In most freshwater fishes, exposure to acidic water decreases rates of active ion uptake and increases rates of passive ion efflux. To survive in low pH conditions, fish must possess mechanisms to maintain net ion flux rates close to zero. In this study, rainbow trout ( Oncorhynchus mykiss) exposed to moderate acidity (pH = 4.5 or 5.5) in hard water (∼180 mg/L CaCO 3 ) demonstrated Na + influx inhibition relative to fish held in control conditions (pH = 8.0). However, Na + efflux rates were significantly lower in fish exposed to low pH, which minimized the net loss of Na + . This represents a relatively novel mechanism to maintain ion balance in low pH for an acid sensitive species, which appeared to be a result of reduced gill paracellular permeability. However, there were no significant differences in the transcript abundance of six different tight junction genes in response to low pH exposure. Overall, we conclude that acid tolerance may be dictated by differences in efflux rates in fishes which, like the rainbow trout, possess acid sensitive mechanisms of ion absorption.

Environmental deoxyribonucleic acid (eDNA) assays represent a non-invasive approach for biomonitoring. To assess Arctic grayling ( Thymallus arcticus, Pallas, 1776) populations in the Little Nahanni watershed of Northwest Territories, Canada, an eDNA assay that has been previously validated for use in fish of the Beringia lineage was used in conjunction with traditional survey methods (i.e., electrofishing). Forty-six 100 m reaches of streams were assessed in August 2015. The assay successfully detected Arctic grayling of the Nahanni lineage, with an eDNA signal recorded at each of the 17 sites in which Arctic grayling were observed by traditional fish surveys, but also at 3 of the 29 sites where Arctic grayling were not observed. The presence of eDNA was related to habitat metrics via Random Forest and correlation analyses. Riffles and water temperature were identified as being predictive of Arctic grayling eDNA abundance; however, no significant relationship between eDNA abundance and biomass proxies (fish abundance and fork length metrics) could be established. The high congruence between traditional approaches and eDNA surveys suggests adoption of the latter method will enhance the temporal and spatial acuity of biomonitoring, thereby improving field assessment of Arctic grayling populations and contributing towards more effective conservation management of this species.

Acidification is a key component of digestion throughout metazoans. The gut digestive fluid of many invertebrates is acidified by the vesicular-type H+-ATPase (VHA). In contrast, vertebrates generate acidic gut fluids using the gastric H+/K+-ATPase (HKA), an evolutionary innovation linked with the appearance of a true stomach that greatly improves digestion, absorption and immune function. Hagfishes are the most basal extant vertebrates, and their mechanism of digestive acidification remains unclear. Herein, we report that the stomachless Pacific hagfish (Eptatretus stoutii) acidify their gut using the VHA, and searches of E. stoutii gut transcriptomes and the genome of a closely related hagfish species (E. burgerii) indicate they lack HKA, consistent with its emergence following the 2R whole-genome duplication. Immunostaining revealed prominent VHA presence in the apical membrane of enterocytes and sub-apical expression of both VHA and soluble adenylyl cyclase. Interestingly, akin to vertebrates, VHA was also observed in immature pancreatic-like zymogen granules and was noticeably absent from the mature granules. Furthermore, isolated gut sacs from fed hagfish demonstrate increased VHA-dependent luminal H+ secretion that is stimulated by the cAMP pathway. Overall, these results suggest that the hagfish gut shares the trait of VHA-dependent acidification with invertebrates, while simultaneously performing some roles of the pancreas and intestine of gnathostomes.

