Transition from ectothermy to endothermy: the development of metabolic capacity in a bird (Gallus gallus)

The evolution of endothermy is one of the most significant events in vertebrate evolution. Adult mammals and birds are delineated from their early ontogenetic stages, as well as from other vertebrates, by high resting metabolic rates and consequent internal heat production. We used the embryonic development of a bird (Gallus gallus) as a model to investigate the metabolic transition between ectothermy and endothermy. Increases in aerobic capacity occur at two functional levels that are regulated independently from each other: (i) upregulation of gene expression; and (ii) significant increases in the catalytic activity of the main oxidative control enzymes. Anaerobic capacity, measured as lactate dehydrogenase activity, is extremely high during early development, but diminishes at the same time as aerobic capacity increases. Changes in lactate dehydrogenase activity are independent from its gene expression. The regulatory mechanisms that lead to endothermic metabolic capacity are similar to those of ectotherms in their response to environmental change. We suggest that the phylogenetic occurrence of endothermy is restricted by its limited selective advantages rather than by evolutionary innovation.     

Energetics, body size, and the limits to endothermy

The scaling rate of metabolism with respect to body mass is analysed. Scaling of heat production implies that scaling also exists between temperature regulation and body mass. Most vertebrates follow a Kleiber relation down to a "critical mass, below which the scaling of metabolism must be changed to ensure the maintenance of endothermy. Such an adjustment is found interspecifically in birds and mammals, and is found intraspecifically in mammals during post-natal growth. If the Kleiber scaling relation is maintained below the critical mass, mammals and birds shiR from endothermic temperature regulation (above critical mass) to endothermy with obligatory torpor (below critical mass). If the Kleiber relation is followed to masses far below the critical mass, ectothermy results. Critical mass varies inversely with the level of energy expenditure, which therefore accounts for the fact that most mammals and birds are endotherms and most reptiles and fish are ectotherms. The same relationship permits the facultative endothermy found in some insects and plants.
The scaling relations existing among rate of metabolism, endothermy, and body mass can be written as a modification of the Kleiber relation. This analysis suggests that any organism, irrespective of phylogenetic position, can be endothermic at any body size, if its rate of metabolism is high enough, or can be endothermic with any rate of metabolism, if it is large enough. Consequently, it is difficult to distinguish minimal endothermy from inertial homoiothermy in animals having a large mass. The boundary conditions for effective endothermy are similar to the relationship described between metabolism and mass in the evolution of endothermy through a decrease in mass in the phylogeny of mammals. Even though endothermy may evolve with an increase in mass, its perfection may always require an evolutionary decrease in mass.     

On the relation between basal and maximum metabolic rate in mammals

Basal and maximum metabolic rates, measured by oxygen consumption, for 18 species of wild mammals have been obtained from a search of literature records. The mass exponent of the allometric regression equation for maximum metabolic rate is significantly higher than that for BMR (0.841 and 0.745, respectively; P less than 0.05) in the group of animals examined. No significant correlation between mass-independent basal and maximum metabolic rates has been found. These results do not support the 'aerobic capacity' model of the origin of endothermy.     

Endothermy in African platypleurine cicadas: the influence of body size and habitat (Hemiptera: Cicadidae)

