Aerobic Energy Production Research Articles

Inhibition by aluminum ion of NAD- and NADP-dependent isocitrate dehydrogenases from yeast

1. 1. NADP-dependent isocitrate dehydrogenase from yeast was potently inhibited by aluminum ion competitively with respect to the substrate isocitrate, and noncompetitively with the other substrate NADP. K i value was determined to be 0.43 μM. 2. 2. Aluminum ion acted as only a weak allosteric inhibitor of yeast NAD-dependent isocitrate dehydrogenase toward isocitrate, and as a noncompetitive inhibitor toward NAD. 3. 3. Inhibition by aluminum ion of NADP- and NAD-isocitrate dehydrogenases can reduce the aerobic energy production in yeast, and may contribute to the biological toxicity of aluminum in ecosystems and human life.

A procedure for selecting mammalian cells with an impairment in oxidative phosphorylation

The mitochondrial encephalomyopathies in man are characterized by heterogeneous defects leading to an impairment in the pathway of aerobic energy production. As a means of investigating the molecular and genetic mechanisms underlying these disorders we have developed a procedure for selecting mammalian cell lines with features resembling the human pathological phenotypes. The principles of the selection is the use of a fluroscent amphiphilic dye, 2,4-(dimethylamino)-1-styrylmethylpyridiniumiodine, a cation showing two main features. Firstly, it is accumulated by mitochondria to an extent correlated with the magnitude of the electrochemical gradient of protons across the mitochondrial inner membrane. Secondly, upon irradiation with UV light, it gives rise to formation of free radicals, which inflict damage to the cell. Mutant cells with an impairment in oxidative phosphorylation will have more chance to survive than wild type cells. The selection procedure was applied to a stock of mutagenized Chinese hamster ovary cells. After subcloning of the cells which survived the selection procedure, twenty-six independent clones were isolated. Eighteen of the clones had a partial deficiency of cytochrome c oxidase ranging from 39 to 60% of the activity in control cells. The properties of two of the clones are described. One clone has been cultured under non-selective conditions for at least 12 months with retention of the partial deficiency of cytochrome c oxidase.

Is intravenous glucose the only treatment for neonatal hypoglycaemia? : J.M. Hawdon and M.P. Ward Platt, Department of Child Health, University of Newcastle upon Tyne, U.K.

The unusual energy metabolism of elasmobranchs is characterized by limited or absent fatty acid oxidation in cardiac and skeletal muscle and a great reliance on ketone bodies and amino acids as oxidative fuels in these tissues. Other extrahepatic tissues in elasmobranchs rely on ketone bodies and amino acids for aerobic energy production but, unlike muscle, also appear to possess a significant capacity to oxidize fatty acids. This organization of energy metabolism is reflected by relatively low plasma levels of non-esterified fatty acids (NEFA) and by plasma levels of the ketone body ß-hydroxybutyrate that are as high as those seen in fasted mammals. The preference for ketone body oxidation rather than fatty acid oxidation in muscle of elasmobranchs under routine conditions is opposite to the situation in teleosts and mammals. Carbohydrates appear to be utilized as a fuel source in elasmobranchs, similar to other vertebrates. Amino acid- and lipid-fueled ketogenesis in the liver, the lipid storage site in elasmobranchs, sustains the demand for ketone bodies as oxidative fuels. The liver also appears to export NEFA and serves a buoyancy role. The regulation of energy metabolism in elasmobranchs and the effects of environmental factors remain poorly understood. The metabolic organization of elasmobranchs was likely present in the common ancestor of the Chondrichthyes ca. 400 million years ago and, speculatively, it may reflect the ancestral metabolism of jawed vertebrates. We assess hypotheses for the evolution of the unusual energy metabolism of elasmobranchs and propose that the need to synthesize urea has influenced the utilization of ketone bodies and amino acids as oxidative fuels.

Altered metabolic response of iron-deficient women during graded, maximal exercise

Metabolic responses during a standardized, progressive, maximal work capacity test on a cycle ergometer were studied in 11 women, mean age 28 (SEM 2) years, at admission to the study, after their body iron stores were depleted by diet, phlebotomy and menstruation for about 80 days and after iron repletion by diet for about 100 days, including daily iron supplementation (0.9 mmol iron as ferrous sulfate) for the last 14 days of repletion. Iron depletion was characterized by a decline (P less than 0.05) in hemoglobin, ferritin and body iron balance. Iron repletion, including supplementation, increased (P less than 0.05) hemoglobin, ferritin and iron balance. No changes were observed in cardiovascular and ventilatory responses or peak oxygen uptake. Iron depletion was associated with a reduced (P less than 0.05) rate of oxygen utilization, total oxygen uptake and aerobic energy expenditure, and elevated (P less than 0.05) peak respiratory exchange ratio and post-exercise concentration of lactate. Reduction of body iron stores without overt anemia affects exercise metabolism by reducing total aerobic energy production and increasing glycolytic metabolism.

