The objectives of the present study were to develop methods for conducting chronic toxicity tests with juvenile mussels under flow‐through conditions and to determine the chronic toxicity of copper and ammonia to juvenile mussels using these methods. In two feeding tests, two‐month‐old fatmucket (Lampsilis siliquoidea) and rainbow mussel (Villosa iris) were fed various live algae or nonviable algal mixture for 28 d. The algal mixture was the best food resulting in high survival (≥90%) and growth. Multiple copper and ammonia toxicity tests were conducted for 28 d starting with two‐month‐old mussels. Six toxicity tests using the algal mixture were successfully completed with a control survival of 88 to 100%. Among copper tests with rainbow mussel, fatmucket, and oyster mussel (Epioblasma capsaeformis), chronic value ([ChV], geometric mean of the no‐observed‐effect concentration and the lowest‐observed‐effect concentration) ranged from 8.5 to 9.8 μg Cu/L for survival and from 4.6 to 8.5 μg Cu/L for growth. Among ammonia tests with rainbow mussel, fatmucket, and wavy‐rayed lampmussel (L. fasciola), the ChV ranged from 0.37 to 1.2 mg total ammonia N/L for survival and from 0.37 to 0.67 mg N/L for growth. These ChVs were below the U.S. Environmental Protection Agency 1996 chronic water quality criterion (WQC) for copper (15 μg/L; hardness 170 mg/L) and 1999 WQC for total ammonia (1.26 mg N/L; pH 8.2 and 20°C). Results indicate that toxicity tests with two‐month‐old mussels can be conducted for 28 d with >80% control survival; growth was frequently a more sensitive endpoint compared to survival; and the 1996 chronic WQC for copper and the 1999 chronic WQC for total ammonia might not be adequately protective of the mussel species tested. However, a recently revised 2007 chronic WQC for copper based on the biotic ligand model may be more protective in the water tested.

Adult rare minnow (Gobiocypris rarus) were exposed to 0, 1, 5, and 25 ng/l (nominal concentrations) of 17a-ethynylestradiol (EE 2 ) and 3, 10, and 30 lg/l (nominal concentrations) of 4-nonylphenol (NP) under flow-through conditions for a period of 28 d. Low mortality was observed at 5 and 25 ng/l EE 2 and the growth of fish reduced significantly at 25 ng/l EE 2 compared to controls. However, the gonadosomatic indices (GSI) of male fish were significantly higher in 1 ng/l EE 2 treatments and in 10 and 30 lg/l NP treatments (p < 0.05). Renal somatic indices (RSI) of male fish in EE 2 treatments were significantly higher than those in controls (p < 0.05). In contrast, significantly decreased GSI and RSI of female fish could only be observed in 5 and 25 ng/l EE 2 treatments (p < 0.05). Hepatosomatic indices (HSI) of male fish were significantly higher in 25 ng/l EE 2 treatments. However, significantly increased of HSI of female fish could only be observed in 1 ng/l EE 2 treatments. Plasma vitellogenin (VTG) induction could be observed in males after exposed to different concentrations of EE 2 and NP, and plasma VTG concentrations in females exposed to 5 and 25 ng/l EE 2 were also significantly higher than in controls (p < 0.05). At level higher than 5 ng/l EE 2 or 30 lg/l NP, hepatic tissue and renal tissue impairment of males could be observed. The pathological male liver was associated with a hypertrophy of hepatocytes and damages to cellar structure and accumulated eosinophilic material. Renal tissue showed different pathological effects which was reflected by accumulated eosinophilic material, hemorrhages within the kidney tubules and hypertrophy of the tubular epithelia. Also at these levels of exposure, feminization of male fish could be noticed and parts of males manifested the testis-ova phenomenon. Ovaries of female rare minnow in 25 ng/l EE 2 treatment group were degenerated. Therefore when exposed to EE 2 and NP even at environmental observed concentrations, adverse effects could occur in the reproductive system of adult fishes. The observed hepatic tissue and renal tissue impairment should be due to the induction and accumulation of VTG in organs, especially in males.

Abstract. Assessment of contaminant impacts to federally identified endangered, threatened and candidate, and stateidentified endangered species (collectively referred to as "listed" species) requires understanding of a species' sensitivities to particular chemicals. The most direct approach would be to determine the sensitivity of a listed species to a particular contaminant or perturbation. An indirect approach for aquatic species would be application of toxicity data obtained from standard test procedures and species commonly used in laboratory toxicity tests. Common test species (fathead minnow, Pimephales promelas; sheepshead minnow, Cyprinodon variegatus; and rainbow trout, Oncorhynchus mykiss) and 17 listed or closely related species were tested in acute 96-hour water exposures with five chemicals (carbaryl, copper, 4-nonylphenol, pentachlorophenol, and permethrin) representing a broad range of toxic modes of action. No single species was the most sensitive to all chemicals. For the three standard test species evaluated, the rainbow trout was more sensitive than either the fathead minnow or sheepshead minnow and was equal to or more sensitive than listed and related species 81% of the time. To estimate an LC50 for a listed species, a factor of 0.63 can be applied to the geometric mean LC50 of rainbow trout toxicity data, and more conservative factors can be determined using variance estimates (0.46 based on 1 SD of the mean and 0.33 based on 2 SD of the mean). Additionally, a low-or no-acute effect concentration can be estimated by multiplying the respective LC50 by a factor of approximately 0

