The relationship between cellular protein content and selenium accumulation in freshwater microalgae.

Variability in the bioconcentration of selenium (Se) by primary producers at the base of the food web results in uncertainty in predictions of bioaccumulation and ecological risk to higher trophic level organisms. Water chemistry, speciation of Se, and periphyton community composition have all been suggested as factors that contribute to variability in bioconcentration by primary producers; however, the role of physiological composition of periphyton species in influencing the bioconcentration of Se has not been previously evaluated. To determine if a relationship exists between algal protein content and Se accumulation, Parachlorella kessleri, Chlorella vulgaris, and Raphidocelis subcapitata were exposed to Se (as selenate) and analyzed for total protein and tissue Se content in the exponential and stationary growth phases. Protein content and Se accumulation in R. subcapitata in the stationary phase were also measured under two light intensities. No relationship between cellular protein content and Se accumulation was found for algae in the exponential phase; however, a strong relationship was found in the stationary phase among species and for R. subcapitata under differing light intensities. Absolute Se accumulations by P. kessleri, C. vulgaris, and R. subcapitata in the stationary phase were statistically different; however, the concentrations of Se in protein were similar across species. These results suggest that cellular protein content in microalgae influences Se bioconcentration and that algal protein content may improve Se bioaccumulation modeling in food webs. Integr Environ Assess Manag 2024;00:1-10. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Maternal Transfer and Effects of Selenium on Early Life Stage Development of Redside Shiner (Richardsonius balteatus).

Maternal transfer of selenium (Se) to developing fish eggs during vitellogenesis can cause larval deformity and mortality. Previous studies have shown wide variation among fish species in both the magnitude of maternal transfer (exposure) and the egg Se concentration causing effects (sensitivity). We studied maternal transfer and effects of Se on early life stage development, survival, and growth of redside shiner (Richardsonius balteatus), a small-bodied cyprinid that has been reported to have relatively high ovary:muscle Se concentration ratios. Gametes were collected from lentic areas in southeast British Columbia (Canada) with a range of dietary Se concentrations related to weathering of waste rock from coal mining. Eggs were fertilized and reared in the laboratory from hatch to the onset of exogenous feeding. Larvae were assessed for survival, length, weight, Se-characteristic deformities, and edema. Eggs from a total of 56 females were collected, with egg Se concentrations from 0.7 to 28 mg/kg dry weight. Maternal transfer varied among sites, with egg:muscle Se concentration ratios ranging from <1 to >4. We also found that sampling residual ovaries can overestimate Se concentrations in ripe eggs by up to a factor of 5.7. A correlation between larval weight and egg Se concentration was identified, although the relationship was weak (r2 < 0.1) and appeared to be a site effect. No other relationships were observed between larval endpoints and egg Se concentrations up to the highest concentration tested, indicating that the effects threshold for this species may be >28 mg/kg dry weight in eggs. These data indicate that redside shiner is less sensitive to maternally transferred Se than most other tested fish species. Environ Toxicol Chem 2023;42:2350-2357. © 2023 SETAC.

An Evaluation of Molybdenum Toxicity to the Oligochaete, Tubifex tubifex, and Early-Life Stages of Brown Trout, Salmo trutta.

Limited data are available describing the aquatic toxicity of molybdenum in freshwater environments, making it difficult to assess the aquatic risk to freshwater organisms. In order to increase available information on the aquatic toxicity of molybdenum, a 96-h LC50 test with the oligochaete Tubifex tubifex and an 85-day development test using brown trout, Salmo trutta, were conducted. The T. tubifex test resulted in an LC50 value of 2782 mg/L. No adverse effects were observed on brown trout survival or length in the concentrations tested, however an IC10 value for growth (wet weight) was determined to be 202 mg/L. Whole body fish tissue concentrations for molybdenum increased in all treatment concentrations tested, although bioconcentration factors decreased at greater exposure concentrations, and ranged from 0.13 at an exposure concentration of 20 mg/L to 0.04 at an exposure of 1247 mg/L. A body burden of 26.0 mg/kg was associated with reduced wet weight.

Lentic, lotic, and sulfate-dependent waterborne selenium screening guidelines for freshwater systems.

