The ecotoxicological recovery of Ely Creek and tributaries (Lee County, VA) after remediation of acid mine drainage.

The Ely Creek watershed (Lee County, VA) was determined in 1995 to be the most negatively affected by acid mine drainage (AMD) within the Virginia coalfield. This determination led the US Army Corps of Engineers to design and build passive wetland remediation systems at two major AMD seeps affecting Ely Creek. This study was undertaken to determine if ecological recovery had occurred in Ely Creek. The results indicate that remediation had a positive effect on all monitoring sites downstream of the remediated AMD seeps. At the site most impacted by AMD, mean pH was 2.93 prior to remediation and improved to 7.14 in 2004. Benthic macroinvertebrate surveys revealed that one AMD influenced site had increased taxa richness from zero taxa in 1997 to 24 in 2004. While in situ testing of Asian clams resulted in zero survival at five of seven AMD influenced sites prior to remediation, some clams survived at all sites after. Clam survival was found to be significantly less than upstream references at only two sites, both downstream of un-mitigated AMD seeps in 2004. An ecotoxicological rating (ETR) system that combined ten biotic and abiotic parameters was developed as an indicator of the ecological status for each study site. A comparison of ETRs from before and after remediation demonstrated that all sites downstream of the remediation had experienced some level of recovery. Although the remediation has improved the ecological health of Ely Creek, un-mitigated AMD discharges are still negatively impacting the watershed.

Preliminary results of laboratory toxicity tests with the mayfly, Isonychia bicolor (Ephemeroptera: Isonychiidae) for development as a standard test organism for evaluating streams in the Appalachian coalfields of Virginia and West Virginia.

Selecting the most appropriate test species for sediment and water column assays has been a primary goal for ecotoxicologists. Standard test organisms and established test guidelines exist, but the USEPA-recommended species may not be the most sensitive organisms to anthropogenic inputs. This paper describes preliminary results of toxicity tests with the mayfly, Isonychia bicolor (Ephemeroptera). Results suggested that Isonychia were moderately sensitive to NaCl after 96 h with an average LC(50) value of 3.10 g NaCl per liter. This value decreased after 7 days of exposure, resulting in a mean LC(50) value of 1.73 g NaCl per liter. When exposed to a coal-mine-processed effluent, Isonychia generated LC(50) values that ranged from 13% to 39% effluent. I. bicolor were more sensitive to the coal processing effluent than Ceriodaphnia dubia with conductivity lowest observable effects concentration (LOEC) values for mayfly survivorship that ranged from 1,508 to 4,101 microS/cm, while LOEC values for C. dubia reproduction ranged from 2,132 to 4,240 microS/cm.

Coal-mine hollow fill and settling pond influences on headwater streams in southern West Virginia, USA.

The influences of coal-mine hollow fills and associated settling ponds in three headwater streams were assessed in southern West Virginia, USA. Fill drainage waters had elevated conductivities and metal concentrations, compared to a regional reference. Benthic macroinvertebrate richness was not affected consistently by the hollow fill drainages, relative to a regional reference, although a more tolerant community, lacking in Ephemeroptera taxa at most locations, was evident. Collector-filterer populations were elevated at monitoring stations directly below the settling ponds, indicating that the ponds' presence influenced macroinvertebrate community structure by means of organic enrichment. Corbicula fluminea growth was enhanced in monitoring locations directly below the settling ponds, also an apparent result of organic enrichment. Results of acute water column toxicity testing with Ceriodaphnia dubia, sediment chronic toxicity testing with Daphnia magna, and in-situ ecotoxicological assessments with C. fluminea demonstrated no mortality or toxic influence at most of the sites tested below the ponds. The settling ponds appear to serve as sinks in collecting some, but not all, trace metals.

Chlorine toxicity to early life stages of freshwater mussels (Bivalvia: Unionidae).

Chlorine (Cl) is a highly toxic, widely used halogen disinfectant that is present in point-source pollution discharges from wastewater treatment plants and industrial facilities. The U.S. Environmental Protection Agency freshwater criteria for Cl are 19 microg total residual Cl (TRC)/L as a maximum 1-h average concentration and 11 microg TRC/L as a maximum 4-d average; however, toxicological data for unionids were not used in these calculations. To address this void in the data, we conducted acute tests with glochidia from several species and 21-d bioassays with three-month-old Epioblasma capsaeformis and three-, six-, and 12-month-old Villosa iris juveniles. The 24-h lethal concentration 50 values for glochidia were between 70 and 220 [Lg TRC/L, which are 2.5 to 37 times higher than those reported in other studies for cladocerans. Significant declines in growth and survivorship were observed in the 21-d test with E. capsaeformis at 20 microg TRC/L. Lowest-observed-adverse-effects concentrations in bioassays with juvenile V. iris were higher (30-60 microg TRC/L) but showed a significant trend of declining toxicity with increased age. Although endpoints were above water quality criteria, the long life spans of unionids and potential implications of chronic exposure to endangered juvenile mussels still warrant concern.

