Sublethal Toxicity of 17 Per- and Polyfluoroalkyl Substances with Diverse Structures to Ceriodaphnia dubia, Hyalella azteca, and Chironomus dilutus.

Seven-day sublethal toxicity tests were performed with the freshwater invertebrates Ceriodaphnia dubia, Hyalella azteca, and Chironomus dilutus to determine the effects of per- or polyfluorinated alkyl substances (PFAS) of varying chain length within four classes: perfluoroalkyl carboxylic acids (PFCAs), perfluoroalkyl sulfonic acids (PFSAs), perfluoroalkane sulfonamides, and fluorotelomer sulfonic acids. In general, toxicity increased with increasing chain length, but the slopes of these relationships varied markedly by species and chemical class. The toxicity of individual PFCAs was similar among species. The toxicity of PFSAs was similar to PFCAs for C. dubia and H. azteca, whereas PFSAs were much more toxic to C. dilutus, with median effect concentrations (EC50s) as low as 0.022 mg perfluorooctane sulfonate (PFOS)/L and 0.012 mg perfluorononane sulfonate (PFNS)/L. Despite the high sensitivity to PFOS and PFNS, C. dilutus was not very sensitive to structurally similar fluorotelomer sulfonates (6:2 and 8:2). Perfluoroalkane sulfonamides were the most toxic class tested among all species (e.g., EC50s of 0.011 and 0.017 mg perfluorooctane sulfonamide/L for C. dilutus and H. azteca, respectively). The differences in toxicity among species and chemical classes suggest that mechanisms of PFAS toxicity may differ as a function of both. Environ Toxicol Chem 2024;43:359-373. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

Acute Toxicity of Major Geochemical Ions to Fathead Minnows (Pimephales promelas): Part B-Modeling Ion Toxicity.

Mathematical models are presented for the acute median lethal concentrations of major geochemical ions (Na+ , K+ , Ca2+ , Mg2+ , Cl- , SO4 2- , HCO3 - /CO3 2- ) to fathead minnows (Pimephales promelas), based on an extensive series of experiments presented in a companion article. Toxicity relationships across different dilution waters, individual salts, and salt mixtures suggest six independent mechanisms of toxicity to consider in modeling efforts, including Mg/Ca-specific toxicity, osmolarity-related toxicity, SO4 -specific toxicity, K-specific toxicity, effects of high pH/alkalinity, and a multiple ion-related toxicity at low Ca distinct from the other mechanisms. Models are evaluated using chemical activity-based exposure metrics pertinent to each mechanism, but concentration-based alternative models that are simpler to apply are also addressed. These models are compared to those previously provided for Ceriodaphnia dubia, and various issues regarding their application to risk assessments are discussed. Environ Toxicol Chem 2022;41:2095-2106. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Acute Toxicity of Major Geochemical Ions to Fathead Minnows (Pimephales Promelas): Part A-Observed Relationships for Individual Salts and Salt Mixtures.

The results of a series of experiments on the acute toxicity of major geochemical ions (Na+ , K+ , Ca2+ , Mg2+ , Cl- , SO4 2- , HCO3 - /CO3 2- ) to fathead minnows (Pimephales promelas) are reported. Tests of individual major ion salts in various dilution waters demonstrated that the toxicities of Na, Mg, and K salts decrease as the overall ion content of the dilution water increases. For Na and Mg salts, this is attributable to Ca content as previously reported for Ceriodaphnia dubia. For K salts, the cause is unclear, but it is not due to Na as reported for C. dubia. In an unregulated test at high pH (9.3), NaHCO3 was also found to be twice as toxic compared to when the pH was reduced to 8.4. Experiments with binary salt mixtures indicated the existence of multiple independent mechanisms of action. These include K-specific toxicity and Ca/Mg-specific toxicity previously reported for C. dubia, but also apparent toxicities related to SO4 and to high pH/alkalinity in CO3 /HCO3 -dominated exposures. Previous work with C. dubia also suggested a general ion toxicity involving all ions that was correlated with osmolarity. For fathead minnow, similar correlations were observed, but multiple mechanisms were indicated. At higher Ca, this general toxicity could be attributable to osmotic effects, but at lower Ca, osmolarity may be more a covariate than a cause, with this toxicity being related to a combined effect of ions other than via osmolarity. Environ Toxicol Chem 2022;41:2078-2094. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

The effects of arsenic speciation on accumulation and toxicity of dietborne arsenic exposures to rainbow trout.

