2,4-Dichlorophenol biotransformation using immobilized marine halophilic Bacillus subtilis culture and laccase enzyme: application in wastewater treatment.

BACKGROUND: 2,4-Dichlorophenol (2,4-DCP) is a very toxic aromatic compound for humans and the environment and is highly resistant to degradation. Therefore, it is necessary to develop efficient remediation and cost-effective approaches to this pollutant. Microbial enzymes such as laccases can degrade phenols, but limited information is known about immobilized bacterial laccase and their reuse. METHODS: Immobilization of marine halophilic Bacillus subtilis AAK cultures via entrapment and adsorption techniques and degradation of different phenolic compounds by immobilized cells were estimated. Partial purification and immobilization of laccase enzymes were carried out. In addition, the biodegradation of 2,4-DCP and others contaminated by wastewater was investigated. RESULTS: Immobilization of cells and partially purified laccase enzymes by adsorption into 3% alginate increased 2,4-DCP biotransformation compared with free cells and free enzymes. In addition, the reuse of both the immobilized culture and laccase enzymes was evaluated. The highest removal of 2,4-DCP from pulp and paper wastewater samples inoculated by immobilized cells and the immobilized enzyme was 90% and 95%, respectively, at 50 h and 52 h of incubation, compared to free cells and free enzyme. CONCLUSION: The results of this study have revealed the immobilization of a biocatalyst and its laccase enzyme as a promising technique for enhancing the degradation of 2,4-DCP and other toxic phenolic and aromatic compounds. The reuse of the biocatalyst and its laccase enzyme enabled the application of this cost-effective bioremediation strategy.

Copper Concentrations in the Upper Columbia River as a Limiting Factor in White Sturgeon Recruitment and Recovery.

Currently there is little natural recruitment of white sturgeon (Acipenser transmontanus) in the Upper Columbia River located in British Columbia, Canada and Washington, USA. This review of life history, physiology, and behavior of white sturgeon, along with data from recent toxicological studies, suggest that trace metals, especially Cu, affect survival and behavior of early life stage fish. Sturgeon free embryos, first feeding embryos, and mixed feeding embryos utilize interstitial spaces between gravel. Although concentrations of Cu in the water column of the Upper Columbia River are typically less than US water quality criteria defined to protect aquatic life, samples at the sediment-water interface were as large as 24 µg/L and exceed the criteria. Toxicological studies reviewed here demonstrate mortality, loss of equilibrium, and immobility at Cu concentrations of 1.5 to <16 µg/L and reduced swimming activity was documented at 0.88 to 7 µg/L. Contaminated invertebrates and slag particles provide other routes of exposure. These additional routes of exposure can cause indirect effects from starvation due to potential lack of prey items and ingestion of contaminated prey or slag particles. The lack of food in stomachs during these critical early life stages may coincide with a threshold "point of no return" at which sturgeon will be unable to survive even if food becomes available following that early time frame. These findings become especially important as work progresses to enhance white sturgeon recruitment in the Upper Columbia River. To date, decisions against including trace metals as a factor in sturgeon recovery have focused on surface-water concentrations and measurements of lethality (LC50) to establish threshold concentrations for sturgeon sensitivity. However, information provided here suggests that measurements from the sediment-water interface and effect concentrations (EC50) be considered with white sturgeon life history characteristics. These data support minimizing Cu exposure risk to enhance a successful white sturgeon recovery effort. Integr Environ Assess Manag 2020;16:378-391. Published 2020. This article is a US Government work and is in the public domain inthe USA.

Multiple approaches to surface water quality assessment provide insight for small streams experiencing oil and natural gas development.

