Toxicological effects and bioaccumulation in the freshwater clam (Corbicula fluminea) following exposure to trivalent arsenic

Contamination of aquatic environments by arsenic is a serious worldwide problem. The main objective of this work was to evaluate the response of a freshwater clam (Corbicula fluminea) to arsenic (As III) exposure and infer its potential as a biological indicator of contamination. Metallothioneins (MTs) were used as indicators of metalloid toxicity in combination with an histological and histochemical evaluation. After a period of acclimatization in the laboratory, 50 C. fluminea (0.4 g +/- 0.1) were exposed to different nominal concentrations of arsenic (100, 300, 500, and 1000 microg L(-1)) for 7 days. The concentration of total As III in the water and in the tissues of the organisms was determined by atomic absorption spectrometry, and MTs were quantified through differential pulse polarography. Results suggest that the organisms exposed to the concentrations of 300 and 1000 microg As L(-1) accumulated the highest levels of As III in the tissues (17 +/- 9 and 15 +/- 3 microg g(-1) distilled water, respectively), which was confirmed through histochemical analysis. An apparent induction of MTs was also observed in the organisms exposed to As III, suggesting that C. fluminea possesses some capacity for arsenic regulation. The results suggest that the induction of MTs may be of high interest as a biomarker for arsenic contamination in aquatic environments, and confirms the potential of C. fluminea as a biological indicator.     

Dynamic features of ecophysiological response of freshwater clam to arsenic revealed by BLM-based toxicological model

The objective of this study was to use a quantitative process-based damage assessment model (DAM) associated with biotic ligand model (BLM) to examine the ecophysiological responses of freshwater clam Corbicula fluminea to waterborne arsenic. We carried out a 14-day exposure experiment to obtain bioaccumulation parameters and a 7-day acute toxicity bioassay to obtain survival data. To investigate the survival dynamics, we examined 2 key parameters characterizing bioaccumulation and damage regulation: capacity to eliminate body arsenic burden and reversible ability to recover the damage. Results show that uptake rate constant of 2.075 ± 0.442 (mean ± SE) ml g⁻¹ d⁻¹ during uptake phase and elimination rate constant was estimated to be 0.1995 ± 0.022 d⁻¹. The derived bioconcentration factor of 10.401 ml g⁻¹ suggests that arsenic has a high potential for bioaccumulation in C. fluminea. Our results show that a negative association between bioaccumulation and the fraction of arsenic binding in biotic ligand at 50% mortality, indicating that C. fluminea with higher arsenic binding in gill biotic ligand at 50% mortality level gives a lower capacity to accumulate bioavailable arsenic. We found a linearly positive correlation between elimination rate and recovery rate constants. Yet, a potential tradeoff between ability to eliminate arsenic and ability to recover the damage is not found. We showed that an ecophysiological significance of C. fluminea exposed to arsenic can be revealed by the elimination–recovery regime. This research may also provide mechanistic insights into the development of biomonitoring organism such as C. fluminea mimicking metal bioaccumulation in a real situation.     

A dose-based modeling approach for accumulation and toxicity of arsenic in tilapia Oreochromis mossambicus