The effects on juvenile rainbow trout survival, growth, food consumption, and food conversion efficiency from dietborne exposures to inorganic arsenic (arsenite, arsenate) and to the organoarsenicals monomethylarsonate (MMA), dimethylarsinate (DMA), and arsenobetaine (AsB) were investigated in two experiments: (1) a 28-d exposure using live diets of oligochaete worms separately exposed via water to these five arsenic compounds and (2) a 56-d exposure using pellet diets prepared from commercial fish food to which arsenite, MMA, or DMA were added. In the live diet experiment, the degree to which worms could be contaminated with the organoarsenicals was limited by toxicity to the worms and other experimental constraints, so that their toxicity relative to inorganic arsenic could not be fully established, but AsB was concluded to have low toxicity, consistent with published results for mammals. For the pellet diet experiment, MMA and DMA were found to be at least an order of magnitude less toxic than inorganic As on the basis of concentration in the diet, as well as much less toxic on the basis of accumulation in the fish. The need to consider speciation in aquatic risk assessments for arsenic was further demonstrated by tissue analyses of three macroinvertebrate species from a mining-impacted stream, which showed large variations in both total arsenic and the relative amounts of inorganic and organic arsenic. Additionally, although effects of arsenic on trout appear to be well correlated with inorganic arsenic, worms were found to be more sensitive to waterborne DMA than to inorganic arsenic, showing that low toxicity of organoarsenicals cannot be assumed for all aquatic organisms. Various difficulties in evaluating and applying studies on dietborne exposures and fish growth are also discussed.

The relative importance of waterborne and dietborne As exposure on survival and growth of juvenile fathead minnows.

The survival and growth of juvenile fathead minnows were investigated at various combinations of waterborne exposure to arsenate and of dietborne exposure to oligochaete worms which had been exposed to inorganic arsenic. Previous work with rainbow trout established that dietborne arsenic can reduce fish growth at environmentally relevant concentrations and could be more important than waterborne exposures. This was found to be less true for fathead minnows, which were less sensitive to dietborne exposures than rainbow trout, while being as or more sensitive to waterborne exposures. When assessed on the basis of accumulation of total As by the fish, further differences between fathead minnows and rainbow trout were evident. Fathead minnows accumulated relatively more arsenic from water versus diet than trout, and the accumulations at which growth effects occurred in minnows were different for dietborne and waterborne exposure, whereas they were the same for trout. These results suggest complex relationships involving arsenic speciation, toxicokinetics, and toxicodynamics, and underscore a need for care in relating effects information to real-world exposures. The present study also demonstrated the challenges in testing and interpreting growth effects in long-term exposures to fish, because the expression of toxicity can be confounded by the relationship of fish growth to size, the feeding regime, and wet weight versus dry weight relationships.

Chronic toxicity of major ion salts and their mixtures to Ceriodaphnia dubia.

In previous work we intensively studied the acute responses of the cladoceran Ceriodaphnia dubia to major geochemical ions (Na+ , K+ , Ca2+ , Mg2+ , Cl- , SO42- , and HCO3- /CO32- ), culminating in the development of models to predict acute toxicity from ionic composition. To quantitatively evaluate whether the toxicological behavior of major ions observed for C. dubia extends to chronic toxicity, we conducted 58 chronic toxicity tests with individual major salts and binary mixtures thereof. Chronic responses paralleled those demonstrated previously for acute exposure, specifically 1) similar relative toxicity of individual salts; 2) different Na salts showing similar potency when exposure is expressed as osmolarity; 3) toxicity of Mg, Ca, and K salts related to cation activity; 4) decreased toxicity of Na and Mg salts when Ca activity is increased at less than toxic concentrations; 5) additive behavior for salt mixtures sharing a common cation; and 6) independent behavior for salt mixtures with dissimilar cations, except Mg/Ca mixtures which appeared additive. Acute-to-chronic ratios were fairly consistent among salts, with values of approximately 1.8 for acute 50% lethal concentration (LC50) to chronic 50% effect concentration (EC50) and 2.8 for LC50/EC20 when expressed on an activity basis. Adjusting the previous acute toxicity model for acute-to-chronic ratios yielded chronic models that predict chronic toxicity within the range of intertest variability. Because these models are informed by a wide range of ion mixtures, they should provide robust assessment tools for natural waters enriched with major ions. Environ Toxicol Chem 2019;38:769-783. © Published 2018 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

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.