The ecotoxicological recovery of Ely Creek and tributaries (Lee County, VA) after remediation of acid mine drainage.

The Ely Creek watershed (Lee County, VA) was determined in 1995 to be the most negatively affected by acid mine drainage (AMD) within the Virginia coalfield. This determination led the US Army Corps of Engineers to design and build passive wetland remediation systems at two major AMD seeps affecting Ely Creek. This study was undertaken to determine if ecological recovery had occurred in Ely Creek. The results indicate that remediation had a positive effect on all monitoring sites downstream of the remediated AMD seeps. At the site most impacted by AMD, mean pH was 2.93 prior to remediation and improved to 7.14 in 2004. Benthic macroinvertebrate surveys revealed that one AMD influenced site had increased taxa richness from zero taxa in 1997 to 24 in 2004. While in situ testing of Asian clams resulted in zero survival at five of seven AMD influenced sites prior to remediation, some clams survived at all sites after. Clam survival was found to be significantly less than upstream references at only two sites, both downstream of un-mitigated AMD seeps in 2004. An ecotoxicological rating (ETR) system that combined ten biotic and abiotic parameters was developed as an indicator of the ecological status for each study site. A comparison of ETRs from before and after remediation demonstrated that all sites downstream of the remediation had experienced some level of recovery. Although the remediation has improved the ecological health of Ely Creek, un-mitigated AMD discharges are still negatively impacting the watershed.

Bioassessment of an Appalachian headwater stream influenced by an abandoned arsenic mine.

Recent debate concerning the modification of safe drinking water standards for arsenic (As) has led to increased awareness of the risks As poses to both humans and the environment. However, few studies have examined the effects of As on the diversity and composition of aquatic assemblages in streams. Benthic macroinvertebrate surveys, chemical analysis of water column and sediment, and laboratory toxicity tests were conducted to assess effects of an abandoned As mine on a headwater stream, and to determine the primary component of toxicity. The average 48-hr LC50 value for Daphnia magna was 4316 microg As/L, and the average 96-hr LC50 value for Lepidostoma spp. was 2138 microg As/L. Reproduction was significantly reduced for D. magna at concentrations > or =312 microg As/L in water column laboratory bioassays, and for treatments in bioassays with sediments containing elevated As (> or =2630 mg/kg). These results support the findings of the in-stream benthic macroinvertebrate survey as the density and percent Ephemeroptera + Plecoptera, + Trichoptera (EPT) were substantially lower at sites downstream of the mine compared to upstream reference sites. Results of bioassays comparing the toxicity of As-contaminated site water and upstream reference water spiked with As salts suggest that As is the primary component of toxicity impacting the stream. Measured As concentrations at downstream sites were above the recommended Criterion Maximum Concentration of 340 microg As/L and Criterion Continuous Concentration of 150 microg As/L for protection of aquatic life published by the United States Environmental Protection Agency. At the study site, elevated As concentrations likely prevent recruitment of benthic macroinvertebrates and recovery of the perturbed headwater stream.

Acute and chronic toxicity of mercury to early life stages of the rainbow mussel, Villosa iris (Bivalvia: Unionidae).

Mercury (Hg) contamination is receiving increased attention globally because of human health and environmental concerns. Few laboratory studies have examined the toxicity of Hg on early life stages of freshwater mussels, despite evidence that glochidia and juvenile life stages are more sensitive to contaminants than adults. Three bioassays (72-h acute glochidia, 96-h acute juvenile, and 21-d chronic juvenile toxicity tests) were conducted by exposing Villosa iris to mercuric chloride salt (HgCl2). Glochidia were more sensitive to acute exposure than were juvenile mussels, as 24-, 48-, and 72-h median lethal concentration values (LC50) for glochidia were >107, 39, and 14 microg Hg/L, respectively. The 24-, 48-, 72-, and 96-h values for juveniles were 162, 135, 114, and 99 microg Hg/L, respectively. In the chronic test, juveniles exposed to Hg treatments > or = 8 microg/L grew significantly less than did control organisms. The substantial difference in juvenile test endpoints emphasizes the importance of assessing chronic exposure and sublethal effects. Overall, our study supports the use of glochidia as a surrogate life stage for juveniles in acute toxicity tests. However, as glochidia may be used only in short-term tests, it is imperative that an integrated approach be taken when assessing risk to freshwater mussels, as their unique life history is atypical of standard test organisms. Therefore, we strongly advocate the use of both glochidia and juvenile life stages for risk assessment.

