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Holly J. Puglis

Publications and source records attributed to Holly J. Puglis.

17 recordsLinked to original sources

Effects of carbamazepine to visual function in early life stage fish

The frequent detection of pharmaceuticals and personal care products (PPCPs) in the environment raises concern for aquatic systems. Carbamazepine (CBZ), an antiepileptic drug, is among the most detected PPCP globally, with concentrations in surface water exceeding those that induce toxicity to aquatic organisms. Non-targeted transcriptomic profiling was conducted in zebrafish ( Danio rerio ) larvae exposed to 0, 1, 5, 10, or 50 μg/L CBZ from 2 h post fertilization (hpf) through hatching, and then sampled at 48, 72, or 144 hpf. Transcriptomic profiles were annotated and characterized with in silico bioinformatic software to assess top enriched pathways and identify targets of environmentally relevant concentrations of CBZ and anchor molecular effects to higher levels of biological organization. Based on this analysis, CBZ was predicted to impair visual perception and sensory system development. The number of eye saccades, determined with a visually mediated behavioral assay, optokinetic response, was significantly reduced in 144 hpf larvae exposed to concentrations as low as 1 μg/L CBZ. These results indicate that environmentally relevant concentrations of CBZ may target and impact processes involved in visual function in fish.

Environmental Research

Are behavioural ecotoxicity endpoints relevant at the population level? Evidence-based insights for environmental protection

A substantial body of evidence exists demonstrating that exposure to environmental contaminants can alter animal behavior. Moreover, methodological and technological advancements, as well as increasing standardization, mean that behavioral ecotoxicity studies are more rigorous and reliable than ever before. Despite this, behavioral data are still seldom used in the risk assessment and regulation of chemicals. This is partly due to a lack of clarity among some stakeholders about whether changes in behavior at the individual level result in population-level outcomes. To address this, we first consider the state of evidence within the field of behavioral ecotoxicology linking individual-level behavioral alterations with population-level consequences. We then assess the evidence from behavioral ecology and other neighboring fields that supports this link. Further, we evaluate whether some behavioral endpoints are more easily tied to population-level changes than others. In this regard, we propose combining insights from two complementary ecological frameworks─the functional trait framework and the limiting traits framework─to evaluate which behaviors should be prioritized in ecotoxicological research and regulatory efforts. We contend that the link between behavioral changes and population-level outcomes is evident, with behavioral endpoints representing a highly valuable yet so far underutilized line of evidence in applied environmental protection.

Environmental Science and Technology

Environmental persistence and toxicity of weathered wildland fire retardants to rainbow trout

Long-term fire retardants are employed to combat and control wildfires by altering the way fuels burn, and they continue to decrease fire intensity after water in the retardant solution has evaporated. After application, fire retardants may persist on dry stream beds or in riparian habitats before precipitation events flush the retardant into intermittent streams. We exposed juvenile (30–60 days post swim-up) rainbow trout ( Oncorhynchus mykiss ) to fire retardants weathered for 7–56 days on different substrates (duff, gravel, high organic content soil, and low organic content soil) under static conditions for 96 h to evaluate the potential toxicity of two current-use long-term fire-retardant (LC95A-R and MVP-Fx) products. Trout mortality was greater in LC95A-R treatments compared to MVP-Fx due to higher concentrations of LC95A-R in the applied product than MVP-Fx at the same application rate. Underlying substrate affected fire-retardant toxicity, with 31% higher average mortality for products applied to duff and gravel compared to soil. Differences in mortality across substrates and products after weathering may be attributed to differences in the mix ratio of applied product and interactions of product chemistries with underlying substrate. These interactions resulted in elevated ionic concentrations of the overlying water in duff and gravel treatments. Trout mortality decreased 15% for products weathered 56 days compared to 7 days. Our results suggest that long-term fire retardants may persist in the environment and that underlying substrate may alter the toxicity of these products upon entrance into an intermittent stream.

