased assimilation by the clams of Cr and Zn, but not of Cd. A significant positive relationship was found between the metal assimilation efficiency (AE) and the metal partitioning in the exchangeable fraction of sediments for Cd and Zn. The assimilation of metals was negatively dependent on the metal distribution in the reducible (Fe oxide) fraction, indicating that Fe oxide reduced metal bioavailability to the clams. Metal AEs measured with artificially prepared sediments were comparable to those measured with natural sediments. The AEs were significantly higher in artificial sediment without Fe oxide. Humic acid coating appeared to have no major and consistent influence on metal assimilation by the clams. The AEs measured for pure inorganic particles were much higher than those measured for sediments or artificially prepared sediments, except for the Fe oxide particles, indicating that digestion may behave differently for pure particles. Metals bound to artificially synthesized acid volatile sulfide were much less bioavailable to the clams. The AEs were as low as 4% for Cd and 7% for Zn, whereas the AE for Cr was somewhat similar to those measured for other types of particles. When all particle types (natural sediments, artificial sediment, and pure particles) were considered, a significant relationship was found between Cd AE and its desorption in seawater. This study has demonstrated that several geochemical fractions of the sediments, particularly the exchangeable, Fe oxide, and sulfide fractions, are important in determining metal bioavailability from sediments to clams.

Recent studies have quantified extensively metal assimilation efficiency from ingested food sources in aquatic invertebrates. Metal absorption efficiency (alpha) from the dissolved phase is analogous to metal assimilation efficiency, but it remains poorly defined and quantified. In this study, the alpha of four trace metals [Cd, Cr(VI), Se(IV), and Zn] was determined in three species of marine bivalves (green mussel [Perna viridis], black mussel [Septifer virgatus], and clam [Ruditapes philippinarum]). Individual bivalves were first measured for their clearance rates, followed by measurements of the metal influx rate, after which the metal alpha and the uptake rate constant (Ku) were then computed. Among the four metals considered, the highest Ku and alpha were found for Zn, followed by Cd > Cr(VI) > Se(IV). The Ku values were comparable between the two mussels but were 1.8- to 3.3-fold lower in the clams. Interspecific difference in metal Ku was strongly related to, but intraspecific difference in Ku was not affected by, the bivalve's clearance rate. Interspecific difference in metal alpha was smaller than the metal Ku and was independent of the clearance rate, whereas the intraspecific difference in metal alpha correlated with the individual variations of the clearance rate. Within each bivalve species, a significant negative correlation was found between the metal alpha and the clearance rate, implying that an individual pumping a greater amount of water was coupled to a lower alpha. Significant correlation between the alpha of four metals was also documented in all three species of bivalves. Thus, metal bioavailability from the aqueous phase was directly related to the physiological conditions of the animals. Both the aqueous chemistry and the physiology of the animals can be important in affecting metal bioavailability from the dissolved phase.

The green mussel Perna viridis and the clam Ruditapes philippinarum have been frequently used as biomonitors of coastal contamination in subtropical and tropical waters, yet the physiological processes controlling metal uptake in these bivalves are unknown. Assimilation efficiency (AE) is an important physiological parameter quantifying metal bioavailability from ingested food. The authors determined the AEs of Cd, CR, and Zn in these bivalves feeding on five species of phytoplankton and one natural section. The influences of the cytoplasmic distribution of metals in the algal cells and the digestive physiology of bivalves on metal AEs were also examined. Among the three metals, Zn was generally assimilated at the highest efficiency, i.e., 21 to 36% in the mussels and 29 to 59% in the clams. Cr was the least assimilated metal, with AEs being 10 to 16% in the mussels and 11 to 24% in the clams. The AEs of Cd and Zn in the clams were 1.8 to 4.7 and 1.1 to 1.9 times higher, respectively, than the AEs in the mussels. Assimilation efficiencies of Cr were, however, comparable between the mussels and the clams. A positive significant relationship between the metal AE and the percent of metals in the algalmore » cytoplasm was found only for Cd in the clams, suggesting that Cd fractionation in the algal cells influenced its assimilation. No significant relationship, however, was found for other metals in both bivalves. A significant relationship between Cr-assimilation efficiency and gut passage time (GPT) was documented in the mussels, indicating a higher assimilation when Cr was retained longer in the gut. There was also significant correlation of metal AEs among the three metals, which were probably subjected to the same digestive pathway in the bivalves. Their study demonstrated that both the green mussels and the clams were able to accumulate metals from ingested food source, and food quality appeared to have different effects on metal assimilation in different bivalve species.« less