The platypleurine cicadas have a wide distribution across Africa and southern Asia. We investigate endothermy as a thermoregulatory strategy in 11 South African species from five genera, with comparisons to the lone ectothermic platypleurine we found, in an attempt to ascertain any influence that habitat and/or body size have on the expression of endothermy in the platypleurine cicadas. Field measurements of body temperature (T(b)) show that these animals regulate T(b) through endogenous heat production. Heat production in the laboratory elevated T(b) to the same range as in animals active in the field. Maximum T(b) measured during calling activity when there was no access to solar radiation ranged from 13.2 degrees to 22.3 degrees C above ambient temperature in the five species measured. The mean T(b) during activity without access to solar radiation did not differ from the mean T(b) during diurnal activity. All platypleurines exhibit a unique behavior for cicadas while warming endogenously, a temperature-dependent telescoping pulsation of the abdomen that probably functions in ventilation. Platypleurines generally call from trunks and branches within the canopy and appear to rely on endothermy even when the sun is available to elevate T(b), in contrast to the facultative endothermy exhibited by New World endothermic species. The two exceptions to this generalization we found within the platypleurines are Platypleura wahlbergi and Albanycada albigera, which were the smallest species studied. The small size of P. wahlbergi appears to have altered their thermoregulatory strategy to one of facultative endothermy, whereby they use the sun when it is available to facilitate increases in T(b). Albanycada albigera is the only ectothermic platypleurine we found. The habitat and host plant association of A. albigera appear to have influenced the choice of ectothermy as a thermoregulatory strategy, as the species possesses the metabolic machinery to elevate to the T(b) range observed in the endothermic species. Therefore, size and habitat appear to influence the expression of thermoregulatory strategies in African platypleurine cicadas.     

The evolution of endothermy and its diversity in mammals and birds

Many elements of mammalian and avian thermoregulatory mechanisms are present in reptiles, and the changes involved in the transition to endothermy are more quantitative than qualitative. Drawing on our experience with reptiles and echidnas, we comment on that transition and on current theories about how it occurred. The theories divide into two categories, depending on whether selection pressures operated directly or indirectly on mechanisms producing heat. Both categories of theories focus on explaining the evolution of homeothermic endothermy but ignore heterothermy. However, noting that hibernation and torpor are almost certainly plesiomorphic (=ancestral, primitive), and that heterothermy is very common among endotherms, we propose that homeothermic endothermy evolved via heterothermy, with the earliest protoendotherms being facultatively endothermic and retaining their ectothermic capacity for "constitutional eurythermy." Thus, unlike current models for the evolution of endothermy that assume that hibernation and torpor are specialisations arising from homeothermic ancestry, and therefore irrelevant, we consider that they are central. We note the sophistication of thermoregulatory behavior and control in reptiles, including precise control over conductance, and argue that brooding endothermy seen in some otherwise ectothermic Boidae suggests an incipient capacity for facultative endothermy in reptiles. We suggest that the earliest insulation in protoendotherms may have been internal, arising from redistribution of the fat bodies that are typical of reptiles. We note that short-beaked echidnas provide a useful living model of what an (advanced) protoendotherm may have been like. Echidnas have the advantages of endothermy, including the capacity for homeothermic endothermy during incubation, but are very relaxed in their thermoregulatory precision and minimise energetic costs by using ectothermy facultatively when entering short- or long-term torpor. They also have a substantial layer of internal dorsal insulation. We favor theories about the evolution of endothermy that invoke direct selection for the benefits conferred by warmth, such as expanding daily activity into the night, higher capacities for sustained activity, higher digestion rates, climatic range expansion, and, not unrelated, control over incubation temperature and the benefits for parental care. We present an indicative, stepwise schema in which observed patterns of body temperature are a consequence of selection pressures, the underlying mechanisms, and energy optimization, and in which homeothermy results when it is energetically desirable rather than as the logical endpoint.     

Climate variability and the energetic pathways of evolution: the origin of endothermy in mammals and birds

