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Archives of Environmental Contamination and Toxicology (v.48, #2)

Assessing Contaminant Sensitivity of Endangered and Threatened Aquatic Species: Part I. Acute Toxicity of Five Chemicals by F. J. Dwyer; F. L. Mayer; L. C. Sappington; D. R. Buckler; C. M. Bridges; I. E. Greer; D. K. Hardesty; C. E. Henke; C. G. Ingersoll; J. L. Kunz; D. W. Whites; T. Augspurger; D. R. Mount; K. Hattala; G. N. Neuderfer (pp. 143-154).

Assessment of contaminant impacts to federally identified endangered, threatened and candidate, and state-identified endangered species (collectively referred to as “listed” species) requires understanding of a species’ sensitivities to particular chemicals. The most direct approach would be to determine the sensitivity of a listed species to a particular contaminant or perturbation. An indirect approach for aquatic species would be application of toxicity data obtained from standard test procedures and species commonly used in laboratory toxicity tests. Common test species (fathead minnow, Pimephales promelas; sheepshead minnow, Cyprinodon variegatus; and rainbow trout, Oncorhynchus mykiss) and 17 listed or closely related species were tested in acute 96-hour water exposures with five chemicals (carbaryl, copper, 4-nonylphenol, pentachlorophenol, and permethrin) representing a broad range of toxic modes of action. No single species was the most sensitive to all chemicals. For the three standard test species evaluated, the rainbow trout was more sensitive than either the fathead minnow or sheepshead minnow and was equal to or more sensitive than listed and related species 81% of the time. To estimate an LC50 for a listed species, a factor of 0.63 can be applied to the geometric mean LC50 of rainbow trout toxicity data, and more conservative factors can be determined using variance estimates (0.46 based on 1 SD of the mean and 0.33 based on 2 SD of the mean). Additionally, a low- or no-acute effect concentration can be estimated by multiplying the respective LC50 by a factor of approximately 0.56, which supports the United States Environmental Protection Agency approach of multiplying the final acute value by 0.5 (division by 2). When captive or locally abundant populations of listed fish are available, consideration should be given to direct testing. When direct toxicity testing cannot be performed, approaches for developing protective measures using common test species toxicity data are available.

Assessing Contaminant Sensitivity of Endangered and Threatened Aquatic Species: Part II. Chronic Toxicity of Copper and Pentachlorophenol to Two Endangered Species and Two Surrogate Species by J. M. Besser; N. Wang; F. J. Dwyer; F. L. Mayer Jr.; C. G. Ingersoll (pp. 155-165).

Early life-stage toxicity tests with copper and pentachlorophenol (PCP) were conducted with two species listed under the United States Endangered Species Act (the endangered fountain darter, Etheostoma fonticola, and the threatened spotfin chub, Cyprinella monacha) and two commonly tested species (fathead minnow, Pimephales promelas, and rainbow trout, Oncorhynchus mykiss). Results were compared using lowest–observed effect concentrations (LOECs) based on statistical hypothesis tests and by point estimates derived by linear interpolation and logistic regression. Sublethal end points, growth (mean individual dry weight) and biomass (total dry weight per replicate) were usually more sensitive than survival. The biomass end point was equally sensitive as growth and had less among-test variation. Effect concentrations based on linear interpolation were less variable than LOECs, which corresponded to effects ranging from 9% to 76% relative to controls and were consistent with thresholds based on logistic regression. Fountain darter was the most sensitive species for both chemicals tested, with effect concentrations for biomass at ≤ 11 μg/L (LOEC and 25% inhibition concentration [IC25]) for copper and at 21 μg/L (IC25) for PCP, but spotfin chub was no more sensitive than the commonly tested species. Effect concentrations for fountain darter were lower than current chronic water quality criteria for both copper and PCP. Protectiveness of chronic water-quality criteria for threatened and endangered species could be improved by the use of safety factors or by conducting additional chronic toxicity tests with species and chemicals of concern.

Acute Toxicity of Six Freshwater Mussel Species (Glochidia) to Six Chemicals: Implications for Daphnids and Utterbackia imbecillis as Surrogates for Protection of Freshwater Mussels (Unionidae) by C. D. Milam; J. L. Farris; F. J. Dwyer; D. K. Hardesty (pp. 166-173).

