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Fish gills are complex organs that have direct contact with the environment and perform numerous functions including gas exchange and ion regulation. Determining if gill morphometry can change under different environmental conditions to maintain and/or improve gas exchange and ion regulation is important for understanding if gill plasticity can improve survival with increasing environmental change. We assessed gill morphology (gas exchange and ion regulation metrics), hematocrit and gill Na+/K+ ATPase activity of wild-captured blackside darter (Percina maculata), greenside darter (Etheostoma blennioides), and johnny darter (Etheostoma nigrum) at two temperatures (10 and 25 °C) and turbidity levels (8 and 94 NTU). Samples were collected August and October 2020 in the Grand River to assess temperature differences, and August 2020 in the Thames River to assess turbidity differences. Significant effects of temperature and/or turbidity only impacted ionocyte number, lamellae width, and hematocrit. An increase in temperature decreased ionocyte number while an increase in turbidity increased lamellae width. Hematocrit had a species-specific response for both temperature and turbidity. Findings suggest that the three darter species have limited plasticity in gill morphology, with no observed compensatory changes in hematocrit or Na+/K+ ATPase activity to maintain homeostasis under the different environmental conditions. [Display omitted] • Temperature and turbidity caused minimal changes in gill morphology. • There are species-specific differences in hematocrit. • Closely related species have differences in gill morphology. [ABSTRACT FROM AUTHOR]

Production of sterile fishes through artificial retention of a third set of chromosomes (triploidy) is a sustainable alternative for aquaculture since it reduces escapee pressure on wild populations. However, these fishes have reduced survival in stressful conditions and in response to infection. In this study, the impact of Vibrio anguillarum infection on diploid and triploid Chinook salmon (Oncorhynchus tshawytscha) was investigated to identify if there was any significant immune regulation by microRNAs (miRNA). Small RNAs from hindgut, head kidney, and spleen were sequenced to determine if miRNA transcript abundance was altered due to ploidy and infection in nine-month old full-sibling diploids and triploids. All three tissues had differentially expressed miRNA prior to infection, indicating subtle changes in epigenetic regulation due to increased ploidy. Additionally, miRNA were altered by infection, but there was only a difference in spleen miRNA expression between diploids and triploids at three days of infection. Furthermore, one miRNA (ssa-miR-2188-3p) was confirmed as having an altered response to infection in triploids compared to diploids, implicating potential immune dysregulation due to increased ploidy. The miRNAs identified in this study are predicted to target immune pathways, providing evidence for their importance in regulating responses to pathogens. This study is the first to investigate how increased ploidy alters miRNA expression in response to infection. Additionally, it provides evidence for epigenetic dysregulation in triploid fishes, which may contribute to their poor performance in response to stress. [Display omitted] • MicroRNA have altered expression due to Vibrio infection in Chinook salmon. • Triploid Chinook have altered miRNA expression in hindgut, head kidney, and spleen. • Spleen microRNA profile is different between diploids and triploids after infection. • miR-2188-3p responds differently to infection in diploids and triploids. • microRNA identified in this study are predicted to regulate immune function. [ABSTRACT FROM AUTHOR]

Oxygen availability varies among aquatic environments, and oxygen concentration has been demonstrated to drive behavioral, metabolic, and genetic adaptations in numerous aquatic species. MicroRNAs (miRNAs) are epigenetic modulators that act at the interface of the environment and the transcriptome and are known to drive plastic responses following environmental stressors. An area of miRNA that has remained underexplored is the sex specific action of miRNAs following hypoxia exposure and its effects as gene expression regulator in fishes. This study aimed to identify differences in mRNA and miRNA expression in the F 1 generation of zebrafish (Danio rerio) at 1 hpf after either F 0 parental male or female were exposed to 2 weeks of continuous (45 %) hypoxia. In general, F 1 embryos at 1 hpf demonstrated differences in mRNA and miRNAs expression related to the stressor and to the specific sex of the F 0 that was exposed to hypoxia. Bioinformatic pathway analysis of predicted miRNA:mRNA relationships indicated responses in known hypoxia signaling and mitochondrial bioenergetic pathways. This research demonstrates the importance of examining the specific male and female contributions to phenotypic variation in subsequent generations and provides evidence that there is both maternal and paternal contribution of miRNA through eggs and sperm. Male and female zebrafish contributions to phenotypic variation in a subsequent generation, following hypoxia exposure, shows that there is both maternal and paternal contribution of miRNA through eggs and sperm. [Display omitted] • Male and female F 0 distinct tolerance to hypoxia influences F 1 microRNA expression. • F 0 male hypoxia exposure increases F 1 hif-1α expression. • There is both maternal and paternal contribution of miRNA through eggs and sperm. • Bioinformatic predicts hypoxia signaling and mitochondrial bioenergetic pathway. [ABSTRACT FROM AUTHOR]