We proposed an approach to relate metal toxicity to the concentrations of arsenic (As) in specific target organs of tilapia Oreochromis mossambicus. The relationships among As exposure, uptake, accumulation, and toxicity of tilapia were investigated using kinetic and dynamic modeling. The biouptake rate of waterborne As through the gills of fish was dependent on exposure concentrations, in that the relationship was well described by incorporating Michaelis-Menten type uptake kinetics. The fitted bioaffinity parameter and limiting uptake flux were 3.07 +/- 2.21 microg/mL(-1) (mean +/- SD) and 2.17 +/- 0.38 microg/mL(-1)/d(-1), respectively, suggesting that a low As binding affinity of tilapia gills, yet a relatively high binding capacity was obtained. The toxicity of As was analyzed by determining the lethal exposure concentration associated with a mortality of 50% (LC50) at different integration times. Our results demonstrate that 96-h and incipient LC50s for tilapia are 28.68 (95% CI: 15.98-47.38) and 25.55 microg/mL(-1), respectively. The organ-specific internal residue associated with 50% mortality was estimated by combining the model-predicted toxicokinetic parameters and the area-under-curve (AUC)-based time-integrated concentration toxicity model. A physiologically based toxicokinetic model was constructed to elucidate the principle mechanisms that account for the observed data and to predict the kinetics of As in tilapia under different water exposure scenarios. We employed the Hill equation model to predict the organ-specific dose-response relationships. We used the liver as a surrogate of target sites to assess the As toxicity to tilapia because of its higher sensitivity to As toxic effects. The predicted mortalities never reach 50% when the tilapia were exposed to waterborne As <2 microg/mL(-1). The predicted mortality is, however, slightly higher than the observed values before the 10th day in that the profile reached the 70% maximum mortality, which is comparable to the observed data when the tilapia were exposed to 4 microg/mL(-1). Our results show that a dose-based toxicokinetic and toxicodynamic modeling approach successfully links metal exposure to bioavailability, bioaccumulation, and toxicity, under variable exposure scenarios.     

Bioaccumulation and toxicity of 4-nonylphenol (4-NP) and 4-(2-dodecyl)-benzene sulfonate (LAS) in Lumbriculus variegatus (Oligochaeta) and Chironomus riparius (Insecta)

The discharge of surfactants, such as 4-nonylphenol (4-NP) and linear alkylbenzene sulfonates (LAS), into water bodies leads to accumulation of the chemicals in the sediments and may thus pose a problem to benthic organisms. To study the bioaccumulation of surfactants, Oligochaeta worm Lumbriculus variegatus was exposed to sediment-spiked, [14C]-labeled 4-NP and 4-(2-dodecyl)-benzene sulfonate (C12-LAS) in three different sediments (S1-S3). The sediments were characterized for organic carbon (OC) content and particle size distribution. The acute toxicity was examined by exposing L. variegatus and three to four instar Chironomus riparius (Insecta) larvae in water-only exposure to 4-NP and LAS at different concentrations. After 48-h exposure, lethal water concentrations (LC50) and lethal body residues (LBR50) were estimated using liquid scintillation counting. Chronic toxicity was evaluated in two different sediments by exposing first instar C. riparius larvae to sediment-spiked chemicals at different concentrations. After 10 days, the sublethal effects of surfactants were observed by measuring wet weight and head capsule length. Finally, another 10-day test was set up in order to measure the LAS body residues associated with sublethal effects in C. riparius in S2 sediment. The bioaccumulation test revealed that the bioaccumulation of both 4-NP and LAS increased as the sediment organic matter content decreased. It is assumed that the chemical binding to organic material decreased chemical bioavailability. The acute toxicity tests showed that L. variegatus was more tolerant of 4-NP, and C. riparius was more tolerant of LAS when based on water exposure concentration. The LBR-estimates revealed, however, that L. variegatus tolerated clearly higher tissue residues of both chemicals compared with C. riparius. Both chemicals had sublethal effects on C. riparius growth in sediment exposure, reducing larvae wet weight and head capsule size. 4-NP, however, showed an irregular dose-response pattern. The characteristics of the exposure media affected the bioaccumulation potential of both chemicals. Thus, exposure concentrations offered no prediction of body residue, and therefore it is proposed that organism body residue offered a more accurate dose-metric for chemical exposure than the chemical concentration of the environment.     

Laboratory and field assessment of uranium trophic transfer efficiency in the crayfish Orconectes limosus fed the bivalve C. fluminea

At present, ecotoxicological information regarding the impact of natural uranium (U) on freshwater ecosystems via the trophic contamination route is scarce. We generated an experimental trophic food chain involving the prey species, Corbicula fluminea, and a predator, Orconectes limosus, for a 10-day and a 30-day feeding periods (food ration: one whole soft body/day/crayfish). We studied the efficiency of U trophic transfer and the distribution of U in the predator. During the test, we varied the quantity of dietary U (from beforehand contaminated bivalves at concentrations ranging from 0.9+/-0.1 to 20.2+/-9 microg/g fw provided to each crayfish over the 10 days) applying a daily feeding rate equal to 3.9+/-0.8% fw. The efficiency of U trophic transfer from clams to crayfish varied between 1 and 13% depending on the prey exposure modalities. Accumulation of U was observed in the digestive gland but also in gills, in the muscle, and in the molt of the crayfish after trophic exposure treatments. Under high-level exposure conditions, the digestive gland was the main target-organ, however a significant accumulation was also observed in the stomach. With regard to low levels of trophic exposure, accumulation of U in gills, in the stomach, and in the digestive gland was of the same order of magnitude. Longer exposure period which incorporated a crayfish molt, resulted in a decrease of trophic transfer ratio and a modified U tissue distribution.     