Acute (24-h) toxicity tests were used in this study to compare lethality responses in early life stages (glochidia) of six freshwater mussel species, Leptodea fragilis, U. imbecillis, Lampsilis cardium, Lampsilis siliquoidea, Megalonaias nervosa, and Ligumia subrostrata, and two standard test organisms, Ceriodaphnia dubia and Daphnia magna. Concentrations of carbaryl, copper, 4-nonylphenol, pentachlorophenol, permethrin, and 2,4-D were used in acute exposures to represent different chemical classes and modes of action. The relative sensitivities of species were evaluated by ranking their LC₅₀ values for each chemical. We used these ranks to determine the extent to which U. imbecillis (one of the most commonly used unionids in toxicity tests) was representative of the tolerances of other mussels. We also calculated geometric mean LC₅₀s for the families Unionidae and Daphnidae. Rankings of these data were used to assess the extent to which Daphnidae can be used as surrogates for freshwater mussels relative to chemical sensitivity. While no single chemical elicited consistently high or low toxicity estimates, carbaryl and 2,4-D were generally the least toxic to all species tested. No species was always the most sensitive, and Daphnidae were generally protective of Unionidae. Utterbackia imbecillis, while often proposed as a standard unionid mussel test species, did not always qualify as a sufficient surrogate (i.e., a substitute organism that often elicits similar sensitivity responses to the same contaminant exposure) for other species of mussels, since it was usually one of the more tolerant species in our rankings. U. imbecillis should be used as a surrogate species only with this caution on its relative insensitivity.

Assessing Contaminant Sensitivity of Endangered and Threatened Aquatic Species: Part III. Effluent Toxicity Tests by F. J. Dwyer; D. K Hardesty; C. E. Henke; C. G. Ingersoll; D. W. Whites; T. Augspurger; T. J. Canfield; D. R. Mount; F. L. Mayer (pp. 174-183).

Toxicity tests using standard effluent test procedures described by the U.S. Environmental Protection Agency were conducted with Ceriodaphnia dubia, fathead minnows (Pimephales promelas), and seven threatened and endangered (listed) fish species from four families: (1) Acipenseridae: shortnose sturgeon (Acipenser brevirostrum); (2) Catostomidae; razorback sucker (Xyrauchen texanus); (3) Cyprinidae: bonytail chub (Gila elegans), Cape Fear shiner (Notropis mekistocholas) Colorado pikeminnow (Ptychocheilus lucius), and spotfin chub (Cyprinella monacha); and (4) Poecillidae: Gila topminnow (Poeciliopsis occidentalis). We conducted 7-day survival and growth studies with embryo-larval fathead minnows and analogous exposures using the listed species. Survival and reproduction were also determined with C. dubia. Tests were conducted with carbaryl, ammonia—or a simulated effluent complex mixture of carbaryl, copper, 4-nonylphenol, pentachlorophenol and permethrin at equitoxic proportions. In addition, Cape Fear shiners and spotfin chub were tested using diazinon, copper, and chlorine. Toxicity tests were also conducted with field-collected effluents from domestic or industrial facilities. Bonytail chub and razorback suckers were tested with effluents collected in Arizona whereas effluent samples collected from North Carolina were tested with Cape Fear shiner, spotfin chub, and shortnose sturgeon. The fathead minnow 7-day effluent test was often a reliable estimator of toxic effects to the listed fishes. However, in 21 % of the tests, a listed species was more sensitive than fathead minnows. More sensitive species results varied by test so that usually no species was always more or less sensitive than fathead minnows. Only the Gila topminnow was consistently less sensitive than the fathead minnow. Listed fish species were protected 96% of the time when results for both fathead minnows and C. dubia were considered, thus reinforcing the value of standard whole-effluent toxicity tests using those two species. If the responses of specific listed species are important for management decisions, our study supports the value in developing culture and testing procedures for those species.

Application of an Enzyme-Linked Immunosorbent Assay for the Analysis of Paraquat in Human-Exposure Samples by M. E. Koivunen; S. J. Gee; E. -K. Park; K. Lee; M. B. Schenker; B. D. Hammock (pp. 184-190).

