Trace Elements and Consequent Ecological Risks in Mining-Influenced Streams of Appalachia.

Appalachian (eastern USA) coal surface mines fracture geologic materials, causing release of both major ions and trace elements to water via accelerated weathering. When elevated above natural background, trace elements in streams may produce adverse effects on biota via direct exposure from water and sediment and via dietary exposure in food sources. Other studies have found elevated water concentrations of multiple trace elements in Appalachia's mining-influenced streams. Except for Se, trace-element concentrations in abiotic and biotic media of Appalachian mining-influenced streams are less well known. We analyzed environmental media of headwater streams receiving alkaline waters from Appalachian coal mines for eight trace elements (Al, As, Cd, Cu, Ni, Sr, V, and Zn) and assessed the potential consequent ecological risks. Streamwater, particulate media (sediment, biofilm, leaf detritus), and benthic macroinvertebrates (primary consumers, secondary consumers, crayfish) were sampled from six mining-influenced and three reference streams during low-flow conditions in two seasons. Dissolved Cu, Ni, and Sr were higher in mining-influenced streams than in reference streams; Ni, Sr, and Zn in fine sediments and Ni in macroinvertebrates were also elevated relative to reference-stream levels in samples from mining-influenced streams. Seasonal ratios of mining-influenced stream concentrations to maximum concentrations in reference streams also demonstrated mining-influenced increases for several elements in multiple media. In most media, concentrations of several elements including Ni were correlated positively. All water-column dissolved concentrations were below protective levels, but fine-sediment concentrations of Ni approached or exceeded threshold-effect concentrations in several streams. Further study is warranted for several elements (Cd, Ni, and Zn in biofilms, and V in macroinvertebrates) that approached or exceeded previously established dietary-risk levels. Environ Toxicol Chem 2023;42:2651-2665. © 2023 SETAC.

Influence of Test Method Variables on Sensitivity of Neocloeon Triangulifer to a Reference Toxicant in Short-Term, Effluent Style Evaluations.

Recent literature has demonstrated the sensitivity of mayflies to environmental contaminants. However, to date, there are no methods approved by the US Environmental Protection Agency for using sensitive insects like mayflies in whole-effluent toxicity or receiving water toxicity tests. The parthenogenetic mayfly Neocloeon triangulifer has been shown to be amenable to continuous culture in the laboratory, and methods have been described for its use in both acute and chronic toxicity studies. The goal of the present study was to investigate aspects of N. triangulifer testing and culturing methods that might require adjustment so that they are applicable for testing effluents and receiving waters in a short-term exposure. To this end, the influence of organism age, test duration, and test temperature on sensitivity to NaCl as a reference toxicant were tested (concentrations ranging from 182 to 2489 mg/L). Further studies were conducted to assess the utility of commercially available diets and the influence of nutrient amendment of water on organism growth and sensitivity. Seven-day NaCl tests started with less than 24-h-old larvae were similar in sensitivity to 14-day and full life chronic tests, and were much more sensitive than those started with 7-day-old organisms. Reducing test temperature from 25 °C to 22 °C had a minor influence on culture timing, and little impact on sensitivity to NaCl. In other experiments, reconstituted test water supplemented with nutrients to potentially improve in-test food quality had minimal effect on growth at 7 days and did not significantly alter acute sensitivity to NaCl relative to unamended reconstituted water. A suitable commercially available, ready-to-feed diet substitute for cultured diatoms was not found. Testing N. triangulifer in effluents or receiving waters with the methods recommended will complement similar methods for Ceriodaphnia dubia. Environ Toxicol Chem 2022;41:2758-2768. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Combined effects of copper, nickel, and zinc on growth of a freshwater mussel (Villosa iris) in an environmentally relevant context.

