Partitioning of respiration between the gills and air-breathing organ in response to aquatic hypoxia and exercise in the pacific tarpon, Megalops cyprinoides

The evolution of air-breathing organs (ABOs) is associated not only with hypoxic environments but also with activity. This investigation examines the effects of hypoxia and exercise on the partitioning of aquatic and aerial oxygen uptake in the Pacific tarpon. The two-species cosmopolitan genus Megalops is unique among teleosts in using swim bladder ABOs in the pelagic marine environment. Small fish (58-620 g) were swum at two sustainable speeds in a circulating flume respirometer in which dissolved oxygen was controlled. For fish swimming at 0.11 m s(-1) in normoxia (Po2 = 21 kPa), there was practically no air breathing, and gill oxygen uptake was 1.53 mL kg(-0.67) min(-1). Air breathing occurred at 0.5 breaths min(-1) in hypoxia (8 kPa) at this speed, when the gills and ABOs accounted for 0.71 and 0.57 mL kg(-0.67) min(-1), respectively. At 0.22 m s(-1) in normoxia, breathing occurred at 0.1 breaths min(-1), and gill and ABO oxygen uptake were 2.08 and 0.08 mL kg(-0.67) min(-1), respectively. In hypoxia and 0.22 m s(-1), breathing increased to 0.6 breaths min(-1), and gill and ABO oxygen uptake were 1.39 and 1.28 mL kg(-0.67) min(-1), respectively. Aquatic hypoxia was therefore the primary stimulus for air breathing under the limited conditions of this study, but exercise augmented oxygen uptake by the ABOs, particularly in hypoxic water.     

Respiratory responses of the air-breathing fish Hoplosternum littorale to hypoxia and hydrogen sulfide

The present study analyzes the respiratory responses of the neotropical air-breathing fish Hoplosternum littorale to graded hypoxia and increased sulfide concentrations. The oxygen uptake (VO2), critical O2 tension (PcO2), respiratory (fR) and air-breathing (fRA) frequencies in response to graded hypoxia were determined for fish acclimated to 28 degrees C. H. littorale was able to maintain a constant VO2 down to a PcO2 of 50 mm Hg, below which fish became dependent on the environmental O2 even with significant increases in fR. The fRA was kept constant around 1 breath h(-1) above 50 mm Hg and increased significantly below 40 mm Hg, reaching maximum values (about 4.5 breaths h(-1)) at 10 mm Hg. The lethality to sulfide concentrations under normoxic and hypoxic conditions were also determined along with the fRA. For the normoxic fish the sulfide lethal limit was about 70 microM, while in the hypoxic ones this limit increased to 87 muM. The high sulfide tolerance of H. littorale may be attributed to the air-breathing capability, which is stimulated by this compound.     

The emersion response of the Australian Yabby Cherax destructor to environmental hypoxia and the respiratory and metabolic responses to consequent air-breathing

The Australian Yabby Cherax destructor voluntarily emerges from water to breathe air with increased frequency as water PO₂ decreases. When the water PO₂ declined below 2.7 kPa the crayfish spent >50% of time breathing air. The respiratory gas transport, acid-base, ionic and energetic status were quantified in simulations of this emersion behaviour to determine the benefits that the crayfish may gain from switching to air-breathing. C. destructor initially showed an elevated O₂ uptake rate on emerging from hypoxic water, but after 1 h the O₂ uptake rate was not different from that of crayfish in normoxic water. During 3 h of air breathing, subsequent to 2.7 kPa aquatic hypoxia, the haemolymph PO₂ increased while oxygen content was essentially unchanged, although cardiac output increased 5-fold. The haemolymph PCO₂ increased from 0.44 to 1.21 kPa after 3 h while the CO₂ content increased from 3.47 to 8.66 mmol · l⁻¹ and the pH decreased from 7.73 to 7.57 after 1 h in air. In air C. destructor eventually achieved an O₂ uptake rate similar to that achieved in water. A general hyperglycaemia occurred without anaerobiosis. In air-breathing C. destructor, small changes in lactate appear to offset the decrease in haemocyanin-O₂ affinity caused by acid Bohr shift. During air-breathing, decreased haemocyanin-O₂ affinity assisted in maintaining O₂ diffusion into the tissues, but the ATP content of the tail muscle decreased so that after 3 h in air the energy charge was only 0.59. The data are consistent with a specific depression of the Emden-Meyerhof pathway, preventing either lactate formation or oxidative phosphorylation in the tail muscle, despite a concomitant glycogenolysis. Accepted: 26 February 1998     

