Evaluation of Sublethal Toxicity of Nitrite to a Suite of Aquatic Organisms in Support of the Derivation of a Chronic Environmental Water Quality Benchmark.

Nitrite is a naturally-occurring inorganic compound that occurs in aquatic environments as an intermediary between nitrate and ammonia in the nitrogen cycle. It is a contaminant of potential concern resulting from anthropogenic activities in some cases. While the acute toxicity of nitrite has been characterized in previous studies, its sublethal toxicity is less understood. To determine the sublethal toxicity of nitrite on freshwater organisms, a suite of organisms was tested including: two salmonids (Oncorhynchus mykiss and O. kisutch), an alga (Pseudokirchneriella subcapitata), an aquatic macrophyte (Lemna minor), and three invertebrates (Ceriodaphnia dubia, Chironomus dilutus, and Neocloeon triangulifer). Test organisms were exposed to nitrite concentrations ranging between 0.02 and 1.28 mg/L nitrite (NO2-N). The toxicity tests were conducted according to procedures specified in standardized methods, allowing for the estimation of multiple endpoints for each test species. Species sensitivity distributions (SSDs) were generated using endpoints from the toxicity testing results, as well as data from previous studies, from which water chemistry approximated that used in the tests (i.e., < 5 mg/L chloride, an important toxicity-modifying factor for nitrite). The mayfly, N. triangulifer, was the most sensitive species, followed by the two salmonids (which represented the second and third most sensitive species), although they were not as sensitive to nitrite exposure as reported in previous studies. The fifth percentile hazard concentration (HC5) generated from the SSD could be used for derivation of regulatory benchmarks and threshold values for site-specific aquatic risk assessments.

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.

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.