The hydrophobic surface of plastics adsorbs hydrophobic persistent organic pollutants (POP) such as Perfluorooctanoic acid (PFOA). The potential for hydrophobic nanoparticles such as titanium dioxide (TiO2) to associate with PFOA and alter accumulation rates has not been investigated. Nanoparticles form ecocorona by adsorption of multiple constituents in water, but few studies have examined if this results in differences in the rate of PFOA accumulation in freshwater animals. We demonstrate the PFOA associates with the hydrophobic surfaces of nano-sized TiO2 particles and this increases the rate of uptake of PFOA into Daphnia magna. Accumulation of PFOA in daphnia was measurement over multiple concentrations, flux times and particle sizes using a radiotracer-based method (C-14-labelled PFOA). Our results show that TiO2 NPs have a high sorption capacity for PFOA and PFOA sorption decreased aggregation of TiO2 as evidenced by a decrease in average hydrodynamic diameter, decreased zeta potential and increased polydispersity index. Uptake of PFOA at 10 mu g/L was found to be 45 % higher in the presence of 500 mu g/L of 5 nm TiO2 compared to control PFOA alone uptake. Potentiation of PFOA uptake using 25 nm TiO2 NPs was 25 % higher than control PFOA alone. PFOA alone (0.5 mg/L) reduced metabolic oxygen consumption (MO2) in daphnia by 52 %, but exposure to (100 mg/L) 5 nm TiO2 NPs sorbed with (0.5 mg/L) PFOA decreased metabolic oxygen consumption (MO2) by similar to 88 %. These findings show that TiO2 nanoparticles act as vectors for hydrophobic organic pollutant accumulation and significantly potentiate PFOA accumulation and toxicity in aquatic organisms.

ABSTRACT Sponges (phylum Porifera) are an early diverging animal lineage without nervous and muscular systems, and yet they are able to produce coordinated whole-body contractions in response to disturbances. Little is known about the underlying signalling mechanisms in coordinating such responses. Previous studies demonstrated that sponges respond specifically to chemicals such as l-glutamate and γ-amino-butyric acid (GABA), which trigger and prevent contractions, respectively. Genes for purinergic P2X-like receptors are present in several sponge genomes, leading us to ask whether ATP works with glutamate to coordinate contractions in sponges as it does in other animal nervous systems. Using pharmacological approaches on the freshwater sponge Ephydatia muelleri, we show that ATP is involved in coordinating contractions. Bath application of ATP caused a rapid, sustained expansion of the excurrent canals in a dose-dependent manner. Complete contractions occurred when ATP was added in the presence of apyrase, an enzyme that hydrolyses ATP. Application of ADP, the first metabolic product of ATP hydrolysis, triggered complete contractions, whereas AMP, the subsequent metabolite, did not trigger a response. Blocking ATP from binding and activating P2X receptors with pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) prevented both glutamate- and ATP-triggered contractions, suggesting that ATP works downstream of glutamate. Bioinformatic analysis revealed two P2X receptor sequences, one of which groups with other vertebrate P2X receptors. Altogether, our results confirm that purinergic signalling by ATP is involved in coordinating contractions in the freshwater sponge.

The freshwater sponge, Ephydatia muelleri, lacks a nervous or endocrine system and yet it exhibits a coordinated whole-body action known as a “sneeze” that can be triggered by exposure to l-glutamate. It is not known how l-glutamate is obtained by E. muelleri in sufficient quantities (i.e., 70 µM) to mediate this response endogenously. The present study tested the hypothesis that l-glutamate can be directly acquired from the environment across the body surface of E. muelleri. We demonstrate carrier mediated uptake of two distinct saturable systems with maximal transport rates (Jmax) of 64.27 ± 4.98 and 25.12 ± 1.87 pmols mg−1 min−1, respectively. The latter system has a higher calculated substrate affinity (Km) of 2.87 ± 0.38 µM compared to the former (8.75 ± 1.00 µM), indicative of distinct systems that can acquire l-glutamate at variable environmental concentrations. Further characterization revealed potential shared pathways of l-glutamate uptake with other negatively charged amino acids, namely d-glutamate and l-aspartate, as well as the neutral amino acid L-alanine. We demonstrate that l-glutamate uptake does not appear to rely on exogenous sodium or proton concentrations as removal of these ions from the bathing media did not significantly alter uptake. Likewise, l-glutamate uptake does not seem to rely on internal proton motive forces driven by VHA as application of 100 nM of the VHA inhibitor bafilomycin did not alter uptake rates within E. muelleri tissues. Whether the acquired amino acid is used to supplement feeding or is stored and accumulated to mediate the sneeze response remains to be determined.