Historic, current, and future oil and natural gas development can affect water quality in streams flowing through developed areas. We compared small stream drainages in a semiarid landscape with varying amounts of disturbance from oil and natural gas development to examine potential effects of this development on surface water quality. We used physical, chemical, and biological approaches to assess water quality and found several potential avenues of degradation. Surface disturbance likely contributed to elevated suspended sediment concentrations and spill history likely led to elevated stream polycyclic aromatic hydrocarbon concentrations. In combination, these environmental stressors could explain the loss of aquatic macroinvertebrate taxon at sites highly affected by oil and natural gas development. Our results provide insight into advantages and disadvantages of approaches for assessing surface water quality in areas affected by oil and natural gas development. Integr Environ Assess Manag 2019;00:000-000. © 2019 SETAC.

Restoration of impaired ecosystems: An ounce of prevention or a pound of cure? Introduction, overview, and key messages from a SETAC-SER workshop.

A workshop on Restoration of Impaired Ecosystems was held in Jackson, Wyoming, in June 2014. Experts from Australia, Canada, Mexico, the United Kingdom, and the United States in ecotoxicology, restoration, and related fields from both the Society of Environmental Toxicology and Chemistry and the Society for Ecological Restoration convened to advance the practice of restoring ecosystems that have been contaminated or impaired from industrial activities. The overall goal of this workshop was to provide a forum for ecotoxicologists and restoration ecologists to define the best scientific practices to achieve ecological restoration while addressing contaminant concerns. To meet this goal, participants addressed 5 areas: 1) links between ecological risk assessment and ecological restoration, 2) restoration goals, 3) restoration design, 4) monitoring for restoration effectiveness and 5) recognizing opportunities and challenges. Definitions are provided to establish a common language across the varied disciplines. The current practice for addressing restoration of impaired ecosystems tends to be done sequentially to remediate contaminants, then to restore ecological structure and function. A better approach would anticipate or plan for restoration throughout the process. By bringing goals to the forefront, we may avoid intrusive remediation activities that close off options for the desired restoration. Participants realized that perceived limitations in the site assessment process hinder consideration of restoration goals; contaminant presence will influence restoration goal choices; social, economic, and cultural concerns can factor into goal setting; restoration options and design should be considered early during site assessment and management; restoration of both structure and function is encouraged; creative solutions can overcome limitations; a regional focus is imperative; monitoring must occur throughout the restoration process; and reciprocal transfer of knowledge is needed among theorists, practitioners, and stakeholders and among varied disciplines.

Transforming ecosystems: When, where, and how to restore contaminated sites.

Chemical contamination has impaired ecosystems, reducing biodiversity and the provisioning of functions and services. This has spurred a movement to restore contaminated ecosystems and develop and implement national and international regulations that require it. Nevertheless, ecological restoration remains a young and rapidly growing discipline and its intersection with toxicology is even more nascent and underdeveloped. Consequently, we provide guidance to scientists and practitioners on when, where, and how to restore contaminated ecosystems. Although restoration has many benefits, it also can be expensive, and in many cases systems can recover without human intervention. Hence, the first question we address is: "When should we restore contaminated ecosystems?" Second, we provide suggestions on what to restore-biodiversity, functions, services, all 3, or something else--and where to restore given expected changes to habitats driven by global climate change. Finally, we provide guidance on how to restore contaminated ecosystems. To do this, we analyze critical aspects of the literature dealing with the ecology of restoring contaminated ecosystems. Additionally, we review approaches for translating the science of restoration to on-the-ground actions, which includes discussions of market incentives and the finances of restoration, stakeholder outreach and governance models for ecosystem restoration, and working with contractors to implement restoration plans. By explicitly considering the mechanisms and strategies that maximize the success of the restoration of contaminated sites, we hope that our synthesis serves to increase and improve collaborations between restoration ecologists and ecotoxicologists and set a roadmap for the restoration of contaminated ecosystems.

Ecological risks of shale oil and gas development to wildlife, aquatic resources and their habitats.

