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Archives of Environmental Contamination and Toxicology (v.52, #1)

Ecotoxicity Monitoring of Hydrocarbon-Contaminated Soil During Bioremediation: A Case Study by Tomáš Hubálek; Simona Vosáhlová; Vít Matějů; Nora Kováčová; Čeněk Novotný (pp. 1-7).

The ecotoxicity of hydrocarbon-contaminated soil originating from a brownfield site was evaluated during a 17-month biodegradation pilot test. The initial concentration of total petroleum hydrocarbons (TPHs) in the soil was 6380 μg/g dry weight. An amount of 200 kg soil was inoculated with 1.5 L of the bacterial preparation GEM-100 containing Pseudomonas sp. and Acinetobacter sp. strains (5.3 × 10¹⁰ CFU.mL⁻¹) adapted to diesel fuel. The concentration of TPHs in the soil decreased by 65.5% after bioremediation. Different organisms such as the bacterium Vibrio fischeri, terrestrial plants Sinapis alba, Lactuca sativa, and Hordeum vulgare, the water plant Lemna minor, the earthworm Eisenia fetida, and the crustacean Heterocypris incongruens were used for ecotoxicity evaluation. The highest toxicity was detected in the first period of bioremediation. However, certain toxic effects were detectable during the whole bioremediation process. The contact tests with plants, earthworms, and crustaceans were the most sensitive of all of the bioassays. Therefore, the contact tests performed directly on soil samples were shown to be a better tool for ecotoxicity evaluation of hydrocarbon-contaminated soil than the tests performed on soil elutriates. The ecotoxicity measured by the responses of the tests did not always correlate with the decrease in TPH concentrations in the soil during bioremediation.

Oxidative Degradation of BPA Using TiO₂ in Water, and Transition of Estrogenic Activity in the Degradation Pathways by Kei Nomiyama; Teiji Tanizaki; Toyokazu Koga; Koji Arizono; Ryota Shinohara (pp. 8-15).

The oxidative degradation behavior of bisphenol A (BPA) using titanium dioxide (TiO₂) in water was investigated. The main purposes were to clarify the relationship with estrogenic activity from the degradation pathways demonstrated by oxidation of BPA in water. Approximately 99% of the BPA decomposed within 300 min, and gas chromatography (GC) mass spectrometry (MS) and liquid chromatography (LC) MS analysis revealed many intermediates during the degradation process. Intermediates by decomposition of BPA, such as hydroxylated-BPA (OH-BPA), carboxylic intermediates, phenolic intermediates, and other intermediates produced by the cleavage of a benzene ring were identified and quantified. Estrogenic activities of the degradation pathways of the BPA in water were assessed by using a constructed yeast two-hybrid assay system for human estrogen receptor α (hERα) and Japanese medaka fish (Oryzias latipes) estrogen receptor α (medERα). Estrogenic activity for hERα and medERα was reduced to less than 20% of the initial activity for BPA after 240 min of UV irradiation. However, estrogenic activity for medERα was increased by 110% from the initial activity for BPA at 60 min of UV irradiation. It was estimated that medERα assay was more sensitive for BPA and the intermediates than was the hERα assay. From these findings, we estimate that the intermediates by the oxidation of BPA have the behaviors of xenoestrogen to the aquatic wildlife in the environment.

Role of Simulated Acid Rain on Cations, Phosphorus, and Organic Matter Dynamics in Latosol by Da-Jiong Ling; Jia-En Zhang; Ying Ouyang; Qian-chun Huang (pp. 16-21).