Exposure to alkaline water (pH > 9.0) is physiologically challenging for fish, yet our understanding of the physiology of alkaline tolerance in fishes is limited to a small number of ihighly specialized species. This study aimed to characterize mechanisms of alkaline tolerance in brook stickleback (Culaea inconstans), a fish species with a broad pH habitat range, including highly alkaline waters such as Buffalo Lake (pH = 9.2) in Alberta, Canada. Stickleback from Buffalo Lake and a neutral reference lake (Buck Lake; pH = 8.2) were collected from the wild and acclimated to common conditions (pH = 8.0) for at least 2 months. Both populations were then exposed to alkaline conditions (pH = 9.5), resulting in a significant decrease in survival (14% by 7 d of exposure) in Buck Lake fish, but no mortality in Buffalo Lake stickleback. In a 4-d exposure to alkaline water, fish from both populations experienced characteristic inhibitions of ammonia excretion followed by subsequent recovery, in conjunction with an accumulation of ammonia within the body. However, no differences were observed between populations. Analysis of tissue Na+ and Cl- content showed a more pronounced decrease in Cl- in Buck Lake fish, suggesting that tighter regulation of Cl- homeostasis and/or acid-base balance may be an important feature of alkaline tolerance. RNA-sequencing analysis highlighted large differences in gene expression between the alkaline and neutral lake populations, and in response to alkaline exposure. Few of these changes in the expression involved genes known to be associated with nitrogen, ion, or acid-base balance. These data indicate that alkaline tolerance is higher in brook stickleback resident to an alkaline lake than those sourced from a neutral lake, a trait that may be related to differences in physiological and transcriptomic responses to alkaline exposure. ### Competing Interest Statement The authors have declared no competing interest.

The mechanism(s) of sodium, chloride and pH regulation in teleost fishes has been the subject of intense interest for researchers over the past 100 years. The primary organ responsible for ionoregulatory homeostasis is the gill, and more specifically, gill ionocytes. Building on the theoretical and experimental research of the past, recent advances in molecular and cellular techniques in the past two decades have allowed for substantial advances in our understanding of mechanisms involved. With an increased diversity of teleost species and environmental conditions being investigated, it has become apparent that there are multiple strategies and mechanisms employed to achieve ion and acid-base homeostasis. This review will cover the historical developments in our understanding of the teleost fish gill, highlight some of the recent advances and conflicting information in our understanding of ionocyte function, and serve to identify areas that require further investigation to improve our understanding of complex cellular and molecular machineries involved in iono- and acid-base regulation.

The effects of plastic pollution on marine organisms is of growing concern. The hydrophobic surface of plastics adsorbs organic contaminants and can alter the rate of chemical uptake in fishes. Per-fluorinated organic chemicals such as Perfluorooctanoic acid (PFOA) are highly hydrophobic toxic chemicals that adsorb to hydrophobic surfaces. We hypothesized that the presence of nano-sized plastic particles adsorbs PFOA and alter both the physical-chemical properties of the plastics and also enhance PFOA uptake into organisms. Using radiolabelled 14C-PFOA, we measured direct unidirectional uptake of PFOA in juvenile Pacific Oysters (Magallana gigas) at different (0.025, 0.50, and 0.100 mg/L) concentrations, for different exposure periods (1, 2, 4, and 6 h) and investigated whether varying concentrations (0.1, 0.5, 1 mg/L) of either 500 nm or 20 nm polystyrene nanoparticles (PS-NPs) differentially altered the uptake rate of PFOA. Our results demonstrate that PFOA adsorbs to the surface of PS-NPs, altering PS-NP behaviour in solution and significantly increases the rate of uptake of PFOA in exposed Pacific oysters. PFOA uptake at 0.1 mg/L was increased 2.3-fold in the presence of 1 mg/L 500 nm PS-NP and 3.2-fold in the presence of 1 mg/L 20 nm PS-NP. In a separate study to examine if PS NPs potentiate the biochemical response to PFOA, both 500 and 20 nm PS-NP at 100 mg/L increased the 1 mg/L PFOA-induced oxidative stress by 2.5-fold and 3-fold respectively. These findings demonstrate that nanoplastics as co-contaminants in marine systems are able to adsorb PFOA and significantly potentiate its uptake and toxicity.