Trace metals rarely contaminate freshwaters independently, hence regulatory limits based on single-metal toxicity may be underprotective of aquatic life. This could be especially the case for rare and sensitive fauna like freshwater mussels, such as those suppressed in the Clinch and Powell Rivers in eastern USA where trace metals are long-term contaminants but at concentrations below regulatory limits. We hypothesized metal mixtures may be exerting combined effects on mussels, resulting in greater toxicity than would be predicted based on single-metal exposures. To test that hypothesis, we conducted two experiments exposing juvenile rainbow mussels (Villosa iris) for 42 days to dissolved copper, nickel, and zinc, individually and in three-metal mixtures, in an environmentally-relevant context of water with chemistry (hardness 155 mg/L as CaCO3, dissolved organic carbon 1.7-2.3 mg/L, pH 8.4) similar to that of the Clinch River, which receives alkaline mine drainage. We used a toxic unit approach, selecting test concentrations based on literature values for the lower of 28-day survival or growth (length) effect concentrations for Villosa iris or Lampsilis siliquoidea (fatmucket). Our first experiment confirmed survival and growth effects when acute and chronic water quality criteria, respectively, are approached and/or exceeded. Our second experiment, at lower concentrations, showed no effects on survival but combined effects on growth were evident: a mixture of Cu, Ni, and Zn (7.2 ± 1.2, 65.3 ± 6.1, 183 ± 32 µg/L, respectively) inhibited growth (dry weight) by 95% versus 73%, 74%, and 83% inhibition for single-metal exposures to Cu, Ni, and Zn of similar concentration (8.0 ± 1.1, 63.5 ± 4.8, 193 ± 31 µg/L, respectively). Furthermore, a mixture of Cu, Ni, and Zn with individual concentrations 21%, 29%, and 37% of their water quality criteria (3.4 ± 1.2, 21.8 ± 1.8, and 62.1 ± 8.4 µg/L, respectively) inhibited growth (dry weight) by 61% relative to controls. Our observation of combined effects suggests that regulatory limits based on single-metal toxicity may be underprotective of freshwater mussels when multiple metals are present.

Comparison of benthic macroinvertebrate assessment methods along a salinity gradient in headwater streams.

Benthic macroinvertebrate community assessments are used commonly to characterize aquatic systems and increasingly for identifying their impairment caused by myriad stressors. Yet sampling and enumeration methods vary, and research is needed to compare their abilities to detect macroinvertebrate community responses to specific water quality variables. A common assessment method, rapid bioassessment, uses subsampling procedures to identify a fixed number of individual organisms regardless of total sample abundance. In contrast, full-enumeration assessments typically allow for expanded community characterization resulting from higher numbers of identified organisms within a collected sample. Here, we compared these two sampling and enumeration methods and their abilities to detect benthic macroinvertebrate response to freshwater salinization, a common stressor of streams worldwide. We applied both methods in headwater streams along a salinity gradient within the coal-mining region of central Appalachia USA. Metrics of taxonomic richness, community composition, and trophic function differed between the methods, yet most metrics exhibiting significant response to SC for full-enumeration samples also did for rapid bioassessment samples. However, full-enumeration yielded taxonomic-based metrics consistently more responsive to the salinization gradient. Full-enumeration assessments may potentially provide more complete characterization of macroinvertebrate communities and their response to increased salinization, whereas the more cost-effective and widely employed rapid bioassessment method can detect community alterations along the full salinity gradient. These findings can inform decisions regarding such tradeoffs for assessments of freshwater salinization in headwater streams and highlight the need for similar research of sampling and enumeration methodology in other aquatic systems and for other stressors.

Modeling the Bioavailability of Nickel and Zinc to Ceriodaphnia dubia and Neocloeon triangulifer in Toxicity Tests with Natural Waters.

We studied biotic ligand model (BLM) predictions of the toxicity of nickel (Ni) and zinc (Zn) in natural waters from Illinois and Minnesota, USA, which had combinations of pH, hardness, and dissolved organic carbon (DOC) more extreme than 99.7% of waters in a nationwide database. We conducted 7-day chronic tests with Ceriodaphnia dubia and 96-hour acute and 14-day chronic tests with Neocloeon triangulifer and estimated median lethal concentrations and 20% effect concentrations for both species. Toxicity of Ni and Zn to both species differed among test waters by factors from 8 (Zn tests with C. dubia) to 35 (Zn tests with N. triangulifer). For both species and metals, tests with Minnesota waters (low pH and hardness, high DOC) showed lower toxicity than Illinois waters (high pH and high hardness, low DOC). Recalibration of the Ni BLM to be more responsive to pH-related changes improved predictions of Ni toxicity, especially for C. dubia. For the Zn BLM, we compared several input data scenarios, which generally had minor effects on model performance scores (MPS). A scenario that included inputs of modeled dissolved inorganic carbon and measured Al and Fe(III) produced the highest MPS values for tests with both C. dubia and N. triangulifer. Overall, the BLM framework successfully modeled variation in toxicity for both Zn and Ni across wide ranges of water chemistry in tests with both standard and novel test organisms. Environ Toxicol Chem 2021;40:3049-3062. © 2021 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Acute and Chronic Toxicity of Sodium Nitrate and Sodium Sulfate to Several Freshwater Organisms in Water-Only Exposures.

