Toxicity of HC Orange No. 1 to Daphnia magna, zebrafish (Brachydanio rerio) embryos, and goldfish (Carassius auratus)

HC Orange No. 1 (HCO1; 2-nitro-4′-hydroxydiphenylamine) (CAS No. 54381-08-7) is used as a color additive in hair dyes and can be released into aquatic environments in wastewater. In this paper, the effects of HCO1 on aquatic organisms were studied using a battery of toxicological tests. These included measuring immobilization of Daphnia magna, inhibition of zebrafish embryo development, and acute lethality in zebrafish and goldfish, which are different species belonging to different trophic levels. HCO1 was toxic to all of the organisms studied. In our experiments, HCO1 remarkably restrained the mobility of D. magna, which may cause subsequent death. The EC₅₀ value for restrained the mobility of D. magna at 48 h was 1.54 mg HCO1 l⁻¹. In addition, HCO1 showed toxicity in zebrafish and goldfish, where LC₅₀values at 96 h were 4.04 and 5.37 mg l⁻¹, respectively. The results also indicated that HCO1 remarkably retarded the development of zebrafish embryos, which may cause embryo abnormality and even lethality. The most sensitive toxicological endpoint in the development of the embryos was failure to hatch, which had an EC₅₀ of 0.19 mg HCO1 l⁻¹. These results indicated that HCO1 is a potential teratogen to zebrafish embryos. In addition, as HCO1 concentrations increased, the outcomes of each of these toxicity tests changed in a concentration-dependent manner. Together, the results revealed that HCO1 appears to be toxic to multiple different species of aquatic organisms. The EC₅₀ (LC₅₀) values contain sufficient discriminatory power for risk assessment of HCO1 in aquatic environments. Based on the present results, more efficient risk assessment procedures for HCO1 will be designed in the future, integrating more flexible testing methods into the testing schemes that employ only the necessary tools for each case.     

Combined effects of cadmium and composted manure to aquatic organisms

To evaluate the interactive toxicity of cadmium (Cd) and composted manure to aquatic organisms 96 h static bioassays were conducted in the laboratory with fry of common carp (Cyprinus carpio), copepod (Diaptomusforbesi) and oligochaete worm (Branchiura sowerbyi). Five concentrations of composted manure (0, 0.25, 0.5, 1.0 and 6.7 g/l) were prepared from the aquatic weed, Pistia stratiotes and each of them was combined with several concentrations of Cd to determine 96 h LC-50 values of Cd for the test organisms. Addition of composted manure, irrespective of concentration, significantly reduced the LC-50 value of Cd to the copepod and common carp fry while it increased the LC-50 value of Cd to the worm. Increased susceptibility of the worm to combined treatment of composted manure and small concentrations of Cd could be revealed only from the dose mortality curve. Results of acute toxicity bioassays were different from the results of bioassays conducted with small concentrations of Cd. Worms, exposed to 2.5 mg/l Cd, accumulated more Cd than did the carp fry and copepod. Accumulation of Cd by worms was increased by the addition of 6.7 g/l composted manure while it decreased in the carp fry and copepod. Food consumption rate of common carp fingerling was significantly reduced relative to the control by exposure to 2.5 mg/l Cd. No change in feeding rate was observed when Cd was combined with composted manure (6.7 g/l).     

Short-term toxicity of pyraclofos used as a blackfly larvicide on non-target aquatic fauna in a tropical environment

Among the many larvicides tested for the control of s.l. larvae, the vector of human onchocerciasis in West Africa, pyraclofos proved to be 100% effective at 100 μg × L⁻¹ for 10 min in river, with a carry of 20 km at 100 m³ × sec⁻¹. Tests were then performed both in laboratory and field conditions to evaluate its toxicity on the non-target aquatic fauna. In experimental short-term gutter tests, the detachment of the total benthic insects was 35% at 100 μg × L⁻¹ for 10 min against 17% for temephos at the same dose and 59% for chlorphoxim at 50μg × L⁻¹ for 10 min. , and were the most affected organisms. The treatment of a river resulted in a considerable detachment of the same taxonomic groups, plus Orthocladiinae. On the other hand, investigations conducted in tanks showed that the 24-hr LC₅₀ for is 150 μg × L⁻¹ and that for 170 μg × L⁻¹, values which are not very different from the operational dose of the larvicide (100 μg × L⁻¹ for 10 min.). Nevertheless, in a river, no fish mortality was recorded. Based on fish LC₅₀ and drift of benthic insects, pyraclofos at 100 μg × L⁻¹ was judged to be less toxic to aquatic fauna in the short term than permethrin and carbosulfan.     