There is consensus that fish are the most sensitive aquatic organisms to selenium (Se) and that Se concentrations in fish tissue are the most reliable indicators of potential toxicity. Differences in Se speciation, biological productivity, Se concentration, and parameters that affect Se bioavailability (e.g., sulfate) may influence the relationship between Se concentrations in water and fish tissue. It is desirable to identify environmentally protective waterborne Se guidelines that, if not exceeded, reduce the need to directly measure Se concentrations in fish tissue. Three factors that should currently be considered in developing waterborne Se screening guidelines are 1) differences between lotic and lentic sites, 2) the influence of exposure concentration on Se partitioning among compartments, and 3) the influence of sulfate on selenate bioavailability. Colocated data sets of Se concentrations in 1) water and particulates, 2) particulates and invertebrates, and 3) invertebrates and fish tissue were compiled; and a quantile regression approach was used to derive waterborne Se screening guidelines. Use of a regression-based approach for describing relationships in Se concentrations between compartments reduces uncertainty associated with selection of partitioning factors that are generally not constant over ranges of exposure concentrations. Waterborne Se screening guidelines of 6.5 and 3.0 µg/L for lotic and lentic water bodies were derived, and a sulfate-based waterborne Se guideline equation for selenate-dominated lotic waters was also developed. Environ Toxicol Chem 2017;36:2503-2513. © 2017 SETAC.

Evaluation of the effect of water type on the toxicity of nitrate to aquatic organisms.

A suite of acute and chronic toxicity tests were conducted to evaluate the sensitivity of freshwater organisms to nitrate (as sodium nitrate). Acute exposures with rainbow trout (Onchorhynchus mykiss) and amphipods (Hyalella azteca), as well as chronic exposures with H. azteca (14-d survival and growth), midges (Chironomus dilutus; 10-d survival and growth), daphnids (Ceriodaphnia dubia; 7-d survival and reproduction), and fathead minnows (Pimephales promelas; 7-d survival and growth) were used to determine sublethal and lethal effect concentrations. Modification of nitrate toxicity was investigated across a range of ionic strengths, created through the use of very soft water, and standard preparations of synthetic soft, moderately-hard and hard dilution waters. The most sensitive species tested were C. dubia and H. azteca, in soft water, with reproduction and growth IC25 values of 13.8 and 12.2 mg/L NO3-N, respectively. All of the organisms exposed to nitrate demonstrated significantly reduced effects with increasing ionic strength associated with changes in water type. Possible mechanisms responsible for the modifying effect of increasing major ion concentrations on nitrate toxicity are discussed.

Dietary selenium disrupts hepatic triglyceride stores and transcriptional networks associated with growth and Notch signaling in juvenile rainbow trout.

Dietary Se has been shown to adversely affect adult fish by altering growth rates and metabolism. To determine the underlying mechanisms associated with these observations, we measured biochemical and transcriptomic endpoints in rainbow trout following dietary Se exposures. Treatment groups of juvenile rainbow trout were fed either control Lumbriculus variegatus worms or worms cultured on selenized yeast. Selenized yeast was cultured at four nominal doses of 5, 10, 20 or 40mg/kg Se dry weight (measured dose in the worms of 7.1, 10.7, 19.5, and 31.8mg/kgSedw respectively) and fish were fed for 60days. At 60 d, hepatic triglycerides, glycogen, total glutathione, 8-isoprostane and the transcriptome response in the liver (n=8/group) were measured. Fish fed the nominal dose of 20 and 40mg/kg Se dry weight had lower body weight and a shorter length, as well as lower triglyceride in the liver compared to controls. Evidence was lacking for an oxidative stress response and there was no change in total glutathione, 8-isoprostane levels, nor relative mRNA levels for glutathione peroxidase isoforms among groups. Microarray analysis revealed that molecular networks for long-chain fatty acid transport, lipid transport, and low density lipid oxidation were increased in the liver of fish fed 40mg/kg, and this is hypothesized to be associated with the lower triglyceride levels in these fish. In addition, up-regulated gene networks in the liver of 40mg/kg Se treated fish included epidermal growth factor receptor signaling, growth hormone receptor, and insulin growth factor receptor 1 signaling pathways. These molecular changes are hypothesized to be compensatory and related to impaired growth. A gene network related to Notch signaling, which is involved in cell-cell communication and gene transcription regulation, was also increased in the liver following dietary treatments with both 20 and 40mg/kg Se. Transcriptomic data support the hypothesis that dietary Se increases the expression of networks for growth-related signaling cascades in addition to those related to fatty acid synthesis and metabolism. We propose that the disruption of metabolites related to triglyceride processing and storage, as well as gene networks for epidermal growth factor and Notch signaling in the liver, represent key molecular initiating events for adverse outcomes related to growth and Se toxicity in fish.