Depuration kinetics and persistence of the cyanobacterial toxin microcystin-LR in the freshwater bivalve Unio douglasiae

We carried out uptake and depuration experiments in the laboratory to investigate the effects of temperature (15 degrees C and 25 degrees C) on the depuration kinetics and persistence of a cyanobacterial toxin, microcystin-LR (MCYST-LR), in a freshwater bivalve, Unio douglasiae. Bivalves were fed toxic Microcystis cells in the 15-day uptake experiment and nontoxic diatoms in the following 15-day depuration experiment. Each bivalve's hepatopancreas was lyophilized and extracted with a butanol:methanol:water solution for analysis of MCYST-LR by high-performance liquid chromatography. The toxin in the organ accumulated rapidly after the beginning of the uptake experiment and reached approximately steady-state conditions on day 5 at concentrations of 130 +/- 11 microg g(-1) dry weight at 15 degrees C and 250 +/- 40 microg g(-1) at 25 degrees C. In the depuration experiments MCYST-LR was eliminated asymptotically from the tissue. The values of the depuration rate constant (k(d)), calculated with a first-order one-compartment model, were found to be 0.142 +/- 0.044 day(-1) at 15 degrees C and 0.226 +/- 0.046 day(-1) at 25 degrees C. The depuration Q(10) value from 15 degrees C to 25 degrees C equaled 1.6. This study was the first to reveal the kinetics of depuration for MCYST-LR in a bivalve. The results show that MCYST-LR may be eliminated slowly in autumn and winter and persist in the tissue until spring. Thus, in terms of toxicokinetics, the risk to people of being poisoned by bivalves would increase if toxic blooms occur in autumn.     

Effects of waterborne uranium on survival, growth, reproduction and physiological processes of the freshwater cladoceran Daphnia magna

Acute uranium toxicity (48 h immobilisation test) for Daphnia magna was determined in two different exposure media, differing in pH and alkalinity. LC(50) varied strongly between media, from 390+/-40 microgL(-1)U at pH 7 to 7.8+/-3.2 mgL(-1)U at pH 8. According to the free ion activity model uranium toxicity varies as a function of free uranyl concentration. This assumption was examined by calculating uranium speciation in our water conditions and in those reported in the literature. Predicted changes in free uranyl concentration could not solely explain observed differences in toxicity, which might be due to a competition or a non-competitive inhibition of H(+) for uranium transport and/or the involvement of other bioavailable chemical species of uranium. Chronic effects of uranium at pH 7 on mortality, ingestion and respiration, fecundity and dry mass of females, eggs and neonates were investigated during 21-day exposure experiments. A mortality of 10% was observed at 100 microgL(-1)U and EC(10) for reproduction was 14+/-7 microgL(-1)U. Scope for growth was affected through a reduction in feeding activity and an increase in oxygen consumption at 25 microgL(-1)U after 7 days of exposure. This had strong consequences for somatic growth and reproduction, which decreased, respectively, by 50% and 65% at 50 microgL(-1)U after 7 days and at 25 microgL(-1)U after 21 days. Uranium bioaccumulation was quantified and associated internal alpha dose rates from 2.1 to 13 microGyh(-1) were estimated. Compared to the toxicity of other alpha-emitting radionuclides and stable trace metals, our results confirmed the general assumption that uranium chemical toxicity predominates over its radiotoxicity.     