Relationship between electron donor and microorganism on the dechlorination of carbon tetrachloride by an anaerobic enrichment culture

An investigation involving the supplement of different concentrations of substrates and microorganisms was carried out under anaerobic condition to assess the feasibility of bioremediation of carbon tetrachloride (CCl₄) with the amendment of low concentrations of auxiliary substrate and microorganisms. The concentrations of substrate and microorganisms ranged from 10 to 100 mg/l and from 3.7 × 10⁴ to 3.7 × 10⁶ cell/ml, respectively. The biotransformation rate of CCl₄ increased progressively with the increase in the concentrations of the substrate and microorganisms. In the low biomass-amended system (3.7 × 10⁴cells/ml), 28–71% and 57–96% of CCl₄ removals were exhibited when 10–100 mg/l of acetate or glucose was supplemented, respectively, whereas nearly complete degradation of CCl₄ was observed in the heavily inoculated systems (3.7 × 10⁶ cells/ml). An addition of electron donor in the low microbial activity batches enhanced greater efficiency in dechlorination than in the high microbial activity batches. The second-order rate constants ranged from 0.0059 to 0.0092 l/mg/day in high biomass input system, while a two- to four-fold increase in rate constant was obtained in the low microbial activity system. This study indicates that biomass was the more important environmental parameter than substrate affecting the fate of CCl₄. The addition of auxiliary substrates was effective only in low biomass-amended batches (0.56 mg-VSS/l) and diminished inversely with the increase of microbial concentration.     

Effects of pH on the toxicity of cadmium, copper, lead and zinc to Folsomia candida Willem, 1902 (Collembola) in a standard laboratory test system

EC₅₀s for cadmium, copper, lead and zinc were determined for juvenile production of Folsomia candida at pH6.0, 5.0 and 4.5 in a standard laboratory test system. In contrast to most previous studies where metal toxicity was increased at low pHs, in our experiments there was no clear relationship between soil acidity and EC_{50-reproduction} in this species. The EC_{50-reproduction} values (μg g⁻¹) for cadmium and zinc were similar at all three pHs (pH6.0: Cd 590, Zn 900; pH5.0: Cd 780, Zn 600; pH4.5: Cd 480, Zn 590). In contaminated field sites adjacent to primary zinc smelters, zinc is invariably present in soils at concentrations of at least 50 times that of cadmium Thus deleterious effects of mixtures of these metals on populations of Collembola in such sites can be attributed to zinc rather than cadmium.     

Effect of clay and organic matter type on the ecotoxicity of zinc and cadmium to the potworm Enchytraeus albidus

Clay and organic matter are considered as important parameters influencing bioavailability and ecotoxicity of metals in soils. As there exists a large variation in the type of clay and organic matter in field soils, the quantity of these variables alone may not be good indicators of metal bioavailability. To test this hypothesis, toxicity experiments with the potworm Enchytraeus albidus were conducted in artificial soils with three types of clay and seven types of organic matter, while the soil pH was kept constant. The 14d LC₅₀ of zinc and cadmium varied from 83.0 to 1140 mg Zn/kg D.W. and from 55.2 to 704 mg Cd/kg D.W., respectively, depending on the type of clay and organic matter that were used. Simultaneous measurements of the cation exchange capacity showed that this soil parameter is a better indicator of the bioavailability as it takes into account the type of clay and organic matter as well as other adsorption phases such as metal oxyhydroxides.     

