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Comparison of methods for conducting marine and estuarine sediment porewater toxicity tests—extraction, storage, and handling techniques

A series of studies was conducted to compare different porewater extraction techniques and to evaluate the effects of sediment and porewater storage conditions on the toxicity of pore water, using assays with the sea urchin Arbacia punctulata . If care is taken in the selection of materials, several different porewater extraction techniques (pressurized squeezing, centrifugation, vacuum) yield samples with similar toxicity. Where the primary contaminants of concern are highly hydrophobic organic compounds, centrifugation is the method of choice for minimizing the loss of contaminants during the extraction procedure. No difference was found in the toxicity of pore water obtained with the Teflon® and polyvinyl chloride pressurized extraction devices. Different types of filters in the squeeze extraction devices apparently adsorbed soluble contaminants to varying degrees. The amount of fine suspended particulate material remaining in the pore water after the initial extraction varied among the methods. For most of the sediments tested, freezing and thawing did not affect the toxicity of porewater samples obtained by the pressurized squeeze extraction method. Pore water obtained by other methods (centrifugation, vacuum) and frozen without additional removal of suspended particulates by centrifugation may exhibit increased toxicity compared with the unfrozen sample. The toxicity of pore water extracted from refrigerated (4°C) sediments exhibited substantial short-term (days, weeks) changes. Similarly, sediment pore water extracted over time from a simulated amphipod solid-phase toxicity test changed substantially in toxicity. For the sediments tested, the direction and magnitude of change in toxicity of pore water extracted from both refrigerated and solid-phase test sediments was unpredictable.

Archives of Environmental Contamination and Toxico

Laboratory toxicity and benthic invertebrate field colonization of Upper Columbia River sediments: Finding adverse effects using multiple lines of evidence

From 1930 to 1995, the Upper Columbia River (UCR) of northeast Washington State received approximately 12 million metric tons of smelter slag and associated effluents from a large smelter facility located in Trail, British Columbia, approximately 10 km north of the United States–Canadian border. Studies conducted during the past two decades have demonstrated the presence of toxic concentrations of heavy metals in slag-based sandy sediments, including cadmium, copper, zinc, and lead in the UCR area as well as the downstream reservoir portion of Lake Roosevelt. We conducted standardized whole-sediment toxicity tests with the amphipod Hyalella azteca (28-day) and the midge Chironomus dilutus (10-day) on 11 samples, including both UCR and study-specific reference sediments. Metal concentrations in sediments were modeled for potential toxicity using three approaches: (1) probable effects quotients (PEQs) based on total recoverable metals (TRMs) and simultaneously extracted metals (SEMs); (2) SEMs corrected for acid-volatile sulfides (AVS; i.e., ∑SEM − AVS); and (3) ∑SEM − AVS normalized to the fractional organic carbon (f oc ) (i.e., ∑SEM − AVS/f oc ). The most highly metal-contaminated sample (∑PEQ TRM = 132; ∑PEQ SEM = 54; ∑SEM − AVS = 323; and ∑SEM − AVS/ foc = 64,600 umol/g) from the UCR was dominated by weathered slag sediment particles and resulted in 80% mortality and 94% decrease in biomass of amphipods; in addition, this sample significantly decreased growth of midge by 10%. The traditional ∑AVS – SEM, uncorrected for organic carbon, was the most accurate approach for estimating the effects of metals in the UCR. Treatment of the toxic slag sediment with 20% Resinex SIR-300 metal-chelating resin significantly decreased the toxicity of the sample. Samples ∑SEM − AVS > 244 was not toxic to amphipods or midge in laboratory testing, indicating that this value may be an approximate threshold for effects in the UCR. In situ benthic invertebrate colonization studies in an experimental pond (8-week duration) indicated that two of the most metal-contaminated UCR sediments (dominated by high levels of sand-sized slag particles) exhibited decreased invertebrate colonization compared with sand-based reference sediments. Field-exposed SIR-300 resin samples also exhibited decreased invertebrate colonization numbers compared with reference materials, which may indicate behavioral avoidance of this material under field conditions. Multiple lines of evidence (analytical chemistry, laboratory toxicity, and field colonization results), along with findings from previous studies, indicate that high metal concentrations associated with slag-enriched sediments in the UCR are likely to adversely impact the growth and survival of native benthic invertebrate communities. Additional laboratory toxicity testing, refinement of the applications of sediment benchmarks for metal toxicity, and in situ benthic invertebrate studies will assist in better defining the spatial extent, temporal variations, and ecological impacts of metal-contaminated sediments in the UCR system.

