Hypoxia Tolerance In Fish Research Articles

Fish models to explore epigenetic determinants of hypoxia-tolerance.

The occurrence of environmental hypoxia in freshwater and marine aquatic systems has increased over the last century and is predicted to further increase with climate change. As members of the largest extant vertebrate group, freshwater fishes, and to a much lesser extent marine fishes, are vulnerable to increased occurrence of hypoxia. This is important as fishes render important ecosystem services and have important cultural and economic roles. Evolutionarily successful, fishes have adapted to diverse aquatic freshwater and marine habitats with different oxygen conditions. While some fishes exhibit genetic adaptions to tolerate hypoxia and even anoxia, others are limited to oxygen-rich habitats. Recent advances in molecular epigenetics have shown that some epigenetic machinery, especially histone- and DNA demethylases, is directly dependent on oxygen and modulates important transcription-regulating epigenetic marks in the process. At the post-transcriptional level, hypoxia has been shown to affect non-coding microRNA abundance. Together, this evidence adds a new molecular epigenetic basis to study hypoxia tolerance in fishes. Here, we review the documented and predicted changes in environmental hypoxia in aquatic systems and discuss the diversity and comparative physiology of hypoxia tolerance in fishes, including molecular and physiological adaptations. We then discuss how recent mechanistic advances in environmental epigenetics can inform future work probing the role of oxygen-dependent epigenetic marks in shaping organismal hypoxia-tolerance in fishes with a focus on within- and between-species variation, acclimation, inter- and multigenerational plasticity, and multiple climate-change stressors. We conclude by describing the translational potential of this approach for conservation physiology, ecotoxicology, and aquaculture.

Whether hypoxia tolerance improved after short-term fasting is closely related to phylogeny but not to foraging mode in freshwater fish species.

The combined stresses of fasting and hypoxia are common events during the life history of freshwater fish species. Hypoxia tolerance is vital for survival in aquatic environments, which requires organisms to down-regulate their maintenance energetic expenditure while simultaneously preserving physiological features such as oxygen supply capacity under conditions of food deprivation. Generally, infrequent-feeding species who commonly experience food shortages might evolve more adaptive strategies to cope with food deprivation than frequent-feeding species. Thus, the present study aimed to test whether the response of hypoxia tolerance in fish to short-term fasting (2 weeks) varied with different foraging modes. Fasting resulted in similar decreases in maintenance energetic expenditure and similar decreases in Pcrit and Ploe between fishes with different foraging modes, whereas it resulted in decreased oxygen supply capacity only in frequent-feeding fishes. Furthermore, independent of foraging mode, fasting decreased Pcrit and Ploe in all Cypriniformes and Siluriformes species but not in Perciformes species. The mechanism for decreased Pcrit and Ploe in Cypriniformes and Siluriformes species is at least partially due to the downregulated metabolic demand and/or the maintenance of a high oxygen supply capacity while fasting. The present study found that the effect of fasting on hypoxia tolerance depends upon phylogeny in freshwater fish species. The information acquired in the present study is highly valuable in aquaculture industries and can be used for species conservation in the field.

Evolution of Key Oxygen-Sensing Genes Is Associated with Hypoxia Tolerance in Fishes.