Bioaccumulation of anthropogenic contaminants by detritivorous fish in the Río de la Plata estuary: 1-aliphatic hydrocarbons

The temporal variability and bioaccumulation dynamics of C_12–25 n-alkanes, isoprenoids and unresolved aliphatic hydrocarbons (UCM) were studied in a detritivorous fish (Sábalo: Prochilodus lineatus) collected from 1999 to 2005 in the sewage impacted Buenos Aires coastal area. Fish muscles contain huge amounts of n-C_12–25 (165 ± 93, 70 ± 48 or 280 ± 134 μg g⁻¹, dry, fresh and lipid weight, respectively) and UCM (931 ± 560, 399 ± 288 and 1567 ± 802 μg g⁻¹) reflecting the chronic bioaccumulation of fossil fuels from sewage particulates. On a temporal basis, lipid normalized aliphatic concentrations peaked by the end of 2001–2002 during the rainiest period over the last four decades (1750 vs. 1083 ± 4.6 mm in 1999, 2004 and 2005), reflecting an enhanced exposition due to massive anthropogenic fluxes from Metropolitan Buenos Aires in wet years. The hydrocarbon composition in fish muscles is enriched in n-C_15–17 and isoprenoids relative to a fresh crude oil and settling particulates, with fresher signatures during the 2001–2002 maxima. Fish/settling material bioaccumulation factors (BAFs: 0.4–6.4 dry weight or 0.07–0.94 lipid-organic carbon) plotted against K_ow showed a parabolic pattern maximizing at n-C_14–18 and isoprenoids. The optimal bioaccumulation window corresponds to highly hydrophobic (log K_ow: 7.2–9.9), intermediate-size C_14–18 n-alkanes and C_15–20 isoprenoids (MW: 198–282; length: 17.9 to 25.4 Å) with melting points ranging from −19.8 to 28 °C. The uptake efficiency is inversely correlated to melting points and increased from 75% for n-C₂₅ to above 90% for n-C_14–15 and isoprenoids.     

Experimental determination of the kinetic rate law for the oxidation of perchloroethylene by potassium permanganate

Flushing soils contaminated with trichloroethylene (TCE) and perchloroethylene (PCE) with a permanganate (MnO₄⁻) solution has been shown to reduce the solvent content of the soil. Experiments were performed to quantify the rate at which KMnO₄ oxidizes aqueous solutions of PCE over a range of concentrations. In a series of homogeneous reactors, aqueous phase PCE concentrations were monitored over time in nine experimental trials with excess oxidant concentrations ranging from 5 to 30 g/l. Analysis of the data was performed to quantify the oxidation reaction order with respect to PCE and KMnO₄ and the reaction rate constant. The reaction between PCE and KMnO₄ was determined to be first-order with respect to both PCE and KMnO₄ with an overall specific reaction rate coefficient of 2.45±0.65 M⁻¹ min⁻¹.     

Impacts of quicklime (CaO) on the toxicity of copper (CuSO4, 5H2O) to fish and fish food organisms

Static bioassays of 96 h duration were conducted in the laboratory using fry of common carp (Cyprinus carpio), adult tubificid worm (Branchiura sowerbyi) and adult copepod plankton (Cyclops viridis) to determine LC50 values of Cu and CaO to these organisms and effects of interaction between Cu and CaO. Ninety-six hour LC(50) values of Cu to fry of common carp, worm and copepod were found to be 1.40 mgl(-1), 0.08 mgl(-1) and 0.03 mgl(-1) respectively. CaO up to 500 mgl(-1) did not produce any mortality of the fry of common carp up to 96 h. But 96 h LC50 values of CaO to worm and copepod were 83.00 mgl(-1) and 27.80 mgl(-1) respectively. When common carp fry, worm and the copepod were exposed to respective LC50 dose of Cu in presence of varying concentration of CaO, mortality of the organisms significantly reduced and was found inversely correlated with the doses of CaO [y = 48.36-0.807x, r = -0.99 (n = 7) for fish; y = 44.46-0.146x, r = -0.97 (n = 7) for worm; y = 49.46-0.66x, r = -0.99 (n = 7) for the copepod]. The present results indicate that CaO is non-toxic to fish and is capable of reducing the toxicity of Cu to fish while CaO and Cu are antagonistic to each other for the worm and the copepod. Potential of using CaO as antitoxic agent for Cu in water is discussed.     