Coal mining activity is one of the major sources of elevated selenium concentration in aquatic systems; however, little is known about its uptake mechanisms in aquatic organisms. Here, we demonstrate that high waterborne selenite uptake in the water flea, Daphnia magna, is likely mediated by a phosphate transporter. Concentrations of selenium that exceed regulatory guidelines have been associated with coal mining activities and have been linked to detrimental effects on aquatic ecosystems and the organisms therein. Although the major route of selenium uptake in macroinvertebrates is via the diet, the uptake of waterborne selenite (HSeO3-), the prominent form at circumneutral pH, can be an important contributor to selenium body burden and thus selenium toxicity. In the current study, radiolabelled selenite (Se75) was used to characterize the mechanism of selenite uptake in the water flea, Daphnia magna. The concentration dependence (1-32 mu M) of selenite uptake was determined in 1-hour uptake assays in artificial waters that independently varied in bicarbonate, chloride, sulphate, phosphate and selenate concentrations. At concentrations representative of those found in highly contaminated waters, selenite uptake was phosphate-dependent and inhibited by foscarnet, a phosphate transport inhibitor. At higher concentrations, selenite uptake was dependent on waterborne bicarbonate concentration and inhibited by the bicarbonate transporter inhibitor DIDS (4,4 '-diisothiocyano-2,2 '-stilbenedisulfonic acid). These findings suggest that concentrations of phosphate in coal mining-affected waters could alter selenite uptake in aquatic organisms and could ultimately affect the toxic impacts of selenium in such waters.

Nanoplastics possess unique characteristics (e.g., high surface area/volume ratio) that enhance the adsorption of organic chemicals onto their surface. Their occurrence raises human health and ecotoxicological concerns, as pollutants bound to nanoplastics can have a larger effect than they would on their own. This study assessed the sorption of plant protection products (glyphosate and methyl parathion), an antidepressant (fluoxetine), a perfluorochemical (perfluorooctanoic acid [PFOA]), and a polycyclic aromatic hydrocarbon (phenanthrene) onto commercially available carboxylated polystyrene (PS) nanoplastics (NPs, 500 and 20 nm). Based on the calculated sorption coefficients (K-d, L/kg), the sequence of chemicals displaying the highest to lowest affinity toward PSNPs is fluoxetine > phenanthrene > methyl parathion > PFOA > glyphosate, with 20 nm PS showing a higher potential to sorb organic chemicals. Cationic (fluoxetine) and hydrophobic (phenanthrene) substances were more amenable to sorption, whereas negatively charged and more hydrophilic ones (i.e., PFOA and glyphosate) showed poor sorption. pH influenced sorption for all target chemicals except phenanthrene. Sorption capacity was further reduced in water spiked with natural organic matter and in tertiary-treated wastewater effluent. Overall, our work enhances the understanding of how representative organic chemicals sorb onto nanoplastics and provides quantitative information (i.e., K-d) on future simulations of nanoplastics' fate and transport.