Low dissolved oxygen (hypoxia) is recognized as a major threat to aquatic ecosystems worldwide. Because oxygen is paramount for the energy metabolism of animals, understanding the functional and genetic drivers of whole-animal hypoxia tolerance is critical to predicting the impacts of aquatic hypoxia. In this study, we investigate the molecular evolution of key genes involved in the detection of and response to hypoxia in ray-finned fishes: the prolyl hydroxylase domain (PHD)-hypoxia-inducible factor (HIF) oxygen-sensing system, also known as the EGLN (egg-laying nine)-HIF oxygen-sensing system. We searched fish genomes for HIFA and EGLN genes, discovered new paralogs from both gene families, and analyzed protein-coding sites under positive selection. The physicochemical properties of these positively selected amino acid sites were summarized using linear discriminants for each gene. We employed phylogenetic generalized least squares to assess the relationship between these linear discriminants for each HIFA and EGLN and hypoxia tolerance as reflected by the critical oxygen tension (Pcrit) of the corresponding species. Our results demonstrate that Pcrit in ray-finned fishes correlates with the physicochemical variation of positively selected sites in specific HIFA and EGLN genes. For HIF2A, two linear discriminants captured more than 90% of the physicochemical variation of these sites and explained between 20% and 39% of the variation in Pcrit. Thus, variation in HIF2A among fishes may contribute to their capacity to cope with aquatic hypoxia, similar to its proposed role in conferring tolerance to high-altitude hypoxia in certain lineages of terrestrial vertebrates.

Effect of Scoparia dulcis Extract on Lipid Oxidation in Fish Feed, Growth Performance, and Hypoxia Tolerance in Juvenile Jian Carp (Cyprinus carpio var. Jian).

Lipid oxidation and hypoxia can lead to oxidative damage in aquatic animals. This study explored the effects of Scoparia dulcis extracts (SDE) on lipid oxidation, fish growth performance, digestive ability, antioxidant capacity, and hypoxia tolerance ability. The results showed that SDE decreased malonaldehyde (MDA), conjugated diene (CD), and peroxide value (PO) in the linoleic acid and linolenic acid as well as in fish feed. Broken-line analysis revealed that the optimal acetone extract of S. dulcis (AE) supplements was 4.02, 4.01, and 4.01 g kg-1 determined from PO, CD, and MDA, respectively. Dietary AE supplementation increased feed intake and specific growth rate and activities of amylase, trypsin, and lipase as well as alkaline phosphatase in fish hepatopancreas and gut. Polynomial regression analysis showed that optimal dietary AE supplement was 3.61 g kg-1 diet determined from weight gain. Furthermore, dietary AE supplementation decreased MDA content and increased glutathione content and the activities of glutathione peroxidase, catalase, superoxide dismutase, and glutathione reductase in fish digestive organs, gills, erythrocytes, and muscle. Dietary AE supplementation increased durative time (DT) and oxygen consumption rate (OCR) under hypoxia condition. Based on polynomial regression analysis, optimal dietary AE supplements were 4.73 and 4.60 g kg-1 diet determined from DT and OCR for hypoxia tolerance in fish, respectively. According to our current research, SDE's antioxidant capacity may be attribute to their phenolic chemicals.

Comparative analysis of glucose and fructose tolerance in two marine fishes: effects on insulin secretion and acute hypoxia tolerance

Carbohydrates are a common and economical energy source in animal feeds. However, most fish show a persistent postprandial hyperglycemia after intake of a high-carbohydrate diet. Unfortunately, the mechanism of glucose metabolism in fish is still unclear. In the present study, tiger puffer (Takifugu rubripes) and turbot (Scophthalmus maximus) were intraperitoneally injected or orally administered with glucose or fructose (500 mg/kg body weight) to evaluate the ability of fish to utilize carbohydrates. Afterwards, serum glucose, fructose, pyruvate, insulin levels, and acute hypoxia tolerance were measured. Our results showed increased serum glucose level and then decreased post intraperitoneal injection with glucose, and reached a peak after 0.5 hours in turbot and 1 hour in tiger puffer. Tiger puffer had significantly lower liver glycogen, serum glucose, fructose, pyruvate, and insulin contents than turbot. Glucose and fructose only induced insulin secretion in turbot, but did not change serum insulin level in tiger puffer. Glucose was a stronger stimulator of insulin than fructose in the two marine species. Both intraperitoneal injection and oral fructose intake increased serum glucose level, while intraperitoneal or oral glucose also increased serum fructose level. Intraperitoneal injection of glucose promoted absorption and utilization of glucose in the blood more effectively than oral glucose intake. In addition, turbot and tiger puffer were intolerant to acute hypoxia, whereas supplementation with glucose or fructose improved hypoxia tolerance in the two marine fishes by activating anaerobic glycolysis. Taken together, our results provide important scientific information for understanding the mechanism for glucose and fructose utilization and improving hypoxia tolerance in fish.