Application of molecular endpoints in early life stage salmonid environmental biomonitoring.

Molecular endpoints can enhance existing whole animal bioassays by more fully characterizing the biological impacts of aquatic pollutants. Laboratory and field studies were used to examine the utility of adopting molecular endpoints for a well-developed in situ early life stage (eyed embryo to onset of swim-up fry) salmonid bioassay to improve diagnostic assessments of water quality in the field. Coastal cutthroat trout (Oncorhynchus clarki clarki) were exposed in the laboratory to the model metal (zinc, 40µg/L) and the polycyclic aromatic hydrocarbon (pyrene, 100µg/L) in water to examine the resulting early life stage salmonid responses. In situ field exposures and bioassays were conducted in parallel to evaluate the water quality of three urban streams in British Columbia (two sites with anthropogenic inputs and one reference site). The endpoints measured in swim-up fry included survival, deformities, growth (weight and length), vitellogenin (vtg) and metallothionein (Mt) protein levels, and hepatic gene expression (e.g., metallothioneins [mta and mtb], endocrine biomarkers [vtg and estrogen receptors, esr] and xenobiotic-metabolizing enzymes [cytochrome P450 1A3, cyp1a3 and glutathione transferases, gstk]). No effects were observed in the zinc treatment, however exposure of swim-up fry to pyrene resulted in decreased survival, deformities and increased estrogen receptor alpha (er1) mRNA levels. In the field exposures, xenobiotic-metabolizing enzymes (cyp1a3, gstk) and zinc transporter (zntBigM103) mRNA were significantly increased in swim-up fry deployed at the sites with more anthropogenic inputs compared to the reference site. Cluster analysis revealed that gene expression profiles in individuals from the streams receiving anthropogenic inputs were more similar to each other than to the reference site. Collectively, the results obtained in this study suggest that molecular endpoints may be useful, and potentially more sensitive, indicators of site-specific contamination in real-world, complex exposure scenarios in addition to whole body morphometric and physiological measures.

A New Toxicity Test Using the Freshwater Copepod Cyclops vernalis.

The cladocerans Ceriodaphnia dubia and Daphnia magna are widely used in environmental toxicity testing and the test methodologies for these species are well developed. However, copepods are a much more abundant contributor to zooplankton in many lakes, but they are not routinely used in toxicity tests. Therefore, we propose toxicity test methods for the freshwater copepod, Cyclops vernalis assessing effects on its survival and growth. A case study is presented in which the proposed test was performed with a range of concentrations of total dissolved solids (TDS) and used as part of a test battery to develop a site-specific water quality objective. C. vernalis was less sensitive to TDS compared to D. magna and C. dubia, but similarly sensitive to an alga, a diatom, a rotifer, a chironomid, and two fish species. No adverse effects were observed on survival or growth of C. vernalis at TDS concentrations up to 1500 mg/L.

Effect of Total Dissolved Solids on Fertilization and Development of Two Salmonid Species.

Some studies have shown that the early life stages of salmonids are particularly sensitive to elevated concentrations of total dissolved solids (TDS). We evaluated the effect of TDS released in treated effluent into Snap Lake (Northwest Territories, Canada) by the Snap Lake Diamond Mine on two salmonids native to Snap Lake: Salvenius namaycush (lake trout) and Thymallus arcticus (Arctic grayling). Exposures encompassed the embryo-alevin-fry early life stages and extended to 142 days for lake trout and 69 days for Arctic grayling. Such extended testing is uncommon with these two species. Two exposures were conducted with each species, one initiated prior to fertilization, and the other subsequent to fertilization. Fertilization, survival, and growth were not adversely affected for either species by TDS at concentrations >1400 mg/L, with the exception of survival of lake trout, which produced an LC20 of 991 mg/L in one test, and >1484 mg/L in the second test. For the specific TDS composition tested, which was dominated by chloride (45 %-47 %) and calcium (20 %-21 %), the early life stages of these two fish species were relatively insensitive. Although some authors have suggested lower TDS regulatory limits for salmonid early life stages, our study indicates that this is not necessary, at least for these two fish species and for the specific ionic composition tested.