Archives of Environmental Contamination and Toxico

Toxicity of wildland fire retardants to rainbow trout in short exposures

Long-term wildland fire retardants are one important tool used to control and suppress wildfires. During suppression activities, these retardants may enter waterbodies; thus, there is a need to understand their potential effects to aquatic biota. We investigated the effect of three current-use wildland fire retardants to juvenile rainbow trout ( Oncorhynchus mykiss) survival in short exposures more realistic to actual intrusion scenarios. Lethal effect concentrations decreased with time and varied among chemicals (LC95A-R > 259-Fx > MVP-Fx). The lowest effect concentrations observed were 2 to 10 times above the threshold used by federal agencies to assess potential impacts to aquatic organisms following a retardant intrusion. These data can be used by resource managers to balance wildfire control with potential environmental impacts of retardant use.

Environmental Toxicology and Chemistry

Ammonia and aquatic ecosystems – A review of global sources, biogeochemical cycling, and effects on fish

The purpose of this review is to better understand the full life cycle and influence of ammonia from an aquatic biology perspective. While ammonia has toxic properties in water and air, it also plays a central role in the biogeochemical nitrogen (N) cycle and regulates mechanisms of normal and abnormal fish physiology. Additionally, as the second most synthesized chemical on Earth, ammonia contributes economic value to many sectors, particularly fertilizers, energy storage, explosives, refrigerants, and plastics. But, with so many uses, industrial N 2 -fixation effectively doubles natural reactive N concentrations in the environment. The consequence is global, with excess fixed nitrogen driving degradation of soils, water, and air; intensifying eutrophication, biodiversity loss, and climate change; and creating health risks for humans, wildlife, and fisheries. Thus, the need for ammonia research in aquatic systems is growing. In response, we prepared this review to better understand the complexities and connectedness of environmental ammonia. Even the term “ammonia” has multiple meanings. So, we have clarified the nomenclature, identified units of measurement, and summarized methods to measure ammonia in water. We then discuss ammonia in the context of the N-cycle, review its role in fish physiology and mechanisms of toxicity, and integrate the effects of human N-fixation, which continuously expands ammonia's sources and uses. Ammonia is being developed as a carbon-free energy carrier with potential to increase reactive nitrogen in the environment. With this in mind, we review the global impacts of excess reactive nitrogen and consider the current monitoring and regulatory frameworks for ammonia. The presented synthesis illustrates the complex and interactive dynamics of ammonia as a plant nutrient, energy molecule, feedstock, waste product, contaminant, N-cycle participant, regulator of animal physiology, toxicant, and agent of environmental change. Few molecules are as influential as ammonia in the management and resilience of Earth's resources.

Science of the Total Environment

Toxicity of wildland fire-fighting chemicals in pulsed exposures to rainbow trout and fathead minnows

Intrusions of fire-fighting chemicals in streams can result from containment and suppression of wildfires and may be harmful to native biota. We investigated the toxicity of seven current-use fire-fighting chemicals to juvenile rainbow trout ( Oncorhynchus mykiss ) and fathead minnows ( Pimephales promelas ) by simulating chemical intrusions under variable field conditions to provide insight on the potential damage these chemicals may cause in waterways. We manipulated water flow rate, water hardness, and concentration of the chemicals in three separate attenuated exposure assays where chemical concentrations decreased throughout the 96-hour exposure period. Concentration of retardant, temperature, duration of chemical exposure, and the number of exposures were manipulated in four pulsed assays where up to one-hour exposures were followed by an observation period in control water to determine delayed toxicity or recovery. Mortality of rainbow trout was higher across treatments at a warmer temperature and also increased with increasing concentration rate, increasing exposure duration, and with sequential exposures across assays. For fathead minnows, mortality increased with increasing concentration of fire retardant and longer exposure durations. Chemical exposure can exert additional stress during wildfire events that may impact stream fishes. Since the ratio of toxic unionized ammonia to ionized ammonia is greater with increasing temperature and pH, future studies could investigate the effects of water temperature and pH on native fishes under environmentally relevant concentrations of fire-fighting chemicals.