Loss of one copy of vhl in zebrafish facilitates hypoxia tolerance

Insufficient oxygen in the water (hypoxia) is one of the limiting factors for high-density culture in modern aquaculture. Hypoxia tolerance in fish has been reported to be related to the hypoxia-inducible factor (HIF)-α-mediated hypoxia signaling pathway. The well-defined function of the von Hippel-Lindau (VHL) tumor suppressor gene VHL is to mediate proteasomal degradation of hydroxylated HIF-α proteins, resulting in the downregulation of hypoxia signaling pathway. However, whether VHL has an impact on hypoxia tolerance is still elusive. In this study, we find that vhl+/- zebrafish have no gross defects in development, growth, reproduction, and movement. In addition, vhl+/- zebrafish also have normal vasculature. However, under hypoxia, vhl+/- zebrafish are more resistant to hypoxia treatment compared with their wildtype siblings. Further assays indicate that the number of erythrocytes is increased and the expression of hypoxia-responsive genes is enhanced in vhl+/- zebrafish compared with their wildtype siblings. These data indicate that loss of one copy of vhl in zebrafish facilitates hypoxia tolerance, revealing a function of vhl in hypoxia tolerance.

Genome-wide association and transcriptome analysis provide the SNPs and molecular insights into the hypoxia tolerance in large yellow croaker (Larimichthys crocea)

The large yellow croaker (Larimichthys crocea) is a commercially important marine fish indigenous to the southeastern coast of China. Cultured L. crocea is frequently subjected to hypoxia which causes huge economic losses to the aquaculture industry. Therefore, it is crucial to elucidate the molecular mechanisms of hypoxia tolerance in L. crocea. Here, we combined a genome-wide association study (GWAS) and transcriptome analysis to reveal single-nucleotide polymorphisms (SNPs) and the mechanisms by which they regulate hypoxia tolerance in L. crocea. We used liquid chips to genotype 400 L. crocea individuals, obtained 120,815 high-quality SNPs, identified seven SNPs with significant suggestive association levels, and found a significant peak on chromosome 1. We then combined the potential candidate genes obtained from GWAS with the differentially expressed genes (DEGs) obtained by transcriptome sequencing. We found that the genes and processes related to glucose transport and metabolism, erythropoiesis, ion regulation, DNA replication and repair, and cell cycle were critical to hypoxia adaptation in L. crocea. We also identified the gene lias that might regulate hypoxia-inducible factor (HIF)-1α. The results of the present study provided insight into the molecular mechanisms of hypoxia tolerance in fish and may facilitate the genetic selection and breeding of hypoxia tolerance in L. crocea.

Cardiac Hypoxia Tolerance in Fish: From Functional Responses to Cell Signals.

Aquatic animals are increasingly challenged by O2 fluctuations as a result of global warming, as well as eutrophication processes. Teleost fish show important species-specific adaptability to O2 deprivation, moving from intolerance to a full tolerance of hypoxia and even anoxia. An example is provided by members of Cyprinidae which includes species that are amongst the most tolerant hypoxia/anoxia teleosts. Living at low water O2 requires the mandatory preservation of the cardiac function to support the metabolic and hemodynamic requirements of organ and tissues which sustain whole organism performance. A number of orchestrated events, from metabolism to behavior, converge to shape the heart response to the restricted availability of the gas, also limiting the potential damages for cells and tissues. In cyprinids, the heart is extraordinarily able to activate peculiar strategies of functional preservation. Accordingly, by using these teleosts as models of tolerance to low O2, we will synthesize and discuss literature data to describe the functional changes, and the major molecular events that allow the heart of these fish to sustain adaptability to O2 deprivation. By crossing the boundaries of basic research and environmental physiology, this information may be of interest also in a translational perspective, and in the context of conservative physiology, in which the output of the research is applicable to environmental management and decision making.