Acute toxicity and bioaccumulation of arsenic in tilapia (Oreochromis mossambicus) from a blackfoot disease area in Taiwan

The general objective of our work was to determine the acute toxicity and bioaccumulation of arsenic (As) in tilapia (Oreochromis mossambicus) from the blackfoot disease (BFD) area in Taiwan. The average concentration of As in pond water ranged from 17.8 to 49 microg L(-1). Acute toxicity tests showed that the As concentration that caused toxicity to tilapia ranged from 69 060 microg As L(-1), in the 24-h toxicity test, to 28 680 microg As L(-1), in the 96-h toxicity test. We measured As concentrations in various tissues of tilapia to identify the affinities of tissues for As. Significant correlations were found among the As concentrations in all tissues. The highest bioconcentration factor (BCF) was found in the intestine (maximum value: 2270). The order of BCFs was: intestine > stomach > liver approximately gill > muscle. Arsenic concentrations in all tissues were allometric, negatively correlating with fish body weight [r(2) = 0.63 +/- 0.045 (mean +/- SE), p < 0.05]. Our results also revealed that As concentrations in muscle tissue were positively correlated with As accumulation in the viscera (r(2) = 0.85, p < 0.05). Significantly higher concentrations of As were obtained in the viscera of tilapia [12.65 +/- 10.17 microg g(-1) dry wt (mean +/- SD)] than in the muscle tissue (3.55 +/- 0.42 microg g(-1) dry wt). Our results suggest that a simple way of reducing the health risk associated with consuming tilapia is to trim and cook the fish properly, that is, removing the viscera of tilapia can greatly reduce the amount of As ingested and consequently reduce the health risks.     

Toxicity of arsenic species to Lemna gibba L. and the influence of phosphate on arsenic bioavailability

The toxicity of arsenic (As) species to Lemna gibba L. and the influence of PO(4) (3-) on As bioavailability and uptake were tested in batch culture. L. gibba were exposed to six test concentrations of NaHAsO(4). 7H(2)O and NaAsO(3), with 0, 0.0136, 13.6, and 40 mg L(-1) KH(2)PO(4). In batch culture As toxicity to L. gibba did not relate linearly to As concentration. The growth rate, related to frond number as recommended by OECD and ISO/DIN, was significantly inhibited in fronds exposed to 20-50 microg L(-1) As(III) compared with fronds exposed to As(V). The growth rate was stimulated when plants were exposed to 50-250 microg L(-1) of both As(III) and As(V). After exposure to 300-800 microg L(-1) growth inhibition was significantly higher for As(III) than for As(V), whereas above 800 microg L(-1) As(V) was inhibited the most. The bioaccumulation of As(III) and As(V) was significantly higher for P-deficient cultures (0.98 +/- 0.08 and 1.02 +/- 0.19 g kg(-1), respectively for 0.0136 mg L(-1) PO(4) (3-)) than for P-sufficient cultures (243 and 343 mg kg(-1) for 40 mg L(-1), respectively). Plants exposed to As(V) had uptake and accumulation values slightly higher than did plants exposed to As(III). No significant differences in bioaccumulation were found between plants exposed to a concentration of As(III) >1 mg L(-1) and those exposed to As(V) at the same concentration. This indicates a direct relationship to P content in the culture. Toxicity may result from the uptake of As(V) instead of PO(4) (3-) as a result of ion competition during uptake because of close thermodynamic properties, which may change the interaction among components in the media. The toxicity pattern is interpreted as a manifestation of changing speciation in the batch culture and of the oxidation of As(III) to As(V) in an oxygen-rich environment.     

Kinetic analysis of uranium accumulation in the bivalve Corbicula fluminea: effect of pH and direct exposure levels