Evaluation of antimicrobial agents for veterinary use in the ecotoxicity test using microalgae

The influence of antimicrobial agents approved as veterinary drugs in Japan on the growth of green algae, Selenastrum capricornutum and Chlorella vulgaris, was studied in accordance with the OECD guidelines for testing chemicals. Among the agents tested, growth inhibitory activity was very varied, i.e. erythromycin showed the strongest activity (EC₅₀, 50% effective concentration, =0.037 mg/l), sulfa drugs had activity to some extent (EC₅₀s of sulfamethoxazole, sulfadiazine, and sulfadimethoxine were 1.5, 2.2, and 2.3 mg/l, respectively), but ampicillin and cefazolin did not inhibit growth (EC₅₀s > 1000 mg/l). We also investigated synergistic effect of combining sulfa drugs with trimethoprim or pyrimethamine, which are commonly used as a combined drug. By adding trimethoprim, the growth inhibitory activity of sulfamethoxazole and sulfadiazine was significantly enhanced. Growth inhibition by sulfa drugs was reduced by the addition of folic acid, indicating that they inhibit folate synthesis in green algae.     

Effects of cadmium and zinc on oxygen consumption, ammonium excretion, and osmoregulation of white shrimp (Litopenaeus vannamei)

White shrimp, Litopenaeus vannamei, a globally important cultured prawn species, is an ideal animal for studying the impairment caused by the effects of heavy metals that are often detected in coastal areas. The main purpose of the present study was to detect the acute toxicity of Cd and Zn to L. vannamei, and to investigate their effects on gill functions, which have not been carried out in this species before, hoping to obtain some important significance. First of all, we examined the acute toxicity of cadmium (Cd) and zinc (Zn) to L. vannamei and obtained 24-, 48-, 72-, and 96-h medium lethal concentration (LC₅₀) values of 2.58, 1.30, 1.14, and 1.07 mg Cd l⁻¹ and 3.98, 2.14, 1.75, and 1.35 mg Zn l⁻¹, respectively. Furthermore, we also found that exposure of shrimp to Cd or Zn caused an inhibition in oxygen consumption of 91.3% and 75.9% lower than that of the control. However, after separate exposure to Cd or Zn, elevations in ammonium excretion were obtained, which were 153.7% and 144.1%, respectively, higher than the control. It is most likely that elevations in ammonium excretion were related to decreased osmotic pressure of shrimp blood. Finally, the distribution of Cd and Zn within gills was also determined.     

The enhancement of photodegradation efficiency using Pt-TiO2 catalyst

The residues from the extraction of lead/zinc (Pb/Zn) ores of most Pb/Zn mines are permanently stored in tailings ponds, which require revegetation to reduce their environmental impact. This can only be done if the main constraints on plant establishment are evaluated. This can readily be done by field and greenhouse studies.

To test this, the properties of different tailings from Lechang Pb/Zn mine located at the north of Guangdong Province in southern China have been studied. Physical and chemical properties including concentrations of metals (Pb, Zn, Cd and Cu) in the tailings and soils collected from different sites have been measured. The results showed that tailings contain low nitrogen (0.016–0.075%), low-organic matter (0.58–1.78%), high salt (3.55–13.85 dS/m), and high total and diethylene–tetramine–pentaacetic acid (DTPA)-extractable metal concentrations (total: 1019–1642 μg g⁻¹ Pb, 3078–6773 μg g⁻¹ Zn, 8–23 μg g⁻¹ Cd, and 85–192 μg g⁻¹ Cu; DTPA-extractable: 59–178 μg g⁻¹ Pb, 21–200 μg g⁻¹ Zn, 0.30–1.5 μg g⁻¹ Cd, and 4.3–12 μg g⁻¹ Cu). Aqueous extracts of tailings/soils (10%, 20% and 30%, w/v) from different sites were prepared for testing their effects on seed germination and root elongation of a vegetable crop Brassica chinensis and a grass species Cynodon dactylon. It was found that root elongation provided a better evaluation of toxicity than seed germination. The ranking of toxicity using root elongation was: high-sulfur tailings>tailingdam>sparsely vegetated tailings>densely vegetated tailings>mountain soil for both plants. This order was consistent with DTPA-extractable Pb contents in the tailings and soils. B. chinensis seedlings were then grown in the mixtures of different proportions of tailings and farm soil for 4 weeks, and the results (dry weights of seedlings) were in line with the root elongation test. All these demonstrated that heavy metal toxicity, especially available Pb, low content of nutrient, and poor physical structure were major constraints on plant establishment and colonization on the Pb/Zn mine tailings.     