Trail

Sediment toxicity identification evaluation (TIE) studies at marine sites suspected of ordnance contamination

A sediment quality assessment survey and subsequent toxicity identification evaluation (TIE) study was conducted at several sites in Puget Sound, Washington. The sites were previously suspected of contamination with ordnance compounds. The initial survey employed sea urchin porewater toxicity tests to locate the most toxic stations. Sediments from the most toxic stations were selected for comprehensive chemical analyses. Based on the combined information from the toxicity and chemical data, three adjacent stations in Ostrich Bay were selected for the TIE study. The results of the phase I TIE suggested that organics and metals were primarily responsible for the observed toxicity in the sea urchin fertilization test. In addition to these contaminants, ammonia was also contributing to the toxicity for the sea urchin embryological development test. The phase II TIE study isolated the majority of the toxicity in the fraction containing nonpolar organics with high log Kow, but chemical analyses failed to identify a compound present at a concentration high enough to be responsible for the observed toxicity. The data suggest that some organic or organometallic contaminant(s) that were not included in the comprehensive suite of chemical analyses caused the observed toxicological responses.

Archives of Environmental Contamination and Toxico

Acute toxicity of lampricides to non-target species of concern in the Lake Champlain watershed

Previous research evaluated the toxicity of the lampricide 4-nitro-3-(trifluoromethyl)phenol (TFM) and the combination of TFM with 1 % niclosamide (TFM:1%Nic) to multiple non-target species in the Laurentian Great Lakes. However, few toxicity studies have been conducted for species of concern in Lake Champlain (NY and VT). We conducted 12-hour flow-through toxicity tests with 4 species of native mussels, 6 species of fish, and 1 amphibian species. All tests included exposure of invasive larval Petromyzon marinus (sea lamprey) and were conducted with concentrations that bracketed the predicted minimum lethal concentration required to control larval sea lamprey. Mussel species’ NOEC, LOEC, LC 25 , and LC 50 values ranged from 1.33 to 2.12, 1.71–2.66, 1.75–3.05, and 2.03–4.84 times field determined LC 99.9 s for sea lamprey (×SL LC99.9 ) in TFM-only toxicity tests, and from 1.36 to 1.70, 1.68–2.03, 1.86–2.10, and 2.35–2.68 × SL LC99.9 for TFM:1%Nic toxicity tests, respectively. Fish species NOEC, LOEC, LC 25 , and LC 50 values ranged from 0.60 to 1.89, 0.73–2.13, 0.72–2.11, and 0.76–2.18 × SL LC99.9 in TFM-only toxicity tests, and from 0.64 to 2.48, 0.85–3.10, 0.74–3.05, and 0.78–3.16 × SL LC99.9 for TFM:1%Nic toxicity tests, respectively. Amphibian species NOEC, LOEC, LC 25 , and LC 50 values ranged from 0.74 to 0.75, 0.85–0.95, 0.83–0.87, and 0.85–0.91 × SL LC99.9 in TFM-only toxicity tests, and from 0.63 to 0.65, 0.80–0.88, 0.77–0.82, and 0.78–0.87 × SL LC99.9 for TFM:1%Nic toxicity tests, respectively. Generally, mussel species were tolerant, fish sensitivities were variable, and the amphibian species was sensitive to TFM and TFM:1%Nic.