The bioaccumulation of natural uranium in the freshwater bivalve Corbicula fluminea was investigated subsequent to the bivalve's experimental waterborne exposures. A first experiment determined the accumulation rate (transfer efficiency, tissular distribution) and subcellular distribution of uranium in organs after over 42 days of uranium exposure (100 microg l(-1); pH 7) and later following 60 days of depuration. Results showed that there was direct transfer of uranium to the bivalve organs ([U]organism/[U]water = 0.16, fresh weight, fw). The highest accumulation levels occurred in the visceral mass and remained constant throughout the exposure duration, although a linear increase in the U concentration in the gills was observed (2.98 +/- 1.3-10.9 +/- 3.7 microg g(-1) between days 2 and 42). A second set of experiments were performed in order to test the influence of the exposure levels (100; 500; 1500 microg l(-1)) and pH (7 and 8.1) on the bioaccumulation capacities. A marked difference of U distribution is observed as a function of exposure levels (gills were favoured in the case of high exposure levels-relative burden: 49.1 +/- 3% (1500 microg l(-1)), whereas the visceral mass presented higher accumulation levels at environmentally relevant U concentrations). Uranium concentration in the insoluble fraction (80%) in the whole body does not depend upon exposure levels in the water column or upon duration. These experiments did not allow any link to be established between the free-metal ion concentration and the bioaccumulation efficiency. Results showed a significant pH effect and indicated a link between the exposure conditions and the distribution of uranium in the bivalve organs.     

Copper uptake and depuration by juvenile and adult Florida apple snails (<Emphasis Type="Italic">Pomacea paludosa</Emphasis>)

The present study characterized copper (Cu) uptake and depuration by juvenile and adult Florida apple snails (Pomacea paludosa) from water, soil, and diet. During a 28-day uptake period, juvenile apple snails were exposed to aqueous Cu and adult apple snails were exposed to Cu-contaminated soil, water, and food. In the follow-up 14-day depuration period, both juvenile and adult apple snails were held in laboratory freshwater with background Cu concentrations <4 μg/l. For juvenile apple snails, whole body Cu concentrations increased with time and reached a plateau after 14 days. The data followed Michaelis–Menten kinetics rather than a one compartment first order kinetics model. The mean Cu bioconcentration factor (BCF) for juvenile apple snails was 1493 and the depuration half-life was 10.5–13.8 days. For adult snails, dietary uptake of Cu resulted in higher bioaccumulation factors (BAFs) compared to uptake from soil. Most of the accumulated Cu was located in soft tissue (about 60% in the viscera and 40% in the foot). The shell contained <1% of the total accumulated copper. Soft tissue is usually consumed by predators of the apple snail. Therefore, the results of the present study show that Cu transfer through the food chain to the apple snail may lead to potential risk to its predators.     

The toxicity and bioaccumulation of cadmium and copper as affected by humic acid

The toxicity of Cd and Cu to daphnids was evaluated over a 42-day exposure period at three humic acid concentrations. The addition of humic acid to test water decreased the acute and chronic toxicity of Cu but increased the acute and chronic toxicity of Cd. However, there was no effect of HA on the bioaccumulation of either metal. At the highest concentration, the presence of humic acid actually resulted in an increase in mean brood sizes of Cu-exposed animals and prevented a reduction in mean brood sizes of Cd-exposed animals.     

Differential effects of arsenic on folate binding protein 2 (Folbp2) null and wild type fibroblasts

Exposure to arsenic results in a wide variety of adverse effects. It has been postulated that one mechanism of arsenic toxicity is disruption of cellular methyl biochemistry. Because dietary folate is required to generate the methyl donor S-adenosyl methionine, we hypothesized that loss of folate binding protein 2 (Folbp2) results in increased susceptibility to arsenic-induced cytotoxicity. Using Folbp2 +/+ and -/- fibroblasts, we determined that Folbp2 null cells display increased sensitivity to arsenic exposure. Folic acid supplementation partially rescues wild type cells from arsenic toxicity, but Folbp2 null cells are not protected. Arsenic inhibits folic acid uptake in Folbp2 null fibroblasts, but not wild type cells; baseline uptake is similar in both cell types. These results support the possibility that arsenic toxicity occurs, in part, by perturbing cellular methyl biochemistry. Furthermore, identification of Folbp2 as a protective protein presents an opportunity to identify populations at increased risk for serious effects of arsenic exposure.     