Cardiorespiratory responses of the facultative air-breathing fish jeju, Hoplerythrinus unitaeniatus (Teleostei, Erythrinidae), exposed to graded ambient hypoxia

The jeju, Hoplerythrinus unitaeniatus, is equipped with a modified part of the swim bladder that allows aerial respiration. On this background, we have evaluated its respiratory and cardiovascular responses to aquatic hypoxia. Its aquatic O2 uptake (V(O2)) was maintained constant down to a critical P(O2) (P(cO2)) of 40 mm Hg, below which V(O2) declined linearly with further reductions of P(iO2). Just below P(cO2), the ventilatory tidal volume (V(T)) increased significantly along with gill ventilation (V(G)), while respiratory frequency changed little. Consequently, water convection requirement (V(G)/V(O2)) increased steeply. The same threshold applied to cardiovascular responses that included reflex bradycardia and elevated arterial blood pressure (P(a)). Aerial respiration was initiated at water P(O2) of 44 mm Hg and breathing episodes and time at the surface increased linearly with more severe hypoxia. At the lowest water P(O2) (20 mm Hg), the time spent at the surface accounted for 50% of total time. This response has a character of a temporary emergency behavior that may allow the animal to escape hypoxia.     

Characterization of carbonic anhydrase and anion exchange in the erythrocytes of bowfin (Amia calva), a primitive air-breathing fish

The purpose of this study was to investigate the characteristics of carbonic anhydrase (CA) and the Cl⁻/HCO₃⁻ exchanger (Band 3; AE1) in the erythrocytes of bowfin (Amia calva), a primitive air-breathing fish, in order to further understand the strategies of blood CO₂ transport in lower vertebrates and gain insights into the evolution of the vertebrate erythrocyte proteins, CA and Band 3. A significant amount of CA activity was measured in the erythrocytes of bowfin (70 mmol CO₂ min⁻¹ ml⁻¹), although it appeared to be lower than that in the erythrocytes of teleost fish. The turnover number (K_cat) of bowfin erythrocyte CA was intermediate between that of the slow type I CA isozyme in agnathans and elasmobranchs and the fast type II CA in the erythrocytes of the more recent teleost fishes, but the inhibition properties of bowfin erythrocyte CA were similar to the fast mammalian CA isozyme, CA II. In contrast to previous findings, a plasma CA inhibitor was found to be present in the blood of bowfin. Bowfin erythrocytes were also found to possess a high rate of Cl⁻/HCO₃⁻ exchange (6 nmol HCO₃⁻ s⁻¹ cm⁻²) that was sensitive to DIDS. Visualization of erythrocyte membrane proteins by SDS-PAGE revealed a major band in the 100 kDa range for the trout, which would be consistent with the anion exchanger. In contrast, the closest major band for the membranes of bowfin erythrocytes was around the 140 kDa range. Taken together, these results suggest that the strategy for blood CO₂ transport in bowfin is probably similar to that in most other vertebrates despite several unique characteristics of erythrocyte CA and Band 3 in these primitive fish.     

The effect of hypoxia on the vulnerability of guppies (Poecilia reticulata,Poeciliidae) to an aquatic predator (Astronotus ocellatus,Cichlidae)

Synopsis When dissolved oxygen concentration was near saturation in a laboratory experiment, guppies that spent (a) more time at the surface and (b) more time moving had a higher probability of being captured by a predatory cichlid fish. With decreasing oxygen concentration surface time and percent time moving increased, but prey risk decreased. In addition, the qualitative correlates of risk changed; under hypoxic conditions, predation risk was lower for prey which spent more time at the surface and in motion. Thus, dissolved oxygen concentration influences both quantitative and qualitative aspects of risk from water-breathing predators.Author to whom reprint requests should be addressed.     

Pump out the volume—The effect of tracheal and subelytral pressure pulses on convective gas exchange in a dung beetle, Circellium bacchus (Fabricus)

Many flightless beetles like the large apterous dung beetle Circellium bacchus, possess a subelytral cavity (SEC) providing an extra air space below the elytra which connects to the tracheal system (TS) via metathoracic and abdominal spiracles. By measuring subelytral and intratracheal pressure as well as body movements and gas exchange simultaneously in a flow-through setup, we investigated the contribution of convection on Circellium respiratory gas exchange.No constriction phase was observed. TS and SEC pressures were always around atmospheric values. During interburst phase open abdominal spiracles and a leaky SEC led to small CO₂-peaks on a continuous CO₂ baseline, driven by intermittent positive tracheal pressure peaks in anti-phase with small negative subelytral pressure peaks caused by dorso-ventral tergite action.Spiracle opening was accompanied by two types of body movements. Higher frequency telescoping body movements at the beginning of opening resulted in high amplitude SEC and TS pressure peaks. High frequency tergite movements caused subelytral pressure peaks and led to a saw tooth like CO₂ release pattern in a burst. We propose that during the burst open mesothoracic spiracles increase the compliance of the subelytral cavity allowing big volumes of tracheal air being pulled out by convection.     