Technological advances in hydraulic fracturing and horizontal drilling have led to the exploration and exploitation of shale oil and gas both nationally and internationally. Extensive development of shale resources has occurred within the United States over the past decade, yet full build out is not expected to occur for years. Moreover, countries across the globe have large shale resources and are beginning to explore extraction of these resources. Extraction of shale resources is a multistep process that includes site identification, well pad and infrastructure development, well drilling, high-volume hydraulic fracturing and production; each with its own propensity to affect associated ecosystems. Some potential effects, for example from well pad, road and pipeline development, will likely be similar to other anthropogenic activities like conventional gas drilling, land clearing, exurban and agricultural development and surface mining (e.g., habitat fragmentation and sedimentation). Therefore, we can use the large body of literature available on the ecological effects of these activities to estimate potential effects from shale development on nearby ecosystems. However, other effects, such as accidental release of wastewaters, are novel to the shale gas extraction process making it harder to predict potential outcomes. Here, we review current knowledge of the effects of high-volume hydraulic fracturing coupled with horizontal drilling on terrestrial and aquatic ecosystems in the contiguous United States, an area that includes 20 shale plays many of which have experienced extensive development over the past decade. We conclude that species and habitats most at risk are ones where there is an extensive overlap between a species range or habitat type and one of the shale plays (leading to high vulnerability) coupled with intrinsic characteristics such as limited range, small population size, specialized habitat requirements, and high sensitivity to disturbance. Examples include core forest habitat and forest specialists, sagebrush habitat and specialists, vernal pond inhabitants and stream biota. We suggest five general areas of research and monitoring that could aid in development of effective guidelines and policies to minimize negative impacts and protect vulnerable species and ecosystems: (1) spatial analyses, (2) species-based modeling, (3) vulnerability assessments, (4) ecoregional assessments, and (5) threshold and toxicity evaluations.

In situ and laboratory toxicity of coalbed natural gas produced waters with elevated sodium bicarbonate.

Some tributaries in the Powder River Structural Basin, USA, were historically ephemeral, but now contain water year round as a result of discharge of coalbed natural gas (CBNG)-produced waters. This presented the opportunity to study field sites with 100% effluent water with elevated concentrations of sodium bicarbonate. In situ experiments, static renewal experiments performed simultaneously with in situ experiments, and static renewal experiments performed with site water in the laboratory demonstrated that CBNG-produced water reduces survival of fathead minnow (Pimephales promelas) and pallid sturgeon (Scaphirhynchus albus). Age affected survival of fathead minnow, where fish 2 d posthatch (dph) were more sensitive than 6 dph fish, but pallid sturgeon survival was adversely affected at both 4 and 6 dph. This may have implications for acute assays that allow for the use of fish up to 14 dph. The survival of early lifestage fish is reduced significantly in the field when concentrations of NaHCO(3) rise to more than 1500 mg/L (also expressed as >1245 mg HCO(3) (-) /L). Treatment with the Higgin's Loop technology and dilution of untreated water increased survival in the laboratory. The mixing zones of the 3 outfalls studied ranged from approximately 800 m to 1200 m below the confluence. These experiments addressed the acute toxicity of effluent waters but did not address issues related to the volumes of water that may be added to the watershed.

Acute toxicity of sodium bicarbonate, a major component of coal bed natural gas produced waters, to 13 aquatic species as defined in the laboratory.

Water produced during coal bed natural gas (CBNG) extraction in the Powder River Structural Basin of Wyoming and Montana (USA) may contain concentrations of sodium bicarbonate (NaHCO3) of more than 3000 mg/L. The authors evaluated the acute toxicity of NaHCO3, also expressed as bicarbonate (HCO3(-)), to 13 aquatic organisms. Of the 13 species tested, 7 had a median lethal concentration (LC50) less than 2000 mg/L NaHCO3, or 1300 mg/L HCO3(-). The most sensitive species were Ceriodaphnia dubia, freshwater mussels (Lampsilis siliquoidea), pallid sturgeon (Scaphirhynchus albus), and shovelnose sturgeon (Scaphirhynchus platorynchus). The respective LC50s were 989 mg/L, 1120 mg/L, 1249 mg/L, and 1430 mg/L NaHCO3, or 699 mg/L, 844 mg/L, 831 mg/L, and 1038 mg/L HCO3(-). Age affected the sensitivity of fathead minnows, even within life stage. Two days posthatch, fathead minnows were more sensitive to NaHCO3 and HCO3(-) compared with 4-d-old fish, even though fish up to 14 d old are commonly used for toxicity evaluations. The authors recommend that ion toxicity exposures be conducted with organisms less than 24 h posthatch to ensure that experiments document the most sensitive stage of development. The results of the present study, along with historical and current research regarding the toxicity of bicarbonate, may be useful to establish regulatory standards for HCO3(-). This article is a US Government work and is in the public domain in the USA.