AMP-activated protein kinase is an enzyme that mediates communication between cellular energy status and diverse effector proteins, particularly those that play roles in determining the metabolic phenotype. By phosphorylating metabolic enzymes, transcriptional regulators and proteins involved in cellular structure, it can modify energy metabolism in both the short term and long term. Its basic features are highly conserved, with homologues in all eukaryotes. Gene and/or genome duplications endowed early vertebrates with paralogs of AMPK subunits, though the nature of their subfunctionalization remains uncertain, even in mammals. While most research focuses on the role of the enzyme in human health, a great deal can be learned from comparative studies targeting non-traditional model animals. Fish, in particular, are interesting models because of the diversity in the metabolic properties and complex relationships between metabolism and environmental challenges. In this review, we examine what is known about AMPK structure and function though the lens of comparative physiology, looking for opportunities to better understand how this vital energy sensor has evolved in animals. [ABSTRACT FROM AUTHOR]

Wastewater effluent is a metabolic stressor to aquatic organisms, though the mechanisms regulating metabolic rate in fish are not fully understood. Changes in metabolism may be regulated by microRNA (miRNA), small RNA molecules that post-transcriptionally regulate target mRNA translation in fish. Nuclear encoded miRNA are present in mammalian mitochondria where they regulate translation of mitochondrial genes, namely subunits for oxidative phosphorylation complexes; though this mechanism has not been identified in fish. This study aimed to identify if miRNA are present in darter (Etheostoma spp.) mitochondria, and if the metabolic stress occurring in darters in the Grand River, Waterloo, is partly regulated by miRNAs supressing translation of target mitochondrial genes. Three species of darters (E. caeruleum; E. nigrum; E. flabellare) were collected from upstream and downstream of the Waterloo wastewater treatment plant, and qPCR analysis confirmed the presence of four miRNA bioinformatically predicted to target mitochondrial mRNAs within the mitochondria, namely let-7a, miR-1, miR-122 and miR-20. E. caeruleum collected from downstream had lower cytochrome c oxidase activity, with a respective higher miR-1 abundance in the mitochondria, while E. nigrum had both a higher miR-20 abundance and cytochrome c oxidase activity downstream. E. flabellare was the only species that exhibited a lower miR-122 abundance downstream, despite no difference in cytochrome c oxidase activity between sites. Overall, this study confirmed the presence of miRNA within the mitochondria of daters, predicted a relationship between miR-1, and miR-20 abundance and cytochrome c oxidase activity, and identified one sex-specific miRNA, miR-20. [Display omitted] • MitomiRs let-7a, miR-1, miR-122 and miR-20 are present within fish mitochondria. • MitomiR miR-20 is sex-specific within rainbow and fantail darters. • MitomiRs may suppress translation of mitochondrial mRNA in darters. • Cytochrome c oxidase activity may be regulated by mitomiRs in darter livers. [ABSTRACT FROM AUTHOR]

Abstract: The role of methionine supplementation in fish metabolism remains largely unexplored. This study investigates the effects of methionine deprivation (MD) on glucose uptake in rainbow trout (Oncorhynchus mykiss) hepatocytes. To this end, primary hepatocytes were incubated in the presence (+M) or the absence (−M) of methionine for 48h and glucose uptake was assessed using a novel non-radioactive, fluorescent-linked enzymatic assay. Evidence indicated that glucose uptake increased under methionine deprivation, primarily due to the increased abundance of membrane bound sodium-glucose transporter 2 (SGLT2), which was likely facilitated by the cellular reduction in [ATP] resulting from increased mitochondrial uncoupling, as supported by elevated transcript levels of uncoupling protein 2a (UCP2a). This study is the first to suggest that the mechanisms responsible for the rapid glucose uptake associated with MR are facilitated by the greater abundance of SGLT2 glucose transporter and mitochondrial uncoupling. [Copyright &y& Elsevier]