Paraquat is a toxic quaternary ammonium compound used as an herbicide around the world. Easy, fast, and inexpensive but sensitive methods are needed to study the effects of long-term, low-level exposure of paraquat on human health. An enzyme-linked immunosorbent assay (ELISA) was used for quantification of paraquat in urine and air-filter samples collected in a human-exposure study among farm workers in Costa Rica. A sample pretreatment consisted of removal of interfering substances using solid-phase extraction resin columns. The precision and accuracy of the method were tested using duplicate spiked urine samples. The correlation between results for blind samples obtained using ELISA and liquid chromatography–mass spectrometry was significant (R² = 0.945 and 0.906 for spiked and field samples, respectively). With an LOQ of 2 ng mL⁻¹, this ELISA method was able to distinguish the exposed from the nonexposed farm workers. For the air-filter analysis, paraquat was extracted by 9M H₂SO₄ at 60°C for 12 hours, and the results obtained by ELISA showed good correlation (R² = 0.918) with the spectrophotometric (256 nm) measurements. Paraquat in acid-stabilized urine samples was very stable, and no significant losses were detected during a 3-month storage at room temperature, at 4°C, or at −20°C.

Use of Daphnia spp. for the Ecotoxicological Assessment of Water Quality in an Agricultural Watershed in South-Central Chile by K. Cooman; P. Debels; M. Gajardo; R. Urrutia; R. Barra (pp. 191-200).

Because of the importance of surface waters from the Chillán River watershed (Chile) for recreation, agricultural irrigation, and the production of drinking water, local concern about river water quality has increased considerably during the last decade. Agricultural and forestry activities in the watershed, characterized by an intensive use of pesticides, are thought to play an important role in the generation of non-point-source pollution, whereas the discharge of urban wastewater from the city of Chillán constitutes a major point source of pollution. In the present investigation, acute and chronic laboratory bioassays using Daphnia spp. were conducted on surface water samples from 17 river stations located throughout the watershed. Sampling occurred on 6 occasions during a 16-month period (2000 to 2001) and included both high and low flow conditions. Almost all toxic effects observed in summer were directly related to the discharge of urban wastewater, whereas toxicity in rural areas was mainly detected during the winter period when rainfall and river flow are high. Toxicity test results were compared with measured physicochemical water-quality data. Mortality and alterations in reproductive success of Daphnia spp. were not consistently reflected in detected chemical pollution. With only one exception (atrazine), detected pesticide concentrations were below known toxicity levels. However, additive and synergistic effects of the presence of a mixture of pesticides could not be excluded as a possible cause of observed toxicity. At several stations, filtering of the water sample led to a strong decrease in toxicity, which suggests the presence of xenobiotics attached to the smaller sediment fraction. Inclusion of sediment chemical analysis and sediment toxicity testing in future work should therefore be encouraged. The presented approach provided information about the adverse effects of human activities on surface water quality in the watershed, not easily obtained from classical monitoring schemes. In specific cases, the approach may represent an economically attractive alternative to physicochemical analyses. Modifications to the proposed methodology should be introduced if the effects of intrastorm and interstorm variability of water quality are to be analyzed.

Identification of the Herbicide Surflan and Its Active Ingredient Oryzalin, a Dinitrosulfonamide, as Xenoestrogens by L. C. Hall; J. M. Rogers; M. S. Denison; M. L. Johnson (pp. 201-208).

Numerous environmental contaminants have been identified as endocrine disruptors (EDs) — substances that alter endocrine homeostasis by interfering with the biological action, production, or pharmacokinetics of endogenous hormones. Xenoestrogens are those EDs whose biological activity is similar to endogenous estrogen. This report presents data that identified Surflan, a proprietary herbicide emulsion, and its active ingredient oryzalin as xenoestrogens. In vitro, Surflan and oryzalin activated an estrogen-inducible reporter gene, and oryzalin competitively displaced 17β-estradiol from the estrogen receptor. In vivo, Surflan and oryzalin induced expression of estrogen-regulated high-molecular-weight choriogenin genes in medaka (Oryzias latipes). These results are consistent with the characteristics of previously identified xenoestrogens and indicate that Surflan and oryzalin have the potential to adversely affect numerous estrogen-regulated biological processes.

Horizontal and Vertical Distribution of Estrogenic Activities in Sediments and Waters from Tokyo Bay, Japan by S. Hashimoto; A. Horiuchi; T. Yoshimoto; M. Nakao; H. Omura; Y. Kato; H. Tanaka; K. Kannan; J. P. Giesy (pp. 209-216).