The chronic toxicity of sodium bicarbonate, a major component of coal bed natural gas produced waters.

Sodium bicarbonate (NaHCO3) is the principal salt in coal bed natural gas produced water from the Powder River Structural Basin, Wyoming, USA, and concentrations of up to 3000 mg NaHCO3/L have been documented at some locations. No adequate studies have been performed to assess the chronic effects of NaHCO3 exposure. The present study was initiated to investigate the chronic toxicity and define sublethal effects at the individual organism level to explain the mechanisms of NaHCO3 toxicity. Three chronic experiments were completed with fathead minnows (Pimephales promelas), 1 with white suckers (Catostomus commersoni), 1 with Ceriodaphnia dubia, and 1 with a freshwater mussel, (Lampsilis siliquoidea). The data demonstrated that approximately 500 mg NaHCO3/L to 1000 mg NaHCO3/L affected all species of experimental aquatic animals in chronic exposure conditions. Freshwater mussels were the least sensitive to NaHCO3 exposure, with a 10-d inhibition concentration that affects 20% of the sample population (IC20) of 952 mg NaHCO3/L. The IC20 for C. dubia was the smallest, at 359 mg NaHCO3/L. A significant decrease in sodium-potassium adenosine triphosphatase (Na(+)/K(+) ATPase) together with the lack of growth effects suggests that Na(+)/K(+) ATPase activity was shut down before the onset of death. Several histological anomalies, including increased incidence of necrotic cells, suggested that fish were adversely affected as a result of exposure to >450 mg NaHCO3/L. This article is a US Government work and is in the public domain in the USA.

Mercury and selenium concentrations in biofilm, macroinvertebrates, and fish collected in the Yankee Fork of the Salmon River, Idaho, USA, and their potential effects on fish health.

The Yankee Fork is a large tributary of the Salmon River located in central Idaho, USA, with an extensive history of placer and dredge-mining activities. Concentrations of selenium (Se) and mercury (Hg) in various aquatic trophic levels were measured in the Yankee Fork during 2001 and 2002. Various measurements of fish health were also performed. Sites included four on the mainstem of the Yankee Fork and two off-channel sites in partially reclaimed dredge pools used as rearing habitat for cultured salmonid eggs and fry. Hg concentrations in whole mountain whitefish and shorthead sculpin ranged from 0.28 to 0.56 µg/g dry weight (dw), concentrations that are generally less than those reported to have significant impacts on fish. Biofilm and invertebrates ranged from 0.05 to 0.43 µg Hg/g dw. Se concentrations measured in biota samples from the Yankee Fork were greater than many representative samples collected in the Snake and Columbia watersheds and often exceeded literature-based toxic thresholds. Biofilm and invertebrates ranged from 0.58 to 4.66 µg Se/g dw. Whole fish ranged from 3.92 to 7.10 µg Se/g dw, and gonads ranged from 6.91 to 31.84 µg Se/g dw. Whole-body Se concentrations exceeded reported toxicological thresholds at three of four sites and concentrations in liver samples were mostly greater than concentrations shown to have negative impacts on fish health. Histological examinations performed during this study noted liver abnormalities, especially in shorthead sculpin, a bottom-dwelling species.

Microbial pollution indicators along the Egyptian coastal waters of Suez and Aqaba Gulfs and Red Sea.