The effect of sulfate on selenate bioaccumulation in two freshwater primary producers: A duckweed (Lemna minor) and a green alga (Pseudokirchneriella subcapitata).

Predicting selenium bioaccumulation is complicated because site-specific conditions, including the ionic composition of water, affect the bioconcentration of inorganic selenium into the food web. Selenium tissue concentrations were measured in Lemna minor and Pseudokirchneriella subcapitata following exposure to selenate and sulfate. Selenium accumulation differed between species, and sulfate reduced selenium uptake in both species, indicating that ionic constituents, in particular sulfate, are important in modifying selenium uptake by primary producers.

Critical predicted no effect concentrations (PNECs) should not be based on a single toxicity test.

Predicted no-effect concentrations (PNECs), which represent the concentration of a substance below which an unacceptable effect most likely will not occur, are widely used for risk assessment and in environmental policy and regulation. They are typically based on single-species laboratory toxicity tests; often, a single test result for the most sensitive endpoints drives the derivation of a PNEC. In the present study, the authors provide a case study emphasizing the importance of determining the reliability of those most sensitive endpoints. Five 21-d Daphnia magna toxicity tests conducted using the same procedures by 2 laboratories gave 20% inhibitory concentration responses to a specific ionic composition of total dissolved solids that varied from 684 mg/L to more than 1510 mg/L. The concentration-response curve was shallow; thus, these differences could have been attributable to chance alone. The authors strongly recommend that the most sensitive endpoints that determine PNECs not be based on a single toxicity test result but rather on the geometric mean of at least 3 test results to adequately assess and bound test variability, especially when the concentration-response curve is shallow.

Development of a strontium chronic effects benchmark for aquatic life in freshwater.

There are no national water-quality guidelines for strontium for the protection of freshwater aquatic life in North America or elsewhere. Available data on the acute and chronic toxicity of strontium to freshwater aquatic life were compiled and reviewed. Acute toxicity was reported to occur at concentrations ranging from 75 mg/L to 15 000 mg/L. The majority of chronic effects occurred at concentrations above 11 mg/L; however, calculation of a representative benchmark was confounded by results from 4 studies indicating that chronic effects occurred at lower concentrations than all other studies, in 2 cases below background concentrations reported for US and European streams. Two of these studies, including 1 reporting effects below background concentrations, were repeated and found not to be reproducible; chronic effects occurred at considerably higher strontium concentrations than in the original studies. Studies with narrow-mouthed toad and goldfish were not repeated; both studies reported chronic effects below background concentrations, and both studies had been conducted by the authors of 1 of the 2 studies that were repeated and shown to be nonreproducible. Studies by these authors (3 of the 4 confounding studies), conducted over 30 yr ago, lacked detail in reporting of methods and results. It is thus likely that repeating the toad and goldfish studies would also have resulted in a higher strontium effects concentration. A strontium chronic effects benchmark of 10.7 mg/L that incorporates the results of additional testing summarized in the present study is proposed for freshwater environments.

Molecular responses to 17ß-estradiol in early life stage salmonids.

Environmental estrogens (EE) are ubiquitous in many aquatic environments and biological responses to EEs in early developmental stages of salmonids are poorly understood compared to juvenile and adult stages. Using 17ß-estradiol (E2) as a model estrogen, waterborne exposures were conducted on early life stage rainbow trout (Oncorhynchus mykiss; egg, alevin, swim-up fry) and both molecular and physiological endpoints were measured to quantify the effects of E2. To investigate developmental stage-specific effects, laboratory exposures of 1 µg/L E2 were initiated pre-hatching as eyed embryos or post-hatching upon entering the alevin stage. High mortality (∼90%) was observed when E2 exposures were initiated at the eyed embryo stage compared to the alevin stage (∼35% mortality), demonstrating stage-specific sensitivity. Gene expression analyses revealed that vitellogenin was detectable in the liver of swim-up fry, and was highly inducible by 1 µg/L E2 (>200-fold higher levels compared to control animals). Experiments also confirmed the induction of vitellogenin protein levels in protein extracts isolated from head and tail regions of swim-up fry after E2 exposure. These findings suggest that induction of vitellogenin, a well-characterized biomarker for estrogenic exposure, can be informative measured at this early life stage. Several other genes of the reproductive endocrine axis (e.g. estrogen receptors and androgen receptors) exhibited decreased expression levels compared to control animals. In addition, chronic exposure to E2 during the eyed embryo and alevin stages resulted in suppressive effects on growth related genes (growth hormone receptors, insulin-like growth factor 1) as well as premature hatching, suggesting that the somatotropic axis is a key target for E2-mediated developmental and growth disruptions. Combining molecular biomarkers with morphological and physiological changes in early life stage salmonids holds considerable promise for further defining estrogen action during development, and for assessing the impacts of endocrine disrupting chemicals in vivo in teleosts.