Elevated nitrate (NO3 ) and sulfate (SO4 ) in surface water are of global concern, and studies are needed to generate toxicity data to develop environmental guideline values for NO3 and SO4 . The present study was designed to fill existing gaps in toxicity databases by determining the acute and/or chronic toxicity of NO3 (tested as NaNO3 ) to a unionid mussel (Lampsilis siliquoidea), a midge (Chironomus dilutus), a fish (rainbow trout, Oncorhynchus mykiss), and 2 amphibians (Hyla versicolor and Lithobates sylvaticus), and to determine the acute and/or chronic toxicity of SO4 (tested as Na2 SO4 ) to 2 unionid mussels (L. siliquoidea and Villosa iris), an amphipod (Hyalella azteca), and 2 fish species (fathead minnow, Pimephales promelas and O. mykiss). Among the different test species, acute NO3 median effect concentrations (EC50s) ranged from 189 to >883 mg NO3 -N/L, and chronic NO3 20% effect concentrations (EC20s) based on the most sensitive endpoint ranged from 9.6 to 47 mg NO3 -N/L. The midge was the most sensitive species, and the trout was the least sensitive species in both acute and chronic NO3 exposures. Acute SO4 EC50s for the 2 mussel species (2071 and 2064 mg SO4 /L) were similar to the EC50 for the amphipod (2689 mg SO4 /L), whereas chronic EC20s for the 2 mussels (438 and 384 mg SO4 /L) were >2-fold lower than the EC20 of the amphipod (1111 mg SO4 /L), indicating the high sensitivity of mussels in chronic SO4 exposures. However, the fathead minnow, with an EC20 of 374 mg SO4 /L, was the most sensitive species in chronic SO4 exposures whereas the rainbow trout was the least sensitive species (EC20 > 3240 mg SO4 /L). The high sensitivity of fathead minnow was consistent with the finding in a previous chronic Na2 SO4 study. However, the EC20 values from the present study conducted in test water containing a higher potassium concentration (3 mg K/L) were >2-fold greater than those in the previous study at a lower potassium concentration (1 mg K/L), which confirmed the influence of potassium on chronic Na2 SO4 toxicity to the minnow. Environ Toxicol Chem 2020;39:1071-1085. © 2020 SETAC.

Impacts to water quality and biota persist in mining-influenced Appalachian streams.

Elevated dissolved major ions (salinization) from surface coal mining are a common impact to central Appalachian headwater streams. Salinization is associated with alterations of benthic macroinvertebrate communities, as many organisms are adapted to the naturally dilute streams of the region. These geochemical and biological alterations have been observed in streams decades after mining, but it remains unclear whether and at what rate water quality and aquatic biota recover after mining. To address this issue, we analyzed temporal trends in specific conductance (SC), ion matrix ratios, and benthic macroinvertebrate communities over an eight-year period in 23 headwater streams, including 18 salinized by surface coal mining. We found strong, negative correlations between SC and diversity of benthic macroinvertebrate communities. Temporal trend analysis demonstrated limited recovery of water chemistry to natural background conditions. Five of the 18 mining-influenced streams exhibited declining SC; however, annual rates of decline in these streams ranged from 1.9% to 3.7% of mean annual SC, suggesting long time periods will be required to reach established benchmark values (ca. 25 years) or values observed in our five reference study streams (ca. 40 years). Similarly, there was limited evidence for recovery of macroinvertebrate community metrics, even in the few mining-influenced streams with decreasing SC. These findings indicate that salinization and its biological effects persist, likely for decades, in central Appalachian headwater streams. Our work also highlights the value of long-term monitoring data for assessing recovery potential of salinized freshwaters, as well as the need for improved understanding of water quality and biological recovery processes and time frames.

Acute and Chronic Toxicity of Nickel and Zinc to a Laboratory Cultured Mayfly (Neocloeon triangulifer) in Aqueous but Fed Exposures.

