Response spectrum of pentachlorobenzene and fluoranthene for Chironomus tentans and Hyalella azteca

The whole-body residues of pentachlorobenzene (PCBz) and fluoranthene (FLU) in Hyalella azteca and Chironomus tentans were determined for a variety of chronic sublethal effects. The endpoints evaluated for H. azteca included 28-d growth and survival and 42-d growth, survival, and reproduction. Adverse effects to C. tentans also were determined at multiple endpoints including 10-d growth, cumulative pupation and emergence, and reproduction. The lowest-observed-effect residue (LOER) based on whole-body residues associated with growth was consistent between compounds and species tested with concentrations ranging from 0.17 to 0.33 micromol/g. For H. azteca, the most sensitive endpoints were growth at 0.23 micromol/g and reproduction at 0.11 micromol/g for PCBz and FLU, respectively. For C. tentans, the most sensitive endpoints were emergence, development and reproduction at 0.02 micromol/g, and development and reproduction at 0.15 micromol/g for PCBz and FLU, respectively. Compared to residues associated with acute lethality, the most sensitive sublethal endpoints were approximately 4 and 60 times lower for PCBz and FLU, respectively. The relative consistency of the sublethal endpoints suggests that body residues can be a valuable tool to evaluate bioaccumulation data as part of a risk assessment to predict adverse effects to biota.     

Interlaboratory evaluation of Hyalella azteca and Chironomus tentans short-term and long-term sediment toxicity tests

Methods for assessing the long-term toxicity of sediments to Hyalella azteca and Chironomus tentans can significantly enhance the capacity to assess sublethal effects of contaminated sediments through multiple endpoints. Sublethal tests allow us to begin to understand the relationship between short-term and long-term effects for toxic sediments. We present an interlaboratory evaluation with long-term and 10-d tests using control and contaminated sediments in which we assess whether proposed and existing performance criteria (test acceptability criteria [TAC]) could be achieved. Laboratories became familiar with newly developed, long-term protocols by testing two control sediments in phase 1. In phase 2, the 10-d and long-term tests were examined with several sediments. Laboratories met the TACs, but results varied depending on the test organism, test duration, and endpoints. For the long-term tests in phase 1, 66 to 100% of the laboratories consistently met the TACs for survival, growth, or reproduction using H. azrteca, and 70 to 100% of the laboratories met the TACs for survival and growth, emergence, reproduction, and hatchability using C. tentans. In phase 2, fewer laboratories participated in long-term tests: 71 to 88% of the laboratories met the TAC for H. azteca, whereas 50 to 67% met the TAC for C. tentans. In the 10-d tests with H. azteca and C. tentans, 82 and 88% of the laboratories met the TAC for survival, respectively, and 80% met the TAC for C. tentans growth. For the 10-d and long-term tests, laboratories predicted similar toxicity. Overall, the interlaboratory evaluation showed good precision of the methods, appropriate endpoints were incorporated into the test protocols, and tests effectively predicted the toxicity of sediments.     

Toxicokinetics of sediment-sorbed benzo[a]pyrene and hexachlorobiphenyl using the freshwater invertebrates Hyalella azteca,Chironomus tentans,and Lumbriculus variegatus

This study investigated the effect of long-term sediment aging on the toxicokinetics of benzo[a]pyrene (BaP) and hexachlorobiphenyl (HCBP) using three freshwater benthic invertebrates. Hyalella azteca, Chironomus tentans, and Lumbriculus variegatus were exposed to BaP- and HCBP-spiked sediments that were aged for 7 d or 1.5 years. The toxicokinetics of the two compounds were determined for each test organism using a two-compartment model. The modeling of BaP was more complex because biotransformation was included within the model. The results of this study showed that the HCBP uptake clearance rates (k(s)) for each species were generally an order of magnitude greater than those determined for BaP and this difference was most likely due to preferential and rapid binding of BaP to sediment particles. Overall, the bioavailability of HCBP in spiked sediments tended to decrease with duration of aging, based on k(s) values and bioaccumulation factors (BAFs). However, the decreases in bioavailability appear to be species specific. Benzo[a]pyrene did not decline in bioavailability for the species tested because it may resist movement into the micropores of the sediment due to its large size. In addition to the bioassays, this article outlines a method for toxicokinetic modeling of biotransformed compounds and methods for statistical comparisons of kinetic parameters (i.e., k(s), k(d)...) and BAF values.     