Environmental Toxicology and Chemistry

Chloride toxicity to native freshwater species in natural and reconstituted prairie pothole waters

Oil and gas extraction in the Prairie Pothole Region (PPR) of the northern USA has resulted in elevated chloride concentrations in ground and surface water due to widespread contamination with highly saline produced water, or brine. The toxicity of chloride is poorly understood in the high hardness waters characteristic of the region. We evaluated the toxicity of chloride to two endemic species, Daphnia magna (water flea) and Lemna gibba (duckweed), exposed in field-collected waters (hardness ~ 3000 mg/L as CaCO 3 ) and reconstituted waters (hardness 370 mg/L as CaCO 3 ) intended to mimic PPR background waters. We also investigated the role of chloride in the toxicity of water reconstituted to mimic legacy brine-contaminated wetlands, using two populations of native Pseudacris maculata (Boreal Chorus Frog). Chloride toxicity was similar in field-collected and reconstituted waters for both D. magna (LC50s 3070–3788 mg Cl −1 /L) and L. gibba (IC50s 2441–2887). Although hardness can ameliorate chloride toxicity at low to high hardness, we did not observe additional protection as hardness increased from 370 to ~ 3000 mg/L. In P. maculata exposures, chloride did not fully explain toxicity. Chloride sensitivity also differed between populations, with mortality at 2000 mg Cl − /L in one population but not the other, and population-specific growth responses. Overall, these results (1) document toxicity to native species at chloride concentrations occurring in the PPR, (2) indicate that very high hardness in the region’s waters may not provide additional protection against chloride and (3) highlight challenges of brine investigations, including whether surrogate study populations are representative of local populations.

Missouri, Montana, North Dakota, Wyoming

Predicting the spatiotemporal exposure of aquatic species to intrusions of fire retardant in streams with limited data

Because fire retardant can enter streams and harm aquatic species including endangered fish, agencies such as the U.S. Forest Service (USFS) must estimate the downstream extent of toxic effects every time fire retardant enters streams (denoted as an “intrusion”). A challenge in estimating the length of stream affected by the intrusion and the exposure time of species in the affected reach is the lack of data typically available on the stream's geometry and flow characteristics. Previously, the USFS estimated the affected reach length assuming instantaneous mixing of the retardant over the reach; however, this approach neglects key river mixing processes. An approach is described that accounts for advection and dispersion of the retardant as well as the downstream growth of the stream. Applied to 13 intrusions documented by the USFS, the new approach shows affected reach lengths range between 8.0 and 362 km; all 13 cases exceeded previous estimates from an instantaneous mixing model. The time that a stationary individual in the affected reach is exposed to concentrations above a pre-defined toxicity threshold (10% of 96-hour LC 50 , for example) ranges from 0.17 to 2.73 h, with all but one case having a maximum exposure time less than 1.5 h. Results from 1152 hypothetical intrusions provided by the USFS confirm that exposure times rarely exceed 5 h. This result suggests that 96-hour tests to determine toxicity (LC 50 ) to various species should be reconsidered. Although the approach described can be improved in several ways, it provides a first estimate of the effects of fire retardant intrusions.

Science of the Total Environment

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.

Washington, British Columbia

Effects of the neonicotinoid insecticide clothianidin on southern leopard frog (Rana sphenocephala) tadpole behavior

Neonicotinoid insecticides are highly water soluble with relatively long half-lives, which allows them to move into and persist in aquatic ecosystems. However, little is known of the impacts of neonicotinoids on non-target vertebrates, especially at sublethal concentrations. We evaluated the effects of the neonicotinoid clothianidin on the behavior of southern leopard frog tadpoles ( Rana sphenocephala ) after a 96-h exposure at 6 concentrations, including 0 (control), 0.375, 0.75, 1.5, 3.0, 6.0 µg/L. We quantified total displacement, mean velocity, maximum velocity, and time spent moving of tadpoles for 1 h post-exposure. Total displacement and mean velocity of tadpoles decreased with clothianidin exposure. Maximum velocity decreased linearly with concentration, but there was no relationship between time spent moving and clothianidin concentration. Our results suggest exposure to clothianidin at sublethal concentrations can affect movement behavior of non-target organisms such as tadpoles.