Freshwater mussel shells as environmental chronicles: geochemical and taphonomic signatures of mercury-related extirpations in the North Fork Holston River, Virginia.

This study utilized freshwater mussel shells to assess mercury (Hg) contamination in the North Fork Holston River that extirpated (caused local extinctions of) a diverse mussel fauna. Shells (n = 366) were collected from five sites situated upstream (two sites), just below (one site), and downstream (two sites) of the town of Saltville, Virginia, where Hg was used to produce chlorine and caustic soda from 1950 to 1972. Shell samples were used to test the (1) utility of geochemical signatures of shells for assessing the spatial variation in Hg levels in the river relative to the contamination source and (2) value of taphonomy (post-mortem shell alteration) for distinguishing sites that differ in extirpation histories. Geochemical signatures of 40 shells, analyzed using atomic absorption spectroscopy, indicated a strong longitudinal pattern. All shells from the two upstream sites had low Hg concentrations (<5-31 microg/kg), shells directly below Saltville had variable, but dramatically higher concentrations (23-4637 microg/kg), and shells from the two downstream sites displayed intermediate Hg levels (<5-115 microg/kg) that declined with distance from Saltville. Two pre-industrial shells, collected at Saltville in 1917, yielded very low Hg estimates (5-6 microg/kg). Hg signatures were consistent among mussel species, suggesting that Hg concentrations were invariant to species type; most likely, highly variable Hg levels, both across sites and through time, overwhelmed any interspecific differences in Hg acquisition. Also, a notable postmortem incorporation of Hg in mussel shells seemed unlikely, as the Hg content was not correlated with shell taphonomy (r= 0.18; p = 0.28). The taphonomic analysis (n = 366) showed that the degree of shell alteration reliably distinguished sites with different extirpation histories. At Saltville, where live mussels have been absent for at least 30 years, shells were most heavily altered and fragmented. Conversely, fresh-looking shells abounded upstream, where reproducing mussel populations are still present. In summary, relic shells offered valuable spatio-temporal data on Hg concentrations in a polluted ecosystem, and shell taphonomic signatures discriminated sites with different extirpation histories. The shell-based strategies exemplified here do not require sampling live specimens and may augment more standard strategies applied to environmental monitoring. The approach should prove especially useful in areas with unknown extirpation and pollution histories.

Effect of cage design on growth of transplanted Asian clams: implications for assessing bivalve responses in streams.

This study was designed to determine whether survivorship and growth of Asian clams (Corbicula fluminea [Müller]) differed significantly between two types of field enclosures. Enclosures were either flexible mesh bags or rigid cages (hereto after referred as bioboxes) designed to homogenize substrate among study sites and accommodate Asian clam feeding mechanisms. For 96d, cages remained at 12 Clinch River (CR), Hurricane Fork (HF), and Dump's Creek (DC) sites upstream and downstream of a coal-fired power plant discharge, coal mining effluent, and coal combustion-related disposal facilities in Carbo, Virginia. Although survivorship was not significantly different between cage types, mean growth of clams in bioboxes was significantly greater overall (p = 0.0157). Despite the difference in growth between the two cage types, both confirmed significant reductions of survivorship and growth directly below the power plant discharge. Additionally, coefficient of variance values for biobox growth data were reduced at most study sites (averages of 16% for bioboxes versus 19% for mesh bags). Our results have implications toward strengthening weight-of-evidence approaches used to link impairment of transplanted bivalves to environmental contaminants. More importantly, these results suggest that ecotoxicological impairment of bivalves transplanted downstream of the coal-fired power plant discharge functioned independently of site-specific substrate provisions.

Sensitivity of juvenile freshwater mussels (Lampsilis fasciola, Villosa iris) to total and un-ionized ammonia.