The effects of the urea-based herbicide linuron on reproductive endpoints in the fathead minnow (Pimephales promelas).

Linuron is a widely used urea-based herbicide that has anti-androgenic activity in both fish and rodents. To further elucidate the potential mode of action (MOA) of linuron on the vertebrate endocrine system, adult male and female fathead minnows were exposed for 21 days to dechlorinated water, a solvent control, 17ß-estradiol (E2; 0.1 µg/L), dihydrotestosterone (DHT; 100 µg/L), linuron (1, 10, 100 µg/L) and one co-treatment of DHT (100 µg/L) and linuron (100 µg/L). There were no effects of linuron on egg hatching, 7 day egg survival, nuptial tubercle formation or gonadal histopathology. Administration of DHT and 1 and 100 µg/L linuron reduced plasma vitellogenin in females, while male plasma vitellogenin were induced after E2 exposure and co-exposure of DHT and linuron. Ovarian mRNA levels were examined for several genes involved in steroidogenesis (e.g. p450scc, cyp19a, star, tspo, hsd17b and hsd11b) and estrogen-mediated responses (esr1, esr2b, esr2a). Only p450scc mRNA was significantly decreased with DHT+linuron co-treatment. Clustering of steroidogenic mRNA transcript expression patterns revealed that patterns for linuron were more similar to E2 compared to DHT. Collectively, this study supports the hypothesis that linuron may not be a pure anti-androgen and may have multiple MOAs that affect vertebrate reproduction.

Triclosan exposure alters postembryonic development in a Pacific tree frog (Pseudacris regilla) Amphibian Metamorphosis Assay (TREEMA).

The Amphibian Metamorphosis Assay (AMA), developed for Xenopus laevis, is designed to identify chemicals that disrupt thyroid hormone (TH)-mediated biological processes. We adapted the AMA for use on an ecologically-relevant North American species, the Pacific tree frog (Pseudacris regilla), and applied molecular endpoints to evaluate the effects of the antibacterial agent, triclosan (TCS). Premetamorphic (Gosner stage 26-28) tadpoles were immersed for 21 days in solvent control, 1.5 µg/L thyroxine (T(4)), 0.3, 3 and 30 µg/L (nominal) TCS, or combined T(4)/TCS treatments. Exposure effects were scored by morphometric (developmental stage, wet weight, and body, snout-vent and hindlimb lengths) and molecular (mRNA abundance using quantitative real time polymerase chain reaction) criteria. T(4) treatment alone accelerated development concomitant with altered levels of TH receptors α and ß, proliferating cell nuclear antigen, and gelatinase B mRNAs in the brain and tail. We observed TCS-induced perturbations in all of the molecular and morphological endpoints indicating that TCS exposure disrupts coordination of postembryonic tadpole development. Clear alterations in molecular endpoints were evident at day 2 whereas the earliest morphological effects appeared at day 4 and were most evident at day 21. Although TCS alone (3 and 30 µg/L) was protective against tadpole mortality, this protection was lost in the presence of T(4). The Pacific tree frog is the most sensitive species examined to date displaying disruption of TH-mediated development by a common antimicrobial agent.

Hepatic protein expression networks associated with masculinization in the female fathead minnow (Pimephales promelas).