Bulletin of Environmental Contamination and Toxico

Behavioral effects of copper on larval white sturgeon

Early–life stage white sturgeon are sensitive to copper (Cu), with adverse behavioral responses observed during previous studies. The objectives of the present study were to quantify the effects of Cu exposure on white sturgeon swimming and feeding behaviors and determine their time to response. Larval sturgeon (1–2, 28, or 35 d posthatch [dph]) were exposed to Cu (0.5–8 μg/L) for 4 to 14 d. Abnormal behavioral changes were observed within the first few days of exposure including loss of equilibrium and immobilization. Digital video tracking software revealed decreased swimming activity with increasing Cu concentration. Significant changes in behavior and mortality occurred at concentrations of Cu between 1 and 8 μg/L. Juvenile white sturgeon, 58 dph, exposed to 12 μg/L Cu consumed 37 to 60% less food than controls after 3 d of exposure. The present results indicate that behavioral endpoints were more sensitive than some standard toxicity test endpoints and can effectively expand the sensitivity of standard toxicity tests for white sturgeon. Swimming behavior was impaired to the extent that survival in the field would likely be jeopardized. Such data would provide managers a useful metric for characterizing the risks of Cu contamination to white sturgeon.

Environmental Toxicology and Chemistry

Potential toxicity of dissolved metal mixtures (Cd, Cu, Pb, Zn) to early life stage white sturgeon (Acipenser transmontanus) in the Upper Columbia River, Washington, United States

The Upper Columbia River (UCR) received historical releases of smelter waste resulting in elevated metal concentrations in downstream sediments. Newly hatched white sturgeon hide within the rocky substrate at the sediment–water interface in the UCR for a few weeks before swim-up. Hiding behavior could expose them to metal contaminants, and metal toxicity could contribute to population declines in white sturgeon over the past 50 years. This study evaluates whether there is a link between the toxicity of dissolved metals across the sediment-water interface in the UCR and the survival of early life stage (ELS) white sturgeon. Toxicity of dissolved metal mixtures is evaluated using a combination of previously collected laboratory and field data and recently developed metal mixture toxicity models. The laboratory data consist of individual metal (Cd, Cu, Pb, and Zn) toxicity studies with ELS white sturgeon. The field data include the chemical composition of surface and pore water samples that were collected across the sediment–water interface in the UCR. These data are used in three metal accumulation and two response models. All models predict low toxicity in surface water, whereas effects concentrations greater than 20% are predicted for 60–72% of shallow pore water samples. The flux of dissolved metals, particularly Cu, from shallow pore water to surface water likely exposes prime ELS sturgeon habitat to toxic conditions.

Washington

Widespread legacy brine contamination from oil production reduces survival of chorus frog larvae

Advances in drilling techniques have facilitated a rapid increase in hydrocarbon extraction from energy shales, including the Williston Basin in central North America. This area overlaps with the Prairie Pothole Region, a region densely populated with wetlands that provide numerous ecosystem services. Historical (legacy) disposal practices often released saline co-produced waters (brines) with high chloride concentrations, affecting wetland water quality directly or persisting in sediments. Despite the potential threat of brine contamination to aquatic habitats, there has been little research into its ecological effects. We capitalized on a gradient of legacy brine-contaminated wetlands in northeast Montana to conduct laboratory experiments to assess variation in survival of larval Boreal Chorus Frogs ( Pseudacris maculata ) reared on sediments from 3 local wetlands and a control source. To help provide environmental context for the experiment, we also measured chloride concentrations in 6 brine-contaminated wetlands in our study area, including the 2 contaminated sites used for sediment exposures. Survival of frog larvae during 46- and 55-day experiments differed by up to 88% among sediment sources (Site Model) and was negatively correlated with potential chloride exposure (Chloride Model). Five of the 6 contaminated wetlands exceeded the U.S. EPA acute benchmark for chloride in freshwater (860 mg/L) and all exceeded the chronic benchmark (230 mg/L). However, the Wetland Site model explained more variation in survival than the Chloride Model, suggesting that chloride concentration alone does not fully reflect the threat of contamination to aquatic species. Because the profiles of brine-contaminated sediments are complex, further surveys and experiments are needed across a broad range of conditions, especially where restoration or remediation actions have reduced brine-contamination. Information provided by this study can help quantify potential ecological threats and help land managers prioritize conservation strategies as part of responsible and sustainable energy development.