Chronic toxicity of the synthetic hormone 17alpha-ethinylestradiol to Chironomus tentans and Hyalella azteca

The chronic toxicity of the synthetic hormone 17alpha-ethinylestradiol (EE2) was investigated in two benthic invertebrates, the midge Chironomus tentans and the freshwater amphipod Hyalella azteca, in life-cycle water-only assays. In C. tentans, a 50% decrease in emergence was observed at a concentration of 1.5 mg/L; emergence was a more sensitive endpoint than survival, growth, or biomass. Reproduction was not significantly affected by EE2 exposure until a concentration of 3.1 mg/L, where emergence, and therefore reproduction, did not occur. In contrast, reproduction was the most sensitive endpoint in H. azteca (50% decrease in reproduction observed at a concentration of 0.36 mg/L). The sensitivity of the F1 generation to EE2 was also investigated with H. azteca, but was not different from the F0 generation. The data from the present study were combined with those from previous 10-d toxicity assays, to derive acute to chronic toxicity ratios (ACRs) for EE2. The ACRs calculated for EE2 were 13 for C. tentans and 16 for H. azteca, indicating that the application factors currently used in ecological risk assessment for the derivation of chronic toxicity are protective and conservative for these organisms. The results of the present study suggest that chronic toxicity was not mediated by disruption of endocrine pathways. Using a hazard quotient approach, the risk associated with sublethal exposure to EE2 was <1 for H. azteca and C. tentans, indicating that adverse effects are not expected, and that environmental exposure to EE2 likely poses a low risk to benthic invertebrates.     

Sediment Toxicity Evaluation for Hexachlorobenzene: Spiked Sediment Tests with Leptocheirus plumulosus, Hyalella azteca, and Chironomus tentans

Hexachlorobenzene (HCB) is a hydrophobic organic chemical that has shown a lack of toxicity in aquatic tests at concentrations up to and exceeding the solubility limit. The equilibrium partitioning approach to deriving sediment quality benchmarks, which assumes that toxicity can be predicted based on contaminant concentrations in interstitial water, predicts that HCB will not produce direct toxicity to benthic invertebrates as a sediment contaminant. However, the potential for toxicity due to direct exposure to sediment-adsorbed HCB has not been thoroughly established. This study evaluated the survival and growth of the estuarine amphipod Leptocheirus plumulosus, the freshwater amphipod Hyalella azteca, and the midge Chironomus tentans (freshwater) following 10-day exposure to sediment spiked with a range of HCB concentrations. H. azteca was tested under both freshwater and estuarine (10 ppt salinity) conditions. No significant toxicity was observed for any test species at the highest test concentration (60 mg/kg normalized to 1% organic carbon). Minimum detectable differences were less than or equal to 20% for three of eight test endpoints. The observed results add to the available weight of evidence indicating a limited potential for HCB-related sediment toxicity to benthic invertebrates. Received: 23 February 1998/Accepted: 8 June 1998     

Toxicity of the explosives 2,4,6-trinitrotoluene,hexahydro-1,3,5-trinitro-1,3,5-triazine,and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine in sediments to Chironomus tentans and Hyalella azteca: low-dose hormesis and high-dose mortality

The toxicity of the explosives 2,4,6-trinitrotoluene (TNT); hexahydro-1,3,5-trinitro-1,3,5-triazine (royal demolition explosive [RDX]); and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (high-melting explosive [HMX]), was evaluated in spiked sediment with two freshwater invertebrates. The midge Chironomus tentans and the amphipod Hyalella azteca demonstrated significant toxic effects after exposure to TNT and its degradation products, 1,3,5-trinitrobenzene (TNB) and 2,4-diamino-6-nitrotoluene (2,4-DANT). Significant reductions in survival of C. tentans exposed to TNT, TNB, and 2,4-DANT were observed at nominal sediment concentrations as low as 200 mg/kg. Hyalella azteca was more sensitive to TNT, TNB, and 2,4-DANT than the midge, where significant reductions in survival were observed at nominal concentrations of 50, 100, and 200 mg/kg, respectively. Survival of the midge and the amphipod was unaffected after exposure to RDX or HMX at the highest concentrations of 1,000 and 400 mg/kg, respectively. Growth of the midge, measured as total weight, was significantly reduced by 2,4-DANT. However, significantly increased growth was observed after exposure to sublethal concentrations of RDX and HMX. Although significant reductions in amphipod survival were observed at high concentrations of TNB, growth was significantly increased at sublethal concentrations. The results of the current investigation suggest that organisms exposed to explosives at contaminated sites may be affected at concentrations less than 25 mg/kg through hormetic growth enhancement and at higher concentrations through increased mortality.     