Seasonal variations in the rates of aquatic and aerial respiration and ammonium excretion of the ribbed mussel, Geukensia demissa (Dillwyn)

Aquatic and aerial rates of oxygen consumption and ammonium excretion of ribbed mussels, Geukensia demissa (Dillwyn), collected from the mid-intertidal zone of a mid-Atlantic salt marsh, were measured under ambient conditions of food, temperature, and salinity over five seasons. Rates of aquatic respiration covaried with body size and season, as the rates were high and strongly related to mussel tissue weight in spring and summer but low and weight independent in winter. There was a significant interannual difference between summer of 1995 and 1996. Rates of aerial respiration also varied seasonally, with high rates of oxygen consumption in summer and low rates in winter. The magnitude of these seasonal variations were greater than those for aquatic respiration, and as a result, the ratio of aerial to aquatic respiration was higher in summer (0.61 and 0.52) than in winter (0.11). This indicates that G. demissa was able to actively regulate aerial respiration, thereby permitting high aerobic metabolism during prolonged periods of air exposure in summer. We hypothesize that such high rates of aerial respiration, during the seasons of high metabolic activity, are required to provide sufficient energy for mussels to facilitate food digestion during air exposure at low tide. Rates of ammonium excretion varied seasonally and increased with mussel weight in all seasons. The atomic ratio of oxygen to nitrogen (O:N), calculated from aquatic respiration vs. ammonium excretion, was significantly lower in autumn (26) than in other seasons (46–60) among which the O:N did not vary significantly.     

The role of the subelytral cavity in respiration in a tenebrionid beetle,Onymacris multistriata (Tenebrionidae: Adesmiini)

This study measured the respiratory patterns in the tenebrionid beetle, Onymacris multistriata, using flow-through respirometry to measure carbon dioxide emission from the mesothoracic spiracles separately and simultaneously with that from around the elytral case. 96% of the total CO(2) emitted was via the mesothoracic spiracles. These spiracles used a discontinuous gas exchange cycle similar to that measured from other tenebrionid beetles. Although the circadian rhythm of the beetles resulted in changes to the period durations and cycle frequencies in the discontinuous gas exchange cycles, the mesothoracic spiracle remained the major site for gas exchange. Thus the subelytral cavity plays a different role in respiration other than the elimination of CO(2) build-up. It is expected that other arid dwelling flightless beetles will also be shown to use the mesothoracic spiracle as the major route for CO(2) emission.     

Impact of acute temperature transition on enzyme activity levels,oxygen consumption,and exogenous fuel utilization in sea raven (Hemitripterus americanus) hearts

Summary The impact of an acute temperature transition between 5 °C and 15 °C on energy metabolism and performance of sea raven (Hemitripterus americanus) heart was assessed. Maximal in vitro activity of hexokinase was 1.2 and 3.7 mol · min^-1 · g^-1 at 5 °C and 15 °C, respectively. Carnitine palmitoyl transferase and carnitine palmitoleoyl transferase activities were 0.07 mol · min^-1 · g^-1 at 15 °C and declined substantially at 5 °C. Oxygen consumption and power output of perfused isolated hearts offered glucose alone as a metabolic fuel decreased significantly between 15 °C and 5 °C. When palmitoleate was included in the perfusion medium, oxygen consumption and power development remained constant between 15 °C and 5 °C, suggesting that glucose alone was not an adequate metabolic fuel at low temperature. However, maximal in vitro activity of HK implied that the catalytic potential at this locus was quite capable of meeting demands of carbon flow, while the maximal in vitro activity of the carnitine acyl CoA transferases implied that fatty acid metabolism should be greatly compromised at low temperatures. In an effort to resolve the contradiction, hearts were perfused with medium containing ¹⁴C-glucose or ¹⁴C-palmitate. Rates of ¹⁴CO₂ production from labelled metabolic fuels could account for only about 2% of the oxygen consumption rates. Most of the label from ¹⁴C-glucose was incorporated into the glycogen and lipid fractions and label from ¹⁴C-palmitate was incorporated into the lipid fraction. The net incorporation rates of label into intracellular pools were temperature insensitive over the range 5–15 °C. The incorporation of exogenous glucose into the lipid fraction suggests that activity of the entire glycolytic pathway was maintained over the temperature range. Thus, the relatively low rate of oxygen consumption of hearts perfused with glucose alone as an exogenous substrate cannot be attributed to a limitation of glucose catabolism. The alternative explanation is that the presence of fatty acids induces an increase in oxygen consumption, especially at 5 °C. It is speculated that this is due to alterations in Ca²⁺ balance.Abbreviations ATPase adenosine triphosphatase - BSA bovine serum albumin - CoA coenzyme A - C palmitoyl T carnitine palmitoyl transferase - CS citrate synthase - HK hexokinase - MO oxygen consumption - PFK phosphofructokinase - PO ₂ oxygen partial pressure     