Acid rain is a serious environmental problem in the world and is of a particular concern in southern China where most of the soils are acidic. This study investigated the dynamics of cations, phosphorus (P), and soil organic matter (SOM) in the Latosol (acidic red soil) from south China under the influences of simulated acid rain (SAR). Laboratory experiments were performed by leaching the soil columns with SAR at pH levels ranging from 2.5 to 7.0 over a 21-day experimental period. Results show that about 34, 46, 20, and 77% of the original exchangeable soil Ca⁺², Mg⁺², K⁺, and Na⁺, respectively, were leached out by the SAR at pH 2.5 after 21 days. Two distinct patterns of the available phosphorus (AP) concentrations were observed: one at pH≤3.5 and the other at pH ≥ 4.0. At pH≤3.5, concentrations of the AP increased from the beginning of the experiments to day 5, then decreased from day 5 to 15, and finally increased from day 15 to the end of the experiments. At pH ≥ 4.0, concentrations of the AP increased consecutively from the beginning of the experiments to day 10 and decreased from day 10 to the end of the experiments. Such a finding is useful for agricultural practices since soil P is one of the most important macronutrients for plant growth. In general, SOM content decreased with time as the Latosol was leached by the SAR at all pH levels. A maximum concentration of soil fulvic acid was found after 15 days of the experiments due to the degradation of the SOM. A multiple regression analysis showed that a very strong relationship was obtained between the soil AP and the other three parameters (i.e., pH, SOM, and sorption P).

Influence of Laboratory Storage on the Organic Contaminant Content and Water-Extractable Ecotoxicological Potential of Soil Samples by Jean-Paul Rila; Adolf Eisentraeger (pp. 22-31).

At present there is a high level of uncertainty about whether or not soil samples, which are required to be toxicologically characterized, could be preserved without affecting their toxicological characteristics. In the existing DIN/ISO standards for soil investigations, there is no documentation on the storage of soil samples after collection for (eco-)toxicological investigations. Furthermore, procedures for receiving justifiable and verifiable results from soil samples are quite vague. Therefore, the stability and changes in the biological effects of different soil contaminants were investigated by varying the storage conditions and the storage times of the soil. The limitations of storing soils could therefore be subsequently outlined. Recommendations for the optimization of storage conditions for back-up soil samples were made. These recommendations have the capability of finding entrance into DIN/ISO standards.

Influence of Phosphate on the Response of Periphyton to Atrazine Exposure by H. Guasch; V. Lehmann; B. van Beusekom; S. Sabater; W. Admiraal (pp. 32-37).

After indications from the literature that nutrient concentrations may modify the toxicity of herbicides to natural periphyton communities, this study aims to provide experimental proof for atrazine. In this microcosm experiment, phosphate (P) addition did not ameliorate atrazine toxicity to periphyton. Three weeks of P addition did not increase atrazine tolerance (measured as EC50 in acute toxicity tests), whereas exposure to atrazine under conditions that were either P-limited or non-P-limited clearly reduced the development of algal biomass. Long-term exposure to atrazine induced tolerance of the community to the herbicide, and this was not influenced by P addition. Tolerance induction in this microcosm experiment has been compared with previously published field data from the same area of study and indicates that tolerance induction by atrazine may take place under atrazine exposure in streams as well as in microcosms.

Response of Pepper Plants (Capsicum annum L.) on Soil Amendment by Inorganic and Organic Compounds of Arsenic by Jiřina Száková; Pavel Tlustoš; Walter Goessler; Daniela Pavlíková; Ernst Schmeisser (pp. 38-46).

The influence of soil contamination by inorganic and organic arsenic compounds on uptake, accumulation, and transformation of arsenic in pepper (Capsicum annum L.) was investigated in greenhouse pot experiments under controlled conditions. Pepper plants were cultivated in substrate amended by aqueous solutions of arsenite, arsenate, methylarsonic acid (MA), and dimethylarsinic acid (DMA) applied individually into cultivation substrate at concentrations of 15 mg As per kg of substrate. The plant availability of the arsenicals increased in the order arsenite = arsenate < MA < DMA. The highest arsenic concentrations were found in roots followed by stems, leaves, and fruits regardless of arsenic compound applied. In the control samples of pepper fruits, As(III), As(V), and DMA were present (25%, 37%, and 39% of the water-extractable arsenic). In control stems + leaves and roots, As(V) was the major compound (63% and 53% in a phosphate buffer extract) followed by As(III) representing 33% and 42%. Additionally, low concentrations (not exceeding 5%) of DMA and MA were detected as well. In all the soils analyzed after the first harvest of pepper fruits, arsenate was the dominating compound followed by arsenite. Methylarsonic acid, methylarsonous acid, and DMA were present at varying concentrations depending on the individual soil treatments. In the treated plants, the arsenic compounds in plant tissues reflected predominantly the extractable portions of arsenic compounds present in soil after amendment, and this pattern was more significant in the first part of vegetation period. The results confirmed the ability of generative parts of plants to accumulate preferably organic arsenic compounds, whereas in the roots and aboveground biomass, mainly inorganic arsenic species are present. Evidently, the source of soil arsenic contamination affects significantly the extractable portions of arsenic compounds in soil and subsequently the distribution of arsenic compounds within the plants.

