Effects of nitrogenous wastes on survival of the Barton Springs salamander (Eurycea sosorum).

The objective of our study was to determine the acute toxicity of 3 common aquatic nitrogenous toxicants to the federally endangered Barton Springs salamander (Eurycea sosorum). Based on our findings, the 96-h median lethal concentrations (96-h LC50) for un-ionized ammonia-N, nitrite-N, and nitrate-N to E. sosorum are 2.0 ± 0.32, 31.7 ± 4.02, and 968.5 ± 150.6 mg/L, respectively. These results establish a benchmark for the tolerance of plethodontid salamanders to these toxicants and indicate that current water quality criteria are adequate for their protection. Environ Toxicol Chem 2017;36:3003-3007. © 2017 SETAC.

Effects of roof and rainwater characteristics on copper concentrations in roof runoff.

Copper sheeting is a common roofing material used in many parts of the world. However, copper dissolved from roof sheeting represents a source of copper ions to watersheds. Researchers have studied and recently developed a simple and efficient model to predict copper runoff rates. Important input parameters include precipitation amount, rain pH, and roof angle. We hypothesized that the length of a roof also positively correlates with copper concentration (thus, runoff rates) on the basis that runoff concentrations should positively correlate with contact time between acidic rain and the copper sheet. In this study, a novel system was designed to test and model the effects of roof length (length of roof from crown to the drip edge) on runoff copper concentrations relative to rain pH and roof angle. The system consisted of a flat-bottom copper trough mounted on an apparatus that allowed run length and slope to be varied. Water of known chemistry was trickled down the trough at a constant rate and sampled at the bottom. Consistent with other studies, as pH of the synthetic rainwater decreased, runoff copper concentrations increased. At all pH values tested, these results indicated that run length was more important in explaining variability in copper concentrations than was the roof slope. The regression equation with log-transformed data (R(2) = 0.873) accounted for slightly more variability than the equation with untransformed data (R(2) = 0.834). In log-transformed data, roof angle was not significant in predicting copper concentrations.

Influence of pH, hardness, dissolved organic carbon concentration, and dissolved organic matter source on the acute toxicity of copper to Daphnia magna in soft waters: implications for the biotic ligand model.

The influence of pH, dissolved organic carbon (DOC) concentration, water hardness, and dissolved organic matter (DOM) source on the acute toxicity of copper were investigated with standardized 48-h Daphnia magna toxicity tests. Toxicity tests were conducted according to a four-factor complete factorial design. Nominal factor levels were as follows: pH 6 and 8; DOC, 2.5 and 10 mg/L; hardness, 10, 20, and 40 mg/L as CaCO3; and two DOM sources (collected from the Black River and Edisto River, SC, USA). The experimental design resulted in 24 different factor level combinations. Results indicated that all factors had significant effects on copper toxicity. Furthermore, a strong interactive effect of DOC concentration and pH was detected. Because the biotic ligand model (BLM) has become a widely used tool for predicting toxicity and interpreting toxicity test results, its performance with these data was evaluated. Seventy percent of BLM predictions were within twofold of the observed median lethal concentrations. However, BLM parameters could be adjusted to improve model performance with this data set. This analysis suggested that in soft waters, the CuOH+ complex binds more strongly with the biotic ligand and that the competitive effect of hardness cations should be increased. The results of the present study may have implications for application of the BLM to some types of surface waters. Furthermore, a comprehensive analysis of BLM performance with all available data should be performed, and necessary updates to model parameters should be made to produce the most robust and widely applicable model.

Influence of copper exposure on whole-body sodium levels in larval fathead minnows (Pimephales promelas).

Because metals such as Cu inhibit ionoregulation, the increased energy requirement to counter passive diffusive losses in soft water may translate into increased sensitivity to metal exposure. We developed a method to determine whole-body Na concentrations of larval fathead minnows (Pimephales promelas) as a physiological indicator of health. This method was used to characterize net rates of Na flux from fish exposed to Cu in the presence of varying levels of hardness and alkalinity. In extremely soft waters (hardness, < or = 10 mg/L as CaCO(3)), larval fish experienced rates of net whole-body Na loss greater than what has been observed in juvenile and adult fish when exposed to Cu at concentrations near the median lethal concentration. Elevating hardness (>10 mg/L as CaCO(3)), however, decreased the apparent kinetics of Na loss caused by Cu exposure, which suggests the process was related to uncompetitive inhibition of Cu by hardness cations. Although the percentage of Na loss associated with mortality in larval fish was similar to that in juvenile and adult fish (30% loss of exchangeable Na pool), larvae reached this level within 12 h of exposure, and it was not representative of the onset of mortality. These results suggested that ionoregulatory measures by themselves are not a conclusive metric for Cu regulation using larval fish. To account for increased sensitivity in low-hardness waters in the development of biotic ligand models, the critical amount of Cu associated with the gill to cause mortality (i.e., the median lethal accumulation value) should be characterized more appropriately as a function of hardness below 20 mg/L as CaCO(3).