Gill ventilation and O2 extraction during graded hypoxia in two ecologically distinct species of flatfish,the flounder (Platichthys flesus) and the plaice (Pleuronectes platessa)

Synopsis Gill ventilation, breathing frequency, breath volume, oxygen extraction from the ventilatory water current and oxygen uptake through the gills were measured in flounder, Platichthys flesus, and plaice, Pleuronectes platessa, at water O₂ tensions ranging from 35 to 155 mm Hg at 10° C. Ventilation volumes were similar in the two species at high water O₂ tension. Exposure to hypoxic water elicited a larger increase in ventilation in the flounder. The per cent extraction of O₂ from water decreased slightly in both species as water O₂ tension was lowered. At comparable levels of ventilation O₂ extraction was higher in flounder. At the higher levels of water O₂ tension, O₂ uptake across the gills of flounder was stable, the critical O₂ tension being between 60 and 100 mm Hg. The plaice behaved as an oxygen conformer over the entire range of O₂ tensions investigated. The superior ability of the flounder in maintaining OZ uptake across the gills during a reduction in water O₂ tension may in part explain why the species, unlike plaice, inhabits very shallow waters with large fluctuations in dissolved oxygen.     

Respiratory responses to temperature and hypoxia in the nonindigenous Brown Mussel, Perna perna (Bivalvia: Mytilidae), from the Gulf of Mexico

The respiratory responses to increasing temperature and progressive hypoxia were examined relative to temperature acclimation in the nonindigenous, brown mussel, Perna perna (Mytilidae) from the Gulf of Mexico. When oxygen uptake rate (V?_O2) was recorded at near full air O₂ saturation, rate-temperature curves for Texas specimens of P. perna were sigmoidal, V?_O2 generally increasing with increasing temperature but becoming suppressed as temperatures approached 10 and 30 °C, corresponding closely to this species' incipient thermal limits. At each tested temperature, V?_O2 did not differ among individuals acclimated to 15, 20, or 25 °C. Lack of thermal acclimation was also reflected in acclimatory Q₁₀ values>1.0 (range=1.34-2.14) recorded across acclimation groups at test temperatures equivalent to acclimation temperature. Low acute respiratory Q₁₀ values in all acclimation groups across 15-20 °C indicated a limited capacity for thermal regulation of V?_O2 within this temperature range. The ability of P. perna to regulate O₂ uptake with progressive hypoxia was temperature-dependent, increasing from poor O₂ regulation at 10 °C to good regulation at 30 °C. The O₂ regulatory ability of P. perna and other open-water mytilids in declining O₂ concentrations does not greatly differ from that of estuarine heterodont bivalves, suggesting that it is not a major factor preventing open-water species, such as P. perna, from invading estuarine environments. However, P. perna's inability to regulate O₂ uptake at temperatures>25 °C combined with its relatively low upper thermal limit of 30 °C will likely prevent it from establishing permanent estuarine populations on Gulf of Mexico shores.     

Diversity of aquatic macrophytes in relation to environmental factors in the Slatina river (Slovakia)

Distribution and plant mass of aquatic macrophytes, and their relation to environmental conditions was studied in the submontane-colline Slatina river in 2004. Diversity of macrophytes was low, only 8 vascular plants, 3 mosses and group Algae filamentosae were found. Myriophyllum spicatum is dominant species, Fontinalis antipyretica, Rhynchostegium riparioides and Algae filamentosae are frequent. Interactions between flow class, bed material, depth of water and the first three mentioned macrophytes, as well as Jungermannia leiantha were detected. Sparganium erectum prefers more antrophogenic conditions and Myriophyllum spicatum prefers the light. According to cluster analysis, three distinct and ecologically well separated parts of the river were identified. Based on Reference index, poor ecological status for the studied part of the Slatina river was estimated.     