Toxicity of uranium, molybdenum, nickel, and arsenic to Hyalella azteca and Chironomus dilutus in water-only and spiked-sediment toxicity tests

A series of laboratory spiked-sediment toxicity tests with the amphipod Hyalella azteca and the midge Chironomus dilutus were undertaken to determine acute and chronic toxicity thresholds for uranium (U), molybdenum (Mo), nickel (Ni), and arsenic (As) based on both whole-sediment (total) and pore water exposure concentrations. Water-only toxicity data were also generated from separate experiments to determine the toxicities of these metals/metalloids under our test conditions and to help evaluate the hypothesis that pore water metal concentrations are better correlated with sediment toxicity to benthic organisms than whole-sediment metal concentrations. The relative toxicity of the four elements tested differed depending on which test species was used and whether whole-sediment or pore water metal concentrations were correlated with effects. Based on measured whole-sediment concentrations, Ni and As were the two most acutely toxic elements to H. azteca with 10-d LC50s of 521 and 532 mg/kg d.w., respectively. Measured pore water concentrations indicated that U and Ni were the two most acutely toxic elements, with 10-d LC50s to H. azteca of 2.15 and 2.05 mg/L, respectively. Based on pore water metal concentrations, the no-observed-effect concentrations (NOECs) for growth were (H. azteca and C. dilutus, respectively) 0.67 and 0.21 mg/L for U, <0.37 and 0.60 mg/L for Ni, and 16.43 and <0.42 mg/L for As. Pore-water lowest-observed-effect concentrations (LOECs) for growth were (H. azteca and C. dilutus, respectively) 2.99 and 0.48 mg/L for U, 0.37 and 2.33 mg/L for Ni, and 58.99 and 0.42 mg/L for As. For U and Ni, results from 96-h water-only acute toxicity tests correlated well with pore water metal concentrations in acutely toxic metal-spiked sediment. This was not true for As where metalloid concentrations in overlying water (diffusion from sediment) may have contributed to toxicity. The lowest whole-sediment LOEC reported here for As was 6.6- and 4-fold higher than the Canadian Council of Ministers of the Environment interim sediment quality guideline and the Canadian Nuclear Safety Commission (CNSC) lowest effect level (LEL), respectively. The lowest whole-sediment LOECs reported here for Ni, U and Mo were 4-, 17.5-, and >260-fold higher, respectively, than the CNSC LELs for these metals/metalloids. Data on pore water metal concentrations in toxic sediment would be a useful addition to future Guidelines documents.     

Exposure-related effects on Cd bioaccumulation explain toxicity of Cd-phenanthrene mixtures in Hyalella azteca

Little is known regarding mixture effects of metals and polynuclear aromatic hydrocarbon (PAHs) under environmentally relevant exposure regimes. Standard U.S. Environmental Protection Agency (U.S. EPA) procedures were applied and extended to test effects of phenanthrene (Phen) on sediment-Cd uptake, aqueous-Cd uptake, and Cd-elimination kinetics in the amphipod Hyalella azteca. In sediment exposures, Phen increased the projected equilibrium-tissue concentration of Cd from 47.2 (36.2-58.3) to 221.1 microg/g ([117.8-324.3], 95% confidence intervals [CI] in parentheses). Although Cd bioaccumulation increased markedly in sediment exposures, dissolved Cd concentrations and physical-chemical parameters indicative of Cd bioavailability were unaffected by Phen. Further, in water-only exposures, Phen had no effect on Cd bioaccumulation or Cd-elimination kinetics. These results indicate that increased Cd bioaccumulation in Cd-Phen mixtures occurred via a sediment-mediated process and was likely a function of increased uptake associated with feeding (i.e., Phen-induced alterations in ingestion and/or digestive processes). Observed increases in H. azteca lethality when exposed to Cd-Phen mixtures in sediment, but not in water-only exposures, likely resulted from increased Cd bioaccumulation rate rather than a true toxicological synergism. Thus, apparent synergisms and antagonisms may result from exposure-mediated effects in sediment that are unrelated to toxicological interactions. Implications of these findings regarding sediment-quality assessment and suggestions for future studies are discussed.     