Preliminary Examination of Short-Term Cellular Toxicological Responses of the Coral Madracis mirabilis to Acute Irgarol 1051 Exposure by C. Downs; A. Downs (pp. 47-57).

Irgarol 1051 is an s-triazine herbicide formulated with Cu2O in antifouling paints. Recent studies have shown that Irgarol 1051 inhibits coral photosynthesis at environmentally relevant concentrations, consistent with its mode of action as a photosystem II inhibitor. Related toxicologic effects of this herbicide on coral cellular physiology have not yet been investigated. We used cellular diagnostics to measure changes in 18 toxicologic cellular parameters in endosymbiotic algal (dinoflagellate) and cnidarian (host) fractions of the common branching coral Madracis mirabilis associated with in vivo 8- and 24-hour exposures to a nominal initial Irgarol 1051 concentration of 10 μg L⁻¹. Responses measured were (1) xenobiotic response, which includes total and dinoflagellate multixenobiotic resistance (MXR), cnidarian cytochrome (CYP) P450−3 and P450-6 classes, cnidarian, and dinoflagellate glutathione-s-transferase (GST); (b) oxidative damage and response, which includes cnidarian and dinoflagellate Cu/Zn and Mn superoxide dismutase (SOD), cnidarian and dinoflagellate glutathione peroxidase (GPx), cnidarian catalase, and total protein carbonyl); (3) metabolic homeostasis, which includes chloroplast and invertebrate small heat-shock proteins (sHsp), cnidarian protoporphyrinogen oxidase IX (PPO), cnidarian ferrochelatase, and cnidarian heme oxygenase; and (4) protein metabolic condition, which includes cnidarian and dinoflagellate heat shock proteins (hsp70 and hsp60), total ubiquitin, and cnidarian ubiquitin ligase. Acute responses to Irgarol 1051 exposure included significant increases in total and dinoflagellate MXR, dinoflagellate Cu/Zn SOD, dinoflagellate chloroplast sHsp, and cnidarian PPO. Irgarol 1051 exposure resulted in decreases in cnidarian GPx, cnidarian ferrochelatase, cnidarian catalase, and cnidarian CYP 450-3 and -6 classes. Related implications of Irgarol 1051 exposure to coral cellular condition are discussed.

Impact of Diuron on Aneuploidy and Hemocyte Parameters in Pacific Oyster, Crassostrea gigas by Karine Bouilly; Marc Bonnard; Béatrice Gagnaire; Tristan Renault; Sylvie Lapègue (pp. 58-63).

Diuron is a substituted urea herbicide used for agricultural and nonagricultural weed control. Its widespread use and relatively slow breakdown led us to analyze its influence on aneuploidy level (lowered chromosome number in a percentage of somatic cells) and hemocyte parameters in Pacific oysters, Crassostrea gigas. Adult oysters were subjected to two diuron concentrations (300 ng L⁻¹ and 3 μg L⁻¹) for 11 weeks. Significantly higher aneuploidy level was observed in diuron-treated oysters compared with the control. Furthermore, the observed impact on aneuploidy persisted to the next generation as offspring exhibited significantly higher aneuploidy levels when their parents had been exposed to diuron. Significant increases in hemocyte parameters (cell mortality, phagocytosis, granulocyte percentage, reactive oxygen species, and lysosome presence) of the adults were also observed after 4 weeks of diuron exposure. The effects observed on oyster aneuploidy level and hemocyte parameters could have serious environmental and practical consequences.

Laboratory Investigation of the Toxicity and Interaction of Pesticide Mixtures in Daphnia magna by Tara K. George; Karsten Liber (pp. 64-72).