Physiologically based pharmacokinetic modeling of arsenic in the mouse

A remarkable feature of the carcinogenicity of inorganic arsenic is that while human exposures to high concentrations of inorganic arsenic in drinking water are associated with increases in skin, lung, and bladder cancer, inorganic arsenic has not typically caused tumors in standard laboratory animal test protocols. Inorganic arsenic administered for periods of up to 2 yr to various strains of laboratory mice, including the Swiss CD-1, Swiss CR:NIH(S), C57Bl/6p53(+/-), and C57Bl/6p53(+/+), has not resulted in significant increases in tumor incidence. However, Ng et al. (1999) have reported a 40% tumor incidence in C57Bl/6J mice exposed to arsenic in their drinking water throughout their lifetime, with no tumors reported in controls. In order to investigate the potential role of tissue dosimetry in differential susceptibility to arsenic carcinogenicity, a physiologically based pharmacokinetic (PBPK) model for inorganic arsenic in the rat, hamster, monkey, and human (Mann et al., 1996a, 1996b) was extended to describe the kinetics in the mouse. The PBPK model was parameterized in the mouse using published data from acute exposures of B6C3F1 mice to arsenate, arsenite, monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA) and validated using data from acute exposures of C57Black mice. Predictions of the acute model were then compared with data from chronic exposures. There was no evidence of changes in the apparent volume of distribution or in the tissue-plasma concentration ratios between acute and chronic exposure that might support the possibility of inducible arsenite efflux. The PBPK model was also used to project tissue dosimetry in the C57Bl/6J study, in comparison with tissue levels in studies having shorter duration but higher arsenic treatment concentrations. The model evaluation indicates that pharmacokinetic factors do not provide an explanation for the difference in outcomes across the various mouse bioassays. Other possible explanations may relate to strain-specific differences, or to the different durations of dosing in each of the mouse studies, given the evidence that inorganic arsenic is likely to be active in the later stages of the carcinogenic process.     

The key role of metallothioneins in the bivalve Corbicula fluminea during the depuration phase,after in situ exposure to Cd and Zn

An experimental study of the role of metallothioneins (MTs) in Cd and Zn depuration processes in the freshwater bivalve Corbicula fluminea was conducted after in situ exposure on the river Lot (France). Specimens of adult C. fluminea were first transplanted from a lacustrine reference site to a polymetallic polluted station (Bouillac, (B)) for a 42-days' exposure period from September to November 1996. They were then depurated after transfer to the laboratory, and were sub-sampled periodically until May 1997. During the first phase, MT concentrations measured with the Mercury-Saturation Assay were induced for a factor of 3.5 compared with time 0, whereas metal uptake showed accumulation factors of 17 and 4 for Cd and Zn, respectively. During the depuration phase, Cd and Zn concentrations decreased by 18 and 70%, respectively, giving estimated biological half-lives of 500 and 40 days. During the same period, MT concentrations decreased by 37% after transfer under unpolluted conditions, especially between 0 and 3 days, suggesting that MTs play a predominant role in Cd depuration. The quantity of Cd sequestered by the MT fraction, after size-exclusion liquid chromatography, represents on average 40% of the total Cd bioaccumulated in the soft body of the molluscs, compared with only 4-9% for total accumulated Zn. This essential metal was principally bound to low molecular weight proteins, which represented 20% of total Zn. Furthermore, it was observed that MTs had a key role in Cd remanence in the bivalves, and it was also reported that other proteins or small peptides were involved in the depuration of Zn.     