The acute toxicity of major ion salts to Ceriodaphnia dubia. III. Mathematical models for mixture toxicity.

Based on previous research on the acute toxicity of major ions (Na+ , K+ , Ca2+ , Mg2+ , Cl- , SO42- , and HCO3- /CO32- ) to Ceriodaphnia dubia, a mathematical model was developed for predicting the median lethal concentration (LC50) for any ion mixture, excepting those dominated by K-specific toxicity. One component of the model describes a mechanism of general ion toxicity to which all ions contribute and predicts LC50s as a function of osmolarity and Ca activity. The other component describes Mg/Ca-specific toxicity to apply when such toxicity exceeds the general ion toxicity and predicts LC50s as a function of Mg and Ca activities. This model not only tracks well the observed LC50s from past research used for model development but also successfully predicts LC50s from new toxicity tests on synthetic mixtures of ions emulating chemistries of various ion-enriched effluents and receiving waters. It also performs better than a previously published model for major ion toxicity. Because of the complexities of estimating chemical activities and osmolarity, a simplified model based directly on ion concentrations was also developed and found to provide useful predictions. Environ Toxicol Chem 2018;37:247-259. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

The acute toxicity of major ion salts to Ceriodaphnia dubia. II. Empirical relationships in binary salt mixtures.

Many human activities increase concentrations of major geochemical ions (Na+1 , K+1 , Ca+2 , Mg+2 , Cl-1 , SO4-2 , and HCO3-1 /CO3-2 ) in freshwater systems, and can thereby adversely affect aquatic life. Such effects involve several toxicants, multiple toxicity mechanisms, various ion interactions, and widely varying ion compositions across different water bodies. Previous studies of individual salt toxicities have defined some useful relationships; however, adding single salts to waters results in atypical compositions and does not fully address mixture toxicity. To better understand mechanisms and interactions for major ion toxicity, 29 binary mixture experiments, each consisting of 7 to 8 toxicity tests, were conducted on the acute toxicity of major ion salts and mannitol to Ceriodaphnia dubia. These tests showed multiple mechanisms of toxicity, including: 1) nonspecific ion toxicity, correlated with osmolarity and to which all ions contribute; and 2) cation-dependent toxicities for potassium (K), magnesium (Mg), and calcium (Ca) best related to their chemical activities. These mechanisms primarily operate independently, except for additive toxicity of Mg-dependent and Ca-dependent toxicities. These mixture studies confirmed ameliorative effects of Ca on sodium (Na) and Mg salt toxicities and of Na on K salt toxicity, and further indicated lesser ameliorative effects of Ca on K salt toxicity and Mg on Na salt toxicity. These results provide a stronger basis for assessing risks from the complex mixtures of ions found in surface waters. Environ Toxicol Chem 2017;36:1525-1537. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

The acute toxicity of major ion salts to Ceriodaphnia dubia: I. influence of background water chemistry.