Aquatic insects are poorly represented in water quality criteria, and previous studies have suggested a lack of sensitivity in acute toxicity tests despite observational studies demonstrating the contrary. Our objectives were to determine the toxicity of nickel (Ni) and zinc (Zn) to the mayfly Neocloeon triangulifer in fed acute (96-h) and chronic exposures to estimate aqueous effect concentrations while acknowledging the importance of dietary exposure for these insects. For the chronic tests, we conducted preliminary full-life cycle (~25-30 d) and subchronic (14 d) exposures to compare the relative sensitivity of the 2 test durations under similar conditions (i.e., feeding rates). Observing similar sensitivity, we settled on 14 d as the definitive test duration. Furthermore, we conducted experiments to determine how much food could be added to a given volume of water while minimally impacting dissolved metal recovery; a ratio of food dry mass to water volume (<0.005) achieved this. In the 14-d tests, we obtained a median lethal concentration and most sensitive chronic endpoint of 147 and 23 µg/L dissolved Ni (acute to chronic ratio [ACR] = 6.4), respectively, and 81 (mean value) and 10 µg/L dissolved Zn (ACR = 8.1), respectively. The acute values are orders of magnitude lower than previously published values for mayflies, probably most importantly due to the presence of dietary exposure but also potentially with some influence of organism age and test temperature. Environ Toxicol Chem 2020;39:1196-1206. © 2020 SETAC.

Selenium Bioaccumulation Across Trophic Levels and Along a Longitudinal Gradient in Headwater Streams.

Toxic effects of selenium (Se) contamination in freshwaters have been well documented. However, study of Se contamination has focused on lentic and larger order lotic systems, whereas headwater streams have received little scrutiny. In central Appalachia, surface coal mining is a common Se source to headwater streams, thus providing a useful system to investigate Se bioaccumulation in headwater food chains and possible longitudinal patterns in Se concentrations. Toward that end, we assessed Se bioaccumulation in 2 reference and 4 mining-influenced headwater streams. At each stream, we sampled ecosystem media, including streamwater, particulate matter (sediment, biofilm, leaf detritus), benthic macroinvertebrates, salamanders, and fish, every 400 m along 1.2- and 1.6-km reaches. We compared media Se concentrations within and among streams and evaluated longitudinal trends in media Se concentrations. Selenium concentrations in sampled media were higher in mining-influenced streams compared with reference streams. We found the highest Se concentrations in benthic macroinvertebrates; however, salamanders and fish bioaccumulated Se to potentially harmful levels in mining-influenced streams. Only one stream demonstrated dilution of streamwater Se with distance downstream, and few longitudinal patterns in Se bioaccumulation occurred along our study reaches. Collectively, our results provide a field-based assessment of Se bioaccumulation in headwater food chains, from streamwater to fish, and highlight the need for future assessments of Se effects in headwater streams and receiving downstream waters. Environ Toxicol Chem 2020;39:692-704. © 2020 SETAC.

Are sulfate effects in the mayfly Neocloeon triangulifer driven by the cost of ion regulation?

Elevated major ion concentrations in streams are commonly observed as a consequence of resource extraction, de-icing and other anthropogenic activities. Ecologists report biodiversity losses associated with increasing salinity, with mayflies typically being highly responsive to increases of different major ions. In this study, we evaluated the performance of the mayfly Neocloeon triangulifer reared for its entire larval phase in a gradient of sulfate concentrations. Two natural waters were amended with SO4 as a blend of CaSO4 and MgSO4 and exposures ranged from 5 to 1500 mg l-1 SO4. Survival (per cent successful emergence to the subimago stage) was significantly reduced at the highest SO4 concentration in both waters, while development was significantly delayed at 667 mg l-1 SO4 Final sub-adult body weights were consistent across treatments, except at the highest treatment concentration. Despite evidence for sulfate uptake rates increasing with exposure concentrations and not being saturated at even extremely high SO4 concentrations, total body sulfur changed little in subimagos. Together, these results suggest that elevated SO4 imposes an energetic demand associated with maintaining homeostasis that is manifested primarily as reduced growth rates and associated developmental delays. We identified two genes related to sulfate transport in N. trianguliferThis article is part of the theme issue 'Salt in freshwaters: causes, ecological consequences and future prospects'.

Selenium dynamics in headwater streams of the central Appalachian coalfield.