Removal of pesticides from surface waters by combined physicochemical processes. Part I: Dodine

The simultaneous action of powdered activated carbon and several coagulant agents on the removal of the fungicide dodine from spiked distilled water, was studied. As coagulants, ferric chloride (FeCl₃) and basic polyaluminium chlorosulfate ([Al(OH)_xCl_y(SO₄)_z]_n) were examined, using polyacrylamide, in certain cases, as coagulant aid (polyelectrolyte). The efficiency of dodine removal was investigated with respect to the added amount of powdered activated carbon (PAC), the pH value, as well as the type and dose of coagulant and polyelectrolyte. The experiments were performed applying the standard jar-test procedure. The initial concentration of dodine was 250 μg/L. At this concentration and pH range 5–8 it was found that a dose of 100 mg/L PAC was necessary to achieve more than 98% removal of dodine, whereas lower removal (91–93%) was obtained applying half the dose of PAC under the same conditions. However, when 10–100 mg/L FeCl₃ were simultaneously added with PAC, the removal efficiency increased to >98%, even with the half PAC dose.     

Effect of La3+ on the activities of antioxidant enzymes in wheat seedlings under lead stress in solution culture

In order to improve the plant ability to resist lead stress, effect of 0.05 mg/l La(NO₃)₃ on the activities of catalase (CAT), superoxide dismutase (SOD), the level of malondialdehyde (MDA) in wheat seedlings under lead stress was studied. The effect of La³⁺ on plant growth, chlorophyll content in wheat seedlings after adding 0, 50, 100 mg/l Pb(NO₃)₃ to the nutrient solution for 12 days was observed. The plants were grown in nutrient solution in a strictly controlled climate growth room. Effects of La³⁺ (with La treatment) compared with check groups was evidently observed. The activities of SOD and CAT in root were enhanced 0.45–1.69 times and 33.20–77.77% respectively and MDA content was reduced 11.05–27.49% in root after treatments from the second day till the end of the experiment. The activities of SOD and CAT was found to be increased slightly (P<0.05) and MDA content decreased in shoot and root of wheat seedlings by La³⁺ under lead stress within five days after treatments compared with Pb1 and Pb2 groups. It was assumed that antioxidant enzymes was found to be increased by La(NO₃)₃, the antioxidant potential of the wheat seedlings to resist lead stress enhanced. It is suggested that La³⁺ could be used to resist lead stress at the beginning under stress while the stress was not so serious.     

Study of the chemistry of an acid soil column and of the corresponding leachates after the addition of an anaerobic municipal sludge

A column leaching study was carried out over a period of 77 days to determine the changes in the chemistry of an acid soil and of the corresponding leachates after the addition of an anaerobic sludge (equivalent to 69 Mg DW ha⁻¹). By the end of the experiment, the addition of the sludge to the soil had induced an increase in soil pH (from pH 3.6–4.0 to pH 4.1–4.8), in spite of the pronounced decrease in pH detected in the leachates by day 18 of the experiment. The decrease in pH (down to pH 3.3) occurred at the same time as leachate SO₄ and Fe peaked. Once the acidification attributed to sulphide oxidation ceased, the “liming effect” of the sludge became evident and counteracted further proton production – such as that associated with oxidation of NH₄ – at least for the duration of the study. Concentrations of Zn, Cd, Ni, and to a lesser extent, Pb in leachates displayed pulses at the beginning of the experiment (first 12 days), whereas the concentration of Cu followed a more irregular pattern; the concentrations of these metals never surpassed the European threshold values for drinking water. In contrast, concentrations of NO₃, Mn, and Cr in leachates had increased by the end of the experiment – in parallel with an increase in dissolved organic C (DOC) – and surpassed the European threshold for drinking water. Mineralisation of native soil organic C (SOC) was enhanced by the addition of this N-rich residue, and the organic C mass balance at the end of the experiment was negative. Nitrogen mass balance, although positive, exhibited a loss of 77% of the N added to the system. The results obtained indicate that application of this sewage sludge to a soil with a pH < 5, at the loading rate used here, and without liming (i.e., non fulfilment of the requirements of the present European Directive) may pose a risk in terms of groundwater contamination.     