Effects of acute hypoxia on nutrient metabolism and physiological function in turbot, Scophthalmus maximus.

Acute hypoxia is a common stress in aquaculture, and causes energy deficiency, oxidative damage and death in fish. Many studies have confirmed that acute hypoxia activated hif1α expression, anaerobic glycolysis and antioxidant system in fish, but the effects of acute hypoxia on lipid and protein metabolism, organelle damage, and the functions of hif2α and hif3α in economic fishes have not been well evaluated. In the present study, turbot was exposed to acute hypoxia (2.0 ± 0.5mg/L) for 6h, 12h, and 24h, respectively. Then, the contents of hemoglobin (HB), metabolite, gene expressions of hifα isoforms, energy homeostasis, endoplasmic reticulum (ER) stress, and apoptosis were measured. The results suggested that turbot is intolerant to acute hypoxia and the asphyxiation point is about 1.5mg/L. Acute hypoxia induced perk-mediated ER stress, and increased lipid peroxidation and liver injury in turbot. The blood HB level and liver vegfab expression were increased under hypoxia, which enhances oxygen transport. At hypoxia stress, hif3α, anaerobic glycolysis-related genes expression, and lactate content were increased in the liver, and glycogen was broken down to ensure ATP supply. Meanwhile, hif2α, lipid synthesis-related genes expression, and TG content were increased in the liver, but the lipid catabolism and protein synthesis were suppressed during hypoxia, which reduced the oxygen consumption and ROS generation. Our results systematically illustrate the metabolic and physiological changes under acute hypoxia in turbot, and provide important guidance to improve hypoxia tolerance in fish.

Hypoxia tolerance in fish depends on catabolic preference between lipids and carbohydrates.

Hypoxia is a common environmental stress factor in aquatic organisms, which varies among fish species. However, the mechanisms underlying the ability of fish species to tolerate hypoxia are not well known. Here, we showed that hypoxia response in different fish species was affected by lipid catabolism and preference for lipid or carbohydrate energy sources. Activation of biochemical lipid catabolism through peroxisome proliferator-activated receptor alpha (Pparα) or increasing mitochondrial fat oxidation in tilapia decreased tolerance to acute hypoxia by increasing oxygen consumption and oxidative damage and reducing carbohydrate catabolism as an energy source. Conversely, lipid catabolism inhibition by suppressing entry of lipids into mitochondria in tilapia or individually knocking out three key genes of lipid catabolism in zebrafish increased tolerance to acute hypoxia by decreasing oxygen consumption and oxidative damage and promoting carbohydrate catabolism. However, anaerobic glycolysis suppression eliminated lipid catabolism inhibition-promoted hypoxia tolerance in adipose triglyceride lipase (atgl) mutant zebrafish. Using 14 fish species with different trophic levels and taxonomic status, the fish preferentially using lipids for energy were more intolerant to acute hypoxia than those preferentially using carbohydrates. Our study shows that hypoxia tolerance in fish depends on catabolic preference for lipids or carbohydrates, which can be modified by regulating lipid catabolism.

Body mass and cell size shape the tolerance of fishes to low oxygen in a temperature-dependent manner.