Effects of pedaling speed on the power-duration relationship for high-intensity exercise

Seven males (age = 20.4 +/- 0.3 yr) each performed a total of eight exhaustive exercise bouts (four at 60 rpm and four at 100 rpm) in order to determine the influence of pedaling frequency on the parameters of the power-duration relationship for high-intensity cycle ergometry. The power-endurance time data for each subject at each rpm were fit by nonlinear regression to extract parameters of the hyperbolic: (P - theta PA). t = W', where P = power output, t = time to exhaustion, and theta PA and W' are constants. theta PA (the power asymptote, in watts (W] reflects an inherent characteristic of aerobic energy production during exercise, above which only a finite amount of work (W', in joules) can be performed, regardless of the rate at which the work is performed. theta PA at 60 rpm (235 +/- 8 W) was significantly (15.9 +/- 4.5%, P less than 0.05) greater than theta PA at 100 rpm (204 +/- 11 W), thus confirming our hypothesis that endurance would be compromised while cycling at the higher pedaling frequency. In contrast, W' was not significantly (P greater than 0.05) affected by cadence (16.8 +/- 1.7 kJ at 60 rpm vs 18.9 +/- 2.2 kJ at 100 rpm). Our data are consistent with the implications of previous investigations which demonstrated a greater cardiorespiratory and blood/muscle lactate response during constant-power exercise while cycling at high vs low rpm and indicate that the theoretical maximum sustainable power (i.e., theta PA) during cycle ergometry in untrained males is greater at 60 rpm than at 100 rpm.

Tricarboxylic acid cycle intermediates in human muscle during prolonged exercise

Seven subjects cycled to fatigue [75 +/- 5 (SE) min] at a work load corresponding to approximately 75% of their maximal oxygen uptake. Biopsies were taken from the quadriceps femoris muscle at rest and during exercise. Muscle glycogen decreased from a preexercise level of 445 +/- 33 mmol glucosyl units/kg dry wt to 50 +/- 14 at fatigue. The sum of the measured tricarboxylic acid cycle intermediates (TCAI = malate + citrate + fumarate + oxaloacetate) was 0.49 +/- 0.05 mmol/kg dry wt at rest, increased to 4.41 +/- 0.23 after 5 min of exercise, and then decreased continuously to 3.33 +/- 0.29 and to 2.83 +/- 0.27 mmol/kg dry wt after 40 min of exercise and at fatigue (P less than 0.05 vs. 5 min), respectively. The point of fatigue was characterized by an enhanced deamination of AMP (judged by increase in IMP) and reduced contents (vs. 5 min of exercise) of lactate, pyruvate, and alanine. In contrast, acetylcarnitine (reflects the availability of acetylunits) increased threefold at the onset of exercise and was maintained approximately at this level until fatigue. It is concluded that prolonged exercise to fatigue at moderate work loads results in glycogen depletion, energy deficiency (increased AMP deamination), reduced levels of three-carbon compounds and TCAI (compared with the initial phase of exercise) but in maintained levels of acetylunits. The present data indicate that carbohydrate depletion may impair aerobic energy production by reducing the level of TCAI.

Mechanical work and efficiency in treadmill running at aerobic and anaerobic thresholds

Mechanical work, mechanical power, energy consumption and mechanical efficiency were studied in constant-speed treadmill running of 5 min at seven different exercises around aerobic (AerT) and anaerobic (AnT) thresholds. The true efficiency of concentric (positive) mechanical work and gross efficiency of the whole body in seven male subjects were calculated. The total mechanical work was calculated from film through the translational, potential and rotational energy states as the sum of the changes of all the mechanical energy states in all body segments allowing energy transfer between segments and from energy state to state. The total energy consumption was measured by combining aerobic and anaerobic energy production in resting and working conditions. When the speed of the treadmill was increased from the velocity of 10 km h-1 (2.8 m s-1) to 22 km h-1 (6.1 m s-1), the concentric mechanical work per one step increased from 129 +/- 45 J to 228 +/- 82 J (P less than 0.01). Oxygen consumption increased from 2.22 +/- 0.27 1 min-1 to 4.47 +/- 0.24 1 min-1. The amount of blood lactate increased from 0.94 +/- 0.53 mmol l-1 at the lowest speed to 9.90 +/- 2.89 mmol l-1 at the highest speed (P less than 0.001). The true efficiency of concentric work decreased from 74 +/- 14% to 56 +/- 8% (P less than 0.05). At the speed of the AerT, the economy of running, the vertical rise of different body segments and mechanical efficiency of positive work were high. The highest gross efficiency was found at the running speed between the AerT and AnT.