The present study evaluated the performance and variability in acute toxicity tests with glochidia and newly transformed juvenile mussels using the standard methods outlined in American Society for Testing and Materials (ASTM). Multiple 48-h toxicity tests with glochidia and 96-h tests with juvenile mussels were conducted within a single laboratory and among five laboratories. All tests met the test acceptability requirements (e.g., >or=90% control survival). Intralaboratory tests were conducted over two consecutive mussel-spawning seasons with mucket (Actinonaias ligamentina) or fatmucket (Lampsilis siliquoidea) using copper, ammonia, or chlorine as a toxicant. For the glochidia of both species, the variability of intralaboratory median effective concentrations (EC50s) for the three toxicants, expressed as the coefficient of variation (CV), ranged from 14 to 27% in 24-h exposures and from 13 to 36% in 48-h exposures. The intralaboratory CV of copper EC50s for juvenile fatmucket was 24% in 48-h exposures and 13% in 96-h exposures. Interlaboratory tests were conducted with fatmucket glochidia and juveniles by five laboratories using copper as a toxicant. The interlaboratory CV of copper EC50s for glochidia was 13% in 24-h exposures and 24% in 48-h exposures, and the interlaboratory CV for juveniles was 22% in 48-h exposures and 42% in 96-h exposures. The high completion success and the overall low variability in test results indicate that the test methods have acceptable precision and can be performed routinely.

Concern with the redistribution of contaminants associated with sediment in the upper Mississippi River (UMR) arose after the flood of 1993. This project is designed to evaluate the status of sediments in the UMR and is one article in a series designed to assess the extent of sediment contamination in navigational pools of the river. Companion articles evaluate sediment toxicity and benthic community composition in navigation pools of the river. The objectives of the present study were to: (1) to assess the bioaccumulation of sediment-associated contaminants in the UMR using laboratory exposures with the oligochaete Lumbriculus variegatus, and (2) to compare bioaccumulation in laboratory-exposed oligochaetes to field-collected oligochaetes. Sediment samples and native oligochaetes were collected from 23 navigational pools on the Upper Mississippi River and the Saint Croix River. Contaminant concentrations measured in the L. variegatus after 28-day exposures to sediment in the laboratory were compared to contaminant concentrations in field-collected oligochaetes from the 13 pools where these sediments were collected. Contaminant concentrations were relatively low in sediments and tissues from the pools evaluated. Only polycyclic aromatic hydrocarbons (PAHs) and total polychlorinated biphenyls (PCBs) were frequently measured above detection limits. The majority of the biota-sediment-accumulation factors (BSAFs) for PAHs were within a range of about 1.0 to 2.6, suggesting that the theoretical BSAF value of 1.7 could be used to predict these mean BSAFs with a reasonable degree of certainty. A positive correlation was observed between lipid-normalized concentrations of PAHs detected in laboratory-exposed and field-collected oligochaetes across all sampling locations. Rank correlations for concentrations of individual compounds between laboratory-exposed and field-collected oligochaetes were strongest for benzo(e)pyrene, perylene, benzo(b,k)fluoranthene, and pyrene. About 90% of the paired PAH concentrations in laboratory-exposed and field-collected oligochaetes were within a factor of three of one another indicating laboratory results could be extrapolated to the field with a reasonable degree of certainty.

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The upper Clark Fork River basin in western Montana is widely contaminated by metals from past mining, milling, and smelting activities As part of a comprehensive ecological risk assessment for the upper Clark Fork River, we measured physical and chemical characteristics of surficial sediment samples that were collected from depositional zones for subsequent toxicity evaluations Sampling stations included five locations along the upper 200 km of the river, six locations in or near Milltown Reservoir (about 205 km from the river origin), and two tributary reference sites Concentrations of As, Cd, Cu, Mn, Pb, and Zn decreased from the upper stations to the downstream stations in the Clark Fork River but then increased in all Milltown Reservoir stations to levels similar to uppermost river stations Large percentages (50 to 90%) of the total Cd, Cu, Pb, and Zn were extractable by dilute (3 n) HCl for all samples Copper and zinc accounted for greater than 95% of extractable metals on a molar basis Acid‐volatile sulfide (AVS) concentrations were typically moderate (0 6 to 23 μmol/g) in grab sediment samples and appeared to regulate dissolved (filterable) concentrations of Cd, Cu, and Zn in sediment pore waters Acid volatile sulfide is important in controlling metal solubility in the depositional areas of the Clark Fork River and should be monitored in any future studies Spatial variability within a sampling station was high for Cu, Zn, and AVS, therefore, the potential for toxicity to sediment dwelling organisms may be highly localized