The probabilistic ecological risk assessment-toxic equivalent (PERA-TE) combination approach is a relatively new risk assessment approach used to assess the toxicity and interaction of chemical mixtures. The validity and effectiveness of the PERA-TE combination approach has been tested previously in field microcosm studies using pesticide mixtures. The related laboratory studies described here, using Daphnia magna, were conducted to verify the conclusions made regarding the toxicity and interaction of the mixtures tested in the microcosms. Two types of pesticide mixture were assessed: the first consisted of pesticides with similar modes of action (chlorpyrifos, diazinon, and azinphos-methyl; OP mixture), and the second consisted of pesticides with different modes of action (chlorpyrifos, endosulfan, and trifluralin; CET mixture). Similar to the field studies, PERA-TE mixtures with a predetermined effect assessment criterion (10th centile of acute toxicity effects distributions) and proportional ratio (89:11 for binary mixture and 80:10:10 for ternary mixtures) were tested. Further assessment of the (PERA-) TE approach was achieved by altering the effect assessment criterion (to EC/LC₅₀ point estimates) and the proportional ratio of the pesticides in the mixture (to 50:25:25). Generally, but with some exceptions, basing mixtures on species-specific effect criteria and/or changing the proportional ratio of pesticides in the mixture redistributed the concentration of pesticides in the mixture to produce an equitoxic response. The ability to produce these equitoxic responses supported the conclusions drawn from the field studies: The pesticide toxicity in the OP and CET PERA-TE mixtures were effectively additive. Furthermore, it is shown that these alternative (PERA-) TE mixtures would be suitable to confirm or reject the interaction of chemicals in a PERA-TE mixture.

Impact of Waterborne Copper on the Structure of Gills and Hepatopancreas and Its Impact on the Content of Metallothionein in Juvenile Giant Freshwater Prawn Macrobrachium rosenbergii (Crustacea: Decapoda) by Na Li; Yunlong Zhao; Jian Yang (pp. 73-79).

This study aims to elucidate both the impact of copper on the structure of the gills and hepatopancreas and the induction of metallothionein (MT) during waterborne copper exposure in juvenile Macrobrachium rosenbergii. Structural observations were performed with light microscopy, and the MT titrations were performed with the cadmium saturation assay. The structural changes that occurred in the gills and hepatopancreas appeared to result from copper accumulation, and the degree of damage observed in both tissues was relevant to the elevated waterborne copper concentration. Exposure to copper concentrations ranging from 0.01 mg/L to 0.4 mg/L for 7 days resulted in profound structural changes including the accumulation of hemocytes in the hemocoelic space; swelling and fusion of the lamellae; abnormal gill tips; and hyperplastic, necrotic, and clavate–globate lamellae in the gills. Similarly, hemocytic infiltration in the interstitial sinuses, an increased number of hemocytes, thickening and ruptures of the basal laminae, and necrosis of the tubules were observed in the hepatopancreas. The MT measurements showed no significant differences in MT contents between the control group and the group treated with 0.01 mg/L waterborne copper. The maximum MT content was observed at the level of 0.4 mg/L waterborne copper.

Contaminants in Fish of the Hackensack Meadowlands, New Jersey: Size, Sex, and Seasonal Relationships as Related to Health Risks by Peddrick Weis; Jeffrey T. F. Ashley (pp. 80-89).