Naphthenic acids (NA) are used in a variety of commercial and industrial applications, and are primary toxic components of oil sands wastewater. We investigated developmental and metabolic responses of tadpoles exposed to sub-lethal concentrations of a commercial NA blend throughout development. We exposed Lithobates pipiens tadpoles to 1 and 2 mg/L NA for 75 days and monitored growth and development, condition factor, gonad and liver sizes, and levels of liver glucose, glycogen, lipids and cholesterol following exposure. NA decreased growth and development, significantly reduced glycogen stores and increased triglycerides, indicating disruption to processes associated with energy metabolism and hepatic glycolysis. Effects on liver function may explain reduced growth and delayed development observed in this and previous studies. Our data highlight the need for greater understanding of the mechanisms leading to hepatotoxicity in NA-exposed organisms, and indicate that strict guidelines may be needed for the release of NA into aquatic environments. [Copyright &y& Elsevier]

The action of the immune response in zebrafish (Danio rerio) has been a target of many studies. However, the energetic demands involved in the immune response are poorly understood in ectothermic poikilotherms, such as fish. This research aims to characterize the energetic response of zebrafish to an immune challenge of heat-killed Vibrio anguillarum at 22 °C and 27.5 °C. Zebrafish were either not injected, injected intraperitoneally with 10 μl of saline and Freund's incomplete adjuvant (sham), or heat-killed Vibrio anguillarum & Freund's incomplete adjuvant (1.21 × 1010 cfu/ml). Respirometry was then performed on these zebrafish for a period of 27 h. Following this, spleen was collected for quantitative PCR analysis of the catalytic subunit of AMPK (ampka1 & ampka2), the nuclear factor kappa-light-chain-enhancer of activated B cells (nf-kb), and several cytokines (tnfa, il-1b, il-8, il-10). While there was no increase in oxygen consumption with any treatment at 22 °C, there was a marked 30% increase in oxygen consumption in zebrafish injected with heat-killed Vibrio at 27.5 °C. Furthermore, temperature had a strong effect on the timing of the immune response. At 22 °C, there was a 2–3-fold increase in the cytokines measured associated with heat-killed Vibrio injection, whereas there were no differences found at 27.5 °C. Furthermore, while there was an increase in ampka2 at 22 °C, there was a sharp decrease in ampka2 at 27.5 °C, although the changes in ampka2 transcript abundance could not be solely attributed to heat-killed Vibrio , as there were similar changes associated with the sham group. The results of this study demonstrate some of the first evidence that zebrafish increase routine metabolic rate associated with immune stimulation. Unlabelled Image • There was a 30% increase in routine metabolic rate associated with heat-killed Vibrio anguillarum in zebrafish at 27.5oC • Temperature appeared to play a significant role in the timing of transcriptional expression differences between 22oC and 27.5oC • While heat-killed Vibrio stimulated a response, likely a live pathogen exposure would have been more robust. • This is some of the first evidence that indicates there is a metabolic cost associated with infection in teleost fish. [ABSTRACT FROM AUTHOR]

The hypothalamic-pituitary-interrenal axis is an important regulator of stress and metabolism in teleosts. Cortisol is secreted by the head kidney where it increases gluconeogenesis in the liver to increase circulating glucose levels. MicroRNAs (miRNAs) are small, non-coding RNA molecules that bind to the 3′ untranslated region of specific mRNA to regulate their expression. MicroRNAs can also be secreted into circulation by association with extracellular vesicles (EVs) where they can influence the phenotype of other tissues. In this study, adult rainbow trout were exposed to a 3-minute acute air stress and allowed to recover for 1-, 3-, or 24-h to determine how miRNAs were altered. MicroRNAs measured in this study were chosen based on their high relative abundance in tissues that drive the stress response (miR-21a-3p, let-7a-5p, miR-143-3p) or their role in regulating DNA methylation (miR-29a-3p). In general, miRNAs increased in circulating EVs during the recovery period while decreasing in head kidney and liver at the same timepoints. Predicted targets for these miRNAs were analyzed using KEGG and DAVID functional enrichment analysis. Pathways involved in metabolism and cell signaling were predicted to be upregulated. Future studies can use these results to investigate how pathways are regulated after stress. Overall, our results indicate that miRNAs are regulated during teleost stress responses and could be supporting the cortisol-mediated changes that occur. Unlabelled Image • Rainbow trout were exposed to a 3 min air stressor and recovered for 1, 3 or 24 h. • Specific microRNAs were increased in extracellular vesicles. • Specific microRNAs were decreased in head kidney and liver. • These microRNAs are predicted to target pathways associated with stress. [ABSTRACT FROM AUTHOR]