Coal mining can cause selenium (Se) contamination in US Appalachian streams, but linkages between water-column Se concentrations and Se bioaccumulation within Appalachian headwater streams have rarely been quantified. Using elevated specific conductance (SC) in stream water as an indicator of mining influence, we evaluated relationships between SC and Se concentrations in macroinvertebrates and examined dynamics of Se bioaccumulation in headwater streams. Twenty-three Appalachian streams were categorized into 3 stream types based on SC measurements: 1) reference streams with no coal-mining history; 2) mining-influenced, high-SC streams; and 3) mining-influenced, low-SC streams. Selenium concentrations in macroinvertebrates exhibited strong positive associations with both SC and dissolved Se concentrations in stream water. At 3 streams of each type, we further collected water, particulate matter (sediment, biofilm, leaf detritus), and macroinvertebrates and analyzed them for Se during 2 seasons. Enrichment, trophic transfer, and bioaccumulation factors were calculated and compared among stream types. Particulate matter and macroinvertebrates in mining-influenced streams accumulated high Se concentrations relative to reference streams. Concentrations were found at levels indicating Se to be a potential environmental stressor to aquatic life. Most Se enrichment, trophic transfer, and bioaccumulation factors were independent of season. Enrichment factors for biofilm and sediments and bioaccumulation factors for macroinvertebrate predators varied negatively with water-column Se. Our results increase scientific understanding of Se bioaccumulation processes in Appalachian headwater streams. Environ Toxicol Chem 2018;37:2714-2726. © 2018 SETAC.

Influence of dilution water ionic composition on acute major ion toxicity to the mayfly Neocloeon triangulifer.

Field and laboratory studies have shown that mayflies (Ephemeroptera) tend to be relatively sensitive to elevated major ion concentrations, but little is known about how ionic composition influences these responses. The present study evaluated the acute toxicity of major ion salts to the mayfly Neocloeon triangulifer over a range of background water quality conditions. The mayfly was particularly sensitive to Na2 SO4 , with the median lethal concentration (LC50) of 1338 mg SO4 /L being lower than LC50s reported for 7 other species at that hardness. Increasing hardness of the dilution water from 30 to 150 mg/L (as CaCO3 ) resulted in doubling of LC50s for sodium salts, and an approximately 1.5-fold increase in LC50 for MgSO4 . Potassium salt toxicity was not strongly influenced by hardness, consistent with findings for other species. When hardness was held constant but the Ca to Mg ratio was manipulated, the ameliorative effect on Na2 SO4 and NaCl did not appear as strong as when hardness was varied; but for MgSO4 the amelioration relative to Ca activity was similar between the 2 experiments. The toxicity of K salts to N. triangulifer was similar to Na salts on a millimolar basis, which contrasts with several other species for which K salts have been much more toxic. In addition, the toxicity of KCl to N. triangulifer was not notably affected by Na concentration, as has been shown for Ceriodaphnia dubia. Finally, plotting LC50s in terms of ion activity (Cl, SO4 , Na, Mg, or K) over the range of Ca activities in dilution water resulted in significant positive relationships, with comparable slopes to those previously observed for C. dubia over the same range of Ca activities. Environ Toxicol Chem 2018;37:1330-1339. © 2018 SETAC.

Seasonal pattern of anthropogenic salinization in temperate forested headwater streams.

Salinization of freshwaters by human activities is of growing concern globally. Consequences of salt pollution include adverse effects to aquatic biodiversity, ecosystem function, human health, and ecosystem services. In headwater streams of the temperate forests of eastern USA, elevated specific conductance (SC), a surrogate measurement for the major dissolved ions composing salinity, has been linked to decreased diversity of aquatic insects. However, such linkages have typically been based on limited numbers of SC measurements that do not quantify intra-annual variation. Effective management of salinization requires tools to accurately monitor and predict salinity while accounting for temporal variability. Toward that end, high-frequency SC data were collected within the central Appalachian coalfield over 4 years at 25 forested headwater streams spanning a gradient of salinity. A sinusoidal periodic function was used to model the annual cycle of SC, averaged across years and streams. The resultant model revealed that, on average, salinity deviated approximately ±20% from annual mean levels across all years and streams, with minimum SC occurring in late winter and peak SC occurring in late summer. The pattern was evident in headwater streams influenced by surface coal mining, unmined headwater reference streams with low salinity, and larger-order salinized rivers draining the study area. The pattern was strongly responsive to varying seasonal dilution as driven by catchment evapotranspiration, an effect that was amplified slightly in unmined catchments with greater relative forest cover. Evaluation of alternative sampling intervals indicated that discrete sampling can approximate the model performance afforded by high-frequency data but model error increases rapidly as discrete sampling intervals exceed 30 days. This study demonstrates that intra-annual variation of salinity in temperate forested headwater streams of Appalachia USA follows a natural seasonal pattern, driven by interactive influences on water quantity and quality of climate, geology, and terrestrial vegetation. Because climatic and vegetation dynamics vary annually in a seasonal, cyclic manner, a periodic function can be used to fit a sinusoidal model to the salinity pattern. The model framework used here is broadly applicable in systems with streamflow-dependent chronic salinity stress.