The trace metal content and related safety (health risk) of Hackensack River fish were assessed within the Hackensack Meadowlands of New Jersey, USA. Eight elements were analyzed in the edible portion (i.e., muscle) of species commonly taken by anglers in the area. The white perch collection (Morone americana) was large enough (n = 168) to enable statistically significant inferences, but there were too few brown bullheads and carp to reach definite conclusions. Of the eight elements analyzed, the one that accumulates to the point of being a health risk in white perch is mercury (Hg). Relationships between mercury concentrations and size and with collection season were observed; correlation with lipid content, total polychlorinated biphenyl (PCB) content, or collection site were very weak. Only 18% of the Hg was methylated in October (n = 8), whereas June and July fish (n = 12) had 100% methylation of Hg. White perch should not be considered edible because the Hg level exceeded the “one meal per month” action level of 0.47 μg/g wet weight (ppm) in 32% of our catch and 2.5% exceeded the “no consumption at all” level of 1 μg/g. The larger fish represent greater risk for Hg. Furthermore, the warmer months, when more recreational fishing takes place, might present greater risk. A more significant reason for avoiding white perch is the PCB contamination because 40% of these fish exceeded the US Food and Drug Administration (FDA) action level of 2000 ng/g for PCBs and all white perch exceeded the US Environmental Protection Agency cancer/health guideline (49 ng/g) of no more than one meal/month. In fact, nearly all were 10 times that advisory level. There were differences between male and female white perch PCB levels, with nearly all of those above the US FDA action level being male. Forage fish (mummichogs and Atlantic silversides) were similarly analyzed, but no correlations were found with any other parameters. The relationship of collection site to contaminants cannot be demonstrated because sufficient numbers of game fish could not be collected at many sites at all seasons.

Biosynthetic Capacity of Rainbow Trout (Oncorhynchus mykiss) Interrenal Tissue After Cadmium Exposure by H. M. Lizardo-Daudt; Onkar S. Bains; Christopher R. Singh; Christopher J. Kennedy (pp. 90-96).

The disruption of endocrine system function in wildlife species, including teleosts, by contaminants such as metals is presently of major environmental concern. Recently, it has been shown that cadmium (Cd) exposure results in significant reductions in corticosteroid secretion by fish interrenal steroidogenic cells, likely through an inhibition of intracellular cortisol synthesis. In the present study, the effects of CdCl₂ on unstimulated and stimulated interrenal steroidogenesis in rainbow trout were examined with the intention of furthering an understanding of the site(s) of Cd toxic action. CdCl₂ alone reduced cortisol secretion in minced interrenal tissues to 59% and 55% of control values when exposed to 10 and 100 μM, respectively. Incubation of interrenal tissues with 0.01 IU/mL adrenocorticotropic hormone (ACTH), which activates rate-limiting steps in steroid synthesis, resulted in significant stimulation of steroidogenesis in controls. However, ACTH-stimulated steroidogenesis was reduced when tissues were previously incubated with Cd. Maximal rates of unstimulated cortisol secretion were achieved by augmentation using 5 μM 25-hydroxycholesterol (25-OHC) or 0.8 μL/mL synthetic cholesterol [SyntheChol^®(SC)]. Steroidogenesis augmentation by 25-OHC was significantly reduced in tissues incubated with Cd. Interestingly, cortisol secretion was significantly higher in SC-augmented tissue exposed to 1 and 10 μM Cd when compared to augmented control tissues. The results of this study show that Cd affects both stimulated and unstimulated steroidogenesis in rainbow trout, and that one major site(s) of action of Cd in the cortisol synthesis pathway is likely prior to cytochrome P450 side chain cleavage.

Relationship Between Wetlands and Mercury in Brook Trout by Mark S. Castro; Robert H. Hilderbrand; Joe Thompson; A. Heft; S. E. Rivers (pp. 97-103).

The purpose of this study was to determine if wetlands influence mercury concentrations in brook trout (Salvelinus fontinalis), benthic macroinvertebrates, and stream water. On September 26, 2005, water samples, benthic macroinvertebrates, and brook trout were collected from four streams in western Maryland under low-flow conditions. Water samples were also collected in these four streams under high-flow conditions in January 2006. The watersheds of Blue Lick and Monroe Run did not contain wetlands, but the watersheds of the Upper Savage River (3% of upstream area) and Little Savage River (7% of upstream area) contained wetlands. We found significantly (p = 0.05) higher average total mercury concentration in brook trout from Little Savage River (129 ± 54 ng g⁻¹); intermediate concentrations (66 ± 19 ng g⁻¹) in brook trout from Upper Savage River; and lowest concentrations in brook trout from Blue Lick (28 ± 11 ng g⁻¹) and Monroe Run (23 ± 19 ng g⁻¹). Brook trout in all streams accumulated mercury at the same rate over their lifetimes, but the youngest fish had significantly different mercury concentrations (Little Savage > Upper Savage > Blue Lick = Monroe Run), which may be due to differences in mercury concentrations in the eggs or food for the fry. Mercury concentrations in brook trout were not consistent with mercury concentrations in stream water and benthic macroinvertebrates. The Little Savage River had significantly higher total and methylmercury concentrations in stream water, but mercury concentrations in the other streams and in the benthic macroinvertebrates were not significantly different among streams. The unusually high methylmercury concentrations (0.5 to 2.1 ng L⁻¹) in the Little Savage River may have been caused by production of methylmercury in the pools. The relatively low methylmercury concentrations in the Upper Savage River may be caused by a mercury concentration gradient downstream of the wetland.