Hydraulic fracturing flowback and produced water (HF-FPW) has raised significant concerns owing to its potential impact on aquatic organisms and human health. Understanding the chemical composition of HF-FPW is crucial for developing appropriate management and remediation strategies. Herein, we performed nontarget screening on hydrophobic organic chemicals in the particulate phase of FPW (P-FPW) using gas chromatography-Orbitrap mass spectrometry coupled with cheminformatic analysis. In total, 5807 features were discovered, with 209 annotated with positive confidence levels, which were further classified based on their chemical taxonomy and functional use information. We found that benzenoids dominated the chemical class, followed by hydrocarbons. The annotated chemicals were classified into fragrances, catalysts, antimicrobials, antioxidants, and other classes. Chemical overlap across countries (China vs. Canada) and wells was observed, with most chemicals reaching peak intensity within 24 or 48 h after the initial flowback and gradually decreasing. Approximately two-thirds of the identified or annotated chemicals have not been previously reported or included in existing HF-related databases, indicating expanded chemical coverage by the current screening workflow. A Toxicological Priority Index (ToxPi) scheme, which integrates chemical properties, ecological toxicities, and in vivo exposure potentials, was adopted to prioritize chemicals for further evaluation. Seven chemicals were proposed as prioritized compounds, of which the ester derivative of perfluorobutanoic acid, octacosyl heptafluorobutyrate (confidence level 2), exhibited the highest ToxPi score. Notably, most prioritized substances have limited toxicological data and are beyond the routine monitoring of the HF industry, highlighting significant gaps in our understanding of HF-related chemical content and environmental risk associated with water management.

An important goal of environmental and comparative physiology research is to identify species or populations that may be susceptible to environmental change such as heat wave events that are predicted to become more frequent and intense in the future. This study tested the hypothesis that fishes inhabiting alkaline lakes face significant physiological challenges, which results in reduced thermal tolerance. Brook stickleback (Culaea inconstans) were collected from an alkaline lake (pH 9.3) in Alberta, Canada and held under neutral conditions in the laboratory. Subsequently, fish were acutely exposed (4 d) to neutral (pH 7) or alkaline (pH 9.5) waters at 10 or 25°C. Exposure to alkaline water reduced critical thermal maximum (CTmax) in stickleback by approximately 1°C, but thermal acclimation capacity (“thermal plasticity”) was unaffected by alkaline exposure. Alkaline conditions resulted in physiological disturbances characteristic of exposure to high pH including elevated whole-body ammonia and lactate concentrations. Acute warming to CTmax in alkaline-exposed fish resulted in reductions in whole-body sodium and chloride concentrations. In addition, alkaline exposure compromised recovery from exercise at elevated temperatures. Overall, these results suggest that the physiological disturbances observed in response to alkaline exposure may render fish more susceptible to acute warming, reducing thermal tolerance.

Agri-chemicals such as fungicides are applied in natural settings and hence are exposed to the environment's ultraviolet (UV) light. Recently, many fungicides in commerce are being modified as nano-enabled formulations to increase agricultural productivity and reduce potential off-target effects. The present study investigated the impacts of sunlight-grade UV emission on the effects of either conventional or nano-enabled azoxystrobin (Az or nAz, respectively), a commonly applied agricultural fungicide, on Daphnia magna. Daphnids were exposed to increasing concentrations of Az or nAz under either full-spectrum (Vis) or full-spectrum Vis+UV (Vis+UV) lighting regimes to evaluate LC50s. Az LC50 was calculated at 268.8 and 234.2 μg/L in Vis or Vis+UV, respectively, while LC50 for nAz was 485.6 and 431.0 μg/L under Vis or Vis+UV light, respectively. Daphnids were exposed to 10% LC50 of either Az or nAz under Vis or Vis+UV lighting regime for 48 h or 21 d (acute and chronic, respectively). By 48 h, both Az and nAz reduced O2 consumption and increased TBARS. Heart rate was increased in Az-exposed daphnids but not in nAz groups. Neither of the two chemicals impacted thoracic limb activity. In 21 d exposures, Az significantly reduced biomass production and fecundity, but nAz groups were not significantly different from controls. The results of the present study demonstrate that conventional Az is more toxic to D. magna at lethal and sub-lethal levels in acute and chronic exposures, and sunlight strength UV can potentiate both acute and chronic effects of Az and nAz on D. magna.

