Are reactive oxygen species always bad? Lessons from hypoxic ectotherms.

Oxygen (O2) is required for aerobic energy metabolism but can produce reactive oxygen species (ROS), which are a wide variety of oxidant molecules with a range of biological functions from causing cell damage (oxidative distress) to cell signalling (oxidative eustress). The balance between the rate and amount of ROS generated and the capacity for scavenging systems to remove them is affected by several biological and environmental factors, including oxygen availability. Ectotherms, and in particular hypoxia-tolerant ectotherms, are hypothesized to avoid oxidative damage caused by hypoxia, although it is unclear whether this translates to an increase in ecological fitness. In this Review, we highlight the differences between oxidative distress and eustress, the current mechanistic understanding of the two and how they may affect ectothermic physiology. We discuss the evidence of occurrence of oxidative damage with hypoxia in ectotherms, and that ectotherms may avoid oxidative damage through (1) high levels of antioxidant and scavenging systems and/or (2) low(ering) levels of ROS generation. We argue that the disagreements in the literature as to how hypoxia affects antioxidant enzyme activity and the variable metabolism of ectotherms makes the latter strategy more amenable to ectotherm physiology. Finally, we argue that observed changes in ROS production and oxidative status with hypoxia may be a signalling mechanism and an adaptive strategy for ectotherms encountering hypoxia.

Oxygen consumption rate during recovery from loss of equilibrium induced by warming, hypoxia, or exhaustive exercise in rainbow darter (Etheostoma caeruleum).

Animals routinely encounter environmental (e.g., high temperatures and hypoxia) as well as physiological perturbations (e.g., exercise and digestion) that may threaten homeostasis. However, comparing the relative threat or "disruptiveness" imposed by different stressors is difficult, as stressors vary in their mechanisms, effects, and timescales. We exploited the fact that several acute stressors can induce the loss of equilibrium (LOE) in fish to (i) compare the metabolic recovery profiles of three environmentally relevant stressors and (ii) test the concept that LOE could be used as a physiological calibration for the intensity of different stressors. We focused on Etheostoma caeruleum, a species that routinely copes with environmental fluctuations in temperature and oxygen and that relies on burst swimming to relocate and avoid predators, as our model. Using stop-flow (intermittent) respirometry, we tracked the oxygen consumption rate (MO2) as E. caeruleum recovered from LOE induced by hypoxia (PO2 at LOE), warming (critical thermal maximum, CTmax), or exhaustive exercise. Regardless of the stressor used, E. caeruleum recovered rapidly, returning to routine MO2 within ~3 h. Fish recovering from hypoxia and warming had similar maximum MO2, aerobic scopes, recovery time, and total excess post-hypoxia or post-warming oxygen consumption. Though exhaustive exercise induced a greater maximum MO2 and corresponding higher aerobic scope than warming or hypoxia, its recovery profile was otherwise similar to the other stressors, suggesting that "calibration" to a physiological state such as LOE may be a viable conceptual approach for investigators interested in questions related to multiple stressors, cross tolerance, and how animals cope with challenges to homeostasis.

Science communication in experimental biology: experiences and recommendations.

During the century of Journal of Experimental Biology's existence, science communication has established itself as an interdisciplinary field of theory and practice. Guided by my experiences as a scientist and science writer, I argue that science communication skills are distinct from scientific communication skills and that engaging in science communication is particularly beneficial to early-career researchers; although taking on these dual roles is not without its difficulties, as I discuss in this Perspective. In the hope of encouraging more scientists to become science communicators, I provide: (i) general considerations for scientists looking to engage in science communication (knowing their audience, storytelling, avoiding jargon) and (ii) specific recommendations for crafting effective contributions on social media (content, packaging, engagement), an emerging, accessible and potentially impactful mode of science communication. Effective science communication can boost the work of experimental biologists: it can impact public opinion by incisively describing the consequences of the climate crisis and can raise social acceptance of fundamental research and experiments on animals.

Physiological differences between wild and captive animals: a century-old dilemma.