Effects of a Branched p-Nonylphenol Isomer (4(3′,6′-dimethyl-3′-heptyl)-phenol) on Embryogenesis in Lymnae stagnalis L. by J. O. Lalah; G. F. Severin; K.-W. Schramm; D. Lenoir; A. Behechti; K. Guenther; A. Kettrup (pp. 104-112).

The tertiary branched alkyl-chain isomers of p-nonylphenol (NP) are perceived to have more estrogenic potency than its constituent secondary and primary straight alkyl-chain isomers. Investigations with single tertiary branched isomers of NP can therefore contribute toward the elucidation of the mechanisms of toxicity and estrogenicity of NP. A single tertiary branched alkyl-chain isomer (4(3′,6′-dimethyl-3′-heptyl)-phenol) was used in studies to determine its effects on embryonic growth and mortality in Lymnaea stagnalis L. Egg masses were exposed to the test compound for 20 days in a static waterborne-exposure regime with an average NP concentration of 105 μg/L and water temperature range of 18–20°C. Observations were made under a microscope and pictures were taken with a digital camera to determine the various developmental stages of growth, the duration of growth in each stage, embryo hatchability, and embryo mortality. The isomer was found to cause significant delay in all stages of growth and more significantly in the Morula and Veliger stages. An increase in embryo mortality, from the third day until the end of the experiment, was observed in exposed egg masses compared to controls. The hatching success of embryos was also significantly reduced by exposure, with 81% hatchability in exposed egg masses compared to 93% in the controls, after 18 days of continuous exposure. The encapsulating jelly strand that completely covers the rows of egg masses may have prevented the isomer residues from effectively penetrating into the embryos as shown by the observed low bioconcentration factors of the isomer in egg masses during exposure, resulting in unexpectedly lower observed estrogenic effects. However, this factor was not investigated. In vivo biotransformation of some of the residues of the isomer into catechol metabolites by the embryos during exposure could also result in the reduction of its estrogenic potential. To understand more fully the extent of toxicity and estrogenicity of this isomer, in vitro estrogenic assays are recommended. It would also be necessary to investigate its estrogenic effects on embryo development after in vivo maternal exposure.

2,4-D Butoxyethyl Ester Kinetics in Embryos of Xenopus laevis: The Role of the Embryonic Jelly Coat in Reducing Chemical Absorption by Andrea N. Edginton; Claude Rouleau; Gerald R. Stephenson; Herman J. Boermans (pp. 113-120).

The role of the jelly coat in providing a protective barrier to chemical absorption was studied using the embryos of the amphibian, Xenopus laevis. Embryos with or without a jelly coat were water exposed to the butoxyethyl ester of 2,4-dichlorophenoxyacetic acid (2,4-D BEE) and the rates of uptake, metabolism, distribution, and excretion were determined. The water uptake clearance rates were slower for embryos with a jelly coat (1.5–4.5 ml_water·g _embryo ⁻¹ ·h⁻¹ or 0.040–0.022 ml_water·h⁻¹ per embryo) in comparison to dejellied embryos (14–21 ml_water·g _embryo ⁻¹ ·h⁻¹ 0.0066–0.021 ml_water·h⁻¹ per embryo). This accounted for the much lower residues in embryos with a jelly coat than in dejellied embryos during 8 h of exposure. Despite quantitative differences in uptake, once 2,4-D BEE had entered the embryos, metabolism and distribution were similar between the two test groups. 2,4-D BEE was metabolized to 2,4-dichlorophenoxyacetic acid (2,4-D) with half-lives ranging from 35 to 42 minutes. The radioactive residues, as determined by whole body autoradiography, appeared throughout the embryo with a slight accumulation in the blastocoel. Furthermore, 35% of the radioactive residues were located in the jelly coat and 65% in the developing embryo. Based on a slower 2,4-D elimination in embryos with a jelly coat, the diffusive properties that decreased 2,4-D BEE uptake appeared to similarly decrease elimination of its metabolite. The common practice of removing jelly coats prior to embryonic amphibian toxicity studies, as in the widely used Frog Embryo Teratogenesis Assay–Xenopus (FETAX), is discouraged based on the kinetic differences observed in this study.