Large-scale climate oscillations in earth's history have influenced the directions of evolution, last but not least, through mass extinction events. This analysis tries to identify some unifying forces behind the course of evolution that favored an increase in organismic complexity and performance, paralleled by an increase in energy turnover, and finally led to endothermy. The analysis builds on the recent concept of oxygen-limited thermal tolerance and on the hypothesis that unifying principles exist in the temperature-dependent biochemical design of the eukaryotic cell in animals. The comparison of extant water-breathing and air-breathing animal species from various climates provides a cause-and-effect understanding of the trade-offs and constraints in thermal adaptation and their energetic consequences. It is hypothesized that the high costs of functional adaptation to fluctuating temperatures, especially in the cold (cold eurythermy), cause an increase in energy turnover and, at the same time, mobility and agility. These costs are associated with elevated mitochondrial capacities at minimized levels of activation enthalpies for proton leakage. Cold eurythermy is seen as a precondition for the survival of evolutionary crises elicited by repeated cooling events during extreme climate fluctuations. The costs of cold eurythermy appear as the single most important reason why metazoan evolution led to life forms with high energy turnover. They also explain why dinosaurs were able to live in subpolar climates. Finally, they give insight into the pathways, benefits, and trade-offs involved in the evolution of constant, elevated body temperature maintained by endothermy. Eurythermy, which encompasses cold tolerance, is thus hypothesized to be the "missing link" between ectothermy and endothermy. Body temperatures between 32 degrees and 42 degrees C in mammals and birds then result from trade-offs between the limiting capacities of ventilation and circulation and the evolutionary trend to maximize performance at the warm end of the thermal tolerance window.     

Relation between the O2 supply:demand ratio, MVO2, and adenosine formation in hearts stimulated with inotropic agents

Mooted controllers of adenosine formation in heart are the oxygen supply:demand ratio, myocardial oxygen consumption (MVO2), the cytosolic phosphorylation potential (log[ATP]/[ADP][Pi]). The relationship between these parameters and purine release (adenosine + inosine) into the venous effluent was examined in isovolumic rat hearts perfused at 20 and 12 mL.min-1.g-1 with a glucose containing crystalloid buffer and stimulated with inotropic agents (isoproterenol, norepinephrine, 3-isobutyl-1-methylxanthine, and ouabain). The oxygen supply:demand ratio and MVO2 were continuously determined using an oxygen electrode to monitor oxygen supply and consumption. The phosphorylation potential was calculated from phosphorus metabolite levels determined by 31P-NMR spectroscopy and HPLC analysis. Left ventricular function was assessed as the rate-pressure product. All inotropic agents increased the rate-pressure product, with increases in function being greater in the hearts perfused at 20 mL.min-1.g-1. MVO2 was linearly related to the rate-pressure product at each flow rate; however, the hearts perfused at 20 mL.min-1.g-1 exhibited approximately twofold greater MVO2 values for similar rate-pressure product values. All inotropic agents increased adenosine release into the venous effluent. While there was a significant linear relation between adenosine formation and MVO2 in hearts perfused at both flow rates and stimulated with drugs, the relations differed with adenosine release being approximately fourfold greater in hearts perfused at 12 mL.min-1.g-1 under similar conditions of MVO2. Adenosine formation correlated exponentially with the ratio of oxygen supply:demand under all conditions (r = 0.97) and the relation did not differ significantly between hearts perfused at different rates.(ABSTRACT TRUNCATED AT 250 WORDS)     

Rates of breathing values and kinetics of respiration response in critical patterns of muscular activity in middle and long distance running

In elite runners, the ventilation influx, ventilation debt, and ventilation demand of the exercises were calculated on the basis of the pulmonary respiration dynamics during the maximum workout and recovery. The breathing values proved to closely reproduce the changes in the main parameters of oxygen demand at high intensity and duration of the exercise and can be used for quantification and standardization of exercise loads in sports. Three important factors of the aerobic exchange in the body were found to ensure the high level of the sports achievements in running: (1) general increase in the level of pulmonary ventilation (VE), oxygen demand (VO₂), and release of carbon dioxide (CO₂); (2) intensity of oxygen supply from lungs to the working muscles; (3) the rate of oxygenation (StO₂) and total rate of blood circulation.     