The ions Na+ , K+ , Ca2+ , Mg2+ , Cl- , SO42- , and HCO3- /CO32- (referred to in the present study as "major ions") are present in all freshwaters and physiologically required by aquatic organisms but can increase to harmful levels from a variety of anthropogenic activities. It is also known that the toxicities of major ion salts can vary depending on the concentrations of other ions, and understanding these relationships is key to establishing appropriate environmental limits. The authors present a series of experiments with Ceriodaphnia dubia to evaluate the acute toxicity of 12 major ion salts and to determine how toxicity of these salts varies as a function of background water chemistry. All salts except CaSO4 and CaCO3 were acutely toxic below saturation, with the lowest median lethal concentrations found for K salts. All 10 salts that showed toxicity also showed some degree of reduced toxicity as the ionic content of the background water increased. Experiments that independently varied Ca:Mg ratio, Na:K ratio, Cl:SO4 ratio, and alkalinity/pH demonstrated that Ca concentration was the primary factor influencing the toxicities of Na and Mg salts, whereas the toxicities of K salts were primarily influenced by the concentration of Na. These experiments also indicated multiple mechanisms of toxicity and suggested important aspects of dosimetry; the toxicities of K, Mg, and Ca salts were best related to the chemical activity of the cation, whereas the toxicities of Na salts also reflected an influence of the anions and were well correlated with osmolarity. Understanding these relationships between major ion toxicity and background water chemistry should aid in the development of sensible risk-assessments and regulatory standards. Environ Toxicol Chem 2016;35:3039-3057. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Using an interlaboratory study to revise methods for conducting 10-d to 42-d water or sediment toxicity tests with Hyalella azteca.

Studies have been conducted to refine US Environmental Protection Agency, ASTM International, and Environment Canada standard methods for conducting 42-d reproduction tests with Hyalella azteca in water or in sediment. Modifications to the H. azteca method include better-defined ionic composition requirements for exposure water (i.e., >15 mg/L of chloride and >0.02 mg/L of bromide) and improved survival, growth, and reproduction with alternate diets provided as increased rations over time in water-only or whole-sediment toxicity tests. A total of 24 laboratories volunteered to participate in the present interlaboratory study evaluating the performance of H. azteca in 42-d studies in control sand or control sediment using the refined methods. Improved growth and reproduction of H. azteca was observed with 2 alternate diets of 1) ramped diatoms (Thalassiosira weissflogii) + ramped Tetramin or 2) yeast-cerophyll-trout chow (YCT) + ramped Tetramin, especially when compared with results from the traditional diet of 1.8 mg YCT/d. Laboratories were able to meet proposed test acceptability criteria and in most cases had lower variation in growth or reproduction compared with previous interlaboratory studies using the traditional YCT diet. Laboratory success in conducting 42-d H. azteca exposures benefited from adherence to several key requirements of the detailed testing, culturing, and handling methods. Results from the present interlaboratory study are being used to help revise standard methods for conducting 10-d to 42-d water or sediment toxicity exposures with H. azteca. Environ Toxicol Chem 2016;35:2439-2447. © 2016 SETAC.

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.

Relative sensitivity of an amphipod Hyalella azteca, a midge Chironomus dilutus, and a unionid mussel Lampsilis siliquoidea to a toxic sediment.

The objective of the present study was to evaluate the relative sensitivity of test organisms in exposures to dilutions of a highly toxic sediment contaminated with metals and organic compounds. One dilution series was prepared using control sand (low total organic carbon [TOC; <0.1%, low binding capacity for contaminants]) and a second dilution series was prepared using control sediment from West Bearskin Lake, Minnesota, USA (high TOC [∼10% TOC, higher binding capacity for contaminants]). Test organisms included an amphipod (Hyalella azteca; 10-d and 28-d exposures), a midge (Chironomus dilutus; 20-d and 48-d exposures started with <1-h-old larvae, and 13-d and 48-d exposures started with 7-d-old larvae), and a unionid mussel (Lampsilis siliquoidea; 28-d exposures). Relative species sensitivity depended on the toxicity endpoint and the diluent. All 3 species were more sensitive in sand dilutions than in West Bearskin Lake sediment dilutions. The <1-h-old C. dilutus were more sensitive than 7-d-old C. dilutus, but replicate variability was high in exposures started with the younger midge larvae. Larval biomass and adult emergence endpoints of C. dilutus exhibited a similar sensitivity. Survival, weight, and biomass of H. azteca were more sensitive endpoints in 28-d exposures than in 10-d exposures. Weight and biomass of L. siliquoidea were sensitive endpoints in both sand and West Bearskin Lake sediment dilutions. Metals, ammonia, oil, and other organic contaminants may have contributed to the observed toxicity.