Blood Lead Concentrations in Waterfowl Utilizing Lake Coeur d’Alene, Idaho by Brian L. Spears; James A. Hansen; Daniel J. Audet (pp. 121-128).

The Coeur d’Alene River Basin, Lake Coeur d’Alene, and the Spokane River contain elevated heavy metal concentrations in sediment and water from historical mining and ore processing operations in the Coeur d’Alene Basin. Lead poisoning has been identified as the cause of death in hundreds of waterfowl utilizing wetlands in the floodplain of the Coeur d’Alene River, but little was known about hazards to waterfowl from heavy metal contamination in shallow bays and wetlands of Lake Coeur d’Alene. We examined lake sediment and blood lead concentrations in waterfowl utilizing Lake Coeur d’Alene, Idaho, to evaluate potential lead contamination of waterfowl utilizing the lake. We collected 56 palustrine and 102 lacustrine sediment samples and 61 mallard and 8 wood duck blood samples. Mean lead concentrations from palustrine and lacustrine sediment samples ranged from 14 to 3508 mg/kg dry weight (dw) and from 19 to 5009 mg/kg (dw), respectively. Lead concentrations in palustrine and lacustrine sediment from several Lake Coeur d’Alene bays were higher than those in lake reference areas and were higher than Bunker Hill Superfund Site target cleanup levels and suggested site-specific toxicity thresholds for swans. Mean blood lead from mallard and wood ducks sampled from Lake Coeur d’Alene bays were within lead toxicity ranges for waterfowl associated with clinical and severe clinical lead poisoning. We also collected 19 Canada goose and 3 mallard fecal samples to evaluate exposure through sediment ingestion. Waterfowl using Lake Coeur d’Alene appear to be exposed to lead by ingesting contaminated lake sediment. Our model predicts a sediment lead effects range of 147–944 mg/kg (dw) and mortality effects level of 1652 mg/kg (dw) for mallards utilizing Lake Coeur d’Alene. The locations of Harrison Slough, Powderhorn Bay, and Cottonwood Bay at the mouth of the Coeur d’Alene River and Blackwell Island and Cougar Bay near the Spokane River outflow of Lake Coeur d’Alene were the areas of greatest concern for waterfowl exposure to lead contaminated sediment.

Mercury Exposure and Effects on Cavity-Nesting Birds from the Carson River, Nevada by Christine M. Custer; Thomas W. Custer; Elwood F. Hill (pp. 129-136).

Mercury (Hg) concentrations were 15–40 times higher in the eggs and livers of tree swallows (Tachycineta bicolor) and house wrens (Troglodytes aedon) that nested along the Carson River at and below Dayton, Nevada than in the same species above the mining-impacted areas. Hg contamination was mainly the result of processing mills in the 1800s that used Hg to separate gold and silver from ore. The exposure pattern of tree swallows and house wrens along the Carson River was consistent with their trophic status (i.e., lower levels in liver tissue of aquatic insectivores than in piscivorous birds nesting nearby). Even though they are aquatic insectivores, tree swallows and house wrens were exposed to the same amount of Hg as piscivores in the Florida Everglades; this indicated the extreme level of Hg contamination in the Carson River. Only 70–74% of the eggs hatched. This was less than the nationwide average for these two species that generally hatch ≥85% of eggs. Although the sample size was small, Hg might be impacting reproductive end points in cavity-nesting birds from the Carson River. Other trace elements were present at background concentrations.