Montana, North Dakota, South Dakota

Sensitivity of lake sturgeon ( Acipenser fulvescens ) early life stages to 2,3,7,8-tetrachlorodibenzo- P -dioxin and 3,3′,4,4′,5-pentachlorobiphenyl

The aquatic food web of the Great Lakes has been contaminated with polychlorinated biphenyls (PCBs) since the mid-20th century. Threats of PCB exposures to long-lived species of fish, such as lake sturgeon ( Acipenser fulvescens ), have been uncertain because of a lack of information on the relative sensitivity of the species. The objective of the present study was to evaluate the sensitivity of early–life stage lake sturgeon to 3,3′,4,4′,5-pentachlorobiphenyl (PCB-126) or 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD) exposure. Mortality, growth, morphological and tissue pathologies, swimming performance, and activity levels were used as assessment endpoints. Pericardial and yolk sac edema, tubular heart, yolk sac hemorrhaging, and small size were the most commonly observed pathologies in both TCDD and PCB-126 exposures, beginning as early as 4 d postfertilization, with many of these pathologies occurring in a dose-dependent manner. Median lethal doses for PCB-126 and TCDD in lake sturgeon were 5.4 ng/g egg (95% confidence interval, 3.9–7.4 ng/g egg) and 0.61 ng/g egg (0.47–0.82 ng/g egg), respectively. The resulting relative potency factor for PCB-126 (0.11) was greater than the World Health Organization estimate for fish (toxic equivalency factor = 0.005), suggesting that current risk assessments may underestimate PCB toxicity toward lake sturgeon. Swimming activity and endurance were reduced in lake sturgeon survivors from the median lethal doses at 60 d postfertilization. Threshold and median toxicity values indicate that lake sturgeon, like other Acipenser species, are more sensitive to PCB and TCDD than the other genus of sturgeon, Scaphirhynchus , found in North America. Indeed, lake sturgeon populations in the Great Lakes and elsewhere are susceptible to PCB/TCDD-induced developmental toxicity in embryos and reductions in swimming performance.

Environmental Toxicology and Chemistry

Quantifying fish swimming behavior in response to acute exposure of aqueous copper using computer assisted video and digital image analysis

Behavioral responses of aquatic organisms to environmental contaminants can be precursors of other effects such as survival, growth, or reproduction. However, these responses may be subtle, and measurement can be challenging. Using juvenile white sturgeon ( Acipenser transmontanus ) with copper exposures, this paper illustrates techniques used for quantifying behavioral responses using computer assisted video and digital image analysis. In previous studies severe impairments in swimming behavior were observed among early life stage white sturgeon during acute and chronic exposures to copper. Sturgeon behavior was rapidly impaired and to the extent that survival in the field would be jeopardized, as fish would be swept downstream, or readily captured by predators. The objectives of this investigation were to illustrate protocols to quantify swimming activity during a series of acute copper exposures to determine time to effect during early lifestage development, and to understand the significance of these responses relative to survival of these vulnerable early lifestage fish. With mortality being on a time continuum, determining when copper first affects swimming ability helps us to understand the implications for population level effects. The techniques used are readily adaptable to experimental designs with other organisms and stressors.

Journal of Visualized Experiments

Acute sensitivity of white sturgeon ( Acipenser transmontanus ) and rainbow trout ( Oncorhynchus mykiss ) to copper, cadmium, or zinc in water-only laboratory exposures

The acute toxicity of cadmium, copper, and zinc to white sturgeon ( Acipenser transmontanus ) and rainbow trout ( Oncorhynchus mykiss ) were determined for 7 developmental life stages in flow-through water-only exposures. Metal toxicity varied by species and by life stage. Rainbow trout were more sensitive to cadmium than white sturgeon across all life stages, with median effect concentrations (hardness-normalized EC50s) ranging from 1.47 µg Cd/L to 2.62 µg Cd/L with sensitivity remaining consistent during later stages of development. Rainbow trout at 46 d posthatch (dph) ranked at the 2nd percentile of a compiled database for Cd species sensitivity distribution with an EC50 of 1.46 µg Cd/L and 72 dph sturgeon ranked at the 19th percentile (EC50 of 3.02 µg Cd/L). White sturgeon were more sensitive to copper than rainbow trout in 5 of the 7 life stages tested with biotic ligand model (BLM)-normalized EC50s ranging from 1.51 µg Cu/L to 21.9 µg Cu/L. In turn, rainbow trout at 74 dph and 95 dph were more sensitive to copper than white sturgeon at 72 dph and 89 dph, indicating sturgeon become more tolerant in older life stages, whereas older trout become more sensitive to copper exposure. White sturgeon at 2 dph, 16 dph, and 30 dph ranked in the lower percentiles of a compiled database for copper species sensitivity distribution, ranking at the 3rd (2 dph), 5th (16 dph), and 10th (30 dph) percentiles. White sturgeon were more sensitive to zinc than rainbow trout for 1 out of 7 life stages tested (2 dph with an biotic ligand model–normalized EC50 of 209 µg Zn/L) and ranked in the 1st percentile of a compiled database for zinc species sensitivity distribution.