Aerobic metabolism generates 15–20 times more energy (ATP) than anaerobic metabolism, which is crucial in maintaining energy budgets in animals, fueling metabolism, activity, growth and reproduction. For ectothermic water‐breathers such as fishes, low dissolved oxygen may limit oxygen uptake and hence aerobic metabolism. Here, we assess, within a phylogenetic context, how abiotic and biotic drivers explain the variation in hypoxia tolerance observed in fishes. To do so, we assembled a database of hypoxia tolerance, measured as critical oxygen tensions (P crit) for 195 fish species. Overall, we found that hypoxia tolerance has a clear phylogenetic signal and is further modulated by temperature, body mass, cell size, salinity and metabolic rate. Marine fishes were more susceptible to hypoxia than freshwater fishes. This pattern is consistent with greater fluctuations in oxygen and temperature in freshwater habitats. Fishes with higher oxygen requirements (e.g. a high metabolic rate relative to body mass) also were more susceptible to hypoxia. We also found evidence that hypoxia and warming can act synergistically, as hypoxia tolerance was generally lower in warmer waters. However, we found significant interactions between temperature and the body and cell size of a fish. Constraints in oxygen uptake related to cellular surface area to volume ratios and effects of viscosity on the thickness of the boundary layers enveloping the gills could explain these thermal dependencies. The lower hypoxia tolerance in warmer waters was particularly pronounced for fishes with larger bodies and larger cell sizes. Previous studies have found a wide diversity in the direction and strength of relationships between P crit and body mass. By including interactions with temperature, our study may help resolve these divergent findings, explaining the size dependency of hypoxia tolerance in fish.

Acclimation to warmer temperature reversibly improves high-temperature hypoxia tolerance in both diploid and triploid brook charr, Salvelinus fontinalis

Rising temperature leads to reduced oxygen solubility and therefore increases the risk of exposure to harmful hypoxic condition for fish in their natural aquatic environments and in aquaculture. The goal of this study was to determine whether acclimation to warmer temperature can improve high-temperature hypoxia tolerance in fish, using sibling diploid and triploid brook charr as the experimental model. Triploid fish are used for aquaculture and fisheries management because they are sterile, but they are known to have reduced thermal and hypoxia tolerance compared to conventional diploids. Fish were pre-acclimated to either 15 °C (optimum temperature for diploids) or 18 °C and then assessed for high-temperature hypoxia tolerance by rapidly increasing temperature to pre-determined levels (up to 30 °C), holding fish at these temperatures for one hour, and then using compressed nitrogen to drive oxygen out of the water. Hypoxia tolerance was expressed as both the oxygen tension at loss of equilibrium and the time taken to reach this endpoint following the start of the trial. Acclimation to 18 °C improved hypoxia tolerance at high temperatures but this advantage was lost after reacclimation to 15 °C. Although 18 °C acclimation improved the hypoxia tolerance of triploids, it remained inferior to that of diploids under identical test conditions. Somatic energy reserves (estimated as condition factor and hepatosomatic index), cardiac output (relative ventricular mass) and oxygen carrying capacity of the blood (hemoglobin concentration and hematocrit) did not markedly affect high-temperature hypoxia tolerance.

Coming up for air

Coming up for air

Cardiac Transcriptomics Reveals That MAPK Pathway Plays an Important Role in Hypoxia Tolerance in Bighead Carp (Hypophthalmichthys nobilis).