Endocrine-disrupting chemicals with estrogenic activity (e.g., alkylphenols) have been detected in coastal Japan. We aimed to determine estrogenic activity in extracts of river water, seawater, sediments, and sediment cores from Tokyo Bay by in vitro gene expression assay. Fifty-one of 57 extracts had some estrogenic activity. E₂ equivalents (ng E₂ equivalents per gram dry weight or per liter above the limit of detection) in river water samples ranged from 0.70 to 4.01 ng/L; in seawater samples from 0.34 to 2.52 ng/L; and in surface sediments from 2.07 to 12.1 ng/g. The relationship between salinity and estrogenic activity in water samples suggested that fresh water is one source of environmental estrogens in Tokyo Bay. Fractionation of sediment extracts showed that the highest estrogenic activity was observed in the midpolar fraction. The observed activities were compared with activities mediated by known concentrations of nonylphenol, bisphenol-A, estrone, and 17β-estradiol. In sediment collected near the sewage treatment plants, the estrogenic activity of the midpolar fraction could be explained about 34% by nonylphenol and estrone contained in this fraction. Core sediment measurements detected estrogenic activity from as far back as the 1960s. The regulations on the industrial wastewater in early 1970s would be one of the main reasons for the lower estrogenic activity in the upper section of the sediment core. The high estrogenic activities as measured in water and sediment samples from Tokyo might be restricted to certain coastal areas.

Reproductive Endocrine Disruption in a Sentinel Species (Chrysemys picta) on Cape Cod, Massachusetts by M. T. Rie; N. Kitana; K. A. Lendas; S. J. Won; I. P. Callard (pp. 217-224).

Freshwater turtles (Chrysemys picta) were collected from two sites on Cape Cod, MA. One site (Moody Pond), adjacent to the Massachusetts Military Reservation (MMR), was considered potentially impacted by toxic agents deriving from contaminant point sources on the MMR. The second (reference) site (Washburn Pond), to the east of the MMR, was considered not impacted by these pollutants and was chosen as a control site. Plasma estradiol 17 β and vitellogenin were significantly lower in female turtles from Moody Pond. Ovarian follicular analysis indicated a significant decrease in the >16.00-mm follicular cohort in Moody Pond female turtles compared with Washburn Pond animals. Although testicular weight was lower at the Moody Pond site, histology, plasma testosterone, and sperm number were similar to these parameters in Washburn Pond animals. The data suggest that in Moody Pond, the reproductive capacity of turtles may be negatively affected by contaminants from the MMR.

Differential Uptake and Transport of Trivalent and Hexavalent Chromium by Tumbleweed (Salsola kali) by J. L. Gardea-Torresdey; G. de la Rosa; J. R. Peralta-Videa; M. Montes; G. Cruz-Jimenez; I. Cano-Aguilera (pp. 225-232).

Experiments were conducted to determine the differential absorption of Cr species by tumbleweed (Salsola kali) as well as the effect of this heavy metal on plant growth and nutrient uptake. Tumbleweed seeds were grown in an agar-based media containing different concentrations of either Cr(III) or Cr(VI). The results demonstrated that the uptake of Cr was influenced by the Cr concentration in the growth medium and the speciation of this heavy metal. When supplied in the hexavalent form, the concentration of Cr in the different plant parts (2900, 790, and 600 mg kg⁻¹ for roots, stems, and leaves, respectively) was between 10 and 20 times higher than the amounts found when Cr was supplied in the trivalent form. In addition, it was found that in most of the experiments, Cr(III) exhibited more toxic effects on tumbleweed plants than Cr(VI). The size of roots of plants grown in 20 mg L⁻¹ Cr(III) were significantly smaller (p < 0.05) than those grown in 20 mg L⁻¹ Cr(VI). Plants exposed to 20 mg L⁻¹ Cr(III) produced shoots significantly shorter (p < 0.05) compared with the size of control plants and with those grown in 20 mg L⁻¹ Cr(VI). In addition, the absorption of macronutrients and microelements was in general lower when the plants were grown in the medium containing Cr(III). The amounts of Cr concentrated in the aerial plant parts under experimental conditions may indicate tumbleweed as a new option for the phytoremediation of Cr-contaminated soil.

Bioaccumulation and Toxicity of Dissolved Heavy Metals from the Guadalquivir Estuary After the Aznalcóllar Mining Spill Using Ruditapes philippinarum by M. Laura Martín-Díaz; Julián Blasco; Marisa González de Canales; Diego Sales; T. Ángel DelValls (pp. 233-241).