New York, Vermont

Comparative toxicity of aircraft deicers, runway deicers, and road salt in winter airport runoff

Aircraft deicer and anti-icer formulations and airfield pavement deicer formulations are commonly used at airports that experience freezing precipitation. In addition, road salt is often applied in urban areas surrounding airports. This mixture of deicers in receiving streams can be toxic to aquatic organisms. A 17-year study of deicers and toxicity in runoff samples from sites in and near Milwaukee Mitchell International Airport included bioassay testing of Ceriodaphnia dubia , Pimephales promelas , Raphidocelis subcapitata , and Aliivibrio fischeri concurrently with quantification of freezing-point depressants that represent presence of the various deicers. Propylene glycol was monitored to represent aircraft deicers and anti-icers, acetate and formate were monitored to represent airfield pavement deicers, and chloride was monitored to represent road salt. Toxicity endpoints (e.g., lethal concentrations and inhibitive concentrations) were compiled for 19 airport deicer products and sodium chloride. Comparison of freezing-point depressant concentrations with toxicity endpoints indicated that propylene glycol, acetate, and chloride exceeded endpoint concentrations in multiple samples: more than 70% of 492 runoff samples exceeded at least one endpoint concentration with exceedances of propylene glycol and acetate-based deicer endpoints as well as road salt endpoints. Validation of these results were observed with measured toxic effects on C. dubia , P. promelas , and R. subcapitata in bioassays conducted on undiluted samples: samples with multiple deicers exceeding endpoint thresholds had greater toxicity than samples with individual deicer endpoint exceedances, and samples with no endpoint exceedances typically exhibited less toxicity than samples with only one endpoint exceedance. A comparison of additivity of toxicity quotients with bioassay results fit a log-logistic regression curve, indicating that the condition of tested organisms degraded with increasing deicer presence. These findings suggest that concurrent reduction of runoff from aircraft, airfield pavement, and road salt deicers will be needed to eliminate aquatic toxicity in adjacent watersheds.

Wisconsin

Modelling interactions of toxicants and density dependence in wildlife populations

1. A major challenge in the conservation of threatened and endangered species is to predict population decline and design appropriate recovery measures. However, anthropogenic impacts on wildlife populations are notoriously difficult to predict due to potentially nonlinear responses and interactions with natural ecological processes like density dependence. 2. Here, we incorporated both density dependence and anthropogenic stressors in a stage-based matrix population model and parameterized it for a density-dependent population of peregrine falcons Falco peregrinus exposed to two anthropogenic toxicants [dichlorodiphenyldichloroethylene (DDE) and polybrominated diphenyl ethers (PBDEs)]. Log-logistic exposure–response relationships were used to translate toxicant concentrations in peregrine falcon eggs to effects on fecundity. Density dependence was modelled as the probability of a nonbreeding bird acquiring a breeding territory as a function of the current number of breeders. 3. The equilibrium size of the population, as represented by the number of breeders, responded nonlinearly to increasing toxicant concentrations, showing a gradual decrease followed by a relatively steep decline. Initially, toxicant-induced reductions in population size were mitigated by an alleviation of the density limitation, that is, an increasing probability of territory acquisition. Once population density was no longer limiting, the toxicant impacts were no longer buffered by an increasing proportion of nonbreeders shifting to the breeding stage, resulting in a strong decrease in the equilibrium number of breeders. 4. Median critical exposure concentrations, that is, median toxicant concentrations in eggs corresponding with an equilibrium population size of zero, were 33 and 46 μg g −1 fresh weight for DDE and PBDEs, respectively. 5. Synthesis and applications. Our modelling results showed that particular life stages of a density-limited population may be relatively insensitive to toxicant impacts until a critical threshold is crossed. In our study population, toxicant-induced changes were observed in the equilibrium number of nonbreeding rather than breeding birds, suggesting that monitoring efforts including both life stages are needed to timely detect population declines. Further, by combining quantitative exposure–response relationships with a wildlife demographic model, we provided a method to quantify critical toxicant thresholds for wildlife population persistence.