Simple SummaryAcute hypoxia treatment was performed in juvenile bighead carp (Hypophthalmicthys nobilis) by decreasing water O2. The results showed that blood lactate and serum glucose increased significantly under hypoxia stress, and some differentially expressed genes were identified among hypoxia tolerant, hypoxia sensitive, and normoxia control groups. Differentially expressed genes between hypoxia tolerant and hypoxia sensitive groups were mainly involved in mitogen-activated protein kinase (MAPK) signaling, insulin signaling, apoptosis, tight junction, and adrenergic signaling in cardiomyocytes pathways, of which MAPK signaling pathway played a key role in cardiac tolerance to hypoxia in bighead carp. These results provide a basis for understanding the physiological and molecular mechanisms underlying hypoxia response in fish and a guide for future genetic breeding programs for hypoxia resistance in bighead carp.As aquatic animals, fishes often encounter various situations of low oxygen, and they have evolved the ability to respond to hypoxia stress. Studies of physiological and molecular responses to hypoxia stress are essential to clarify genetic mechanisms underlying hypoxia tolerance in fish. In this study, we performed acute hypoxia treatment in juvenile bighead carp (Hypophthalmicthys nobilis) by decreasing water O2 from 6.5 mg/L to 0.5 mg/L in three hours. This hypoxia stress resulted in a significant increase in blood lactate and serum glucose. Comparisons of heart transcriptome among hypoxia tolerant (HT), hypoxia sensitive (HS), and normoxia control (NC) groups showed that 820, 273, and 301 differentially expressed genes (DEGs) were identified in HS vs. HT, NC vs. HS, and NC vs. HT (false discovery rate (FDR) < 0.01, Fold Change> 2), respectively. KEGG pathway enrichment showed that DEGs between HS and HT groups were mainly involved in mitogen-activated protein kinase (MAPK) signaling, insulin signaling, apoptosis, tight junction and adrenergic signaling in cardiomyocytes pathways, and DEGs in MAPK signaling pathway played a key role in cardiac tolerance to hypoxia. Combined with the results of our previous cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis of hypoxia stress in this species, such genes as stbp2, ttn, mapk, kcnh, and tnfrsf were identified in both studies, representing the significance of these DEGs in hypoxia tolerance in bighead carp. These results provide insights into the understanding of genetic modulations for fish heart coping with hypoxia stress and generate basic resources for future breeding studies of hypoxia resistance in bighead carp.

Rising CO2 saves lives

Anthropogenic climate change is a multi-faceted problem. Rising CO2 levels in the atmosphere cause a warming of global temperatures, acidify the oceans and may ultimately deplete aquatic oxygen, leading to hypoxia. By itself, any one of these conditions can pose a real challenge to an animal, but how the stressors interact is still largely unexplored. Aquatic hypoxia may be especially intricate and, unlike the hypoxic conditions that one might experience when climbing a mountain, is almost always caused by organismal respiration in the water and, thus, is associated with an additional, simultaneous increase in CO2. The linkage between oxygen and CO2 levels in water has long been known, but how fish are affected by the combined challenge is poorly studied.To address the issue, Daniel Montgomery and his colleagues at the University of Exeter, UK, set out to test the effect of rising CO2 on the hypoxia tolerance of European seabass. They measured the oxygen consumption of the fish while lowering the available oxygen in the water; the point at which the animal can no longer extract oxygen from the depleted water and their oxygen consumption starts to decline is a popular indicator of hypoxia tolerance in fish. Knowing that higher levels of CO2 tend to acidify the blood of fish – compromising the ability of haemoglobin to bind oxygen and hindering the fish's ability to extract oxygen from the water – Montgomery then supplemented one of the treatments with a simultaneous increase in CO2, expecting that the combined challenge would render the fish more susceptible to hypoxia.However, contrary to these predictions, CO2-treated fish excelled at extracting oxygen from the water and outperformed the low-CO2 group in terms of hypoxia tolerance. When Montgomery took a closer look at the fish's blood, he found no difference in blood pH between the treatments, indicating a remarkable capacity of the seabass to counteract the acidifying effect of elevated CO2; surely this is good news for fish that must cope with global rises in CO2. In addition, fish in the rising CO2 treatment had a higher haemoglobin–oxygen affinity compared with the low-CO2 group – a surprising finding, given the common blood pH in the two groups – and this may explain their improved ability to tolerate hypoxia. Haemoglobin is housed within red blood cells, which may actively alter their internal environment to modulate oxygen transport. Whether this is part of the mechanism by which CO2-exposed seabass increased their haemoglobin–oxygen affinity remains to be seen.Studying the effects of climate change is complicated as numerous factors may interact to challenge an organism. Understandably, researchers often choose to study single factors in isolation, as this simplifies the interpretation of the data. However, Montgomery and colleagues show that overlooking interaction effects may limit our ability to predict the response of animals to climate change in the wild. In European seabass it appears that the combined condition of a decreasing oxygen level and an increasing CO2 level in the water, as is found in nature, improves hypoxia tolerance. To account for such effects in future work, current methods to assess hypoxia tolerance in fish should be revised to control for, or at least measure and report, CO2 levels in the water. Rising CO2 levels receive a lot of bad press and, in the context of climate change, rightfully so. However, at least for seabass that find themselves gasping for oxygen, the natural rise in CO2 that comes with aquatic hypoxia may actually save lives.