Environmental risk assessment for trisodium [S,S]-ethylene diamine disuccinate, a biodegradable chelator used in detergent applications

Environmental safety data are presented for [S,S]-Ethylene Diamine Disuccinate ([S,S]EDDS), a new, biodegradable, strong transition metal chelator. An environmental risk assessment for its use in detergent applications, which takes into account the chelating properties of [S,S]-EDDS, is proposed.

A property of [S,S]-EDDS that distinguishes it from other strong transition metal chelators is its, “ready” and transparent (no recalcitrant metabolites) biodegradation profile. Because its sorption to activated sludge solids is low ( Kp of 40 1/kg), removal of [S,S]EDDS during sewage treatment, which is greater than 96% as determined by the Continuous Activated Sludge test , is mainly ascribed to biodegradation. At projected use volumes in detergent applications [S,S] - EDDS predicted steady-state concentration in rivers leaving the mixing zone will be below 5 pg/I due to rapid biodegradation. [S,S]-EDDS exhibits low toxicity to fish and Daphnia ( both EC₅₀s> 1000 mg/l). By contrast, due to limitation of the algal test for chelators apparent toxicity was observed (EC₅₀ = 0.290 mg/l, NOEC - No observable Effect Concentration = 0.125 mg/l). Schowanek et al. [1] demonstrated that this is not toxicity sensu stricto but a chelation effect of trace metals in the test medium and of resulting essential nutrients limitation. This requires specific attention when the results of algal toxicity are to be extrapolated to a field situation to perform realistic risk assessment. Metal speciation calculations, using MINEQL+, show that at the predicted environmental concentrations of [S,S] - EDDS (1–5 μg/l), such a chelation effect would be insignificant. These calculations allow to estimate the NOEC for chelation effects in the field to be in the range of 0.250-0.500 mg/l, depending on the background water chemistry. These values are well above the laboratory NOEC.

An environmental risk assessment was performed using the EUSES (1.0) program. EUSES is currently the EU recommended tool for conducting risk assessments (TGD 1995). It was applied to estimate the river water and soil concentrations from production, formulation and private use life stages. The estimated PEC/PNEC ratio in all relevant environmental compartments is smaller than 1, indicating “no immediate concern” at the anticipated usage level.     

Biodegradation of nonylphenol in sewage sludge

We investigated the effects of various factors on the aerobic degradation of nonylphenol (NP) in sewage sludge. NP (5 mg/kg) degradation rate constants (k₁) calculated were 0.148 and 0.224 day⁻¹ for the batch experiment and the bioreactor experiment, respectively, and half-lives (t_1/2) were 4.7 and 3.1 days, respectively. The optimal pH value for NP degradation in sludge was 7.0 and the degradation rate was enhanced when the temperature was increased and when yeast extract (5 mg/l) and surfactants such as brij 30 or brij 35 (55 or 91 μM) were added. The addition of aluminum sulfate (200 mg/l) and hydrogen peroxide (1 mg/l) inhibited NP degradation within 28 days of incubation. Of the microorganism strains isolated from the sludge samples, we found that strain CT7 (identified as Bacillus sphaericus) manifested the best degrading ability.     