Laboratory-based research dominates the fields of comparative physiology and biomechanics. The power of lab work has long been recognized by experimental biologists. For example, in 1932, Georgy Gause published an influential paper in Journal of Experimental Biology describing a series of clever lab experiments that provided the first empirical test of competitive exclusion theory, laying the foundation for a field that remains active today. At the time, Gause wrestled with the dilemma of conducting experiments in the lab or the field, ultimately deciding that progress could be best achieved by taking advantage of the high level of control offered by lab experiments. However, physiological experiments often yield different, and even contradictory, results when conducted in lab versus field settings. This is especially concerning in the Anthropocene, as standard laboratory techniques are increasingly relied upon to predict how wild animals will respond to environmental disturbances to inform decisions in conservation and management. In this Commentary, we discuss several hypothesized mechanisms that could explain disparities between experimental biology in the lab and in the field. We propose strategies for understanding why these differences occur and how we can use these results to improve our understanding of the physiology of wild animals. Nearly a century beyond Gause's work, we still know remarkably little about what makes captive animals different from wild ones. Discovering these mechanisms should be an important goal for experimental biologists in the future.

Upstream migrant sea lamprey (Petromyzon marinus) show signs of increasing oxidative stress but maintain aerobic capacity with age.

Following the parasitic juvenile phase of their life cycle, sea lamprey (Petromyzon marinus) mature into a reproductive but rapidly aging and deteriorating adult, and typically die shortly after spawning in May or June. However, pre-spawning upstream migrant sea lamprey can be maintained for several months beyond their natural lifespan when held in cold water (∼4-8 °C) under laboratory conditions. We exploited this feature to investigate the interactions between senescence, oxidative stress, and metabolic function in this phylogenetically ancient fish. We investigated how life history traits and mitochondria condition, as indicated by markers of oxidative stress (catalase activity, lipid peroxidation) and aerobic capacity (citrate synthase activity), changed in adult sea lamprey from June to December after capture during their upstream spawning migration. Body mass but not liver mass declined with age, resulting in an increase in hepatosomatic index. Both effects were most pronounced in males, which also tended to have larger livers than females. Lamprey experienced greater oxidative stress with age, as reflected by increasing activity of the antioxidant enzyme catalase and increasing levels of lipid peroxidation in liver mitochondrial isolates over time. Surprisingly, the activity of citrate synthase also increased with age in both sexes. These observations implicate mitochondrial dysfunction and oxidative stress in the senescence of sea lamprey. Due to their unique evolutionary position and the technical advantage of easily delaying the onset of senescence in lampreys using cold water, these animals could represent an evolutionary unique and tractable model to investigate senescence in vertebrates.

Physiological and behavioural strategies of aquatic animals living in fluctuating environments.

Shallow or near-shore environments, such as ponds, estuaries and intertidal zones, are among the most physiologically challenging of all aquatic settings. Animals inhabiting these environments experience conditions that fluctuate markedly over relatively short temporal and spatial scales. Living in these habitats requires the ability to tolerate the physiological disturbances incurred by these environmental fluctuations. This tolerance is achieved through a suite of physiological and behavioural responses that allow animals to maintain homeostasis, including the ability to dynamically modulate their physiology through reversible phenotypic plasticity. However, maintaining the plasticity to adjust to some stresses in a dynamic environment may trade off with the capacity to deal with other stressors. This paper will explore studies on select fishes and invertebrates exposed to fluctuations in dissolved oxygen, salinity and pH. We assess the physiological mechanisms these species employ to achieve homeostasis, with a focus on the plasticity of their responses, and consider the resulting physiological trade-offs in function. Finally, we discuss additional factors that may influence organismal responses to fluctuating environments, such as the presence of multiple stressors, including parasites. We echo recent calls from experimental biologists to consider physiological responses to life in naturally fluctuating environments, not only because they are interesting in their own right but also because they can reveal mechanisms that may be crucial for living with increasing environmental instability as a consequence of climate change.

Niclosamide Is a Much More Potent Toxicant of Mitochondrial Respiration than TFM in the Invasive Sea Lamprey (Petromyzon marinus).