California

Toxicities of several pesticides to two species of Cladocerans

Laboratory bioassays were conducted with some chlorinated hydrocarbon insecticides, organophosphate insecticides, other insecticides and acaricides, and with herbicides to determine their relative toxicities and immobilization values for two species of daphnids, Daphnia pulex and Simocephalus serrulatus. Both species are satisfactory bioassay organisms for the determination of a wide variety of pesticides, with D. pulex being the more sensitive. The organophosphate insecticides were generally more toxic than the chlorinated hydrocarbons to both species. DDVP was the most toxic compound investigated. DDT was the most toxic chlorinated hydrocarbon tested, and lindane the least. There was a wide range in the toxicity of hydrocarbons to D. pulex, with 48-hour EC 50 values ranging from 0.36 to 460 ppb. DDT was 2.9 times more toxic at 50 F than at 80 F. Malathion was 8.8 times more toxic at 50 F than at 70 F to S. serrulatus. Endrin was 12 times more toxic than dieldrin to D. pulex. DDT was 2.6 times more toxic at 60 F to first-instar organisms up to 18 hours old than to 7-day-old organisms.

Transactions of the American Fisheries Society

Influence of multiple water-quality characteristics on copper toxicity to fathead minnows (Pimephales promelas)

Water quality influences the bioavailability and toxicity of copper to aquatic organisms. Understanding the relationships between water-quality parameters and copper toxicity may facilitate the development of site-specific criteria for water quality and result in better protection of aquatic biota. Many studies have examined the influence of a single water-quality parameter on copper toxicity, but the interactions of several characteristics have not been well studied in low-hardness water. The goal of the present research was to examine the interactions among water-quality characteristics and their effects on copper toxicity to larval fathead minnows (Pimephales promelas). The effects of dissolved organic carbon (DOC) concentration, DOC source, pH, and hardness on acute copper toxicity were determined using a complete factorially designed experiment. Hardness, pH, DOC, and interaction of pH and DOC all significantly affected copper toxicity. A predictive model based on these data described 88% of the variability in copper toxicity. This model also explained 58% of the variability in copper toxicity for an independent dataset of South Carolina (USA) waters. The biotic ligand model underpredicted the acute copper toxicity to fathead minnows when compared with observed values.

Environmental Toxicology and Chemistry

Sensitivity of mottled sculpins (Cottus bairdi) and rainbow trout (Onchorhynchus mykiss) to acute and chronic toxicity of cadmium, copper, and zinc

Studies of fish communities of streams draining mining areas suggest that sculpins ( Cottus spp.) may be more sensitive than salmonids to adverse effects of metals. We compared the toxicity of zinc, copper, and cadmium to mottled sculpin ( C. bairdi ) and rainbow trout ( Onchorhynchus mykiss ) in laboratory toxicity tests. Acute (96-h) and early life-stage chronic (21- or 28-d) toxicity tests were conducted with rainbow trout and with mottled sculpins from populations in Minnesota and Missouri, USA, in diluted well water (hardness = 100 mg/L as CaCO 3 ). Acute and chronic toxicity of metals to newly hatched and swim-up stages of mottled sculpins differed between the two source populations. Differences between populations were greatest for copper, with chronic toxicity values (ChV = geometric mean of lowest-observed-effect concentration and no-observed-effect concentration) of 4.4 μg/L for Missouri sculpins and 37 μg/L for Minnesota sculpins. Cadmium toxicity followed a similar trend, but differences between sculpin populations were less marked, with ChVs of 1.1 μg/L (Missouri) and 1.9 μg/L (Minnesota). Conversely, zinc was more toxic to Minnesota sculpins (ChV = 75 μg/L) than Missouri sculpins (chronic ChV = 219 μg/L). Species-average acute and chronic toxicity values for mottled sculpins were similar to or lower than those for rainbow trout and indicated that mottled sculpins were among the most sensitive aquatic species to toxicity of all three metals. Our results indicate that current acute and chronic water quality criteria for cadmium, copper, and zinc adequately protect rainbow trout but may not adequately protect some populations of mottled sculpins. Proposed water quality criteria for copper based on the biotic ligand model would be protective of both sculpin populations tested.