Metabolism and disposition of 2,3,7,8-tetrachlorodibenzo-p-dioxin in guinea pigs

Marked interspecies variability exists in the acute toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), with the guinea pig being the mammalian species most sensitive to the acute toxicity of TCDD. The metabolism and disposition of TCDD was investigated in guinea pigs for 45 days following a single exposure to purified [3H]TCDD (0.56 microgram/kg, ip). Guinea pigs included in the toxicokinetic study gained body weight, maintained a normal relative body composition, and exhibited no gross signs of toxicity during the 45-day study. Approximately 36% of the dose of TCDD-derived 3H remained in the adipose tissue at 45 days following exposure to [3H]TCDD, while the liver, pelt, and skeletal muscle and carcass each contained about 7% of the administered dose. Although most of the TCDD-derived radioactivity in liver, kidney, perirenal adipose tissue, and skeletal muscle represented unchanged TCDD, from 4 to 28% of the 3H was associated with metabolites of TCDD. This unexpected finding suggests that TCDD metabolites are not efficiently excreted from guinea pigs. The urinary and fecal excretion of TCDD-derived radioactivity followed apparent first-order kinetics, with an elimination half-life of 93.7 +/- 15.5 days (mean +/- SD). HPLC analysis of urine and bile from [3H]TCDD-treated guinea pigs showed that all of the radioactivity represented metabolites of TCDD, indicating that these routes of elimination are dependent on prior metabolism of TCDD. However, 70 to 90% of the radioactivity in fecal samples was found to represent unmetabolized TCDD throughout the 45-day excretion study. The presence of TCDD in feces and its absence in bile suggest that the fecal excretion of unchanged TCDD resulted from the direct intestinal elimination of the lipophilic toxin. Furthermore, the cumulative excretion of TCDD-derived radioactivity over 45 days indicated that 74.3% of the 3H was excreted in feces as unchanged TCDD, while 25.7% of the 3H was excreted in urine and feces as TCDD metabolites. Thus, TCDD is primarily eliminated unchanged in the feces of guinea pigs, indicating that the metabolism of TCDD does not play a major role in the ultimate elimination of the toxin from the guinea pig. This may in part explain the relatively long excretion half-life for TCDD in the guinea pig and may contribute to the remarkable sensitivity of the guinea pig to the acute toxicity of TCDD.     

Kinetics of radiolabelled silver uptake and depuration in the gills of rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla): the influence of silver speciation.

We examined the influence of speciation on the kinetics of silver uptake and depuration in the gills of two freshwater fish, the rainbow trout (Oncorhynchus mykiss) which has high branchial Na(+) and Cl(-) uptake rates and is relatively sensitive to silver, and the European eel (Anguilla anguilla, yellow stage) which has low ion uptake rates and is relatively resistant to silver. Fish previously acclimated to the appropriate chloride level were exposed to 110mAgNO(3) (1.3 microg l(-1), sublethal) for 24 h in synthetic softwater with either low (10 microM) or high (1200 microM) chloride concentration, and then followed over a subsequent 67-day post-exposure period in silver-free water of the same chloride content. The exposures were therefore mainly to the free ion, Ag(+) in the low chloride water versus mainly to the neutral aqueous complex, AgCl(aq) in the high chloride water. In trout, but not in eel, water chloride is known to protect against physiological disturbances and toxicity caused by Ag(+). In both fish species, at both chloride levels, silver uptake exhibited complex kinetics. Gill silver loading occurred slowly until 6 h, then rose greatly to a peak at 12 h, followed by significant net depuration thereafter during continued exposure. By 24 h, net gill loading was three- to fivefold greater from AgCl(aq) than from Ag(+) exposure in both species, and threefold greater in trout than in eel under both conditions, with trout holding a lower fraction of the whole body burden in their gills. During the post-exposure period, depuration of silver from the gills occurred rapidly in trout, but very slowly in eel, such that gill silver burdens were greater in eel throughout the 67-day period on both an absolute and relative basis (e.g. 35% of whole body burden in eel versus <3% in trout at day 8). The kinetics of depuration were described by two phase exponential models, with break points between the fast and slow phases at 1 and 15 days for trout and eel, respectively. We conclude that speciation affects not only uptake rates but also the kinetics of depuration. When silver is loaded from AgCl(aq) it is clearly more labile than from Ag(+) exposures, with 1.6-1.8-fold greater loss rates during the fast phases in both species. Differences in branchial silver uptake between eel and trout correlate well with differences in acute toxicity, but are not as large as differences in ion uptake rates. The complex time-dependent patterns of gill loading, and the higher loading from AgCl(aq) than from Ag(+), mean that gill total silver burden is not an appropriate endpoint for biotic ligand modelling.     

Influence of intracellular toxin concentrations on cylindrospermopsin bioaccumulation in a freshwater gastropod (Melanoides tuberculata).