Environmental Toxicology and Chemistry

Acute and chronic sensitivity of white sturgeon ( Acipenser transmontanus ) and rainbow trout ( Oncorhynchus mykiss ) to cadmium, copper, lead, or zinc in laboratory water-only exposures

White sturgeon (Acipenser transmontanus) are experiencing poor recruitment in the trans boundary reach of the upper Columbia River in eastern Washington State. Limited toxicity data indicated that early life stages of white sturgeon are sensitive to metals. In acute 4-day (d) exposures with larval white sturgeon, previous studies have reported that the 4-day median lethal concentrations (LC50) based on biotic ligand model (BLM) normalization for copper were below the U.S. Environmental Protection Agency national recommended acute water-quality criterion. In previously published chronic 66-d exposures starting with newly fertilized eggs of white sturgeon, 20-percent lethal effect concentrations (LC20s) for copper, cadmium, or zinc generally were within a factor of two of the chronic values of the most sensitive fish species in the databases of the U.S. Environmental Protection Agency water-quality criteria (WQC) for the three metals. However, there were some uncertainties in the chronic exposures previously performed with white sturgeon, including (1) low control survival (37 percent), (2) more control fish tested in each replicate compared to other treatments, (3) limited replication of treatments (n=2), (4) lack of reported growth data (such as dry weight), and (5) wide dilution factors for exposure concentrations (6- to 8-fold dilutions). The U.S. Environmental Protection Agency concluded that additional studies are needed to generate more toxicity data to better define lethal and sublethal toxicity thresholds for metals for white sturgeon. The objective of the study was to further evaluate the acute and chronic toxicity of cadmium, copper, lead, or zinc to early life stages of white sturgeon in water-only exposures. Toxicity tests also were performed with commonly tested rainbow trout (Oncorhynchus mykiss) under similar test conditions to determine the relative sensitivity between white sturgeon and rainbow trout to these metals. Toxicity data generated from this study were used to evaluate the sensitivity of early life stages of white sturgeon and rainbow trout relative to data published for other test organisms. Toxicity data generated from this study also were used to evaluate the level of protection of U.S. Environmental Protection Agency WQC or Washington State water-quality standards (WQS) for copper, zinc, cadmium, or lead to white sturgeon inhabiting the upper Columbia River. Chapter A of this report summarizes the results of acute toxicity tests performed for 4 d with white sturgeon and rainbow trout exposed to copper, cadmium, or zinc. Chapter B of this report summarizes the results of chronic toxicity tests performed for as many as 53 days with white sturgeon or rainbow trout exposed to copper, cadmium, zinc, or lead. Appendixes to the report are available at http://pubs.usgs.gov/sir/2013/5204 . Supporting documentation for chapter A toxicity testing is provided in appendix 1. Supporting documentation for chapter B toxicity testing is provided in Appendix 2. Supporting documentation on analysis of water chemistry for chapter A and chapter B is provided in appendix 3 and 4. The rationale for applying corrections to measured copper and zinc values in water samples from some of the toxicity tests performed in chapter A is provided in appendix 5. A summary of dissolved organic carbon measurement variability and implications for biotic ligand model normalization for toxicity data summarized in chapter A and chapter B are provided in appendix 6. An evaluation of an interlaboratory comparison of analyses for dissolved organic carbon in water from the U.S. Geological Survey Columbia Environmental Research Center and University of Saskatchewan is provided in appendix 7. Finally, appendix 8 provides a summary of retesting of white sturgeon in 2012 to determine if improved survival of sturgeon would affect copper effect concentrations in 24-d copper exposures started with newly hatched larvae, and to evaluate the effect of light intensity or temperature on the response of newly hatched larvae during a 25-d study.

Scientific Investigations Report