Assimilation efficiencies of contaminants from ingested food are critical for understanding chemical accumulation and trophic transfer in aquatic invertebrates. Assimilation efficiency is a first-order physiological parameter that can be used to systematically compare the bioavailability of different contaminants from different foods. The various techniques used to measure contaminant assimilation efficiencies are reviewed. Pulse-chase feeding techniques and the application of gamma-emitting radiotracers have been invaluable in measuring metal assimilation efficiencies in aquatic animals. Uniform radiolabeling of food is required to measure assimilation, but this can be difficult when sediments are the food source. Biological factors that influence contaminant assimilation include food quantity and quality, partitioning of contaminants in the food particles, and digestive physiology of the animals. Other factors influencing assimilation include the behavior of the chemical within the animal's gut and its associations with different geochemical fractions in food particles. Assimilation efficiency is a critical parameter to determine (and to make predictions of) bioaccumulation of chemicals from dietary exposure. Robust estimates of assimilation efficiency coupled with estimates of aqueous uptake can be used to determine the relative importance of aqueous and dietary exposures. For bioaccumulation of metals from sediments, additional studies are required to test whether metals bound to the acid-volatile sulfide fraction of sediments can be available to benthic deposit-feeding invertebrates. Most assimilation efficiency studies have focused on chemical transfer in organisms at the bottom of the food chain; additional studies are required to examine chemical transfer at higher trophic levels.

The authors explore the consequences of consumer-resource interactions for bioaccumulation in two aquatic systems (cadmium accumulation in Dreissena polymorpha and polychlorinated biphenyl accumulation in calanoid copepods). They explicitly link the feeding and growth rates of consumers to the abundance of resources under a variety of assumptions about the nature of the interactions between them. The models are parameterized using field and laboratory data, and predictions are quantitatively compared with field-measured distributions of tissue concentrations. Different assumptions about consumer-resource interactions result in different predicted distributions of tissue concentrations and illustrate the way these interactions constrain the bioaccumulation of contaminants. Linking feeding and growth rates to resource abundances will be important whenever these abundances change over time. User-friendly software will make these ideas accessible to nontheoreticians.

We review recent progress in understanding the trophic transfer of silver (Ag) in marine herbivores, especially mussels that have been extensively used as biomonitors of coastal contamination. A bioenergetic-based kinetic model is invaluable in predicting the trophic transfer and bioaccumulation of Ag in aquatic animals. Critical parameters that need to be quantified in predicting trophic transfer include Ag assimilation efficiency (AE) from ingested food particles, animal feeding rates, and Ag efflux rates. Silver AEs in marine herbivores are generally low (< 30%). Assimilation efficiencies from ingested sediments tend to be lower than those from ingested phytoplankton. Various biological and chemical factors, including Ag distribution in phytoplankton cytoplasm, gut passage time, importance of intracellular versus extracellular digestion, and metal desorption at lowered pHs typical of invertebrate guts, all influence Ag assimilation from ingested particles. Many experimental studies show that uptake from the dissolved phase exceeds uptake via ingestion in the overall Ag bioaccumulation in aquatic animals. However, these results are probably not predictive of field situations due to their simplistic experimental conditions in which fluctuations of feeding conditions of animals and physicochemistry of Ag are not considered. In mussels, the kinetic model predicts that either the solute or particulate pathway can dominate Ag overall uptake in nature, and this is dependent on Ag partition coefficients for suspended particles and Ag AE. Silver is the only metal that varies substantially in the importance of different uptake pathways due to its very high particle reactivity and high uptake rate from the dissolved phase. Total suspended solids (TSS) loads can sharply affect Ag bioaccumulation in mussels because high TSS loads can dilute Ag concentrations in both dissolved and particulate phases. Processes affecting Ag trophic transfer and bioaccumulation are discussed.