Toxicity of two pulsed metal exposures to Daphnia magna: relative effects of pulsed duration-concentration and influence of interpulse period.

Aquatic organisms living in surface waters experience fluctuating contaminant exposures that vary in concentration, duration, and frequency. This study characterized the role of pulsed concentration, pulsed duration, and the interval between pulses on the toxicity of four metals (Cu, Zn, Se, and As) to Daphnia magna. During 21-d toxicity tests, neonatal D. magna were exposed to single or double pulses. Pulsed concentrations and durations ranged from 32 to 6000 microg/L and 8 to 96 h, respectively. Intervals between two pulses ranged from 24 to 288 h. Mortality, growth, and reproduction were characterized for exposures. For single-pulse exposures of Cu and As, metal concentration had a stronger effect on survival of D. magna than did pulsed duration: pulses with 2X concentration and 1Y duration resulted in more mortality than did pulses with 1X concentration and 2Y duration. In contrast, effects of pulsed duration were stronger than metal concentration for Zn. However, the effects of duration and concentration were similar for Se. The relative effects of pulsed concentration and duration found in the present study revealed that the common method using area under the curve (AUC = concentration x duration) may not always accurately estimate environmental risk from metals (e.g., for Cu, Zn, As). In addition, the occurrence of delayed mortality in the present study revealed that using continuous exposure bioassays might underestimate metal toxicity to aquatic biota. For double-pulse exposures, the toxicity of the second pulse was influenced by the first pulse for all four metals. This influence was dependent on the pulsed concentration and duration and the interval between pulses. Further, toxicity caused by the second pulse decreased as the time between the exposures increased. For all four metals, there existed an interval great enough that the toxicity of the two pulses was independent. This would result in less toxicity for multiple exposures than continuous exposures with the same total exposure duration. The interval time at which the effects of the two pulses were independent increased with increasing concentration. Growth and cumulative reproduction of D. magna over 21 d were not significantly affected by pulsed exposures examined in the present study, indicating recovery of the organisms.

An integrated model describing the toxic responses of Daphnia magna to pulsed exposures of three metals.

Some toxicology research in which toxicant exposures are continual (pulsed) rather than continuous have been reported. A number of toxicity models have been developed for pulsed and continuous exposures. Most of these models were developed based on one- or two-compartment, first-order toxicokinetics and were calibrated with organic compounds. In the present study, the relationship between mortality (after 21 d) of Daphnia magna in response to pulsed and continuous exposures to Cu, Zn, and Se was used to develop a model that integrated the effects of single and multiple pulsed metal exposures based on first-order uptake and depuration kinetics. Mortality was a function of exposure concentration, duration, and recovery time between exposures. The model was successfully validated using an independent data set. It is applicable to risk assessment and, potentially, may be incorporated with other models (e.g., the biotic ligand model) to predict the toxicity of pulsed metal exposures under a range of environmental conditions.

Whole-body sodium concentration in larval fathead minnows (Pimephales promelas) during and following copper exposure.

This research used whole-body sodium concentration to characterize exposure and recovery of larval fathead minnows (Pimephales promelas) from acute pulsed copper exposures. Whole-body sodium was chosen because the acute mechanism of copper toxicity to fishes is putative disruption of ion regulation, resulting in a loss of sodium and eventually leading to mortality. Whole-body sodium response in larval fathead minnows exposed to copper was both concentration and duration dependent. The loss of sodium to approximately 70% of control levels occurred within 12 h of exposure. Organisms demonstrated an ability to recover whole-body sodium within 48 h after exposure to concentrations below 0.47 microM Cu2+ for 3, 6, or 9 h. However, at higher concentrations, organisms required more than 48 h to recover. Whole-body sodium concentrations and mortality for all continuous exposures were strongly correlated. These results may facilitate development of a physiologically based model to predict the response of organisms to copper in receiving streams.

Evaluation of acute copper toxicity to larval fathead minnows (Pimephales promelas) in soft surface waters.