MicroRNA (miRNA) are short, non-coding RNA that act by downregulating targeted mRNA transcripts. Only recently have they been used as endpoints in studies of aquatic toxicology. The purpose of this study was to determine the effect of an antidepressant contaminant, venlafaxine (VFX), and increased temperature on specific microRNA levels in zebrafish (Danio rerio) reproductive tissue. Adult zebrafish were exposed to one of four conditions; control, 1 μg/L VFX (VFX), 32 °C (Temp), or 1 μg/L VFX + 32 °C (VFX & Temp) for 21 days. Half of the fish were returned to control conditions for a 21-day recovery period. RT-qPCR was performed to measure relative abundances of several miRNAs known to respond to antidepressant exposure: dre-miR-22b-3p, dre-miR-301a, dre-miR-140-5p, dre-let-7d-5p, dre-miR-210-5p, and dre-miR-457b-5p. After the exposure period, dre-miR-22b-3p and dre-miR-301a showed a significant downregulation in response to all treatments. In contrast, after the recovery period, there were no significant differences in microRNA abundance. These altered microRNA are predicted to target several genes, including phosphofructokinase, and are associated with ovarian pathologies. Combined, we have shown that VFX and increased water temperature alter miRNA abundances in zebrafish reproductive tissue, an effect correlated with a functional stress response and cell cycle dysregulation. Unlabelled Image • Fish stress due to pharmaceutical and climate change exposure needs to be measured. • MicroRNA are altered in response to increased water temperature and venlafaxine. • Altered microRNA are associated with ovarian pathologies and the stress response. • Specific microRNA can be used as biomarkers of environmental stress. [ABSTRACT FROM AUTHOR]

Abstract: The antibacterial properties of silver nanoparticles (AgNPs) are widely exploited in a variety of medical and consumer products. AgNPs in these products can be released into the aquatic environment, however, the potential toxicity of AgNPs to organisms, including fish, is yet to be fully understood. The present study aimed to investigate the effects of the early life exposure to AgNPs on the hypothalamic–pituitary–interrenal (HPI) axis-mediated stress response in zebrafish (Danio rerio) larvae and adults. Zebrafish embryos were treated with AgNPs (0.5μg/mL) or Ag+ (0.05μg/mL) starting at 2h post fertilization (hpf). At 96hpf the larvae were either subjected to a swirling stress and euthanized, or raised to adulthood (10months) in silver-free water and then net-stressed, euthanized, and sampled. Whole-body basal or stress-induced cortisol levels in larvae were not affected by either AgNPs or Ag+; however, the transcript levels of corticotropin releasing factor (CRF), CRF-binding protein (CRF-BP), CRF-receptor 2 (CRF-R2), and pro-opiomelanocortin (POMCb) were significantly decreased by Ag+. The ability of the adult fish to release cortisol in response to a stressor was also not affected, although the transcript levels of CRF, CRF-BP, and CRF-R1 in the telencephalon were differentially affected in fish exposed to Ag+ as embryos. This is the first study that investigated the potential endocrine-disrupting effects of AgNPs during the early life stages and although AgNPs or Ag+ did not affect the ability of zebrafish to elevate cortisol levels in response to a stressor, the effects on transcript levels by Ag+ should be investigated further since CRF does not solely regulate the HPI axis but is also implicated in other physiological processes. [Copyright &y& Elsevier]