Influence of chloride on the chronic toxicity of sodium nitrate to Ceriodaphnia dubia and Hyalella azteca.

While it has been well established that increasing chloride concentration in water reduces the toxicity of nitrite to freshwater species, little work has been done to investigate the effect of chloride on nitrate toxicity. We conducted acute and chronic nitrate (as sodium nitrate) toxicity tests with the cladoceran Ceriodaphnia dubia and the amphipod Hyalella azteca (chronic tests only) over a range of chloride concentrations spanning natural chloride levels found in surface waters representative of watersheds of the Great Lakes Region. Chronic nitrate toxicity test results with both crustaceans were variable, with H. azteca appearing to be one of the more sensitive invertebrate species tested and C. dubia being less sensitive. While the variability in results for H. azteca were to an extent related to chloride concentration in test water that was distinctly not the case for C. dubia. We concluded that the chloride dependent toxicity of nitrate is not universal among freshwater crustaceans. An additional sodium chloride chronic toxicity test with the US Lab strain of H. azteca in the present study suggested that when present as predominantly sodium chloride and with relatively low concentrations of other ions, there is a narrow range of chloride concentrations over which this strain is most fit, and within which toxicity test data are reliable.

Selection of food combinations to optimize survival, growth, and reproduction of the amphipod Hyalella azteca in static-renewal, water-only laboratory exposures.

Although standardized sediment toxicity testing methods have been developed for the amphipod Hyalella azteca, no standardized chronic water-only toxicity testing methods have been established. Furthermore, optimal feeding and water quality conditions for culturing and toxicity testing with this species remained unclear. The objective of the present study was to determine the food or combination of foods that best promotes survival, growth, and reproduction of the US Lab strain of Hyalella azteca under 42-d, water-only, static-renewal testing conditions. The authors conducted 7 42-d control (no toxicant) tests with various combinations of food (including Tetramin, yeast-cereal leaves-trout chow, diatoms, wheatgrass, alfalfa, and maple leaves) and substrate types (clean "unconditioned" Nitex screens vs "conditioned" Nitex screens that were colonized by live biofilms). Over all treatments, survival ranged from 18% to 96%, dry weight per individual from 0.084 mg to 1.101 mg, and reproduction from 0 young/female to 28.4 young/female. Treatments that included Tetramin tended to result in better performance than those that did not. In particular, treatments that included Tetramin and either conditioned screens or diatoms consistently had high survival, weight, and reproduction values as well as low variability among replicates (measured as coefficient of variation). A ramped Tetramin plus diatom suspension feeding regime appears to have the greatest potential to produce consistently good performance across laboratories using static-renewal systems. Environ Toxicol Chem 2016;35:2407-2415. © 2016 SETAC.

Effects of environmentally relevant mixtures of major ions on a freshwater mussel.

The Clinch and Powell Rivers (Virginia, USA) support diverse mussel assemblages. Extensive coal mining occurs in both watersheds. In large reaches of both rivers, major ion concentrations are elevated and mussels have been extirpated or are declining. We conducted a laboratory study to assess major ion effects on growth and survival of juvenile Villosa iris. Mussels were exposed to pond water and diluted pond water with environmentally relevant major ion mixtures for 55 days. Two treatments were tested to mimic low-flow concentrations of Ca(2+), Mg(2+), [Formula: see text] , [Formula: see text] , K(+) and Cl(-) in the Clinch and Powell Rivers, total ion concentrations of 419 mg/L and 942 mg/L, respectively. Mussel survival (>90%) and growth in the two treatments showed little variation, and were not significantly different than in diluted pond water (control). Results suggest that major ion chronic toxicity is not the primary cause for mussel declines in the Clinch and Powell Rivers.