The hardness-based regulatory approach for Cu prescribes an extrapolation of the toxicity-versus-hardness relationship to low hardness (< or =50 mg/L as CaCO3). Hence, the objective of the present research was to evaluate the influences of water quality on acute Cu toxicity to larval fathead minnow (Pimephales promelas) in low-hardness surface waters. Seasonal water sampling was conducted at 24 sites throughout South Carolina, USA, to determine the site-specific influences of soft surface-water conditions on acute Cu toxicity. Concurrent toxicity tests in laboratory water, matched for hardness and alkalinity (modified method), also were conducted to allow calculation of water-effect ratios (WERs). In addition, tests were conducted at recommended hardness levels (recommended method) for comparison of WER methodology in soft water. Surface-water conditions (average+/-standard deviation, n = 53) were hardness of 16+/-8 mg/L as CaCO3, alkalinity of 18+/-11 mg/L as CaCO3, and dissolved organic carbon of 6+/-4 mg/L. Dissolved Cu 48-h median lethal concentration (LC50) values varied nearly 45-fold across the dataset and greater than four-fold at individual sites. Spatial (p < 0.0001) and seasonal (p = 0.026) differences among LC50 values were determined for eight sites that had multiple toxicity results for one year. All modified WERs were greater than 1.0, suggesting that the site waters were more protective of Cu toxicity than the matched laboratory water. Some WERs generated using recommended methods were less than 1.0, suggesting limited site-specific protection. Based on these observations, extrapolation of the hardness-based equation for Cu at 50 mg/L or less as CaCO3 would adequately protect fathead minnow populations in soft surface waters. The WER results presented here demonstrate the inconsistency between hardness-based criteria and the methodology for deriving site-specific water-quality criteria in low-hardness waters.

Physiological basis for large differences in resistance to nitrite among freshwater and freshwater-acclimated euryhaline fishes.

Uptake of environmental NO2- by most freshwater fishes occurs at the gills where NO2- is actively transported into the blood by the Cl- uptake pathway. Some freshwater fishes do not concentrate NO2- in their plasma, regardless of environmental NO2- exposure and exhibit a high degree of resistance to NO2-. Recent studies indicate that freshwater-adapted killifish (Fundulidae: Fundulus heteroclitus) and European eel (Anguillidae: Anguilla anguilla) have no or minimal Cl- uptake activity at the gills relative to most freshwater fishes; rather, Cl- requirements are met in other ways (probably dietary). We hypothesized that different rates of Cl- uptake by the gill may explain the observed differences in NO2- uptake and consequent toxicity among freshwater fishes. Cl- influx rates of channel catfish (Ictaluridae: Ictalurus punctatus), a species that concentrates NO2- in the plasma and is sensitive to NO2-, and bluegill (Centrarchidae: Lepomis macrochirus), a species that does not concentrate NO2- in the plasma and is resistant to NO2-, were determined over a range of environmental Cl- concentrations. Channel catfish actively transported chloride into the plasma (Km = 155.6+/-101.2 micromol/L Cl-; Jmax = 414.9+/-51.4 nmol/g/h; +/-SEM). In contrast, bluegill exhibited no observable Cl- uptake. We placed our results and previously reported results in a phylogenetic context and concluded that differences in Cl- uptake mechanisms among groups of freshwater fishes may explain, in large part, the wide range of sensitivity to environmental NO2-. NO2- uptake determinations may also prove to be an easy screening method when studying the phylogenetic distribution and nature of Cl- uptake mechanisms in the gills of fishes.

Changes in water quality after addition of sea salts to fresh water: implications during toxicity testing.

Sea salts (seven brands from six commercially-available sources) were dissolved in water to develop 30 g/l solutions, and selected water quality characteristics were then monitored for 96 h. One or more water quality characteristics changed significantly during the 96 h period in six of the reconstituted sea salts. Measured characteristics of sea water diluted to 30 g/l demonstrated no changes during the observation period. The sea salts from different sources also demonstrated differences in absolute concentrations of some characteristics measured. Application of the Biotic Ligand Model to predict copper toxicity to the bivalve Mytilus edulis in solutions of the salts tested yielded 96-h median-lethal concentrations that ranged from 2 to 13 microg/l. Since water quality affects toxicity of many environmental pollutants, the source of the sea salt and equilibration time should be considered when planning toxicity tests.

Influence of natural organic matter source on copper toxicity to larval fathead minnows (Pimephales promelas): implications for the biotic ligand model.

The influence of dissolved natural organic matter (NOM) source on copper toxicity was investigated with larval fathead minnows (Pimephales promelas) in reconstituted moderately hard water. Ninety-six-hour static renewal toxicity tests were conducted to investigate an assumption of the biotic ligand model (BLM) that NOM source does not need to be considered to adequately predict copper toxicity. The nine different NOM isolates used in these toxicity tests were chemically well-characterized substances that were obtained by reverse osmosis as part of an NOM typing project based in southern Norway. Three median lethal concentration (LC50) values were estimated for toxicity tests conducted with each NOM, at nominal dissolved organic carbon (DOC) concentrations of 2, 5, and 10 mg/L. Tests also were conducted in dilution waters in which no NOM was added. Regression analyses were conducted to compare NOM-specific (specific NOM source) LC50s versus DOC concentration relationships to each other, as well as to the overall LC50 versus DOC concentration relationship. Statistical differences were found regarding the effects of NOM source on copper toxicity. Similar analyses were conducted with humic acid (HA) concentrations and spectral absorbance, and differences in the effect of NOM source on copper toxicity were similarly concluded. These results do not support the assumption that copper toxicity can be adequately predicted by utilizing DOC concentration, regardless of NOM source. Evaluation of relationships between LC50 values and other NOM characteristics revealed that despite significant differences due to NOM source on copper toxicity, DOC and HA concentrations were the most effective parameters in explaining variability in LC50 values. When BLM-predicted LC50 values were compared to observed LC50 values, predicted values showed reasonable agreement with observed values, but some deviations occurred due to NOM source and DOC concentration.