The antidepressant, venlafaxine (VFX), and climate change stressors, such as increased water temperature and decreased dissolved oxygen, are current threats to aquatic environments. This study aimed to determine how microRNAs (miRNAs) and predicted targeted transcripts were altered in livers of zebrafish exposed to these stressors, and livers of their un-exposed F 1 and F 2 offspring. Following a 21 day exposure to multiple stressors (1 μg/L VFX, +5 °C ambient, 50% O 2), then a subsequent 21 day recovery, relative abundances of cyp3a65 , hsp70 , hsp90 , and ppargc1a and miRNAs predicted to target them (miR-142a, miR-16c, miR-181c, and miR-129, respectively) were measured in the liver via quantitative PCR (RT-qPCR). There were significant decreases in miR-142a in the exposed F 0 generation and the exposed F 1 generation. While there were no changes detected in cyp3a65 relative abundance, there was a significant inverse relationship between cyp3a65 and miR-142a. Hsp70 expression significantly increased in the F 1 generation, which persisted to the F 2 generation and the relative abundance of hsp90 significantly increased in all generations. There was a significant reduction in miR-181c in the F 1 generation, but there was no significant relationship between miR-181c and hsp90. Finally, there was a significant decrease in ppargc1a relative abundance in the F 1 generation which was associated with an increase in miR-129. Combined, these results suggest that parental exposure to multiple, environmentally relevant stressors can confer transcriptional and epigenetic responses in the F 1 and F 2 generations, although identifying which stressor is a driving force becomes unclear. Unlabelled Image • Adult zebrafish were exposed to 21 days of +5 °C, hypoxia (50%), and 1 μg/L venlafaxine. • Zebrafish were returned to control conditions and bred to F 1 and F 2 generations. • Inverse relationships were found between miR-142a and cyp3a65 & miR-129 and ppargc1a. • Hsp70 and hsp90 were significantly increased in the F 1 and F 2 generations. • Multi-stressor exposure confers mRNA and microRNA changes in future generations. [ABSTRACT FROM AUTHOR]

Fish live in continuous contact with various stressors and antigenic material present within their environments. The impact of stressors associated with wastewater-exposed environments on fish has become of particular interest in toxicology studies. The objectives of this study were to examine potential effects of wastewater treatment plant (WWTP) effluent-associated stressors on innate cytokine expression within the gills of darter species (Etheostoma spp.), using both field and laboratory approaches. Male and female darters (rainbow, greenside, fantail, and johnny darters) were collected upstream and downstream of the Waterloo WWTP in the Grand River, Ontario. Gill samples were collected from fish in the field and from a second subset of fish brought back to the laboratory. Laboratory fish were acutely exposed (96-h) to an environmentally relevant concentration of venlafaxine (1.0 μg/L), a commonly prescribed antidepressant. To assess the impacts of these stressors on the innate immunity of darters, the expression of key innate cytokines was examined. Minor significant effects on innate cytokine expression were observed between upstream and downstream fish. Moderate effects on cytokine expression were observed in venlafaxine-exposed fish compared to their control counterparts however, changes were not indicative of a biologically significant immune response occurring due to the exposure. Although the results of this study did not display extensive impacts of effluent and pharmaceutical exposure on innate cytokine expression within the gills, they provide a novel avenue of study, illustrating the importance of examining potential impacts that effluent-associated stressors can have on fundamental immune responses of native fish species. [Display omitted] • WWTP effluent exposure had minor impacts on the expression of examined cytokines. • Venlafaxine exposure significantly impacted expression of several examined cytokines. • Changes in the expression of studied cytokines may not be biologically significant. • Provides important framework for future studies examining WWTP effluents and immunity. [ABSTRACT FROM AUTHOR]

The objective of this study was to assess the effects of municipal wastewater treatment plant effluent on the energetics and stress response of rainbow darter ( Etheostoma caeruleum ). Male and female rainbow darter were collected upstream and downstream of the Waterloo WWTP in the Grand River watershed, ON, Canada. To assess the effects of wastewater treatment plant effluent on whole-body and tissue specific metabolic capacity, closed-chamber respirometry and muscle-enzyme activity analyses were performed. Plasma cortisol was also collected from fish before and after an acute air-exposure stressor to evaluate the cortisol stress response in fish exposed to additional stressors. Male and female rainbow darter collected downstream of the effluent had higher oxygen consumption rates, while differences in enzyme activities were primarily associated with sex rather than collection site. No impairment in the cortisol stress response between downstream and upstream fish was observed, however baseline cortisol levels in female fish from the downstream site were significantly higher compared to other baseline groups. Stress-induced cortisol levels were also higher in female fish from both sites when compared to their male counterparts. Overall, this study demonstrates that chronic exposure to WWTP effluent impacts whole-body metabolic performance. This study was also able to demonstrate that sex-differences are a key determinant of various metabolic changes in response to physiological stress, thereby, providing a novel avenue to be considered and further explored. [ABSTRACT FROM AUTHOR]