Arctic grayling (Thymallus arcticus) is a salmonid fish of significant conservation value. However, conservation efforts are hindered by a lack of fundamental information regarding details such as current population distribution, migratory patterns, and natal habitats. In the current study, we designed, optimized, and field- and laboratory-validated an environmental DNA (eDNA) and environmental RNA (eRNA) assay for Arctic grayling biomonitoring. Using an in silico approach, a robust species-specific eDNA assay was generated, and filtering and extraction protocols were optimized for maximal eDNA yield. A Preserve, Precipitate, Lyse, Precipitate, and Purify (PPLPP) extraction method generated 70-fold higher eDNA yields than a column-based approach. Species-specificity relative to co-occurring salmonid fish was validated, and no significant amplification was noted for rainbow trout, brook trout, or mountain whitefish. Shedding rates of eDNA were around eight to nine times higher than those of eRNA, although the two types of nucleic acids decayed at similar rates. Shedding and decay rates were subsequently used to build detection probability models that account for pool size and water exchange rates. These data indicate that eDNA and eRNA are detectable in pools up to 32,500 m(3) in volume and with water flow of less than 0.5 m(3)s(-1) when an Arctic grayling is present. This knowledge can be implemented when designing field sampling strategies. Finally, the assay successfully amplified Arctic grayling eDNA from field-collected samples, with signal strength indicating preferred Arctic grayling habitat or conditions that favored the concentration and retention of eDNA.

The accumulation and tissue distribution of toxicants in aquatic biota can be determinative of their toxic impact to both exposed organisms and their potential human consumers. In the present study, accumulation of the trace metal thallium (Tl) in gill, muscle, plasma, and otoliths of rainbow trout (Oncorhynchus mykiss) following acute (96-h) and subchronic (28-day) waterborne exposures was investigated. Owing to known interactions between Tl and potassium ions (K+ ), plasma and muscle K+ concentrations were also determined. Branchial Tl accumulated in a dose-dependent manner in both acute and subchronic exposures, while plasma Tl was rapidly mobilized to tissues and accumulated only at exposure concentrations of 141 µg L-1 or higher. For muscle tissue, Tl concentrations at 28 days were markedly lower than those at 96 h at comparable exposure concentrations (0.9 µg L-1 ), indicating the presence of mechanisms that act to reduce Tl accumulation over time. However, after acute exposure, muscle Tl reached concentrations that, if consumed, would exceed acceptable daily intake values for this element, indicating some risk to human health from the consumption of fish from waters heavily contaminated with Tl. Otoliths showed Tl concentrations that reflected exposure concentration and length, confirming their capacity to provide insight into fish exposure history. No changes in tissue K+ concentrations were observed, suggesting that accumulation of Tl in rainbow trout plasma and muscle does not occur at the expense of K+ homeostasis. In addition to highlighting the capacity of rainbow trout to accumulate Tl to levels that exceed recommended dietary doses to human consumers, the present study provides the first data of tissue-specific Tl accumulation in an important regulatory species. Environ Toxicol Chem 2023;42:1553-1563. © 2023 SETAC.