Influence of water quality and age on nickel toxicity to fathead minnows (Pimephales promelas).

This research characterized the effects of water quality and organism age on the toxicity of nickel (Ni) to fathead minnows (Pimephales promelas) to facilitate the accurate development of site-specific water-quality criteria. Nickel sulfate hexahydrate (NiSO4 x 6H2O) was used as the Ni source for performing acute toxicity tests (median lethal concentration after 96-h exposure [96-h LC50]) with < 1-d-old and 28-d-old P. promelas under varying regimes of hardness, pH, alkalinity, and natural organic matter (NOM). The toxicity of Ni was inversely related to water hardness between hardness values of 20 and 150 mg/L (as CaCO3). Below 30 mg/L alkalinity, Ni toxicity was related to alkalinity. The effect of pH was confounded by hardness and the presence of NOM. In the absence of NOM, the toxicity of Ni increased as pH increased at high hardness and alkalinity. In general, 28-d-old fish were less sensitive than < 1-d-old fish to Ni. This lower sensitivity ranged from 12-fold at low hardness and alkalinity (20 and 4 mg/L, respectively) to 5-fold at high hardness and alkalinity (100 and 400 mg/L, respectively). The presence of NOM (10 mg/L as dissolved organic carbon [DOC]) reduced Ni toxicity by up to 50%, but this effect appeared to be saturated above DOC at 5 mg/L. Incubating Ni with the NOM solution from 1 to 17 days had no effect on Ni toxicity. When using multivariate analysis, the 96-h LC50 for Ni was a function of fish age, alkalinity, hardness, and NOM (96-h LC50 = -0.642 + 0.270(fish age) + 0.005(alkalinity) + 0.018(hardness) + 0.138(DOC)). When using this model, we found a strong relationship between measured and predicted 96-h LC50 values (r2 = 0.94) throughout the treatment water qualities. The biotic ligand model (BLM) did not accurately predict Ni toxicity at high or low levels of alkalinity. Results of our research suggest that the BLM could be improved by considering NiCO3 to be bioavailable.

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.

Influence of dissolved organic matter source on silver toxicity to Pimephales promelas.

In the environment, the formation of organic and inorganic silver complexes can decrease Ag bioavailability (toxicity) to aquatic organisms. However, current water quality regulations do not consider the protective effects of water quality parameters such as dissolved organic carbon (DOC) concentration. To determine the effect of DOC concentration and source on silver toxicity, nine different natural organic matter isolates were used in 96-h static-renewal toxicity tests with fathead minnow (Pimephales promelas). The 96-h dissolved silver median lethal concentrations (LC50) among different sources of dissolved organic matter varied by up to fivefold (4.5-23.3 microg/L). Further, toxicity tests with organic matter from the site with the lowest 96-h LC50 value suggested only limited additional attenuation of silver toxicity when DOC concentration was increased from 5.1 to 14.0 mg/L. With this site excluded, we found little more than a twofold difference among 96-h dissolved Ag LC50s for the remaining sources (10.1-23.3 microg/L). However, significant toxicological differences among sites remained. It was apparent that organic matter from different sources varied both chemically and toxicologically, but no conclusions could be drawn that related compositional variation to observed Ag toxicity for these isolates.

Brain Monoamine Concentrations as Predictors of Growth Inhibition in Channel Catfish Exposed to Ammonia.

Fingerlings of channel catfish Ictalurus punctatus exhibited a significant exposure-dependent decrease in growth (measured by weight gain and increase in total length) and condition factor after 9 weeks of exposure to environmental ammonia. Concentrations of 5-hydroxytryptamine (5-HT) and dopamine in the brain decreased significantly whereas the ratio of 5-hydroxyindoleacetic acid (5-HIAA) to 5-HT increased significantly in exposure-dependent manners. The brain dopamine concentrations and the 5-HIAA : 5-HT ratio collectively explained 88% of the variation in growth due to ammonia exposure. This study demonstrates the potential to predict ammonia-induced inhibition of growth in channel catfish with physiological changes.