The role of leaf and canopy-level gas exchange in the replacement of Quercus virginiana (Fagaceae) by Juniperus ashei (Cupressaceae) in semiarid savannas

Photosynthesis, transpiration, and leaf area distribution were sampled in mature Quercus virginiana and Juniperus ashei trees to determine the impact of leaf position on canopy-level gas exchange, and how gas exchange patterns may affect the successful invasion of Quercus communities by J. ashei. Sampling was conducted monthly over a 2-yr period in 12 canopy locations (three canopy layers and four cardinal directions). Photosynthetic and transpiration rates of both species were greatest in the upper canopy and decreased with canopy depth. Leaf photosynthetic and transpiration rates were significantly higher for Q. virginiana (4.1–6.7 μmol CO₂·m⁻²·s⁻¹ and 1.1–2.1 mmol H₂O·m⁻²·s⁻¹) than for J. ashei (2.1–2.8 μmol CO₂·m⁻²·s⁻¹ and 0.7–1.0 mmol H₂O·m⁻²·s⁻¹) in every canopy level and direction. Leaves on the south and east sides of both species had higher gas exchange rates than leaves on the north and west sides. Although Quercus had a greater mean canopy diameter than Juniperus (31.3 vs. 27.7 m²), J. ashei had significantly greater leaf area (142 vs. 58 m²/tree). A simple model combining leaf area and gas exchange rates for different leaf positions demonstrated a significantly greater total canopy carbon dioxide uptake for J. ashei compared to Q. virginiana (831 vs. 612 g CO₂·tree⁻¹·d⁻¹, respectively). Total daily water loss was also greater for Juniperus (125 vs. 73 Ltree⁻¹·d⁻¹). Differences in leaf gas exchange rates were poor predictors of the relationship between the invasive J. ashei and the codominant Q. virginiana. Leaf area and leaf area distribution coupled with leaf gas exchange rates were necessary to demonstrate the higher overall competitive potential of J. ashei.     

Biodiversity of aquatic gastropods in the Mont St-Michel basin (France) in relation to salinity and drying of habitats

Seventy-seven polder and marsh stations were sampled for aquatic and riparian gastropods in the terrestrial basin of Mont St-Michel from March 1994–March 1995 and during a drought in 1996. In addition, four ponds with riparian coppice (bocage) were investigated. Twenty-one stations, mostly in polders, lacked snails; high values of conductivity, and by implication salinity, appeared to make the habitats unfavourable. Only the euryhaline hydrobid Potamopyrgus antipodarum and the amphibious lymnaeid Lymnaea truncatula survived in areas of high conductivity. In associated lagoons, the maximum level of salinity tolerated by freshwater gastropods was 35. Of the 14 species of gastropods collected, Lymnaea peregra and Anisus leucostoma were the most abundant. Stations had 1–7 species and the average gastropod species richness was 3.32 (±1.67). The canals that were the most susceptible to drying had a community of desiccation-resistant species including L. truncatula, Aplexa hypnorum, A. leucostoma and L. peregra. It appears that gastropod assemblages in the terrestrial basin of Mont St-Michel have achieved their current diversity by surviving strong ecological constraints.     

Response of the chromatophores in relation to the healing of skin wounds in an Indian Major Carp, Labeo rohita (Hamilton)

Chromatophores show significant changes during healing of skin wounds in Labeo rohita (Common Name - Rohu). Wound area can be divided into regions I, II and III. After infliction of wound, skin colour becomes significantly dark by 2 h that is gradually restored by 2 d. In regions II and III at 5 min, epidermal melanophores appear with beaded dendrites. In these regions at 2 h and in region I at 6 h, epidermal melanophores appear small, rounded or irregular shaped having dendritic processes with aggregated melanosomes. Subsequently, melanophores appear having elongated dendrites with dispersed or aggregated melanosomes. At 24 h, clusters of pigmented bodies appear in regions I and II. These bodies increase up to 2 d, and then diminish gradually and disappear by 8 d. Changes in dermal melanophores in region II at 5 min indicate the onset of degeneration. Degenerating melanophores increase up to 12 h, then gradually decline, and disappear by 4 d. Simultaneously, stellate melanophore reappear, gradually increase and appear like control by 8 d. Dermal melanophores in region III at different intervals appear stellate. In region I stellate dermal melanophores appear at 4 d. Stellate melanophores in all regions show different distribution of dispersed or aggregated melanosomes. With the appearance of dermal melanophores, highly refractive, crystalline structures, possibly the refractive platelets of the iridophores, are visualized around them. At subsequent intervals, these are frequently observed. This study provides interesting insights in injury induced changes in chromatophores in fish. The findings could be considered useful in perception of intriguing features in the development of pigment research in future.     