The shore clam Ruditapes philippinarum was used as a biomonitoring organism to measure the potential impact that the mining spill in the Guadalquivir Estuary (SW, Spain) in 1998 may have exerted on local biota. Individuals were exposed to dissolved cadmium, copper, and zinc at concentrations found in local waters after the spill (3 μg · L⁻¹ Cd, 15 μg · L⁻¹ Cu, 700 μg · L⁻¹ Zn) at two salinity values: 10 and 35. Residues of metals were measured in gill and digestive gland, together with metallothioneins in the digestive gland and histopathological lesions in gill, digestive gland, and gonad tissues over time. Heavy metals Zn and Cd associated with the mining spill, were bioaccumulated in clam tissues, associated with the activation of metallothioneins, and related to the histopathological lesions measured at all the clam tissues. The heavy metal Cu not related to the spill was not directly associated with effects measured. The bioaccumulation and adverse effects associated with Cd and Zn were significantly higher at low salinity (10) than at high salinity (35) values.

Comparative Sublethal Toxicity of Nine Pesticides on Olfactory Learning Performances of the Honeybee Apis mellifera by A. Decourtye; J. Devillers; E. Genecque; K. Le Menach; H. Budzinski; S. Cluzeau; M. H. Pham-Delègue (pp. 242-250).

Using a conditioned proboscis extension response (PER) assay, honeybees (Apis mellifera L.) can be trained to associate an odor stimulus with a sucrose reward. Previous studies have shown that observations of conditioned PER were of interest for assessing the behavioral effects of pesticides on the honeybee. In the present study, the effects of sublethal concentrations of nine pesticides on learning performances of worker bees subjected to the PER assay were estimated and compared. Pesticides were tested at three concentrations. The highest concentration of each pesticide corresponded to the median lethal dose value (48-h oral LD50), received per bee and per day, divided by 20. Reduced learning performances were observed for bees surviving treatment with fipronil, deltamethrin, endosulfan, and prochloraz. A lack of behavioral effects after treatment with λ -cyalothrin, cypermethrin, τ -fluvalinate, triazamate, and dimethoate was recorded. No-observed-effect concentrations (NOECs) for the conditioned PER were derived for the studied pesticides. Our study shows that the PER assay can be used for estimating sublethal effects of pesticides on bees. Furthermore, comparisons of sensitivity as well as the estimation of NOECs, useful for regulatory purposes, are possible.

A Comparison of the Toxicity of Synergized and Technical Formulations of Permethrin, Sumithrin, and Resmethrin to Trout by E. A. Paul; H. A. Simonin; T. M. Tomajer (pp. 251-259).

Synthetic pyrethroids often have synergists added to improve effectiveness, yet decisions regarding the use of these pesticides are often based upon toxicity tests using technical material without the synergist, piperonyl butoxide. We conducted toxicity tests with brook trout (Salvelinus fontinalis) and brown trout (Salmo trutta) to compare the toxicity of synergized and technical formulations of permethrin, sumithrin, and resmethrin. We found a significant increase in toxicity in the synergized permethrin formulation using traditional 24, 48, and 96-h tests, relative to tests with the technical formulation. However, there was little difference in toxicity between synergized and technical sumithrin until 48 h had elapsed. Many test fish were strongly intoxicated by either formulation of permethrin or sumithrin, but the synergized formulations of both chemicals affected fish at lower concentrations. Intoxication was potentially severe enough to reduce the survival of these fish in the wild. Following short (6-h) exposures, we also found a larger difference in the number of fish that died or became intoxicated between the synergized and technical formulations of permethrin and sumithrin. Finally, we tested the ability of exposed fish to swim against a current. Fish exposed for 6 h to synergized permethrin and resmethrin had far less swimming stamina than those exposed to technical formulations. We found no difference in the effect on swimming between the synergized and technical formulation of sumithrin. In general, the synergized formulations of these chemicals appeared to cause a faster response than the technical formulations. This response increases the lethal and sublethal impacts of the insecticides. We also found that sumithrin was the least toxic of the three pyrethroids. Since the maximum application rate of sumithrin is half that of the other two pyrethroids, the potential risk to wild trout in streams may be reduced.

Environmental Toxicity Studies Using Chickens as Surrogates for Wildlife: Effects of Vehicle Volume by Jamie C. DeWitt; Erin B. Meyer; Diane S. Henshel (pp. 260-269).