Minnesota, Missouri

Applicability of ambient toxicity testing to national or regional water-quality assessment

Comprehensive assessment of the quality of natural waters requires a multifaceted approach. Based on experimentation designed to monitor responses of organisms to environmental stresses, toxicity testing may have diverse purposes in water quality assessments. These purposes may include identification that warrant further study because of poor water quality or unusual ecological features, verification of other types of monitoring, or assessment of contaminant effects on aquatic communities. A wide variety of toxicity test methods have been developed to fulfill the needs of diverse applications. The methods differ primarily in the full selections made relative to four characteristics: (1) test species, (2) endpoints (acute or chronic), (3) test enclosure type, and (4) test substance (toxicant) that functions as the environmental stress. Toxicity test approachs vary in their capacity to meet the needs of large-scale assessments of existing water quality. Ambient testing is more likely to meet these needs than are the procedures that call for exposure of the test organisms to known concentrations of a single toxicant. However, meaningful interpretation of ambient test results depend on the existence of accompanying chemical analysis of the ambient media. The ambient test substance may be water or sediments. Sediment tests have had limited application, but they are useful because of the fact that most toxicants tend to accumulate in sediments, and many test species either inhabit the sediments or are in frequent contact with them. Biochemical testing methods, which have been developing rapidly in recent years, are likely to be among the most useful procedures for large-scale water quality assessments. They are relatively rapid and simple, and more importantly, they focus on biochemical changes that are the initial responses of virtually all organisms to environmental stimuli. Most species are sensitive to relatively few toxicants and their sensitivities vary as conditions change. One of the most informative approaches for toxicity testing is to combine biochemical tests with other test methods in a ' battery or tests ' that is diversified enough to characterize different types of toxicants and different trophic levels. (Lantz-PTT)

Open-File Report

Pesticide toxicity index for freshwater aquatic organisms, 2nd edition

The U.S. Geological Survey's National Water-Quality Assessment (NAWQA) Program is designed to assess current water-quality conditions, changes in water quality over time, and the effects of natural and human factors on water quality for the Nation's streams and ground-water resources. For streams, one of the most difficult parts of the assessment is to link chemical conditions to effects on aquatic biota, particularly for pesticides, which tend to occur in streams as complex mixtures with strong seasonal patterns. A Pesticide Toxicity Index (PTI) was developed that combines pesticide exposure of aquatic biota (measured concentrations of pesticides in stream water) with acute toxicity estimates (standard endpoints from laboratory bioassays) to produce a single index value for a sample or site. The development of the PTI was limited to pesticide compounds routinely measured in NAWQA studies and to toxicity data readily available from existing databases. Qualifying toxicity data were found for one or more types of test organisms for 124 of the 185 pesticide compounds measured in NAWQA samples, but with a wide range of available bioassays per compound (1 to 232). In the databases examined, there were a total of 3,669 bioassays for the 124 compounds, including 398 48-hour EC50 values (concentration at which 50 percent of test organisms exhibit a sublethal response) for freshwater cladocerans, 699 96-hour LC50 values (concentration lethal to 50 percent of test organisms) for freshwater benthic invertebrates, and 2,572 96-hour LC50 values for freshwater fish. The PTI for a particular sample is the sum of toxicity quotients (measured concentration divided by the median toxicity concentration from bioassays) for each detected pesticide, and thus, is based on the concentration addition model of pesticide toxicity. The PTI can be calculated for specific groups of pesticides and for specific taxonomic groups. Although the PTI does not determine whether water in a sample is toxic to aquatic organisms, its values can be used to rank or compare the toxicity of samples or sites on a relative basis for use in further analysis or additional assessments. The PTI approach may be useful as a basis for comparing the potential significance of pesticides in different streams on a common basis, for evaluating relations between pesticide exposure and observed biological conditions, and for prioritizing where further studies are most needed.