Metabolic tolerance of the cold-water coral Lophelia pertusa (Scleractinia) to temperature and dissolved oxygen change

Lophelia pertusa is the world's most common and widespread framework-forming cold-water coral. It forms deep-water coral reefs and carbonate mounds supporting diverse animal communities on the continental shelf and on seamounts. These recently discovered ecosystems have been damaged by deep-sea fishing and are threatened by predicted shallowing of the aragonite saturation horizon. Despite this, very little is known about the ecophysiology of L. pertusa and its likely response to environmental changes. Here we describe the first study of the respiratory physiology of L. pertusa and the effects of altered temperature and oxygen level. This study shows that L. pertusa can maintain respiratory independence over a range of PO₂ illustrated by a high regulation value (R = 78%). The critical PO₂ value of 9-10 kPa is very similar to the lower values of oxygen concentration recorded in the field. This suggests that oxygen level may be a limiting factor in the distribution of this cold-water coral. L. pertusa survived periods of anoxia (1 h), hypoxia (up to 96 h), but high Q₁₀ values revealed sensitivity to short-term temperature changes (6.5-11 °C). For the first time vital data have been gathered on the physiology of this species that is essential to understand distribution and underpin future climate change studies.     

Storage metabolism in the Pacific oyster (Crassostrea gigas) in relation to summer mortalities and reproductive cycle (west coast of France)

We describe seasonal changes in the biochemical composition of digestive gland, adductor muscle and gonad and surrounding mantle area in Crassostrea gigas from the Western Atlantic coast of France. Seasonality in histology of storage tissues and glycogen storage capacity in isolated vesicular cells were also studied. Proteins, the main muscle components did not contribute to the gametogenetic effort. Glycogen and lipids were stored in the digestive gland, gonad and surrounding mantle area during the wintering period and the gonad and surrounding mantle area represented the main storage compartment supplying the reproductive effort. Gametogenesis in spring and summer was associated with an increase in lipid and protein contents and took place at the expense of glycogen reserves. Histological study of storage tissue in the gonad led us to define four seasonal stages of storage tissue development. In vitro, glycogen storage capacity in isolated vesicular cells was high from November to March and markedly reduced during gametogenesis, decreasing below detectable levels after spawning. This physiological state should be taken into account with relation to summer mortalities occurring in commercial growing areas.     

Oxygen limits heat tolerance and drives heat hardening in the aquatic nymphs of the gill breathing damselfly Calopteryx virgo (Linnaeus, 1758)

Thermal limits in ectotherms may arise through a mismatch between O₂ supply and demand. At higher temperatures, the ability of their cardiac and ventilatory activities to supply O₂ becomes insufficient to meet their elevated O₂ demand. Consequently, higher levels of O₂ in the environment are predicted to enhance heat tolerance, while reductions in O₂ are expected to reduce thermal limits. Here, we extend previous research on thermal limits and oxygen limitation in aquatic insect larvae and report critical upper temperatures in nymphs of the damselfly Calopteryx virgo (Linnaeus, 1758) exposed to different levels of O₂. In addition, we explore the potential for a mechanistic link between O₂ conditions and thermal plasticity by exposing nymphs to two consecutive extreme heat events, using different levels of O₂ in the second exposure. As predicted, hypoxia severely lowered critical temperatures. However, thermal tolerance was not improved under hyperoxia. Damselfly nymphs may be precluded to take advantage of hyperoxia if O₂ uptake and delivery is controlled locally near the caudal gills where most of the gas exchange occurs. The same asymmetrical effects of hypoxia and hyperoxia on heat tolerance in terrestrial insects could be similarly explained if tracheal opening and/or ventilation are not centrally regulated. Prior exposure to hypoxia enhanced critical thermal maxima in subsequent heat exposures and hyperoxia negated this hardening effect, indicating potential for oxygen-driven heat hardening in these aquatic insects. Our study provides broad confirmation for oxygen limitation as a key mechanism setting upper thermal limits, pointing to a vital role for heat shock proteins in reducing O₂ requirements by slowing down rates of protein denaturation.