Copper toxicity in Bristol Bay headwaters: Part 1-Acute mortality and ambient water quality criteria in low-hardness water.

The world-class Alaskan Bristol Bay salmon fishery and vast deposits of copper (Cu) and other metals in the watershed warrant further investigation into the potential toxicity of Cu to salmonids under the low water-hardness conditions that occur in the watershed. Therefore we investigated the acute toxicity of Cu to rainbow trout (Oncorhynchus mykiss) and fathead minnows (Pimephales promelas) in low-hardness water (∼ 30 mg/L as CaCO3 ) formulated in the laboratory and collected from the Bristol Bay watershed. The median lethal concentration (LC50) for rainbow trout exposed to Cu in low-hardness laboratory water was 16 µg Cu/L (95% confidence intervals [CIs]: 12, 21; dissolved Cu, filtered to 0.45 µm). The LC50 values for fathead minnows exposed to Cu in low-hardness laboratory water or site water were 29 and 79 µg Cu/L (95% CIs: 23, 35 and 58, 125; dissolved Cu), respectively. The biotic ligand model (BLM) LC50 estimates for these bioassays were 1.3 to 2.3 times higher than the actual LC50 values. We also calculated and analyzed acute Cu water quality criteria, also known as criterion maximum concentration (CMC), using hardness-based methods and the BLM for water samples collected throughout the Bristol Bay watershed in 2007. Biotic ligand model CMCs ranged from 0.05 to 17.5 µg Cu/L and hardness-based CMCs ranged from 2.3 to 6.1 µg Cu/L for the 65 samples analyzed. Our results show the need for site-specific research and subsequent water quality guidelines in low-hardness aquatic habitats. Environ Toxicol Chem 2019;38:190-197. © 2018 SETAC.

Copper toxicity in Bristol Bay headwaters: Part 2-Olfactory inhibition in low-hardness water.

We investigated the olfactory toxicity of copper (Cu) to rainbow trout in low-hardness (27 mg/L as CaCO3 ) water formulated in the laboratory over a 120-h period using a flow-through design. The fish's response to an alarm cue (e.g., reduction in activity) was recorded to determine the exposure concentrations and durations that inhibited olfactory detection of the cue after 3, 24, 48, and 96 h of Cu exposure and after 24 h of clean water recovery following the 96-h exposure period. Exposures were conducted with a range of Cu concentrations from 0.13 (control) to 7.14 µg Cu/L (dissolved Cu). We observed a dose-dependent response in olfactory inhibition with a 20% reduction in the probability of responding to the alarm cue, relative to controls, at 2.7 and 2.4 µg Cu/L after 24 or 96 h of exposure, respectively. Olfactory inhibition manifested between 3 and 24 h of exposure. Our 24- and 96-h 20% olfactory inhibition estimates fell between the criteria derived using the biotic ligand model (BLM; criterion maximum concentration [CMC] and criterion continuous concentration [CCC] values were 0.63 and 0.39 µg Cu/L, respectively) and water hardness-based criteria (CMC and CCC values were 3.9 and 2.9 µg Cu/L, respectively). Therefore, the hardness-based criteria do not appear to be protective and the BLM-derived criteria do appear to be protective against Cu-induced olfactory inhibition given our test water chemistry. Neither the hardness-based criteria nor the BLM-derived criteria appear to be protective against our estimated Cu behavioral avoidance response concentrations at 24- and 96-h exposures (0.54 and 0.50 µg Cu/L, respectively). Environ Toxicol Chem 2019;38:198-209. © 2018 SETAC.

Changes in white cell estimates and plasma chemistry measurements following oral or external dosing of double-crested cormorants, Phalacocorax auritus, with artificially weathered MC252 oil.

Scoping studies were designed whereby double-crested cormorants (Phalacocorax auritus) were dosed with artificially weathered Deepwater Horizon (DWH) oil either daily through oil injected feeder fish, or by application of oil directly to feathers every three days. Preening results in oil ingestion, and may be an effective means of orally dosing birds with toxicant to improve our understanding of the full range of physiological effects of oral oil ingestion on birds. Blood samples collected every 5-6 days were analyzed for a number of clinical endpoints including white blood cell (WBC) estimates and differential cell counts. Plasma biochemical evaluations were performed for changes associated with oil toxicity. Oral dosing and application of oil to feathers resulted in clinical signs and statistically significant changes in a number of biochemical endpoints consistent with petroleum exposure. In orally dosed birds there were statistically significant decreases in aspartate amino transferase (AST) and gamma glutamyl transferase (GGT) activities, calcium, chloride, cholesterol, glucose, and total protein concentrations, and increases in plasma urea, uric acid, and phosphorus concentrations. Plasma electrophoresis endpoints (pre-albumin, albumin, alpha-2 globulin, beta globulin, and gamma globulin concentrations and albumin: globulin ratios) were decreased in orally dosed birds. Birds with external oil had increases in urea, creatinine, uric acid, creatine kinase (CK), glutamate dehydrogenase (GLDH), phosphorus, calcium, chloride, potassium, albumin, alpha-1 globulin and alpha-2 globulin. Decreases were observed in AST, beta globulin and glucose. WBC also differed between treatments; however, this was in part driven by monocytosis present in the externally oiled birds prior to oil treatment.

Differential developmental toxicity of naphthoic acid isomers in medaka (Oryzias latipes) embryos.

Polycyclic aromatic hydrocarbons (PAHs) are widespread persistent pollutants that readily undergo biotic and abiotic conversion to numerous transformation products in rivers, lakes and estuarine sediments. Here we characterize the developmental toxicity of four PAH transformation products each structural isomers of hydroxynaphthoic acid: 1H2NA, 2H1NA, 2H3NA, and 6H2NA. Medaka fish (Oryzias latipes) embryos and eleutheroembryos were used to determine toxicity. A 96-well micro-plate format was used to establish a robust, statistically significant platform for assessment of early life stages. Individual naphthoic acid isomers demonstrated a rank order of toxicity with 1H2NA>2H1NA>2H3NA>6H2NA being more toxic. Abnormalities of circulatory system were most pronounced including pericardial edema and tube heart. To determine if HNA isomers were AhR ligands, spatial-temporal expression and activity of CYP1A was measured via in vivo EROD assessments. qPCR measurement of CYP1A induction proved different between isomers dosed at respective concentrations affecting 50% of exposed individuals (EC50s). In vitro, all ANH isomers transactivated mouse AhR using a medaka CYP1A promoter specific reporter assay. Circulatory abnormalities followed P450 induction and response was consistent with PAH toxicity. A 96-well micro-plates proved suitable as exposure chambers and provided statistically sound evaluations as well as efficient toxicity screens. Our results demonstrate the use of medaka embryos for toxicity analysis thereby achieving REACH objectives for the reduction of adult animal testing in toxicity evaluations.