Cold season CH4, CO2 and N2O fluxes from freshwater marshes in northeast China

Cold season (winter and thaw) CH₄, CO₂ and N₂O fluxes from freshwater marshes (47°35′N, 133°31′E, Northeast China) were measured, using the static chamber method. The mean CH₄ and CO₂ fluxes from Carex lasiocarpa (Cl) were 0.5 ± 0.19 and 6.23 ± 1.36 mg C m⁻² h⁻¹, respectively, and those from Deyeuxia angustifoli (Da) were 0.18 ± 0.15 and 5.22 ± 2.48 mg C m⁻² h⁻¹, respectively in winter. There was no significant difference between Cl and Da (p > 0.05). The contributions of winter CH₄ fluxes were about 5.5% and 3% in the Cl and Da, respectively. Marshes are an important potential N₂O sink in winter season in northeast China. During thaw, the CH₄ and CO₂ emissions rapidly increased, 4.5–6 times of winter emissions. Wetland became a source of N₂O. Cold season gases flux from northern wetlands play an important role in the seasonal gas exchange.     

Development of a terrestrial vertebrate model for assessing bioavailability of cadmium in the fence lizard (Sceloporus undulatus) and in ovo effects on hatchling size and thyroid function

In the terrestrial environment, standardized protocols are available for measuring the exposure and effects of contaminants to invertebrates, but none currently exist for vertebrates. In an effort to address this, we proposed that developing lizard embryos may be used as a terrestrial vertebrate model. Lizard eggs may be particularly susceptible to soil contamination and in ovo exposure may affect hatchling size, mortality, as well as thyroid function. Toxicant-induced perturbations of thyroid function resulting from in ovo chemical exposure may result in toxicity during the critical perinatal period in reptiles. Fertilized Eastern fence lizard (Sceloporus undulatus) eggs were placed in cadmium (Cd)-spiked expanded perlite (0, 1.48, 14.8, 148, 1480, 14 800 μg Cd/g, nominal concentrations), artificially incubated at 28 °C, and examined daily for mortality. Whole lizard hatchlings as well as failed hatches were homogenized in ethanol and the homogenate was divided for Cd body residue analysis and thyroid hormone (triiodothyronine (T₃) and thyroxine (T₄)) analyses. Acute mortality was observed in the two highest doses (1480 and 14 800 μg Cd/g). Cadmium body residues showed a higher internal concentration with increasing exposure concentration indicating uptake of Cd. There was a decrease in T₃:T₄ ratio at the highest surviving dose (148 μg Cd/g), however, there were no differences observed in hatchling size measured as weight and snout-vent length, or in whole body thyroid hormone levels. In summary, this study has shown Cd amended to a solid phase representing soil (perlite) can traverse the thin, parchment-like shell membrane of the fence lizard egg and bioaccumulate in lizard embryos. We believe this study is a good first step in investigating and evaluating this species for use as a model.     

Cadmium accumulation and Cd-binding proteins in marine invertebrates--a radiotracer study

Tissue and subcellular accumulation of cadmium were studied in different tissues of three marine invertebrates (blue mussel Mytilus edulis, the tunicate Ciona intestinalis and the sea star Asterias rubens) using radioactive ¹⁰⁹Cd as a tracer. The organisms were exposed to 0.05, 2 and 50 μg Cd l⁻¹ for 21 days. Quantitative data were obtained by dissecting, weighing and subsequently measuring radioactivity in organs and tissues. Differences between each exposure and each tissue with regard to the amount of radioactivity and metallothionein (MT) content were evaluated. Obvious interspecies differences in Cd accumulation were observed, as well as differences between tissues of the three species. The highest concentrations of Cd in all exposure treatments were found in the hepatopancreas of M. edulis and body wall of A. rubens. Taking all treatments into account, Cd accumulation in the tunic of C. intestinalis was high compared to other tissues from this species. Over 60% of Cd was present in the S50 fraction in all treatments in all three species. Metallothionein levels were increased at the highest Cd-exposure in all species and tissues, except in branchial pharynx of C. intestinalis where the highest MT level was reached following exposure to 2 μg Cd l⁻¹. The most surprising finding was that even the lowest Cd exposure concentration (0.05 μg Cd l⁻¹) caused MT induction in pyloric caeca of A. rubens, but there was no dose-dependent increase in MT at higher exposure levels.