Oscillatory pattern in oxygen consumption of Hummingbirds

Mammals and birds offer the most conspicuous example of homeothermic endothermy, a metabolic feature that implies maintenance of a constant body temperature along broad ranges of ambient temperature. The concept of homeothermic endothermy has been developed in close association with the terms thermoneutral zone and basal metabolic rate. These two metabolic parameters, however, are not easily estimated in micro-endotherms, a difficulty that might emerge from intrinsic aspects of endothermy in minute animals. To address this issue, we used empirical work derived from theoretical considerations. Our theoretical analysis is based on a model of body temperature control by shifts in metabolic rate, and assumes that micro-endotherms lose heat very quickly due to body size, and exhibit a remarkable capacity to rapidly increase metabolic output. We found that these two metabolic traits can lead to non-equilibrium metabolic rate and body temperature. We then measured metabolic rate and body temperature during euthermia in two species of hummingbirds, and analyzed data using the χ² periodogram statistic and a power spectral analysis. We found long-range correlation in both oxygen consumption and body temperature during euthermia, a finding that suggests non-random 1/f oscillations. A similar pattern was not found in the rat, a much larger endotherm. Hummingbirds, then, do not appear to maintain steady-state metabolic conditions during euthermia. If, as we suggest, this pattern applies to micro-endotherms in general, the traditional concepts of thermoneutral zone and basal rate of metabolism might not apply to these animals.     

The respiratory responses of Carcinus maenas (L.) to changes in environmental salinity

Exposure of Carcinus to a reduction in salinity is accompanied by an increase in oxygen consumption. The highest rates of oxygen consumption are found during the first 2–3 h after a reduction in salinity, after which oxygen consumption then declines, but to a higher than ‘normal’ level which is maintained throughout the period of exposure to low salinity. This augmented rate of oxygen consumption persists even after 3–4 days in water of reduced salinity and indicates that the respiratory rate of Carcinus does not acclimate rapidly to reduced salinity. The increased oxygen consumption is associated with an increase in ventilation volume resulting from an increase in the rate of beating of the scaphognathites, while oxygen utilization remains at a low level. Since the oxygen-transporting properties of the blood of Carcinus show little change under conditions of reduced salinity, the increased oxygen demand of the tissues is met by a rise in cardiac output resulting mainly from an increase in heart rate.     

Development of homeothermy in the diving petrels Pelecanoides urinatrix exsul and P. georgicus, and the Antarctic prion Pachyptila desolata

Body temperatures of South Georgia diving petrel (P. georgicus) chicks increased from about 37.5 degrees C at hatching to between 38.5 and 39 degrees C within two weeks. Temperatures of common diving petrel P. u. exsul chicks averaged 38.8 degrees C after two weeks of age. Burrow temperatures varied between 5 and 10 degrees C. Measurements of oxygen consumption and body temperature indicated that chicks achieve effective endothermy at 5 degrees C after 9 days in P. u. exsul, 5-6 days in P. georgicus, and 0 days in the Antarctic prion (Pachyptila desolata). The maximum mass-specific, cold-induced oxygen consumption of small chicks that we could measure with our apparatus (ca. 5-6 cc O2/g per hr) was achieved at 5-6 days in P. u. exsul, 3 days in P. georgicus, and 0 days in P. desolata. Mass-specific thermal conductance decreased with age and body size in all 3 species, but was highest in P. u. exsul and lowest in P. desolata. Conductance was similar at the age of effective endothermy in all 3 species (ca. 3 J/g per hr per degrees C). The period required for the development of endothermy is related to age-specific changes in both conductance and capacity for heat production and it closely parallels the length of the brooding period. It is suggested that the length of the period of thermal dependence of the chick is related to the distance between feeding areas and the nesting site.     

Biomechanics of Running Indicates Endothermy in Bipedal Dinosaurs

Background

One of the great unresolved controversies in paleobiology is whether extinct dinosaurs were endothermic, ectothermic, or some combination thereof, and when endothermy first evolved in the lineage leading to birds. Although it is well established that high, sustained growth rates and, presumably, high activity levels are ancestral for dinosaurs and pterosaurs (clade Ornithodira), other independent lines of evidence for high metabolic rates, locomotor costs, or endothermy are needed. For example, some studies have suggested that, because large dinosaurs may have been homeothermic due to their size alone and could have had heat loss problems, ectothermy would be a more plausible metabolic strategy for such animals.