In aquatic environments, transformation of pollutants by association with functionalized carbon-based nanoparticles can dramatically change their cycling pathways. The present study quantified the uptake and depuration behavior of cadmium and zinc bound with functionalized fullerene nanoparticles (f-nC(60)) in a freshwater cladoceran, Daphnia magna, in a well-dispersed medium. Metal uptake proceeded with a linear pattern during the 8-h exposure period, and the uptake rate constants (ku) were 1.3-fold to 1.4-fold higher for Cd or comparable for Zn bound with f-nC(60) than those of the free ones. The assimilation efficiencies of Cd and Zn bound with f-nC(60) were significantly enhanced when compared with those metals bound with algal food. Furthermore, the depuration of metals bound with f-nC(60) was relatively slower compared to the depuration of metals bound with carbon nanotubes. A longer exposure to f-nC(60) resulted in an even slower depuration of metals. The authors conclude that metal binding with f-nC(60) as modified nanoparticles could serve as a new pathway for the elevated metal accumulation in Daphnia.

With increasing use of silver nanoparticles (AgNPs), concerns about their potential deleterious effects on aquatic ecosystems have increased. Most previous studies have focused on the toxicity of AgNPs while their bioavailability has been seldom investigated. The present study examined the effects of salinity on the aggregation kinetics as well as the bioavailability of commercial 80-nm citrate-coated AgNPs (c-AgNPs) in the presence or absence of a nonionic surfactant (Tween 20) to marine medaka (Oryzias melastigma). In addition, the uptake of soluble Ag was quantified for comparison and for deducting the uptake of soluble Ag during AgNP exposure by applying a biokinetic model. The authors found that the addition of Tween 20 immediately slowed down the process of aggregation of AgNPs, and an elevated amount of Tween 20 (20 µM) kept AgNPs well dispersed, even in the 30-psu salinity medium. Uptake rate constants (ku ) of AgNPs were less than half the soluble Ag at low salinities (1 psu and 5 psu), while limited bioavailability of c-AgNPs was observed at high salinities (15 psu and 30 psu). However, the Tween 20-stabilized AgNPs (t-AgNPs) were accumulated by medaka at comparable rates as the soluble Ag, indicating the importance of dispersion for bioavailability of AgNPs in a highly ionic environment. The present study provided the first insight of the bioavailability of AgNPs to fish in a high-ionic environment. More studies are needed to gain a full understanding of bioavailability of AgNPs in marine environments.

In the present study, the authors used a supply of naturally contaminated oysters to investigate how the subcellular metal distribution and the metal burden in prey affected the transfer of metals to a marine fish, the grunt Terapon jarbua. The oysters, Crassostrea hongkongensis, each with different contamination histories, were collected and separated into 3 subcellular fractions: 1) metal-rich granules, 2) cellular debris, and 3) a combined fraction of organelles, heat-denatured proteins, and metallothionein-like proteins, defined as the trophically available metal (TAM). These purified fractions showed a wide range of metal concentrations and were fed to the fish for a period of 7 d at a daily comparable feeding rate of 3% of fish body weight. After 7 d exposure, the newly absorbed metals were mainly distributed in the intestine and liver, indicating a significant tissue-specific trophic transfer, especially for Cd and Cu. The trophic transfer factors (TTFs) showed a sequence of cellular debrisTAM metal-rich granules, suggesting the impact of subcellular distribution in prey on metal bioavailability. However, significant inverse relationships between the TTFs and the metal concentrations in diets were also found in the present study, especially for Cd and Zn. The subcellular metal compartmentalization might be less important than the metal concentration in prey influencing the trophic transfer. The authors' results have important implications for bioavailability and environmental assessment of dietary metals.