The utility and determination of Pcrit in fishes.

The critical O2 tension (Pcrit) is the lowest PO2 at which an animal can maintain some benchmark rate of O2 uptake (ṀO2 ). This PO2 has long served as a comparator of hypoxia tolerance in fishes and aquatic invertebrates, but its usefulness in this role, particularly when applied to fishes, has recently been questioned. We believe that Pcrit remains a useful comparator of hypoxia tolerance provided it is determined using the proper methods and hypoxia tolerance is clearly defined. Here, we review the available methods for each of the three steps of Pcrit determination: (1) measuring the most appropriate benchmark ṀO2 state for Pcrit determination (ṀO2,std, the ṀO2 required to support standard metabolic rate); (2) reducing water PO2 ; and (3) calculating Pcrit from the ṀO2 versus PO2 curve. We make suggestions on best practices for each step and for how to report Pcrit results to maximize their comparative value. We also discuss the concept of hypoxia tolerance and how Pcrit relates to a fish's overall hypoxia tolerance. When appropriate methods are used, Pcrit provides useful comparative physiological and ecological information about the aerobic contributions to a fish's hypoxic survival. When paired with other hypoxia-related physiological measurements (e.g. lactate accumulation, calorimetry-based measurements of metabolic depression, loss-of-equilibrium experiments), Pcrit contributes to a comprehensive understanding of how a fish combines aerobic metabolism, anaerobic metabolism and metabolic depression in an overall strategy for hypoxia tolerance.

Acute hypoxia/reoxygenation affects muscle mitochondrial respiration and redox state as well as swimming endurance in zebrafish.

Rapid fluctuations of the oxygen content of both natural and anthropogenic origin are relatively common in freshwater environments. Fish adaptation to these conditions implies tolerance of both low levels of oxygen availability and reoxygenation. Hypoxia tolerance in fish has been widely studied, but the involvement of mitochondria in the response of fish to rapid hypoxia/reoxygenation stress is less known. Zebrafish, a floodplain species, is likely facing significant changes in dissolved oxygen in its natural environment and displays a moderate ability to tolerate hypoxia. In the present study, we report the effects of an acute hypoxia/reoxygenation stress (H/R) protocol on mitochondrial functionality (respiration, complex activities, rate of H2O2 release) and redox state (level of HPs and protein oxidation) of muscle tissue. In parallel, the animal metabolic performance (routine metabolism, nitrogen excretion and swimming performance) was measured. Additionally, the recovery from H/R was tested 20h after treatment. A significant stimulation by H/R of muscle mitochondrial respiration and H2O2 release was observed, which was only in part counteracted by stimulation of the antioxidant system, resulting in an increased level of lipid peroxides and protein carbonyls. In parallel, H/R increased the animal oxygen consumption and urea excretion rate and reduced routine activity. A significant strong reduction of endurance at 80% Ucrit was also observed. Most of the altered parameter did not recover 20h after reoxygenation. These data indicate a significant alteration of zebrafish muscle mitochondrial state after acute H/R, associated with changes in tissue redox state and locomotor performance.