Scientific Investigations Report

Using enzyme bioassays as a rapid screen for metal toxicity

Mine tailings piles and abandoned mine soils are often contaminated by a suite of toxic metals, which were released in the mining process. Traditionally, toxicity of such areas has been determined by numerous chemical methods including the Toxicity Characteristic Leachate Procedure (TCLP) and traditional toxicity tests using organisms such as the cladoceran Ceriodaphnia dubia. Such tests can be expensive and time-consuming. Enzymatic bioassays may provide an easier, less costly, and more time-effective toxicity screening procedure for mine tailings and abandoned mine soil leachates. This study evaluated the commercially available MetPLATE™ enzymatic toxicity assay test kit. The MetPLATE™ assay uses a modified strain of Escherichia coli bacteria as the test organism. Toxicity is defined by the activity of β-galactosidase enzyme which is monitored colorometrically with a 96-well spectrophotometer. The study used water samples collected from North Fork Clear Creek, a mining influenced water (MIW) located in Colorado. A great benefit to using the MetPLATE™ assay over the TCLP is that it shows actual toxicity of a sample by taking into account the bioavailability of the toxicants rather than simply measuring the metal concentration present. Benefits of the MetPLATE™ assay over the use of C. dubia include greatly reduced time for the testing process (∼2 hours), a more continuous variable due to a greater number of organisms present in each sample (100,000+), and the elimination of need to maintain a culture of organisms at all times.

Conference Paper

Development of a new toxic-unit model for the bioassessment of metals in streams

Two toxic-unit models that estimate the toxicity of trace-metal mixtures to benthic communities were compared. The chronic criterion accumulation ratio (CCAR), a modification of biotic ligand model (BLM) outputs for use as a toxic-unit model, accounts for the modifying and competitive influences of major cations (Ca 2+ , Mg 2+ , Na + , K + , H + ), anions (HCO 3 − , CO 3 2− ,SO 4 2− , Cl − , S 2− ) and dissolved organic carbon (DOC) in determining the free metal ion available for accumulation on the biotic ligand. The cumulative criterion unit (CCU) model, an empirical statistical model of trace-metal toxicity, considers only the ameliorative properties of Ca 2+ and Mg 2+ (hardness) in determining the toxicity of total dissolved trace metals. Differences in the contribution of a metal (e.g., Cu, Cd, Zn) to toxic units as determined by CCAR or CCU were observed and attributed to how each model incorporates the influences of DOC, pH, and alkalinity. Akaike information criteria demonstrate that CCAR is an improved predictor of benthic macroinvertebrate community metrics as compared with CCU. Piecewise models depict great declines (thresholds) in benthic macroinvertebrate communities at CCAR of 1 or more, while negative changes in benthic communities were detected at a CCAR of less than 1. We observed a 7% reduction in total taxa richness and a 43% decrease in Heptageniid abundance between background (CCAR = 0.1) and the threshold of chronic toxicity on the basis of continuous chronic criteria (CCAR = 1). In this first application of the BLM as a toxic-unit model, we found it superior to CCU.

Environmental Toxicology and Chemistry

A comparison of the chemical sensitivities between in vitro and in vivo propagated juvenile freshwater mussels: Implications for standard toxicity testing

Unionid mussels are ecologically important and are globally imperiled. Toxicants contribute to mussel declines, and toxicity tests using juvenile mussels—a sensitive life stage—are valuable in determining thresholds used to set water quality criteria. In vitro culture methods provide an efficient way to propagate juveniles for toxicity testing, but their relative chemical sensitivity compared with in vivo propagated juveniles is unknown. Current testing guidelines caution against using in vitro cultured juveniles until this sensitivity is described. Our objective was to evaluate the relative sensitivity of juvenile mussels produced from both in vitro and in vivo propagation methods to selected chemicals. We conducted 96-h acute toxicity tests according to ASTM International guidelines with 3 mussel species and 6 toxicants: chloride, nickel, ammonia, and 3 copper-based compounds. Statistically significant differences between in vitro and in vivo juvenile 96-h median effect concentrations were observed in 8 of 17 tests, and in vitro juveniles were more sensitive in 6 of the 8 significant differences. At 96 h, 4 of the 8 statistically different tests for a given chemical were within a factor of 2, which is the intralaboratory variation demonstrated in a recent evaluation of mussel toxicity tests. We found that although differences in chemical sensitivity exist between in vitro and in vivo propagated juvenile mussels, they are within normal toxicity test variation. Therefore, in vitro propagated juvenile mussels may be appropriate for use in ASTM International-based toxicity testing.