The relationships among the uptake of toxic methylmercury (MeHg) and two important fish physiological processes-respiration and water pumping--in the Nile tilapia (Oreochromis niloticus) were explored in the present study. Coupled radiotracer and respirometric techniques were applied to measure simultaneously the uptake rates of MeHg, water, and oxygen under various environmental conditions (temperature, dissolved oxygen level, and water flow). A higher temperature enhanced MeHg influx and the oxygen consumption rate but had no effect on the water uptake, indicating the influence of metabolism on MeHg uptake. The fish showed a high tolerance to hypoxia, and the oxygen consumption rate was not affected until the dissolved oxygen concentration decreased to extremely low levels (below 1 mg/L). The MeHg and water uptake rates increased simultaneously as the dissolved oxygen level decreased, suggesting the coupling of water flux and MeHg uptake. The influence of fish swimming performance on MeHg uptake was also investigated for the first time. Rapidly swimming fish showed significantly higher uptake rates of MeHg, water, and oxygen, confirming the coupling relationships among respiration, water pumping, and metal uptake. Moreover, these results support that MeHg uptake is a rate-limiting process involving energy. Our study demonstrates the importance of physiological processes in understanding mercury bioaccumulation in fluctuating aquatic environments.

We examined the subcellular partitioning of zinc (Zn) in Daphnia magna both under acute and chronic exposures. In the acute Zn toxicity tests, the daphnids were exposed to different Zn concentrations for 48 h or to one lethal concentration (1,000 µg/L) for different durations (time to death for up to 47 h). Significant mortality of daphnids was observed when the newly accumulated Zn concentration reached a threshold level of approximately 40 µg/g wet weight (or 320 µg/g dry wt), approximately 3.5 times higher than the background tissue concentration (92 µg/g dry wt). Chronic exposure (14 d) to Zn resulted in nonobservable effect on survivorship and growth at newly accumulated tissue concentration of over 40 µg/g wet weight. With increasing Zn acute exposure, more Zn was partitioned into the cellular debris fraction, indicating that this fraction was presumably the first targeted site of binding for Zn upon entering the animals. The importance of other subcellular fractions either decreased accordingly or remained comparable. We found that the metal-sensitive fraction (Zn distribution in the organelles and heat-denatured proteins) did not predict the acute Zn toxicity in Daphnia. During chronic exposure, however, no major change of the subcellular partitioning of Zn with increasing Zn exposure was documented. Zinc was mainly found in the organelles and heat-stable protein fractions during chronic exposure, suggesting that any subcellular repartitioning occurred primarily during acute exposure. Metallothioneins were induced upon chronic Zn exposure, but its induction evidently lagged behind the Zn accumulation. Our present study showed that the subcellular fractionation approach could not be readily used to predict the acute and chronic toxicities of Zn in Daphnia. A tissue-based Zn accumulation approach with a threshold Zn tissue concentration was better in predicting acute Zn toxicity. Environ. Toxicol. Chem. 2010; 29:1841–1848. © 2010 SETAC

Toxicity tests for metals have traditionally focused on selected biomarkers to characterize the biological stress induced by metals in marine organisms. Here nuclear magnetic resonance (NMR)-based metabolomics, a system biology tool, was applied to the marine green mussel, Perna viridis, to investigate the toxicological effects of Cd in both digestive gland and adductor muscle tissues. After Cd exposure for either two or four weeks, there was no significant metabolic change in the mussels exposed to Cd at 2 µg/L. At 20 µg/L, there were major metabolite changes related to amino acids, osmolytes, and energy metabolites. Digestive gland tissue was more sensitive to Cd than adductor muscle tissue. The adductor muscle tissue showed elevated levels of glutamine, glutamate, and lactate, and reduced levels of branched chain amino acids, aspartate, phenylalanine, and tyrosine. Overall, four weeks of Cd exposure produced neurotoxicity and metabolic disturbances and disturbed osmoregulation. These results suggest that the adductor muscle tissue of mussels may be a suitable supplemental biomarker for exposure to toxicants. In addition, the results demonstrate that 1H-NMR-based metabolomic analysis can provide a systematic view of the toxicological effects of metals on mussels, suggesting that it might be employed to investigate the toxicological effects of other marine pollutants. Environ. Toxicol. Chem. 2011; 30:806–812. © 2010 SETAC