Contrasting effects of chloride on growth, reproduction, and toxicant sensitivity in two genetically distinct strains of Hyalella azteca.

The strain of Hyalella azteca (Saussure: Amphipoda) commonly used for aquatic toxicity testing in the United States has been shown to perform poorly in some standardized reconstituted waters frequently used for other test species. In 10-d and 42-d experiments, the growth and reproduction of the US laboratory strain of H. azteca was shown to vary strongly with chloride concentration in the test water, with declining performance observed below 15 mg/L to 20 mg/L. In contrast to the chloride-dependent performance of the US laboratory strain of H. azteca, growth of a genetically distinct strain of H. azteca obtained from an Environment Canada laboratory in Burlington, Ontario, Canada, was not influenced by chloride concentration. In acute toxicity tests with the US laboratory strain of H. azteca, the acute toxicity of sodium nitrate increased with decreasing chloride in a pattern similar not only to that observed for control growth, but also to previous acute toxicity testing with sodium sulfate. Subsequent testing with the Burlington strain showed no significant relationship between chloride concentration and the acute toxicity of sodium nitrate or sodium sulfate. These findings suggest that the chloride-dependent toxicity shown for the US laboratory strain may be an unusual feature of that strain and perhaps not broadly representative of aquatic organisms as a whole.

Full-life chronic toxicity of sodium salts to the mayfly Neocloeon triangulifer in tests with laboratory cultured food.

Although insects occur in nearly all freshwater ecosystems, few sensitive insect models exist for use in determining the toxicity of contaminants. The objectives of the present study were to adapt previously developed culturing and toxicity testing methods for the mayfly Neocloeon triangulifer (Ephemeroptera: Baetidae), and to further develop a method for chronic toxicity tests spanning organism ages of less than 24 h post hatch to adult emergence, using a laboratory cultured diatom diet. The authors conducted 96-h fed acute tests and full-life chronic toxicity tests with sodium chloride, sodium nitrate, and sodium sulfate. The authors generated 96-h median lethal concentrations (LC50s) of 1062 mg Cl/L (mean of 3 tests), 179 mg N-NO3 /L, and 1227 mg SO4 /L. Acute to chronic ratios ranged from 2.1 to 6.4 for chloride, 2.5 to 5.1 for nitrate, and 2.3 to 8.5 for sulfate. The endpoints related to survival and development time were consistently the most sensitive in the tests. The chronic values generated for chloride were in the same range as those generated by others using natural foods. Furthermore, our weight-versus-fecundity plots were similar to those previously published using the food culturing method on which the present authors' method was based, indicating good potential for standardization. The authors believe that the continued use of this sensitive mayfly species in laboratory studies will help to close the gap in understanding between standard laboratory toxicity test results and field-based observations of community impairment.

Elements of concern in fillets of bighead and silver carp from the Illinois River, Illinois.

Efforts to control invasive bighead (Hypophthalmichthys nobilis) and silver carp (H. molitrix) may include harvest for human consumption. We measured concentrations of arsenic (As), mercury (Hg), and selenium (Se) in fillets from silver and bighead carp collected from the lower Illinois River, Illinois, USA, to determine whether concentrations were of health concern and differed by species, size, and location. Concentrations of total As were below detection limits in most bighead (92%) and silver (77%) carp fillets, whereas inorganic As was below detection limits in all samples. Mean Hg concentrations were greater in bighead (0.068 mg kg(-1)) than in silver carp (0.035 mg kg(-1)), and were smallest in carp from the confluence of the Illinois and Mississippi rivers. Mercury concentrations in fillets were positively correlated with body mass in both species. Concentrations of Hg were below the US Food and Drug Administration's (USFDA) action level (1 ppm as methyl-Hg); however, concentrations in some bighead (70%) and silver (12%) carp fell within the range that would invoke a recommendation to limit meals in sensitive cohorts. Mean Se concentrations were greater in silver (0.332 mg kg(-1)) than in bighead (0.281 mg kg(-1)) carp fillets, and were below the 1.5 mg kg(-1) limit for an unrestricted number of meals/month. The mean molar ratio of Se:Hg in fillets was lower in bighead (14.0) than in silver (29.1) carp and was negatively correlated with mass in both species Concentrations of Hg in bighead and silver carp fillets should be considered when assessing the risks associated with the use of these species as a protein source.