BACKGROUND: Nowadays, the Egyptian coasts of the Aqaba and Suez Gulfs, and the Red Sea proper, are under the direct effects of many recreational resorts, urban agglomeration, marine shipping, activity of the phosphate industry, fishing ports, and limited freshwater and sewage surfaces. Therefore, the water, especially those used for recreational activities, must be of a very good quality to be able to increase the national income. OBJECTIVES: To investigate the conventional water-quality bacteria, total coliforms (TC), Escherichia coli (EC), and fecal streptococci (FS), in the Egyptian coastal waters of Suez and Aqaba Gulfs and the Red Sea. MATERIALS AND METHODS: A total of 2372 surface water samples were collected from 42 sampling sites during 12 years (1998-2009) to detect and estimate TC, EC, and FS using the membrane filtration method. RESULTS: On the basis of the national and international bacterial standards, 540 samples (22.8%) out of 2372 were found to exceed the guide values (positive samples) and were not accepted for marine recreational purposes. During the course of the study, Suez Gulf showed the highest positive records of 54 and 96 for TC and EC, respectively, whereas FS recorded 94 and 88 positive samples in the Red Sea and Suez Gulf, respectively. The lowest positive records were found in 1998 and 2009, whereas the highest were in 2000 and 2002-2004. The highest polluted sampling sites were recorded in Su7 (Suez Gulf), Aq2 (Aqaba Gulf), and Re15 (Red Sea), and were generally affected by sewage disposal and/or anthropogenic influences. CONCLUSION AND RECOMMENDATIONS: The most polluted sites were in the Suez Gulf, reaching 238 sites, followed by 194 sites in the Red Sea, whereas the Aqaba Gulf had only 108 polluted sites. Moreover, the most polluted sample locations throughout the study were Su7, Aq2, and Re15, without implementation of corrective actions from authorized organizations. The data of the current study must be taken into consideration by the government for a safer and cleaner seawater in the eastern Egyptian coasts, especially those in which critical limitations in terms of microbial pollution are found.

Trout density and health in a stream with variable water temperatures and trace element concentrations: does a cold-water source attract trout to increased metal exposure?

A history of hard-rock mining has resulted in elevated concentrations of heavy metals in Prickly Pear Creek (MT, USA). Remediation has improved water quality; however, dissolved zinc and cadmium concentrations still exceed U.S. Environmental Protection Agency water-quality criteria. Physical habitat, salmonid density, fish health, and water quality were assessed, and metal concentrations in fish tissues, biofilm, and macroinvertebrates were determined to evaluate the existing condition in the watershed. Cadmium, zinc, and lead concentrations in fish tissues, biofilm, and invertebrates were significantly greater than those at the upstream reference site and an experimental site farther downstream of the confluence. Fish densities were greatest, and habitat quality for trout was better, downstream of the confluence, where water temperatures were relatively cool (16 degrees C). Measures of fish health (tissue metal residues, histology, metallothionein concentrations, and necropsies), however, indicate that the health of trout at this site was negatively affected. Trout were in colder but more contaminated water and were subjected to increased trace element exposures and associated health effects. Maximum water temperatures in Prickly Pear Creek were significantly lower directly below Spring Creek (16 degrees C) compared to those at an experimental site 10 km downstream (26 degrees C). Trout will avoid dissolved metals at concentrations below those measured in Prickly Pear Creek; however, our results suggest that the preference of trout to use cool water temperatures may supersede behaviors to avoid heavy metals.

Influence of in-stream diel concentration cycles of dissolved trace metals on acute toxicity to one-year-old cutthroat trout (Oncorhynchus clarki lewisi).