The trace element thallium (Tl) exerts its toxic effects, at least in part, through its mimicry of potassium (K+) and subsequent impairment of K+ homeostasis. However, the specific nature of this effect remains poorly understood, especially in aquatic biota that are threatened by elevated concentrations of Tl associated with mining and refining effluents. In this study experiments were conducted to mechanistically examine the relationship between Tl and K+ in terms of uptake and toxicity in the regulatory model species Daphnia magna. In one set of experiments the effects of K+, the K+ analog rubidium (Rb+), and generalized K+ channel blocker cesium (Cs+) on Tl-induced acute toxicity were examined. The presence of increasing concentrations of K+ and Rb+ in exposure water reduced waterborne Tl toxicity, indicating that the actions of Tl were mediated at least in part through interactions with K+. However, in the presence of elevated water Cs+, the toxicity of Tl paradoxically increased. Pharmaceuticals with putative blocking actions on K+ channels failed to alter whole-body K+ of control organisms, but in the case of clozapine and chlorpropamide, whole-body K+ status was significantly elevated relative to exposures with Tl alone, which tended to reduce this metric. These data identify inwardly rectifying and voltage gated K+ channels as potential loci of Tl toxicity. Experiments using rubidium (Rb+) as a tracer of K+, showed that waterborne Tl affects the uptake of K+, but the magnitude of inhibition by Tl was not sufficient to explain the effect on whole-body K+. While these data indicate interactions between Tl and K occur at K+ transporters in D magna, they also indicate that environmental levels of K+ are likely to ameliorate toxicity in most natural waters.

Fish mortality is associated with harmful algal blooms, although whether toxicity is related directly to the presence of cyanotoxins or the prevailing water chemistry remains unclear. Similarly, while planktivorous fish may be exposed to toxin through the diet, the hazard posed by waterborne extracellular toxin to carnivorous fish is less well understood. In this study rainbow trout (Oncorhynchus mykiss) were exposed for up to 28 d to waterborne microcystin-LR at nominal concentrations of 1.5 and 50 µg L-1 (measured values 2 and 49 µg L-1, respectively). The former represents the Canadian drinking water guideline, and the latter an elevated environmental level. This study hypothesised that waterborne toxin exposure would specifically impact gill function, and given the importance of this tissue in freshwater fish ion regulation, effects on plasma ions and branchial ion transporter activity would be observed. Microcystin-LR exposure resulted in a significant and persistent hypocalcaemia at the higher exposure concentration, but plasma sodium and branchial activities of the sodium/potassium ATPase, proton ATPase and calcium ATPase enzymes remained unaffected. An in vitro assessment failed to show any effect of microcystin-LR on branchial calcium ATPase activity even at exposure concentrations as high as 1000 µg L-1. A transient increase in hepatic alkaline phosphatase activity was also observed at 49 µg L-1, but there were no effects of toxin exposure on branchial or hepatic lactate dehydrogenase activity. These results suggest that microcystin-LR exposure does not have a general effect on ion regulation, but instead produces a novel and specific impact on calcium metabolism in rainbow trout, although the mechanism underlying this effect remains unknown.

The mechanisms of acute (96-hour) and subchronic (28-day) toxicity of the waterborne trace metal thallium (Tl) to rainbow trout (Oncorhynchus mykiss) were investigated. Specifically, effects on branchial and renal ionoregulatory enzymes (sodium/potassium adenosine triphosphatase [ATPase; NKA] and proton ATPase) and hepatic oxidative stress endpoints (protein carbonylation, glutathione content, and activities of catalase and glutathione peroxidase) were examined. Fish (19-55 g) were acutely exposed to 0 (control), 0.9 (regulatory limit), 2004 (half the acute median lethal concentration), or 4200 (acute median lethal concentration) µg Tl L-1 or subchronically exposed to 0, 0.9, or 141 (an elevated environmental concentration) µg Tl L-1 . The only effect following acute exposure was a stimulation of renal H+ -ATPase activity at the highest Tl exposure concentration. Similarly, the only significant effect of subchronic Tl exposure was an inhibition of branchial NKA activity at 141 µg Tl L-1 , an effect that may reflect the interaction of Tl with potassium ion handling. Despite significant literature evidence for effects of Tl on oxidative stress, there were no effects of Tl on any such endpoint in rainbow trout, regardless of exposure duration or exposure concentration. Elevated basal levels of antioxidant defenses may explain this finding. These data suggest that ionoregulatory perturbance is a more likely mechanism of Tl toxicity than oxidative stress in rainbow trout but is an endpoint of relevance only at elevated environmental Tl concentrations. Environ Toxicol Chem 2024;43:87-96. © 2023 SETAC.