Endocrine disruptors that act via the androgen receptor (AR) are less well studied than environmental estrogens, and there is evidence that treatment with AR agonists can result in masculinization of female fish. In this study, female fathead minnows (FHM) were exposed to the model nonaromatizable androgen 5-alpha dihydrotestosterone (DHT) (100 µg/L), the ureic-based herbicide linuron (LIN) (100 µg/L), and a mixture of DHT and LIN (100 µg/L each) to better characterize androgen action in females. LIN was used because of reports that this chemical has an antiandrogenic mode of action in fish. After 21d, DHT and LIN treatments resulted in a significant depression of plasma vitellogenin (Vtg) and DHT and DHT+LIN increased the prevalence of nuptial tubercles in female FHMs indicating masculinization. Using iTRAQ and an LTQ Orbitrap Velos, ∼2000 proteins were identified in the FHM liver and the number of proteins quantified after exposures was >1200. Proteins that significantly and consistently changed in abundance across biological replicates included prostaglandin E synthase 3, programmed cell death 4a, glutathione S transferases, canopy, selenoprotein U, and ribosomal proteins. Subnetwork enrichment analysis identified that interferon and epidermal growth factor signaling were regulated by DHT and LIN, suggesting that these signaling pathways are correlated to depressed plasma vitellogenin. These data provide novel insight into hepatic protein networks that are associated with the process of masculinization in teleosts.

An aquatic toxicological evaluation of sulfate: the case for considering hardness as a modifying factor in setting water quality guidelines.

Elevated concentrations of sulfate occur commonly in anthropogenically impacted and natural waters. However, water quality guidelines (WQG) have not been developed in many jurisdictions, and chronic toxicity data are scarce for this anion. A variety of test organisms, including species of invertebrate, fish, algae, moss, and an amphibian, were tested for chronic toxicity to develop a robust dataset that could be used to develop WQGs. As an example of how these data might be used to establish guidelines, calculations were performed using two standard procedures: a species sensitivity distribution (SSD) approach, following methods employed in developing Canadian WQGs, and a safety factor approach, according to procedures typically used in the development of provincial WQGs in British Columbia. The interaction of sulfate toxicity and water hardness was evaluated and incorporated into the calculations, resulting in separate values for soft (10-40 mg/L), moderately hard (80-100 mg/L) and hard water (160-250 mg/L). The resulting values were 129, 644, and 725 mg/L sulfate, respectively, following the SSD approach, and 75, 625, and 675 mg/L sulfate, following the safety factor approach.

Chronic toxicity of chloride to freshwater species: effects of hardness and implications for water quality guidelines.

Toxicity tests using nine freshwater species (Ceriodaphnia dubia, Daphnia magna, Oncorhynchus mykiss, Pimephales promelas, Lumbriculus variegatus, Tubifex tubifex, Chironomus dilutus, Hyallela azteca, and Brachionus calyciflorus) were conducted to evaluate their sensitivity to chloride. Acute-to-chronic ratios (ACRs) from these tests indicate the ACR of 7.59 employed by the United States Environmental Protection Agency (U.S. EPA) in deriving its water quality guideline for chloride may be conservative; a revised ACR of 3.50 is presented here. The endpoints used to calculate the ACR included 24-h to 96-h median lethal concentrations (LC50s) for acute tests, and 48-h to 54-d inhibition concentration (ICx) values for growth or reproduction for chronic exposures. Data from the present chronic toxicity tests, and other investigators, were used to propose a water quality guideline for long-term exposure to chloride using a species sensitivity distribution (SSD) approach. The 5th percentile from the SSD was calculated as 307 mg/L and proposed as the water quality guideline. Cladocerans were the most sensitive species in the dataset. Ceriodaphnia dubia was used to evaluate the relationship between water hardness and sensitivity to chloride. A strong relationship was observed and was used to establish a hardness-related equation to modify the proposed water quality guideline on the basis of water hardness, resulting in values ranging from 64 mg/L chloride at 10 mg/L hardness to 388 mg/L chloride at 160 mg/L hardness (as CaCO3). These data suggest that current water quality guidelines for chloride may be overly conservative in water with moderate-to-high hardness, and may not be sufficiently protective under soft-water conditions.

Developmental toxicity of selenium to Dolly Varden char (Salvelinus malma).

Gametes were collected from Dolly Varden char (Salvelinus malma) from waterbodies in a region exposed to mining-related selenium (Se) releases in British Columbia, Canada. Fertilized eggs were incubated in a laboratory and deformities were assessed on newly-hatched alevins using a graduated severity index. No effects were observed on egg or alevin survival or larval weight across the studied exposure range of 5.4 to 66 mg/kg dry weight in egg. Length of some larvae was reduced at the highest egg Se concentrations and a clear residue-response relationship was observed for larval deformity. The egg concentration corresponding to a 10% increase in the frequency of deformity (EC10) was 54 mg/kg dry weight, which is substantially higher than reported for other cold-water fish species.