Environmental Toxicology and Chemistry

Influences of water hardness on chronic toxicity of potassium chloride to a unionid mussel (Lampsilis siliquoidea)

Elevated concentrations of potassium (K) often occur in effluents from wastewater treatment plants, oil and gas production operations, mineral extraction processes, and from other anthropogenic sources. Previous studies have demonstrated that freshwater mussels are highly sensitive to K in acute and chronic exposures, and acute toxicity of K decreases with increasing water hardness. However, little is known about the influence of hardness on the chronic toxicity of K. The objective of this study was to evaluate the chronic toxicity of K (tested as KCl) to a commonly tested unionid mussel (fatmucket , Lampsilis siliquoidea ) at five hardness levels (25, 50, 100, 200, 300 mg/L as CaCO 3 ) representing most surface waters in the United States. Chronic 28-d K toxicity tests were conducted with 3-week-old juvenile fatmucket in the five hardness waters using an ASTM standard method. The maximum acceptable toxicant concentrations (geometric mean of the no-observed-effect concentration and the lowest-observed-effect concentration) increased from 15.1 to 69.3 mg K/L for survival and from 15.1 to 35.8 mg K/L for growth (length and dry weight) and biomass when water hardness was increased from 25 mg/L (soft) to 300 mg/L (very hard). These results provided evidence to support water hardness influence on chronic K toxicity to juvenile fatmucket. However, the chronic effect concentrations based on the more sensitive endpoint (growth or biomass) increased only 2.4-fold from the soft water to the very hard water, indicating that water hardness had limited influence on the chronic toxicity of K to the mussels. These results can be used to establish chronic toxicity thresholds for K across a broad range of water hardness and to derive environmental guideline values for K to protect freshwater mussels and other organisms.

Environmental Toxicology and Chemistry

Benthic invertebrate bioassays with toxic sediment and pore water

The relative sensitivities of bioassays to determine the toxicity of sediments were investigated and three methods of making the sample dilutions required to generate dose-response relationships were compared. The assays studied were: (a) Microtox®, a 15-min assay of Photobacterium phosphoreum bioluminescence inhibition by pore water; (b) 48-h Daphnia magna lethality test in pore water; (c) 10-d subchronic assay of lethality to and reduction of weight gain by Chironomus tentans performed in either whole sediment or pore water; (d) 168-h acute lethality assay of Hexagenia limbata in either whole sediment or pore water. The three methods of diluting sediments were: (a) extracting pore water from the toxic location and dilution with pore water from the control station; (b) diluting whole sediment from the toxic location with control whole sediment from a reference location, then extracting pore water; and (c) diluting toxic, whole sediment with whole sediment from a reference location, then using the whole sediment in bioassays. Based on lethality, H. limbata was the most sensitive organism to the toxicity of Detroit River sediment. Lethality of D. magna in pore water was similar to that of H. limbata in whole sediment and can be used to predict effects of whole sediment toxicity to H. limbata. The concentration required to cause a 50% reduction in C. tentans growth (10-d EC50) was approximately that which caused 50% lethality of D. magna (48-h LC50) and was similar to the toxicity that restricts benthic invertebrate colonization of contaminated sediments. While the three dilution techniques gave similar results with some assays, they gave very different results in other assays. The dose-response relationships determined by the three dilution techniques would be expected to vary with sediment, toxicant and bioassay type, and the dose-response relationship derived from each technique needs to be interpreted accordingly.