The effects of oil induced respiratory impairment on two indices of hypoxia tolerance in Atlantic croaker (Micropogonias undulatus)

The Gulf of Mexico was home to the Deepwater Horizon oil spill, and is also known to exhibit seasonal declines in oxygen availability. Oil exposure in fish is known to impact oxygen uptake through cardiac impairment, which raises questions about the additive effects of these two stressors. Here we explore this question on the Atlantic croaker using two measures of hypoxia tolerance: critical oxygen threshold (Pcrit), and time to loss of equilibrium (LOE). We first demonstrated that 24 h exposure to 10.1 and 23.2 μg l−1 ΣPAH50 significantly impaired oxygen uptake. There was no effect of exposure on Pcrit or LOE. Exposure did result in significantly different repeatability between pre- and post-exposure Pcrit, suggesting that hypoxia tolerant individual may see greater impacts following exposure. These results suggest oil exposure does not have wide scale detrimental outcomes for hypoxia tolerance in fish, yet there may be fine scale impairments of ecological significance.

Sub‐lethal effects on fish provide insight into a biologically‐relevant threshold of hypoxia

Hypoxia (low dissolved oxygen) is a mounting concern for aquatic ecosystems as its prevalence increases with rising anthropogenic nutrient inputs. Hypoxia is most commonly defined as 2.0 mg l–1 of dissolved oxygen, although this level varies widely across studies and agency regulations. Such definitions may be too conservative, as ecologically‐relevant non‐lethal effects (e.g. consumption and growth) of hypoxia on important aquatic species, such as fish, often occur at oxygen levels much higher than 2.0 mg l–1. In addition, many mechanisms that regulate hypoxia tolerance in fish have been proposed, including temperature, habitat, location in the water column, and body size, but there is ongoing debate over which mechanisms are most important. Using a structured meta‐analysis of published studies, we showed consistent, significant negative effects on fish growth and consumption below 4.5 mg l–1. While the total amount of variation explained was generally low, below 4.5 mg l–1 of dissolved oxygen, phylogenetic relationships accounted for most of the explained variation in fish growth. Ecological factors including body size, location in the water column (pelagic, demersal, or benthopelagic), habitat (freshwater, marine, or diadromous), and temperature explained very little of the effect of hypoxia on fish growth and explained only a moderate level of variation in consumption. Our results suggest a dramatically higher threshold for sub‐lethal effects of hypoxia on fish than oxygen levels generally set for regulation purposes, and provide little support for accepted ecological mechanisms thought to influence hypoxia tolerance.

A new analysis of hypoxia tolerance in fishes using a database of critical oxygen level (P crit).

Hypoxia is a common occurrence in aquatic habitats, and it is becoming an increasingly frequent and widespread environmental perturbation, primarily as the result of anthropogenic nutrient enrichment and climate change. An in-depth understanding of the hypoxia tolerance of fishes, and how this varies among individuals and species, is required to make accurate predictions of future ecological impacts and to provide better information for conservation and fisheries management. The critical oxygen level (P crit) has been widely used as a quantifiable trait of hypoxia tolerance. It is defined as the oxygen level below which the animal can no longer maintain a stable rate of oxygen uptake (oxyregulate) and uptake becomes dependent on ambient oxygen availability (the animal transitions to oxyconforming). A comprehensive database of P crit values, comprising 331 measurements from 96 published studies, covering 151 fish species from 58 families, provides the most extensive and up-to-date analysis of hypoxia tolerance in teleosts. Methodologies for determining P crit are critically examined to evaluate its usefulness as an indicator of hypoxia tolerance in fishes. Various abiotic and biotic factors that interact with hypoxia are analysed for their effect on P crit, including temperature, CO2, acidification, toxic metals and feeding. Salinity, temperature, body mass and routine metabolic rate were strongly correlated with P crit; 20% of variation in the P crit data set was explained by these four variables. An important methodological issue not previously considered is the inconsistent increase in partial pressure of CO2 within a closed respirometer during the measurement of P crit. Modelling suggests that the final partial pressure of CO2 reached can vary from 650 to 3500 µatm depending on the ambient pH and salinity, with potentially major effects on blood acid-base balance and P crit itself. This database will form part of a widely accessible repository of physiological trait data that will serve as a resource to facilitate future studies of fish ecology, conservation and management.