Scant information is available regarding the bioaccumulation of cylindrospermopsin (CYN) in aquatic organisms, particularly in invertebrates. This study examined toxin bioconcentration and bioaccumulation in the aquatic snail, Melanoides tuberculata, following exposure to freeze-thawed whole cell extracts and a live Cylindrospermopsis raciborskii culture containing CYN. Both bioconcentration and bioaccumulation were evident, but exposure to toxin in the freeze-thawed solutions resulted in minor tissue contamination compared with that resulting from live C. raciborskii exposure. Thus, whilst CYN uptake resulted from both extracellular and intracellular exposures, the availability of intracellular toxin was critical in affecting tissue CYN values. M. tuberculata did not bioconcentrate CYN into the shell. Bioaccumulation of the analog deoxy-CYN was also recorded. Knowledge of intracellular toxin concentrations may be critical in evaluating the bioaccumulation, ecological and human health risks associated with contaminated systems.     

Biochemical endpoints on juvenile Solea senegalensis exposed to estuarine sediments: the effect of contaminant mixtures on metallothionein and CYP1A induction

Juvenile Solea senegalensis were exposed to fresh sediments from three stations of the Sado estuary (Portugal) in 28-day laboratory assays. Sediments revealed distinct levels of total organic matter, fine fraction, redox potential, trace elements (arsenic, cadmium, chromium, copper, nickel, lead and zinc) and organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls and a pesticide: dichloro diphenyl trichloroethane). Organisms were surveyed for contaminant bioaccumulation and induction of two hepatic biochemical biomarkers: metallothionein (MT) and cytochrome P450 (CYP1A), as potential indicators of exposure to metallic and organic contaminants, respectively. Using an integrative approach it was established that, although bioaccumulation is in general accordance with sediment contamination, lethality and biomarker responses are not linearly dependent of the cumulative concentrations of sediment contaminants but rather of their bioavailability and synergistic effects in organisms. It is concluded that metals and organic contaminants modulate both MT and CYP1A induction and it is suggested that reactive oxygen species may be the link between responses and effects of toxicity.     

Monomethylarsonic acid reductase and monomethylarsonous acid in hamster tissue

The formation of monomethylarsonous acid (MMA(III)) by tissue homogenates of brain, bladder, spleen, liver, lung, heart, skin, kidney, or testis of male Golden Syrian hamsters was assessed using [(14)C]monomethylarsonic acid (MMA(V)) as the substrate for MMA(V) reductase. The mean +/- SEM of MMA(V) reductase specific activities (nanomoles of MMA(III) per milligram of protein per hour) were as follows: brain, 91.4 +/- 3.0; bladder, 61.8 +/- 3.7; spleen, 30.2 +/- 5.4; liver, 29.8 +/- 1.4; lung, 21.5 +/- 0.8; heart, 19.4 +/- 1.5; skin, 14.7 +/- 1.6; kidney, 10.6 +/- 0.4; and testis, 9.8 +/- 0.6. The concentrations of MMA(III) in male Golden Syrian hamster livers were determined 15 h after administration of a single intraperitoneal dose of 145 microCi of [(73)As]arsenate (2 mg of As/kg of body weight). Trivalent arsenic species (arsenite, MMA(III), and dimethylarsinous acid, DMA(III)) were extracted from liver homogenates using carbon tetrachloride (CCl(4)) and 20 mM diethylammonium salt of diethyldithiocarbamic acid (DDDC). Pentavalent arsenicals (arsenate, MMA(V), and dimethylarsinic acid, DMA(V)) remained in the aqueous phase. The organic and the aqueous phases then were analyzed by HPLC. Metabolites of inorganic arsenate present in hamster liver after 15 h were observed in the following concentrations (nanograms per gram of liver +/- SEM): MMA(III), 38.5 +/- 2.9; DMA(III), 49.9 +/- 10.2; arsenite, 35.5 +/- 3.0; arsenate, 118.2 +/- 8.7; MMA(V), 31.4 +/- 2.8; and DMA(V), 83.5 +/- 6.7. This first-time identification of MMA(III) and DMA(III) in liver after arsenate exposure indicates that the significance of arsenic species in mammalian tissue needs to be re-examined and re-evaluated with respect to their role in the toxicity and carcinogenicity of inorganic arsenic.