Methodology/Principal Findings

Here we describe two new biomechanical approaches for reconstructing the metabolic rate of 14 extinct bipedal dinosauriforms during walking and running. These methods, well validated for extant animals, indicate that during walking and slow running the metabolic rate of at least the larger extinct dinosaurs exceeded the maximum aerobic capabilities of modern ectotherms, falling instead within the range of modern birds and mammals. Estimated metabolic rates for smaller dinosaurs are more ambiguous, but generally approach or exceed the ectotherm boundary.

Conclusions/Significance

Our results support the hypothesis that endothermy was widespread in at least larger non-avian dinosaurs. It was plausibly ancestral for all dinosauriforms (perhaps Ornithodira), but this is perhaps more strongly indicated by high growth rates than by locomotor costs. The polarity of the evolution of endothermy indicates that rapid growth, insulation, erect postures, and perhaps aerobic power predated advanced “avian” lung structure and high locomotor costs.     

Effects of temperature and dissolved oxygen on heart rate,ventilation rate and oxygen consumption of spangled perch,Leiopotherapon unicolor (Günther 1859), (Percoidei,Teraponidae)

Summary Heart, ventilation and oxygen consumption rates ofLeiopotherapon unicolor were studied at temperatures ranging from 5 to 35°C, and during progressive hypoxia from 100% to 5% oxygen saturation. Biopotentials recorded from the water surrounding the fish corresponded to ventilation movements, and are thought to originate from the ventilatory musculature. Cardio-respiratory responses to temperature and dissolved oxygen follow the typical teleost pattern, with bradycardia, increased ventilation rate and reduced oxygen consumption occurring during hypoxia. However, ventilation rate did not increase at 15°C and below. Ventilation rate showed a slower response to increasing temperature (normoxic Q₁₀=1.39) than heart rate and oxygen consumption (normoxic Q₁₀=2.85 and 2.38).L. unicolor is unable to survive prolonged hypoxia by utilising anaerobic metabolism, but has a large gill surface area which presumably facilitates oxygen uptake in hypoxic environments. Periodic ventilation during normoxia in restingL. unicolor may improve ventilation efficiency by increasing the oxygen diffusion gradient across the gills.Abbreviations EBG electrobranchiogram - ECG electrocardiogram     

An allometric analysis of oxygen consumption rate and cardiovascular function in the cockroach, Blaberus discoidalis.

The interrelationship of metabolic rate and cardiovascular function has been well documented in vertebrates through allometric analyses. However, similar studies are lacking in insects. Unlike vertebrates, the cardiovascular system of insects does not play a significant role in oxygen transport. A comparison of the interrelationship in insects and vertebrates might provide insight into the nature of the connection between metabolic rate and the cardiovascular system. Oxygen consumption, heart rate and heart dimensions were measured in the nymphs of the cockroach Blaberus discoidalis over a mass range of 0.03-5 g. Oxygen consumption rate scaled with an exponent of 0.83. The relationship between heart rate and body mass scaled negatively, however, it did not appear to be linear. Using measurements of heart widths, abdominal length and heart rate, stroke volume and cardiac output were estimated. Cardiac output appeared to scale linearly with an exponent of 0.85, which was not significantly different from the exponent observed for the rate of oxygen consumption. Thus, the observed similarity between the exponents for oxygen consumption rate and cardiac output in vertebrates also appears to be present in insects.     