Extrapolating results of laboratory bioassays to streams is difficult, because conditions such as temperature and dissolved metal concentrations can change substantially on diel time scales. Field bioassays conducted for 96 h in two mining-affected streams compared the survival of hatchery-raised, metal-naïve westslope cutthroat trout (Oncorhynchus clarki lewisi) exposed to dissolved (0.1-microm filtration) metal concentrations that either exhibited the diel variation observed in streams or were controlled at a constant value. Cadmium and Zn concentrations in these streams increased each night by as much as 61 and 125%, respectively, and decreased a corresponding amount the next day, whereas Cu did not display a diel concentration cycle. In High Ore Creek (40 km south of Helena, MT, USA), survival (33%) after exposure to natural diel-fluctuating Zn concentrations (range, 214-634 microg/L; mean, 428 microg/L) was significantly (p = 0.008) higher than survival (14%) after exposure to a controlled, constant Zn concentration (422 microg/L). Similarly, in Dry Fork Belt Creek (70 km southeast of Great Falls, MT, USA), survival (75%) after exposure to diel-fluctuating Zn concentrations (range, 266-522 microg/L; mean, 399 microg/L) was significantly (p = 0.022) higher than survival (50%) in the constant-concentration treatment (392 microg/L). Survival likely was greater in these diel treatments, both because the periods of lower metal concentrations provided some relief for the fish and because toxicity during periods of higher metal concentrations was lessened by the simultaneous occurrence each night of lower water temperatures, which reduce the rate of metal uptake. Based on the present study, current water-quality criteria appear to be protective for streams with diel concentration cycles of Zn (and, perhaps, Cd) for the hydrologic conditions tested.

Concentrations of metals in water, sediment, biofilm, benthic macroinvertebrates, and fish in the Boulder River watershed, Montana, and the role of colloids in metal uptake.

To characterize the partitioning of metals in a stream ecosystem, concentrations of trace metals including As, Cd, Cu, Pb, and Zn were measured in water, colloids, sediment, biofilm (also referred to as aufwuchs), macroinvertebrates, and fish collected from the Boulder River watershed, Montana. Median concentrations of Cd, Cu, and Zn in water throughout the watershed exceeded the U.S. EPA acute and chronic criteria for protection of aquatic life. Concentrations of As, Cd, Cu, Pb, and Zn in sediment were sufficient in the tributaries to cause invertebrate toxicity. The concentrations of As, Cu, Cd, Pb, and Zn in invertebrates from lower Cataract Creek (63, 339, 59, 34, and 2,410 microg/g dry wt, respectively) were greater than the concentrations in invertebrates from the Clark Fork River watershed, Montana (19, 174, 2.3, 15, and 648 microg/g, respectively), that were associated with reduced survival, growth, and health of cutthroat trout fed diets composed of those invertebrates. Colloids and biofilm seem to play a critical role in the pathway of metals into the food chain and concentrations of As, Cu, Pb, and Zn in these two components are significantly correlated. We suggest that transfer of metals associated with Fe colloids to biological components of biofilm is an important pathway where metals associated with abiotic components are first available to biotic components. The significant correlations suggest that Cd, Cu, and Zn may move independently to biota (biofilm, invertebrates, or fish tissues) from water and sediment. The possibility exists that Cd, Cu, and Zn concentrations increase in fish tissues as a result of direct contact with water and sediment and indirect exposure through the food chain. However, uptake through the food chain to fish may be more important for As. Although As concentrations in colloids and biofilm were significantly correlated with As water concentrations, As concentrations in fish tissues were not correlated with water. The pathway for Pb into biological components seems to begin with sediment because concentrations of Pb in water were not significantly correlated with any other component and because concentrations of Pb in the water were often below detection limits.

Biomonitoring in the Boulder River Watershed, Montana, USA: metal concentrations in biofilm and macroinvertebrates, and relations with macroinvertebrate assemblage.

Portions of the Boulder River watershed contain elevated concentrations of arsenic, cadmium, copper, lead, and zinc in water, sediment, and biota. We measured concentrations of As, Cd, Cu, Pb, and Zn in biofilm and macroinvertebrates, and assessed macroinvertebrate assemblage and aquatic habitat with the objective of monitoring planned remediation efforts. Concentrations of metals were generally higher in downstream sites compared with upstream or reference sites, and two sites contained metal concentrations in macroinvertebrates greater than values reported to reduce health and survival of resident trout. Macroinvertebrate assemblage was correlated with metal concentrations in biofilm and macroinvertebrates. However, macroinvertebrate metrics were significantly correlated with a greater number of biofilm metals (8) than metals in invertebrates (4). Lead concentrations in biofilm appeared to have the most significant impact on macroinvertebrate assemblage. Metal concentrations in macroinvertebrates were directly proportional to concentrations in biofilm, indicating biofilm as a potential surrogate for monitoring metal impacts in aquatic systems.