Domestic chicken embryos are frequently used for avian developmental toxicity studies of polyhalogenated aromatic hydrocarbons, which are often injected into eggs with oil-based vehicles. The volume of toxicant and vehicle injected ranges from relatively low volumes (0.1–0.5 μl/g egg) to relatively high volumes (1.0 μl/g egg and above). Previous research from our laboratory suggested that high volumes of vehicle oil may disrupt normal growth of chicken embryos, possibly from hypoxia-like effects. This analysis explored the potential effects of vehicle volume on developing chicken embryos. We assessed standard measures of mortality, organ growth, body growth, and behavior from chickens developmentally exposed in ovo prior to incubation to low (0.1 μl/g egg) or high volumes (1.0 μl/g egg) of corn oil injected into airsacs or to no injection. The chickens receiving high volumes of oil showed increases in overall embryonic mortality and early embryo mortality compared to chickens receiving low volumes of oil or no injection. The chickens receiving high volumes of oil showed decreased activity during righting reflex, running time, visual discrimination, and olfactory aversion tests, and increased activity during an open-field activity test compared to chickens receiving low volumes of oil or no injection. Somatic endpoints do not appear to be affected by high volume injections. These results suggest that high volumes of vehicle injected into airsacs of eggs may lead to hypoxia-like conditions that increase embryonic mortality and disrupt simple behaviors. However, some effects of volume may diminish when injections are performed later in incubation.

Environmental Toxicity Studies Using Chickens as Surrogates for Wildlife: Effects of Injection Day by Jamie C DeWitt; Erin B Meyer; Diane S Henshel (pp. 270-277).

Domestic chicken embryos are frequently used for avian developmental toxicity studies of polyhalogenated aromatic hydrocarbons, which are often injected into eggs with oil-based vehicles. Injection times range from immediately prior to incubation (embryonic day zero, E0) to after 4 days of incubation (E4) and beyond. Because the majority of organogenesis in chicken embryos occurs during the first 4 days of development, injection after E0 may miss critical, sensitive, developmental periods. We evaluated whether differences in the day of vehicle administration would lead to differences in standard measures of embryotoxicity. We assessed embryotoxicity using mortality, organ somatic indices, teratogenesis, and behavior in hatchling chickens developmentally exposed to a high volume (1.0 μl/g egg) of corn oil vehicle, which was injected into the airsac at E0 or E4. The E0 vehicle group had 76.5% higher overall embryo mortality, embryos died earlier in development, and hatchlings took more than two times longer to right in a righting reflex test compared to the E4 vehicle group. Other behavioral results demonstrated that hatchling chickens from the E0 vehicle group performed differently from their respective no-inject controls, whereas hatchling chickens from the E4 vehicle group did not. The bursal somatic index differed statistically by injection day and weighed 23.7% more in the E0 vehicle group than the E4 vehicle group. These results suggest that the embryonic day of contaminant injection is an important consideration, particularly when using a high volume of vehicle to evaluate developmental toxicity of a contaminant on embryo mortality or behavior.

Effects of Calcium on the Uptake and Elimination of Cadmium and Zinc in Asiatic Clams by J.-W. Qiu; Z.-C. Xie; W.-X. Wang (pp. 278-287).

We examined the effects of Ca, one of the major ions contributing to water hardness, on the uptake and elimination of Cd and Zn in the Asiatic clam Corbicula fluminea, a common bivalve species in the rivers and streams of Southern China. Over the wide range of dissolved Cd (4.5 to 446 nM) and Zn (15.3 to 1529 nM) concentrations, uptake of both metals increased with increasing dissolved metal concentration, showing Michaelis-Menten–type saturation kinetics. At each tested metal concentration, increased Ca concentration led to a significantly lower metal uptake. When the Ca, Cd, Zn concentrations were expressed as free-ion activities, Cd and Zn uptake data fitted the Michaelis-Menten inhibition model well. We also quantified the effects of the two Ca blockers on metal uptake by the clams. Verapamil significantly inhibited the uptake Cd and Zn, but the inhibitive effect of lanthanum on metal uptake was more evident for Cd than for Zn. Ca did not significantly affect the assimilation efficiency of either metal from ingested phytoplankton, nor did it affect the elimination of the two metals during a 1-month depuration period. Our study has shown that Ca inhibited the uptake of metals from water; such an effect could be predicted based on the free-metal ion activities. Ca did not seem to have a direct effect on metal assimilation from food or metal elimination from the bivalve.

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Archives of Environmental Contamination and Toxicology (v.48, #2)