The heart as a working model to explore themes and strategies for anoxic survival in ectothermic vertebrates

Most vertebrates die within minutes when deprived of molecular oxygen (anoxia), in part because of cardiac failure, which can be traced to an inadequate matching of cardiac ATP supply to ATP demand. Cardiac power output (PO; estimated from the product of cardiac output and central arterial pressure and an indirect measure of cardiac ATP demand) is directly related to cardiac ATP supply up to some maximal level during both normoxia (ATP supply estimated from myocardial O(2) consumption) and anoxia (ATP supply estimated from lactate production rates). Thus, steady state PO provides an excellent means to examine anoxia tolerance strategies among ectothermic vertebrates by indicating a matching of cardiac glycolytic ATP supply and demand. Here, we summarize in vitro measurements of PO data from rainbow trout, freshwater turtles and hagfishes to provide a reasonable benchmark PO of 0.7 mW g(-1) for maximum glycolytic potential of ectothermic hearts at 15 degrees C, which corresponds to a glycolytic ATP turnover rate of about 70 nmol ATP g(-1) s(-1). Using this benchmark to evaluate in vivo PO data for hagfishes, carps and turtles, we identify two cardiac survival strategies, which in conjunction with creative waste management techniques to reduce waste accumulation, allow for long-term cardiac survival during anoxia in these anoxia-tolerant species. Hagfish and crucian carp exemplify a strategy of evolving such a low routine PO that routine cardiac ATP demand lies within the range of the maximum cardiac glycolytic potential. Common carp and freshwater turtles exemplify an active strategy of temporarily and substantially decreasing cardiac and whole body metabolism so that PO is below maximum cardiac glycolytic potential during chronic anoxia despite being quite close to this potential under normoxia.     

Alteration in myocardial oxygen balance during exercise after beta-adrenergic blockade in dogs

The effects of beta-adrenergic blockade upon myocardial blood flow and oxygen balance during exercise were evaluated in eight conscious dogs, instrumented for chronic measurements of coronary blood flow, left ventricular pressure, aortic blood pressure, heart rate, and sampling of arterial and coronary sinus venous blood. The administration of propranolol (1.5 mg/kg iv) produced a decrease in heart rate, peak left ventricular (LV) dP/dt, LV (dP/dt/P, and an increase in LV end-diastolic pressure during exercise. Mean coronary blood flow and myocardial oxygen consumption were lower after propranolol than at the same exercise intensity in control conditions. The oxygen delivery-to-oxygen consumption ratio and the coronary sinus oxygen content were also significantly lower. It is concluded that the relationship between myocardial oxygen supply and demand is modified during exercise after propranolol, so that a given level of myocardial oxygen consumption is achieved with a proportionally lower myocardial blood flow and a higher oxygen extraction.     

The emergence of endothermy in the black-footed and Laysan albatrosses

Eggs with pip-holes of the black-footed (Diomedea nigripes) and Laysan (Diomedea immutabilis) albatrosses were exposed to various air temperatures in the range 20–35°C in order to detect signs of incipient endothermy in late embryos. No evidence of endothermy was found. In contrast, the O₂ consumption of most hatchlings increased in response to cooling, the O₂ consumption at an air temperature of 25° C exceeding that between 34 and 35°C by 40%. In a minority of hatchlings this response was not seen. It was suggested that endothermy may develop at some time during the 24 h after hatching.Abbreviations bm body mass - C _total total thermal conductance of tissues and plumage - f respiratory frequency - FE_{O 2} fractional concentration of oxygen in air leaving chamber - FI_{O 2} fractional concentration of oxygen in air entering chamber - T _a an temperature - T _b deep-body temperature - V air-flow rate - VO₂ oxygen consumption     

Compensatory and adaptive rearrangement in the human respiratory system under acute hypoxic conditions