Invasive sea lampreys in the Laurentian Great Lakes are controlled by applying TFM (3-trifluoromethyl-4-nitrophenol) and niclosamide to streams infested with their larvae. Both agents uncouple oxidative phosphorylation in the mitochondria, but TFM specifically targets lampreys, which have a lower capacity to detoxify the lampricide. Niclosamide lacks specificity and is more potent than TFM. However, its greater potency is poorly understood. We tested the hypothesis that niclosamide is a stronger uncoupler of mitochondrial oxidative phosphorylation than TFM by measuring oxygen consumption rates in isolated liver mitochondria exposed to physiologically relevant concentrations of TFM, niclosamide, or their mixture (100 TFM:1 niclosamide) at environmentally relevant temperatures (7, 13, and 25 °C). Niclosamide increased State 4 respiration and decreased the respiratory control ratio (RCR) at much lower concentrations than TFM. Calculations of the relative EC50 values, the amount of TFM or niclosamide required to decrease the RCR by 50%, indicated that niclosamide was 40-60 times more potent than TFM. Warmer temperature did not appear to decrease the sensitivity of mitochondria to niclosamide or TFM, as observed in the intact sea lamprey exposed to TFM in warmer waters. We conclude that the extreme sensitivity of mitochondria to niclosamide contributes to its greater in vivo toxicity in the whole animal.

Rapid and reversible modulation of blood haemoglobin content during diel cycles of hypoxia in killifish (Fundulus heteroclitus).

We investigated whether fish can make dynamic haematological adjustments to support aerobic metabolism during repeated cycles of hypoxia-reoxygenation. Killifish were acclimated to normoxia, constant hypoxia (2 kPa O2), or intermittent cycles of nocturnal hypoxia (12 h of normoxia: 12 h of 2 kPa O2 hypoxia) for 28 days. Normoxia-acclimated fish were sampled in the daytime in normoxia and after exposure to a single bout of nocturnal hypoxia. Each hypoxia acclimation group were sampled at the PO2 experienced during acclimation during both the day and night. All acclimation groups had increased blood haemoglobin content and haematocrit and reduced spleen mass during nocturnal hypoxia compared to normoxic controls. Blood haemoglobin content was negatively correlated with spleen mass at both the individual and group level. Fish acclimated to intermittent hypoxia rapidly reversed these changes during diurnal reoxygenation. The concentrations of haemoglobin, ATP, and GTP within erythrocytes did not vary substantially between groups. We also measured resting O2 consumption rate (MO2) and maximum MO2 (induced by an exhaustive chase) in hypoxia in each acclimation group. Fish acclimated to intermittent hypoxia maintained higher resting MO2 than other groups in hypoxia, comparable to the resting MO2 of normoxia-acclimated controls measured in normoxia. Differences in resting MO2 in hypoxia did not result from variation in O2 transport capacity, because maximal MO2 in hypoxia always exceeded resting MO2. Therefore, reversible modulation of blood haemoglobin content along with metabolic adjustments help killifish cope with intermittent cycles of hypoxia in the estuarine environment.

Hypoxia acclimation alters reactive oxygen species homeostasis and oxidative status in estuarine killifish (Fundulus heteroclitus).

Hypoxia is common in aquatic environments, and exposure to hypoxia followed by re-oxygenation is often believed to induce oxidative stress. However, there have been relatively few studies of reactive oxygen species (ROS) homeostasis and oxidative status in fish that experience natural hypoxia-re-oxygenation cycles. We examined how exposure to acute hypoxia (2 kPa O2) and subsequent re-oxygenation (to 20 kPa O2) affects redox status, oxidative damage and anti-oxidant defenses in estuarine killifish (Fundulus heteroclitus), and whether these effects were ameliorated or potentiated by prolonged (28 days) acclimation to either constant hypoxia or intermittent cycles of nocturnal hypoxia (12 h:12 h normoxia:hypoxia). Acute hypoxia and re-oxygenation led to some modest and transient changes in redox status, increases in oxidized glutathione, depletion of scavenging capacity and oxidative damage to lipids in skeletal muscle. The liver had greater scavenging capacity, total glutathione concentrations and activities of anti-oxidant enzymes (catalase, glutathione peroxidase) than muscle, and generally experienced less variation in glutathiones and lipid peroxidation. Unexpectedly, acclimation to constant hypoxia or intermittent hypoxia led to a more oxidizing redox status (muscle and liver) and it increased oxidized glutathione (muscle). However, hypoxia-acclimated fish exhibited little to no oxidative damage (as reflected by lipid peroxidation and aconitase activity), in association with improvements in scavenging capacity and catalase activity in muscle. We conclude that hypoxia acclimation leads to adjustments in ROS homeostasis and oxidative status that do not reflect oxidative stress, but may instead be part of the suite of responses that killifish use to cope with chronic hypoxia.

Interspecific variation in hypoxia tolerance and hypoxia acclimation responses in killifish from the family Fundulidae.