Toxicity of sixty-three metals and metalloids to Hyalella azteca at two levels of water hardness

The toxicity of all atomically stable metals in the periodic table, excluding Na, Mg, K, and Ca, was measured in one-week exposures using the freshwater amphipod Hyalella azteca in both Lake Ontario, Canada, and soft water (10% Lake Ontario). Metals were added as atomic absorption standards (63 metals), and also as anion salts for 10 metals. Lethal concentrations resulting in 50% mortality (LC50s) were obtained for 48 of the metals tested; the rest were not toxic at 1,000 microg/L. The most toxic metals on a molar basis were Cd, Ag, Pb, Hg, Cr (anion), and Tl, with nominal LC50s ranging from 5 to 58 nmol/L (1 to 58 nmol/L measured). These metals were followed by U, Co, Os, Se (anion), Pt, Lu, Cu, Ce, Zn, Pr, Ni, and Yb with nominal LC50s ranging from 225 to 1,500 nmol/L (88-1,300 nmol/L measured). Most metals were similarly or slightly more toxic in soft water, but Al, Cr, Ge, Pb, and U were >17-fold more toxic in soft water; Pd was less toxic in soft water. Atomic absorption (AA) standards of As and Se in acid had similar toxicity as anions, Sb was more toxic as the AA standard, and Cr and Mn were more toxic as anions. One-week LC50s for H. azteca correlate strongly with three-week LC50s and three-week effect concentrations resulting in 50% reduction in reproduction (EC50s) in Daphnia magna.     

Effects of Calcium, Magnesium, and Sodium on Alleviating Cadmium Toxicity to Hyalella azteca

Received: 14 July 1999/Accepted: 16 November 1999     

Effect of sediment-associated pyrethroids, fipronil, and metabolites on Chironomus tentans growth rate, body mass, condition index, immobilization, and survival

Pyrethroids and fipronil insecticides partition to sediment and organic matter in aquatic systems and may pose a risk to organisms that use these matrices. It has been suggested that bioavailability of sediment-sorbed pesticides is reduced, but data on toxicity of sediment-associated pesticides for pyrethroids and fipronil are limited. In the current study, 10-d sediment exposures were conducted with larval Chironomus tentans for bifenthrin, lambda-cyhalothrin, permethrin, fipronil, fipronil-sulfide, and fipronil-sulfone, the last two being common fipronil metabolites. Sublethal endpoints included immobilization, instantaneous growth rate (IGR), body condition index, and growth estimated by ash-free dry mass (AFDM). Pyrethroid lethal concentrations to 50% of the population (LC50s) were 6.2, 2.8, and 24.5 microg/g of organic carbon (OC) for bifenthrin, lambda-cyhalothrin, and permethrin, respectively; with the former two lower than previously published estimates. Fipronil, fipronil-sulfide, and fipronil-sulfone LC50 values were 0.13, 0.16, and 0.12 microg/g of OC, respectively. Ratios of LC50s to sublethal endpoints (immobilization, IGR, and AFDM) ranged from 0.90 to 9.03. The effects on growth observed in the present study are important because of the unique dipteran life cycle involving pupation and emergence events. Growth inhibition would likely lead to ecological impacts similar to mortality (no emergence and thus not reproductively viable) but at concentrations up to 4.3 times lower than the LC50 for some compounds. In addition, C. tentans was highly sensitive to fipronil and metabolites, suggesting that dipterans may be important for estimating risk and understanding effects of phenylpyrazole-class insecticides on benthic macroinvertebrate communities.     

The role of metabolism in the toxicity of 2,4,6-trinitrotoluene and its degradation products to the aquatic amphipod Hyalella azteca

Toxicological data on the effects of the explosive, 2,4,6-trinitrotoluene (TNT), and its degradation products suggests an unpredictable toxicological response in aquatic organisms. Several studies suggest TNT becomes more toxic as it degrades while others suggest TNT becomes less toxic. This study focused on the toxicity of TNT and several degradation products as well as the role of oxidative metabolism in the toxicity of TNT. The aquatic invertebrate Hyalella azteca was used to evaluate the toxicity of TNT and four of its degradation products. The most reduced degradation product, 2,4-diamino, 6-nitrotoluene (2,4-DANT) was the most toxic to H. azteca. However, 2,4-DANT was only a minor metabolite in H. azteca. The influence of metabolism on the toxicokinetics of TNT was assessed indirectly through the use of a CYP450 inducer and inhibitor. Treatment of organisms with beta-napthoflavone (BNF), a CYP450 inducer, increased the toxicity of TNT and increased the rate of elimination and metabolism of TNT. Similar to BNF, organisms treated with clotrimazole (CTZ), a CYP450 inhibitor, resulted in increased toxicity and TNT metabolism. It is likely the ability to metabolize or bioactivate TNT to a more reactive intermediate plays a significant role in the sensitivity of organisms to TNT.     