The effect of chronic chromium exposure on the health of Chinook salmon (Oncorhynchus tshawytscha).

This study was designed to determine fish health impairment of Chinook salmon (Oncorhynchus tshawytscha) exposed to chromium. Juvenile Chinook salmon were exposed to aqueous chromium concentrations (0-266 microgl(-1)) that have been documented in porewater from bottom sediments and in well waters near salmon spawning areas in the Columbia River in the northwestern United States. After Chinook salmon parr were exposed to 24 and 54 microg Crl(-1) for 105 days, neither growth nor survival of parr was affected. On day 105, concentrations were increased from 24 to 120 microg Crl(-1) and from 54 to 266 microg Crl(-1) until the end of the experiment on day 134. Weight of parr was decreased in the 24/120 microg Crl(-1) treatment, and survival was decreased in the 54/266 microg Crl(-1) treatment. Fish health was significantly impaired in both the 24/120 and 54/266 microg Crl(-1) treatments. The kidney is the target organ during chromium exposures through the water column. The kidneys of fish exposed to the greatest concentrations of chromium had gross and microscopic lesions (e.g. necrosis of cells lining kidney tububules) and products of lipid peroxidation were elevated. These changes were associated with elevated concentrations of chromium in the kidney, and reduced growth and survival. Also, variations in DNA in the blood were associated with pathological changes in the kidney and spleen. These changes suggest that chromium accumulates and enters the lipid peroxidation pathway where fatty acid damage and DNA damage (expressed as chromosome changes) occur to cause cell death and tissue damage. While most of the physiological malfunctions occurred following parr exposures to concentrations > or =120 microg Crl(-1), nuclear DNA damage followed exposures to 24 microg Crl(-1), which was the smallest concentration tested. The abnormalities measured during this study are particularly important because they are associated with impaired growth and reduced survival at concentrations > or =120 microg Crl(-1). Therefore, these changes can be used to investigate the health of resident fish in natural waters with high chromium concentrations as well as provide insight into the mechanisms of chromium toxicity.

Leachability of protein and metals incorporated into aquatic invertebrates: are species and metals-exposure history important?

To partially simulate conditions in fish intestinal tracts, we leached six groups of metals-contaminated invertebrates at pH 2 and pH 7, and analyzed the concentrations of four metals (Cd, Cu, Pb, and Zn) and total protein in the leachates. Four of the groups of invertebrates were benthic macroinvertebrates collected from metals-contaminated rivers (the Clark Fork River in Montana and the Coeur d'Alene River in Idaho, USA); the other two groups of invertebrates (one of which was exposed to metals in the laboratory) were laboratory-reared brine shrimp (Artemia sp.). Additionally, we fractionated the pH 2 leachates using size-exclusion chromatography (SEC). Protein content was 1.3 to 1.4x higher in Artemia than in the benthic macroinvertebrates, and leachability of metals and protein differed considerably among several of the groups of invertebrates. In SEC fractions of the pH 2 leachates from both groups of Artemia, Cu and protein co-eluted; however, Cu and protein did not co-elute in SEC fractions of the leachates from any of the benthic macroinvertebrate groups. Although none of the other three metals co-eluted with protein in any of the pH 2 leachates, one or more of the metals co-eluted with lower-molecular-weight molecules in the leachates from all of the groups of invertebrates. These results suggest fundamental differences in metal-binding properties and protein leachability among some invertebrates. Thus, different invertebrates and different histories of metals exposure might lead to different availability of metals and protein to predators.