Hypoxia is a pervasive stressor in aquatic environments, and both phenotypic plasticity and evolutionary adaptation could shape the ability to cope with hypoxia. We investigated evolved variation in hypoxia tolerance and the hypoxia acclimation response across fundulid killifishes that naturally experience different patterns of hypoxia exposure. We compared resting O2 consumption rate (MO2 ), and various indices of hypoxia tolerance [critical O2 tension (Pcrit), regulation index (RI), O2 tension (PO2 ) at loss of equilibrium (PLOE) and time to LOE (tLOE) at 0.6 kPa O2] in Fundulus confluentus, Fundulus diaphanus, Fundulus heteroclitus, Fundulus rathbuni, Lucania goodei and Lucania parva We examined the effects of chronic (28 days) exposure to constant hypoxia (2 kPa) or nocturnal intermittent hypoxia (12 h normoxia:12 h hypoxia) in a subset of species. Some species exhibited a two-breakpoint model in MO2  caused by early, modest declines in MO2  in moderate hypoxia. We found that hypoxia tolerance varied appreciably across species: F. confluentus was the most tolerant (lowest PLOE and Pcrit, longest tLOE), whereas F. rathbuni and F. diaphanus were the least tolerant. However, there was not a consistent pattern of interspecific variation for different indices of hypoxia tolerance, with or without taking phylogenetic relatedness into account, probably because these different indices are underlain by partially distinct mechanisms. Hypoxia acclimation generally improved hypoxia tolerance, but the magnitude of plasticity and responsiveness to different hypoxia patterns varied interspecifically. Our results therefore suggest that hypoxia tolerance is a complex trait that is best appreciated by considering multiple indices of tolerance.

Distinct metabolic adjustments arise from acclimation to constant hypoxia and intermittent hypoxia in estuarine killifish (Fundulus heteroclitus).

Many fish experience daily cycles of hypoxia in the wild, but the physiological strategies for coping with intermittent hypoxia are poorly understood. We examined how killifish adjust O2 supply and demand during acute hypoxia, and how these responses are altered after prolonged acclimation to constant or intermittent patterns of hypoxia exposure. We acclimated killifish to normoxia (∼20 kPa O2), constant hypoxia (2 kPa) or intermittent cycles of nocturnal hypoxia (12 h:12 h normoxia:hypoxia) for 28 days, and then compared whole-animal O2 consumption rates (MO2 ) and tissue metabolites during exposure to 12 h of hypoxia followed by reoxygenation in normoxia. Normoxia-acclimated fish experienced a pronounced 27% drop in MO2  during acute hypoxia, and modestly increased MO2  upon reoxygenation. They strongly recruited anaerobic metabolism during acute hypoxia, indicated by lactate accumulation in plasma, muscle, liver, brain, heart and digestive tract, as well as a transient drop in intracellular pH, and they increased hypoxia inducible factor (HIF)-1α protein abundance in muscle. Glycogen, glucose and glucose-6-phosphate levels suggested that glycogen supported brain metabolism in hypoxia, while the muscle used circulating glucose. Acclimation to constant hypoxia caused a stable ∼50% decrease in MO2  that persisted after reoxygenation, with minimal recruitment of anaerobic metabolism, suggestive of metabolic depression. By contrast, fish acclimated to intermittent hypoxia maintained sufficient O2 transport to support normoxic MO2 , modestly recruited lactate metabolism and increased MO2  dramatically upon reoxygenation. Both groups of hypoxia-acclimated fish had similar glycogen, ATP, intracellular pH and HIF-1α levels as normoxic controls. We conclude that different patterns of hypoxia exposure favour distinct strategies for matching O2 supply and O2 demand.

Mitochondrial physiology and reactive oxygen species production are altered by hypoxia acclimation in killifish (Fundulus heteroclitus).