Bioactive compounds in the aquatic environment: Studies on the mode of uptake of DDE by the aquatic midge,Chironomus tentans (diptera: Chironomidae)

The mode of uptake of DDE-¹⁴C byChironomus tentans larvae was investigated. There was no difference in the amount of DDE-¹⁴C accumulated by live and dead fourth instar larvae as a function of exposure time. Dead and live larvae were also exposed to an aqueous and substrate source of DDE-¹⁴C contamination and again no differences were found in the amount of DDE-¹⁴C accumulated indicating passive accumulation. Cuticle surface area and DDE-¹⁴C uptake relationships were found to have a high degree of correlation. The amount of DDE-¹⁴C concentrated by the larvae was increased by manipulation of water hardness. Calcium and magnesium ion concentrations in the water were increased and a subsequent increase in DDE-¹⁴C accumulation by the larvae resulted. An adsorption-diffusion mechanism is proposed to account for the mode of uptake and biological concentration capabilities of the midge.     

Effect of low dissolved oxygen on aquatic life stages of the caddisfly Clistoronia magnifica (Limnephilidae)

Embryos, larval stages (instars I–V), pupal stages, and pharate adults of the caddisfly Clistoronia magnifica (Limnephilidae) were exposed to a range of dissolved oxygen (DO) concentrations (0.9–8.3 mg/L) for 4–88 days in the laboratory. Some embryos suspended growth at low DO, resuming growth and hatch when DO was increased. Embryos and larvae all had 96-h EC50 values (50% mortality at 96 h) of about 2.0 mg/L DO. The statistical Effect and No-Effect Thresholds for larvae exposed through two molts from instars I–III were 1.6 and 2.4 mg/L, respectively. At DO concentrations below 4.6 mg/L, egg hatch, larval development, molting success, time of molting, pupation, and adult emergence were delayed.The information in this paper has been funded by the U.S. Environmental Protection Agency. It has been subjected to the Agency's peer and administrative review, and has been approved for publication.     

Effects of Temperature on the Toxicity of M-Parathion, Chlorpyrifos, and Pentachlorobenzene to Chironomus tentans

This study examined the influence of temperature (10, 20, and 30°C) on the acute toxicity and accumulation of two organophosphate (OP) insecticides and a narcotic chemical to the midge (Chironomus tentans). OP insecticides used in this study included chlorpyrifos and m-parathion, and pentachlorobenzene was the chosen narcotic. Chlorpyrifos was the most toxic chemical tested, followed by m-parathion and then pentachlorobenzene. A positive correlation was found between temperature and toxicity for each of the chemicals tested. A reverse trend was noted for total OP insecticide body residues with decreased concentrations found at the higher temperatures. Pentachlorobenzene body residues remained constant at all temperatures. All three chemicals showed increased uptake rates at 20 and 30°C in comparison to 10°C. The noted decrease in midge body residues at the higher temperatures for the OP insecticides was contributed to increased biotransformation and elimination rates at the higher temperatures. Overall, temperature had a greater influence on OP toxicity than for pentachlorobenzene, and this may be due to accelerated biotransformation of the OPs to more toxic o-analog metabolites at the higher temperatures. Accepted: 22 May 1999     

Bioconcentration and depuration of copper, cadmium, and zinc mixtures by the freshwater amphipod Hyalella azteca

The uptake and elimination of copper (Cu), cadmium (Cd), and zinc (Zn) by the amphipod Hyalella azteca during exposure to the metals singly and in various combinations was examined in controlled laboratory experiments. In single metal exposures the accumulation of all metals was rapid and increased with exposure time. Copper elimination was slower compared to that for zinc and for cadmium no elimination was detected after 5 days in clean water. In the two-metal mixtures it appears that the presence of one metal influenced the bioconcentration of the other, since the bioconcentration factor (BCF) for copper was higher in the presence of cadmium than in the presence of zinc and in the case of cadmium, the decrease of K(1) values from cadmium single exposure to the binary and tertiary mixtures suggests possible inhibition of cadmium uptake by the other metals. In the case of the three-metal mixture the situation is less clear, with both increased and decreased BCFs recorded, in comparison to single-metal and two-metal mixtures, suggesting both stimulation and inhibition of metal accumulation.     

Toxicity, residue dynamics, and reproductive effects of phthalate esters in aquatic invertebrates

Aquatic invertebrates were exposed to di-n-butyl and di-2-ethylhexyl phthalate esters in water to determine toxicity, accumulation, and reproductive effects of these compounds. The acute toxicities were low and ranged from 2.1 mg/liter to greater than 32 mg/liter. Residue accumulation was rapid resulting in body residues 70-13,600 times that of the water concentration. Phthalate residues were essentially gone after 10 days in fresh water. A reproductive impairment of 60% occurred in Daphnia magna exposed continuously to 3 μg/liter of di-2-ethylhexyl phthalate.