Evaluation of Developmental Toxicity of Amitraz in Sprague-Dawley Rats by J. C. Kim; J. Y. Shin; Y. S. Yang; D. H. Shin; C. J. Moon; S. H. Kim; S. C. Park; Y. B. Kim; H. C. Kim; M. K. Chung (pp. 137-144).

This study investigated the potential adverse effects of amitraz on the initiation and maintenance of pregnancy in Sprague-Dawley rats as well as its effects on embryo–fetal development after maternal exposure during the entire pregnancy period. Amitraz was administered to pregnant rats by gavage from days 1 to 19 of gestation at dose levels of 0, 3, 10, and 30 mg/kg/day. All dams underwent a caesarean section on day 20 of gestation and their fetuses were examined for any external, visceral, and skeletal abnormalities. At 30 mg/kg, maternal toxicity manifested as an increase in the incidence of abnormal clinical signs and a lower body weight gain and food intake. Developmental toxicity included an increase in the fetal death rate, a decrease in the litter size, and a reduction in the fetal body weight. In addition, there was an increase in the incidence of fetal external, visceral, and skeletal abnormalities. At 10 mg/kg, maternal toxicity observed included a decrease in the body weight gain and a decrease in food intake. In addition, minimal developmental toxicity, including a decrease in the fetal body weight, an increase in the visceral and skeletal aberrations, and a delay in fetal ossification. There were no signs of either maternal toxicity or developmental toxicity at 3 mg/kg. These results show that amitraz administered during the entire pregnancy period in rats is embryotoxic and teratogenic at the maternally toxic dose (i.e., 30 mg/kg/day) and is minimally embryotoxic at a minimally maternally toxic dose (i.e., 10 mg/kg/day). Under these experimental conditions, the no-observed-adverse-effect level of amitraz for both dams and embryo-fetal development is estimated to be 3 mg/kg/day.

Induction of Oxidative Stress in Erythrocytes of Male Rats Subchronically Exposed to a Mixture of Eight Metals Found as Groundwater Contaminants in Different Parts of India by S. H. Jadhav; S. N. Sarkar; M. Aggarwal; H. C. Tripathi (pp. 145-151).

Exposure of animals and humans to different metal components through contaminated drinking water can result in a wide range of adverse clinical conditions. Toxicological consequences arising from the concurrent repeated exposure to multiple metal contaminants are not known. The purpose of the present study was to evaluate the oxidative stress-inducing potential of a mixture of eight metals (arsenic, cadmium, lead, mercury, chromium, nickel, manganese, iron), representative of groundwater contamination in different areas of India, in erythrocytes of male rats subchronically exposed to environmentally relevant doses via drinking water. The selection of these metals, as determined by literature survey of groundwater contamination in India, was primarily based on the frequency of their occurrence and contamination level above World Health Organization maximum permissible limit (MPL) in drinking water. Male albino Wistar rats were exposed to the metal mixture at 0, 1, 10, and 100 times the mode concentrations (the most frequently occurring concentration) of the individual metals in drinking water for 90 days. In addition, one group of rats was also exposed to the mixture at a concentration equal to the MPL of individual components. The oxidative stress in erythrocytes was evaluated by assessing the magnitude of malondialdehyde production and reduced glutathione (GSH) content and the activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and glutathione reductase (GR) after 30, 60, and 90 days of exposure. MPL and 1× dose levels did not cause any changes. The mixture at 10× and 100× doses caused dose- and time-dependent effects. After 30 days, the 10× dose did not cause any changes except increase in SOD activity. The 100× dose increased the activities of SOD, catalase and GR and the GSH level, but caused no alterations in lipid peroxidation (LPO) and GPx activity. After 60 days, the 10× dose did not cause any changes. The 100× dose increased LPO and decreased all the antioxidant parameters, except GSH. After 90 days, both 10× and 100× levels elevated LPO. The 10× dose decreased GSH level and activities of SOD and catalase, but not of GPx and GR, whereas the 100× dose decreased all the antioxidative systems. Overall, the present study demonstrates that the subchronic exposure of male rats to the mixture of metals via drinking water results in induction of oxidative stress and concomitant reduction in antioxidative defense system in erythrocytes at 10 and 100 times the mode concentrations of the individual metals in contaminated groundwater.

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