Many fish encounter hypoxia in their native environment, but the role of mitochondrial physiology in hypoxia acclimation and hypoxia tolerance is poorly understood. We investigated the effects of hypoxia acclimation on mitochondrial respiration, O2kinetics, emission of reactive oxygen species (ROS), and antioxidant capacity in the estuarine killifish ( ITALIC! Fundulus heteroclitus). Killifish were acclimated to normoxia, constant hypoxia (5 kPa O2) or intermittent diel cycles of nocturnal hypoxia (12 h:12 h normoxia:hypoxia) for 28-33 days and mitochondria were isolated from liver. Neither pattern of hypoxia acclimation affected the respiratory capacities for oxidative phosphorylation or electron transport, leak respiration, coupling control or phosphorylation efficiency. Hypoxia acclimation also had no effect on mitochondrial O2kinetics, but ITALIC! P50(the O2tension at which hypoxia inhibits respiration by 50%) was lower in the leak state than during maximal respiration, and killifish mitochondria endured anoxia-reoxygenation without any impact on mitochondrial respiration. However, both patterns of hypoxia acclimation reduced the rate of ROS emission from mitochondria when compared at a common O2tension. Hypoxia acclimation also increased the levels of protein carbonyls and the activities of superoxide dismutase and catalase in liver tissue (the latter only occurred in constant hypoxia). Our results suggest that hypoxia acclimation is associated with changes in mitochondrial physiology that decrease ROS production and may help improve hypoxia tolerance.

Interspecific and environment-induced variation in hypoxia tolerance in sunfish.

Hypoxia tolerance is a plastic trait, and can vary between species. We compared hypoxia tolerance (hypoxic loss of equilibrium, LOE, and critical O2 tension, Pcrit) and traits that dictate O2 transport and metabolism in pumpkinseed (Lepomis gibbosus), bluegill (L. macrochirus), and the naturally occurring hybrid in different acclimation environments (wild versus lab-acclimated fish) and at different temperatures. Wild fish generally had lower Pcrit and lower PO2 at LOE in progressive hypoxia than lab-acclimated fish, but time to LOE in sustained hypoxia (PO2 of 2kPa) did not vary between environments. Wild fish also had greater gill surface area and higher haematocrit, suggesting that increased O2 transport capacity underlies the environmental variation in Pcrit. Metabolic (lactate dehydrogenase, LDH; pyruvate kinase, PK; citrate synthase; cytochrome c oxidase) and antioxidant (catalase and superoxide dismutase) enzyme activities varied appreciably between environments. Wild fish had higher protein contents across tissues and higher activities of LDH in heart, PK in brain, and catalase in brain, liver, and skeletal muscle. Otherwise, wild fish had lower activities for most enzymes. Warming temperature from 15 to 25°C increased O2 consumption rate, Pcrit, PO2 at LOE, and haemoglobin-O2 affinity, and decreased time to LOE, but pumpkinseed had ≥2-fold longer time to LOE than bluegill and hybrids across this temperature range. This was associated with higher LDH activities in the heart and muscle, and lower or similar antioxidant enzyme activities in several tissues. However, the greater hypoxia tolerance of pumpkinseed collapsed at 28°C, demonstrating that the interactive effects of hypoxia and warming temperature can differ between species. Overall, distinct mechanisms appear to underpin interspecific and environment-induced variation in hypoxia tolerance in sunfish.

Distinct physiological strategies are used to cope with constant hypoxia and intermittent hypoxia in killifish (Fundulus heteroclitus).

Many fish encounter hypoxia on a daily cycle, but the physiological effects of intermittent hypoxia are poorly understood. We investigated whether acclimation to constant (sustained) hypoxia or to intermittent diel cycles of nocturnal hypoxia (12 h normoxia:12 h hypoxia) had distinct effects on hypoxia tolerance or on several determinants of O2 transport and O2 utilization in estuarine killifish. Adult killifish were acclimated to normoxia, constant hypoxia, or intermittent hypoxia for 7 or 28 days in brackish water (4 ppt). Acclimation to both hypoxia patterns led to comparable reductions in critical O2 tension and resting O2 consumption rate, but only constant hypoxia reduced the O2 tension at loss of equilibrium. Constant (but not intermittent) hypoxia decreased filament length and the proportion of seawater-type mitochondrion-rich cells in the gills (which may reduce ion loss and the associated costs of active ion uptake), increased blood haemoglobin content, and reduced the abundance of oxidative fibres in the swimming muscle. In contrast, only intermittent hypoxia augmented the oxidative and gluconeogenic enzyme activities in the liver and increased the capillarity of glycolytic muscle, each of which should facilitate recovery between hypoxia bouts. Neither exposure pattern affected muscle myoglobin content or the activities of metabolic enzymes in the brain or heart, but intermittent hypoxia increased brain mass. We conclude that the pattern of hypoxia exposure has an important influence on the mechanisms of acclimation, and that the optimal strategies used to cope with intermittent hypoxia may be distinct from those for coping with constant hypoxia.