This study evaluated the sensitivity of juveniles of two freshwater unionid mussel species (Villosa iris [Lea] and Lampsilis fasciola [Rafinesquel) to un-ionized and total ammonia. Five concentrations of ammonium chloride were tested using 96-h static-renewal toxicity tests at 12 and 20 degrees C. Based on their respective mean 96-h lethal concentration to 50% (LC50s), V. iris (0.11 mg/ L NH3-N) was more sensitive than L. fasciola (0.26 mg/L NH3-N). At 96 h, significant differences in sensitivity to un-ionized ammonia between the two temperatures were not observed for either species. Comparison of LC50s reported for other aquatic organisms to the 96-h LC50s calculated for juvenile L. fasciola and V. iris shows these two mussel species to be among the most sensitive to un-ionized ammonia. Based on reported levels of un-ionized ammonia in the aquatic environment from anthropogenic sources, un-ionized ammonia may be an important limiting toxicological factor to freshwater mussel populations.

Toxicity evaluation of a commercial bioremediation agent mixed with crude oil.

The toxicity and efficacy of a bacteria-based commercial bioremediation agent (CBA) was assessed through bioassays with juvenile inland silverside minnows, Menidia beryllina, and flask studies of oil degradation. Addition of the CBA to weathered Alaska, USA North Slope crude oil (ANS-521) prior to testing increased toxicity of the water-soluble fraction (WSF) of the oil in both chronic (growth) and acute (mortality) toxicity tests. Time-course toxicity assessment of the water-soluble fraction of the CBA/oil combination indicated increases in effect after 7 to 14 d of mixing that coincided with elevated concentrations of both alkanes and aromatics in the WSF. Under controlled laboratory conditions, the CBA significantly enhanced degradation of the oil compared with a treatment with nutrients alone. The alkane fraction was degraded by nearly 100% over a 42-d period while the aromatic fraction was decreased by 70%. While toxicity testing is not currently required to list bioremediation agents on the product schedule of the National Oil and Hazardous Substances Pollution Contingency Plan, the potential interaction between bioremediation agents and oil should be further investigated and listing requirements reassessed as necessary. Recommendations for future investigations of this issue include characterization of temporal trends in toxicity of CBA and oil mixtures, use of multiple test methods (battery testing) when evaluating toxicity, comparative evaluations of indigenous versus product-derived microorganisms in efficacy studies, and the use of a comparable oil between studies to facilitate comparison of efficacy and toxicity data for different products.

Integrative assessment of benthic macroinvertebrate community impairment from metal-contaminated waters in tributaries of the Upper Powell River, Virginia, USA.

Benthic macroinvertebrate communities of the North Fork Powell River (NFP), southwest Virginia, USA, appear to be impacted by aluminum (Al) and iron (Fe) from acid mine drainage (AMD) beyond the zone of pH depression. As part of a watershed restoration project, we used integrative techniques, including water column, sediment, and in situ toxicity tests; sediment and water column chemistry; and habitat assessments, to detect AMD impacts. An analysis of variance, least significant difference post hoc test, and Spearman correlations were used to test the sensitivity of these integrative techniques to detect various (acidic or neutralized) levels of AMD input and to determine the mode of impairment (metal-contaminated sediments or water) to the benthic macroinvertebrate community. Benthic macroinvertebrate indices were the most sensitive endpoint to AMD inputs and were significantly correlated (p < or = 0.05) with water column metal concentrations in in situ and water column toxicity tests. Sediment chemistry and toxicity did not detect AMD impacts and were not significantly correlated with benthic macroinvertebrate indices. These results suggest that the primary mode of impairment to the benthic macroinvertebrate communities beyond the zone of pH depression were waterborne Al and Fe.

Modification of an ecotoxicological rating to bioassess small acid mine drainage-impacted watersheds exclusive of benthic macroinvertebrate analysis.

The modified ecotoxicological rating (METR) approach to synthesizing integrative bioassessment data into a single number ranging from 0 to 100 was found to differentiate between multiple levels of impacts from acid mine drainage (AMD). Our objective was to develop a more cost-effective and time-efficient bioassessment technique than previously used in other large scale ecotoxicological ratings (ETRs) by minimizing the number of parameters required to rank stations to only those most descriptive of the benthic macroinvertebrate community responses to AMD. Nineteen physical, chemical, toxicological, and ecological measurements were made at 38 stations in two adjacent watersheds. The most descriptive parameters were selected through multiple linear regression analysis, bivariate correlation analysis, and one-way analysis of variance. We found that habitat assessment, 30-d in situ Asian clam survival, mean conductivity, and mean total water column concentration of aluminum and manganese were the most descriptive parameters. The METR constructed from these parameters was equally effective at differentiating stations as were two previous published ETRs that incorporated up to 10 parameters, including benthic macroinvertebrate indices. When the METR was applied to a new watershed, the scores were significantly correlated with benthic macroinvertebrate indices for those stations.