Environmental Toxicology and Chemistry

The use of freshwater and saltwater animals to distinguish between the toxic effects of salinity and contaminants in irrigation drain water

Irrigation drain waters entering Stillwater Wildlife Management Area (SWMA) in south-western Nevada contain elevated levels of salinity and several inorganic contaminants (As, B, Cu, Li, Mo, and Sr). Mortalities of fish and waterfowl at the management area are believed to be associated with the poor water quality of the drains. The objective of the present study was to use fresh-water and saltwater animals to distinguish between the toxic effects of salinity and contaminants in effluent samples collected from irrigation drain waters. Static acute effluent tests were conducted with water collected from four sites at SWMA. Animals acclimated or cultured in fresh water (fathead minnows, Pimephales promelas ; amphipods, Hyalella azteca ; cladocerans, Daphnia magna ) and salt water (striped bass, Morone saxatilis ; amphipods, Hyalella azteca ; and cladocerans, Daphnia magna ) were used to separate toxic effects of salinity from the effects of inorganic contaminants in the drain water. One drain water (TJ drain, salinity 19 parts per thousand (grams per liter), osmolality 503 mmol/kg, hardness 3,780 mg/L as CaCO 3 ) was toxic only to freshwater animals and saltwater cultured daphnids; water from a receiving pond (Pintail Bay, salinity 23 g/L, osmolality 542 mmol/kg, hardness 830 mg/L as CaCO 3 ) was toxic to both freshwater and saltwater animals. Acute tests conducted with reconstituted waters representative of the Pintail Bay sample indicated that atypical ion ratios were toxic to striped bass and amphipods, even without the addition of inorganic contaminants. However, the addition of inorganic contaminants representative of the Pintail Bay sample increased the toxicity of this reconstituted water. These findings indicate that the toxicity of the TJ drain sample was related mainly to elevated salinity and that the toxicity of the Pintail Bay sample was a function of inorganic contamination and atypical ion ratios in combination with elevated salinity.

Environmental Toxicology and Chemistry

Acute toxicity of 4-nitrophenol, 2,4-dinitrophenol, terbufos and trichlorfon to grass shrimp (Palaemonetes spp.) and sheepshead minnows (Cyprinodon variegatus) as affected by salinity and temperature

The toxicities of two industrial chemicals (4-nitrophenol and 2,4-dinitrophenol) and two organophosphate insecticides (terbufos and trichlorfon) to juvenile grass shrimp (Palaemonetes spp.) and sheepshead minnows (Cyprinodon vanegatus) were determined by static, 96-h toxicity tests in a factorial design with 12 combinations of salinity and temperature (15, 20, 25, 30ppt x 17, 22, 27°C). Concentrations of the toxicants, including bioconcentradon, were determined as appropriate by gas or liquid chromatography and the use of 14 C-labeled compounds. The 96-h LC50s for 4-nitrophenol ranged from 12 to 31 mg/L and for 2,4-dinitrophenol from 13 to 50 mg/L. Toxicity decreased as salinity increased for 4-nitrophenol and both test organisms. Toxicity decreased as salinity increased for 2,4-dinitrophenol and sheepshead minnows, but toxicity to grass shrimp increased as salinity increased. Toxicity decreased with increased temperature for grass shrimp exposed to 2,4-dinitrophenol and sheepshead minnows exposed to 4-nitrophenol, increased with temperature for sheepshead minnows exposed to 2,4-dinitrophenol, and no change was observed for grass shrimp exposed to 4-nitrophenol. Bioconcentration of phenols in both test organisms increased as concentration increased. The 96-h LC50s for terbufos ranged from 3.4 to 6.6 μg/L and for trichlorfon from 6.3 to 19,300 μg/L. Terbufos and trichlorfon toxicity to grass shrimp and sheepshead minnows increased with increased temperature. BCFs for terbufos were greater in sheepshead minnows than grass shrimp, but were reversed for trichlorfon.

Environmental Toxicology and Chemistry