Parental Males of the Plainfin Midshipman Are Physiologically Resilient to the Challenges of the Intertidal Zone.

The decision of where to rear young is influenced by both the needs of offspring and the costs parents incur in certain rearing environments. Plainfin midshipman fish (Porichthys notatus) provide extended paternal care in rocky intertidal zones, where they experience regular bouts of aquatic hypoxia and air exposure during low-tide events. We investigated the physiological responses of plainfin midshipman males to three conditions for 6 h that simulate what these fish naturally experience during tidal cycles while nesting: normoxia, progressive hypoxia, or air exposure. Hypoxia- and air-exposed fish exhibited shifts in energy metabolites, driven largely by elevated lactate and glucose content and reduced glycogen content in several tissues (muscle, heart, liver, and brain), but the magnitude of these changes was relatively modest. Hematocrit increased most in air-exposed fish relative to normoxia-exposed fish, contributing to an increase in whole-blood hemoglobin concentration. Air exposure reduced swim bladder oxygen content, suggesting that internal O2 stores are drawn on during air exposure. In a second experiment, we found that aquatic surface respiration and gill ventilation frequency increased in hypoxia-exposed fish relative to normoxia-exposed fish. Overall, our results suggest that plainfin midshipman overcome the challenges of the intertidal environment through a variety of physiological strategies and exhibit little physiological disturbance in response to the fluctuating and extreme conditions created by regular low tides.

Metabolic implications of exposure to wastewater effluent in bluegill sunfish.

Effluent from wastewater treatment plants (WWTP) contains a complex mixture of contaminants and is a major worldwide source of aquatic pollution. We examined the effects of exposure to treated effluent from a municipal WWTP on the metabolic physiology of bluegill sunfish (Lepomis macrochirus). We studied fish that were wild-caught or experimentally caged (28 d) downstream of the WWTP, and compared them to fish that were caught or caged at clean reference sites. Survival was reduced in fish caged at the effluent-contaminated site compared to those caged at the reference site. Resting rates of O2 consumption (MO2) were higher in fish from the contaminated site, reflecting a metabolic cost of wastewater exposure. The increases in routine MO2 did not reduce aerobic scope (difference or quotient of maximal MO2 and resting MO2), suggesting that physiological compensations accompanied the metabolic costs of wastewater exposure. Fish exposed to wastewater also had larger hearts and livers. The activity of mitochondrial enzymes (cytochrome c oxidase, citrate synthase) per liver mass was unaltered across treatments, so the increased mass of this organ increased its cumulative oxidative capacity in the fish. Wastewater exposure also reduced glycogen content per liver mass. The effects of caging itself, based on comparisons between fish that were wild-caught or caged at clean sites, were generally subtle and not statistically significant. We conclude that exposure to wastewater effluent invokes a metabolic cost that leads to compensatory physiological adjustments that partially offset the detrimental metabolic impacts of exposure.

The Preference for Social Affiliation Renders Fish Willing to Accept Lower O2 Levels.

Animals are bombarded with information about their environment and must select and interpret the relevant cues to make behavioral adjustments critical to survival. How animals integrate and balance the many signals they receive about their environment is rarely assessed. We investigated how signals from the social and physical environment interact to influence environmental preferences in the endemic Tanganyikan cichlid Neolamprologus pulcher. Specifically, we explored how fish respond to the physiological challenge of declining O2 levels in light of embedded social preferences using a modified shuttle box apparatus to test O2 preferences. In the presence of a conspecific, the average (preferred) partial pressure of oxygen (Po2) and minimum Po2 experienced were significantly lower ([Formula: see text] and [Formula: see text] kPa, respectively) than in trials without a conspecific ([Formula: see text] and [Formula: see text] kPa, respectively). Fish with conspecifics also spent more time in the low Po2 zone of the shuttle box and moved between the high and low Po2 zones less frequently. Hence, O2 preferences were modified, and fish willingly remained in an area of continuously declining O2 availability to associate with a conspecific. The O2 preferences of an individual during social trials correlated with its excess postexercise O2 consumption following an exhaustive chase but not with its aerobic scope, routine O2 consumption rate, or body mass. These results suggest that some aspects of respiratory and metabolic physiology (such as the propensity to use anaerobic metabolism) but not others (such as O2 transport capacity) underpin some variation in social behavior under environmental stress.