While the field of epigenetics is increasingly recognized to contribute to the emergence of phenotypes in mammalian research models across different developmental and generational timescales, the comparative biology of epigenetics in the large and physiologically diverse vertebrate infraclass of teleost fish remains comparatively understudied. The cypriniform zebrafish and the salmoniform rainbow trout and Atlantic salmon represent two especially important teleost orders, because they offer the unique possibility to comparatively investigate the role of epigenetic regulation in 3R and 4R duplicated genomes. In addition to their sequenced genomes, these teleost species are well-characterized model species for development and physiology, and therefore allow for an investigation of the role of epigenetic modifications in the emergence of physiological phenotypes during an organism's lifespan and in subsequent generations. This review aims firstly to describe the evolution of the repertoire of genes involved in key molecular epigenetic pathways including histone modifications, DNA methylation and microRNAs in zebrafish, rainbow trout, and Atlantic salmon, and secondly, to discuss recent advances in research highlighting a role for molecular epigenetics in shaping physiological phenotypes in these and other teleost models. Finally, by discussing themes and current limitations of the emerging field of teleost epigenetics from both theoretical and technical points of view, we will highlight future research needs and discuss how epigenetics will not only help address basic research questions in comparative teleost physiology, but also inform translational research including aquaculture, aquatic toxicology, and human disease. [ABSTRACT FROM AUTHOR]

The Grand River watershed is the largest in southern Ontario and assimilates thirty wastewater treatment plants (WWTP) with varied degrees of treatment. Many WWTPs are unable to effectively eliminate several contaminants of emerging concern (CECs) from final effluent, leading to measurable concentrations in surface waters. Exposures to CECs have reported impacts on oxidative stress measured through antioxidative enzymes (SOD, CAT, GPX). This study focuses on the effects of WWTP effluent on four Etheostoma (Darter) species endemic to the Grand River, by investigating if increased antioxidative response markers are present in darter brains downstream from the effluent outfall compared to an upstream reference site relative to the Waterloo, ON WWTP across two separate years (Oct 2020 and Oct 2021). This was assessed using transcriptional and enzyme analysis of antioxidant enzymes and an enzyme involved in serotonin synthesis, tryptophan hydroxylase (tph). In fall 2020, significant differences in transcript markers were found between sites and sexes in GSD with SOD and CAT showing increased expression downstream, in JD with both sexes showing increased SOD downstream, and an interactive effect for tph in RBD. Changes in transcripts aligned with enzyme activity where interactive effects with sex-related differences were observed in fish collected fall 2020. In contrast, transcripts measured in fall 2021 were increased upstream compared to downstream species in RBD and GSD. This study additionally displayed yearly, species and sex differences in antioxidant responses. Continued investigation on the impacts of CECs in effluent in non-target species is required to better understand WWTP effluent impacts. [Display omitted] • Variability in antioxidant response in non-target species exposed to effluent • Antioxidant sex-differences are observed in darters following effluent exposure. • Influence of yearly variation on the antioxidant response • Effluent tolerance varies between species indicated by morphological differences. [ABSTRACT FROM AUTHOR]

The acute stress response is well-characterized, with rainbow trout as a teleost model for physiological and molecular responses. Air exposure, which stimulates an acute stress response, modulates liver microRNAs in rainbow trout; however, these highly conserved non-coding RNAs that bind to mRNA and repress translation, have never been measured in brook trout and it is unknown how miRNA expression responds following air exposure in this less studied salmonid. Our objective was to characterize the effects of air exposure on rainbow and brook trout liver miRNA expression, as well as the mRNA expression and enzyme activity that the miRNAs are predicted to target. Brook and rainbow trout were sampled pre- and 1-, 3-, and 24-h post- a three-minute air exposure. Plasma cortisol, glucose, and lactate were measured. Relative expression of miR-21a-5p, miR-143-3p, let-7a-5p and relative expression and enzyme activities of five predicted targets (pyruvate kinase, glucokinase, citrate synthase, cytochrome c oxidase, and catalase) were measured in liver. Rainbow and brook trout both had increases in plasma cortisol and lactate, while only rainbow trout had significant post-stress increases in plasma glucose. Furthermore, both trout species had increased miR-143-3p and miR-21a-5p relative expression 24-h post-stress. Four of the five enzymes measured had altered activity following stress. Brook trout miRNAs had inverse relative expression with relative catalase mRNA expression and cytochrome c oxidase enzyme activity, but no relationship was found in rainbow trout. Therefore, we have further characterized the transcriptional and enzymatic response to air exposure in two salmonids. [Display omitted] • Characterized rainbow and brook trout response to 3-min air exposure. • Plasma metabolites increased post-air exposure with some variability. • Inverse relationships between miRNAs and predicted targets in brook trout. • Altered liver miRNAs and enzyme activities are linked to oxidative stress. [ABSTRACT FROM AUTHOR]