Evaluation of PCB bioaccumulation by Lumbriculus variegatus in field-collected sediments.

Review of data from several contaminated sediment sites suggested that biota-sediment accumulation factors (BSAFs) declined with increasing contaminant concentrations in the sediment. To evaluate the consistency and possible causes of this behavior, polychlorinated biphenyl (PCB)-contaminated sediment samples from the Hudson, Grasse, and Fox River Superfund sites were used in sediment bioaccumulation tests with the freshwater oligochaete, Lumbriculus variegatus, with PCB concentrations in interstitial water (IW) quantified using polyoxymethylene passive samplers. Measured BSAFs tended to decrease with increasing PCB concentration in sediment, especially for the more highly chlorinated congeners. Measures of partitioning between sediment, IW, and oligochaetes showed that measured sediment-IW partition coefficients (KTOC ) tended to increase slightly with increasing sediment contamination, whereas the ratio of tissue PCB to IW PCB tended to decrease with increasing concentration in IW. Variation in accumulation among sediments was clearly influenced by bioavailability, as reflected by IW measurements, although the specific cause of varying KTOC was not clear. Calculated partitioning between IW and organism lipid (Klipid ) indicated that accumulation was generally 5 to 10-fold higher than would be predicted if Klipid was approximately equal to the n-octanol-water partition coefficient (KOW ). While affirming previous observations of decreasing BSAFs with increasing PCB contamination, the relatively shallow slope of the observed relationship in the current data may suggest that this concentration dependence is not a major uncertainty in sediment risk assessment, particularly if measurements of PCBs in IW are incorporated.

Evaluating the role of ion composition on the toxicity of copper to Ceriodaphnia dubia in very hard waters.

The mitigating effect of increasing hardness on metal toxicity is reflected in water quality criteria in the United States over the range of 25-400 mgl(-1) (as CaCO(3)). However, waters in the arid west of the US frequently exceed 400 mgl(-1) hardness, and the applicability of hardness-toxicity relationships in these waters is unknown. Acute toxicity tests with Ceriodaphnia dubia were conducted at hardness levels ranging from approximately 300 to 1,200 mgl(-1) using reconstituted waters that mimic two natural waters with elevated hardness: (1) alkaline desert southwest streams (Las Vegas Wash, NV), and (2) low alkalinity waters from a CaSO(4)-treated mining effluent in Colorado. The moderately-alkaline EPA synthetic hard water was also included for comparison. Copper toxicity did not consistently vary as a function of hardness, but likely as a function of other water quality characteristics (e.g., alkalinity or other correlated factors). The hardness equations used in regulatory criteria, therefore, may not provide an accurate level of protection against copper toxicity in all types of very hard waters. However, the mechanistic Biotic ligand model generally predicted copper toxicity within +/-2X of observed EC(50) values, and thus may be more useful than hardness for modifying water quality criteria.

The effect of calcium and magnesium ratios on the toxicity of copper to five aquatic species in freshwater.

While it is generally accepted that water hardness affects copper toxicity, the major ions that contribute to water hardness (calcium [Ca] and magnesium [Mg]) may affect copper toxicity differently. This is important because the Ca:Mg ratio in standard laboratory-reconstituted waters often differs from the ratio in natural surface waters. Copper toxicity was assessed for five different aquatic species: rainbow trout (RBT), fathead minnow (FHM), Ceriodaphnia dubia, Daphnia magna, and an amphipod (Gammarus sp.) under different Ca:Mg ratios (4:0, 3:1, 1:1, 1:3, and 1:4 mass basis) at a common hardness (180 mg/L as CaCO3) and alkalinity (120 mg/L as CaCO3). Copper toxicity increased at lower Ca:Mg ratios for RBT but increased at higher Ca:Mg ratios for D. magna. Fathead minnows (<24 h old) were more sensitive to copper in 1:1 Ca:Mg waters compared to 3:1 Ca:Mg waters. The toxicity of copper did not vary under different Ca:Mg ratios for Gammarus sp., C. dubia, and 28-d-old FHM. The effect of Ca:Mg ratios on copper toxicity changed for D. magna in softer water (90 mg/L as CaCO3) compared with hard water studies.