Relationships between the parameters of external respiration (minute volume and respiration rate) and those of internal, tissue respiration (oxygen consumption, arteriovenous oxygen difference and efficiency of oxygen uptake) were studied during a period of acute hypoxia and upon its completion. The subjects were exposed to hypoxia for 25 min using oxygen-nitrogen hypoxic gas mixtures (HGMs) differing in oxygen content (8 and 12%, HGM-8 and HGM-12, respectively). From the third to the fifth minutes of exposure to HGM-8, the respiration minute volume (RMV) was found to increase by 51 ± 33% as compared to the background value; however, the body’s oxygen consumption (OC) was 35 ± 22% reduced. Afterwards, OC grew to reach, from the 20th to the 25th min of hypoxia, 108 ± 21% of the background value and 181% of the value determined from the third to the fifth minutes of hypoxia. OC growth was accompanied by an insignificant RMV increase (by 12%) as compared to the level determined from the third to the fifth minutes of hypoxia, whereas the efficiency of oxygen uptake from the arterial blood increased by 75% for the same period. RMV growth from the third to the fifth minutes of hypoxia occurred as expense result of a higher breathing depth; at the same time, the respiration rate decreased as compared to the background value. By the period from the 20th to the 25th min of exposure to HGM-8, the respiration rate increased by 21% as compared to the period from the third to the fifth minutes of hypoxia. The efficiency of oxygen uptake from the arterial blood remained higher than the background value for at least 5 min after completion of the exposure to HGM-8. During the same period, the ventilation equivalent, an indicator of the efficiency of external respiration, i.e., of oxygen supply to the body, was significantly lower than the background value. During the exposure to HGM-12, RMV increased to a lesser extent than on exposure to HGM-8, however, the efficiency of oxygen uptake was higher during exposure to HGM-12; therefore, OC was also higher in the latter case. Therefore, the assumption that, during hypoxia, intensified external respiration (ventilatory response) itself compensates oxygen deficiency in inhaled air is revised. Ventilatory response is only a portion of the entire functional system of respiration (both external and tissue respiration). The role of ventilatory response is important for conditioning the tissue respiration rearrangement to eliminate deficiency of oxygen consumption during hypoxia. The retained higher oxygen uptake from the arterial blood during the period after completion of hypoxic treatment testifies to the adaptive implication of changes in tissue respiration; the same is confirmed by a reduced ventilation equivalent after hypoxia, which is indicative of the growing efficiency of external respiration, i.e., of an improved oxygen supply to the body.     

Behavioral inference of diving metabolic rate in free-ranging leatherback turtles

Good estimates of metabolic rate in free-ranging animals are essential for understanding behavior, distribution, and abundance. For the critically endangered leatherback turtle (Dermochelys coriacea), one of the world's largest reptiles, there has been a long-standing debate over whether this species demonstrates any metabolic endothermy. In short, do leatherbacks have a purely ectothermic reptilian metabolic rate or one that is elevated as a result of regional endothermy? Recent measurements have provided the first estimates of field metabolic rate (FMR) in leatherback turtles using doubly labeled water; however, the technique is prohibitively expensive and logistically difficult and produces estimates that are highly variable across individuals in this species. We therefore examined dive duration and depth data collected for nine free-swimming leatherback turtles over long periods (up to 431 d) to infer aerobic dive limits (ADLs) based on the asymptotic increase in maximum dive duration with depth. From this index of ADL and the known mass-specific oxygen storage capacity (To(2)) of leatherbacks, we inferred diving metabolic rate (DMR) as To2/ADL. We predicted that if leatherbacks conform to the purely ectothermic reptilian model of oxygen consumption, these inferred estimates of DMR should fall between predicted and measured values of reptilian resting and field metabolic rates, as well as being substantially lower than the FMR predicted for an endotherm of equivalent mass. Indeed, our behaviorally derived DMR estimates (mean=0.73+/-0.11 mL O(2) min(-1) kg(-1)) were 3.00+/-0.54 times the resting metabolic rate measured in unrestrained leatherbacks and 0.50+/-0.08 times the average FMR for a reptile of equivalent mass. These DMRs were also nearly one order of magnitude lower than the FMR predicted for an endotherm of equivalent mass. Thus, our findings lend support to the notion that diving leatherback turtles are indeed ectothermic and do not demonstrate elevated metabolic rates that might be expected due to regional endothermy. Their capacity to have a warm body core even in cold water therefore seems to derive from their large size, heat exchangers, thermal inertia, and insulating fat layers and not from an elevated metabolic rate.