Reduction in Spinal Cord Postischemic Lactic Acidosis and Functional Improvement with Dichloroacetate

Pyruvate dehydrogenase complex (PDHC) is a major enzyme of glucose metabolism. Dichloroacetate (DCA) is a noncompetitive inhibitor of PDHC kinase, an enzyme that inactivates PDHC. We examined the effects of DCA on extracellular lactate and pyruvate concentration changes and spinal somatosensory evoked potentials (SSEP) in ischemic rabbit spinal cords. In the first group of 26 animals, the aorta was occluded until postsynaptic SSEP waves were completely suppressed for 10 min, a period of ischemia that causes neurologic deficits in 50% of untreated animals. DCA (25 mg/kg) was given to 13 of these animals before ischemia. In the second group of 24 animals, the aorta was occluded until the postsynaptic SSEP waves were absent for 20 min, a period of ischemia that produces paraplegia in 100% of untreated animals. DCA (25 mg/kg) was given to 16 of these animals just before the aortic occlusion was released. After occlusion, extracellular spinal lactate concentrations increased abruptly while pyruvate concentrations fell. Both lactate and pyruvate concentrations reached a plateau during the ischemic period but increased when the aortic balloon was deflated. DCA-treated animals had lower lactate and pyruvate peak concentrations during reperfusion, as well as more rapid and greater recovery of SSEP at 2 h after reperfusion. DCA did not alter spinal metabolism during the ischemia but appeared to produce a more rapid shift to glucose metabolism on reperfusion. Thus, DCA treatment resulted in better electrophysiological recovery after both moderate and severe ischemia, either by reducing lactic acidosis or by increasing the recovery rate of aerobic energy production.

Nucleated red cell function: metabolism and pH regulation

The full extent and apportionment of aerobic and anaerobic contributions to energy transduction for membrane pumps associated with cellular pH regulation are very poorly understood. One way of approaching this problem at the cellular level is by using the nucleated erythrocyte as a model cell. Indeed, the aerobic and anaerobic capacity of salmonid erythrocytes and their β-adrenergic mediated pH regulation offers a model "pH regulating system" for examining cellular strategies of response to acute and (or) chronic changes in oxygen availability. Much of our work has focused on the balance between metabolic energy production and the maintenance of erythrocytic pH through primarily or secondarily active ionic exchange mechanisms at the cell membrane. Upon adrenergic stimulation, a rise in cyclic AMP activates the Na+–H+ exchanger, leading to cell alkalinization and an elevation of intracellular Na+. The increased Na+ evidently stimulates Na+,K+-ATPase activity and the increased ATP consumption is matched with aerobic energy production. The pHi that is subsequently established appears to be set by levels of poly anionic phosphates.

Phosphoinositide metabolism and metabolism-contraction coupling in rabbit aorta.

We tested a hypothesis that metabolism-contraction coupling in vascular smooth muscle is controlled by the rate of delivery of energy to ATP-dependent reactions in the inositol phospholipid transduction system that generate second messengers exerting control on smooth muscle force. Rabbit aorta was contracted by norepinephrine (NOR) under conditions of normoxia and hypoxia (bath PO2 less than 40 mmHg), and changes in inositol phospholipid pool sizes and metabolic flux rates (JF) were determined. JF was determined by labeling free cytosolic myo-inositol by incubation of unstimulated muscle with myo-[3H]inositol and then measuring rates of incorporation of this isotope into inositol phospholipids and inositol phosphates when the muscle was activated by NOR. JF measured during maintenance of NOR-induced force was markedly inhibited during hypoxia to 40-50% of that determined during normoxia; rates of increases in inositol phosphate radioactivities were similarly depressed during NOR activation under hypoxia. The hypoxia-induced decrease in JF was associated with four- to fivefold increase in phosphatidylinositol 4-phosphate (PIP) total pool size, suggesting PIP kinase was inhibited and rate limiting. Total pool sizes of phosphatidylinositol, phosphatidylinositol bisphosphate, and phosphatidic acid were unchanged from values seen during activation under normoxia. These data suggest that activation of inositol phospholipid metabolism, which generates inositol 1,4,5-trisphosphate (IP3) and diacylglycerol, is blunted under conditions where aerobic energy production is inhibited. Data are consistent with "rate-limiting" effects of decreased ATP delivery, or decreased phosphate potential, on PIP kinase and reactions that control resynthesis of phosphatidylinositol.

Oxygen deficit at the onset of submaximal exercise is not due to a delayed oxygen transport.

Six subjects cycled on two occasions for 10 min at power output of 188 +/- 11 W (means +/- SEM), which corresponded to 70 +/- 2% of their maximal oxygen uptake (VO2 max). The exercise intensity was either increased gradually in a stepwise manner over about 15 min (slow transition-S), or increased directly (direct transition-D) to the predetermined power output. Muscle samples from the quadriceps femoris muscle were taken at rest and immediately after exercise in both trials. During exercise with both D and S muscle lactate increased approximately 10 times (P less than 0.01), phosphocreatine decreased about 50% (P less than 0.01) and ADP increased about 20% (P less than 0.05). There were no significant differences between S and D (P greater than 0.05). Furthermore, blood lactate, O2 deficit, O2 debt, and the calculated increase in muscle content of inorganic phosphate (Pi) were all similar between D and S (P greater than 0.05). It is concluded that the O2 deficit and the anaerobic energy utilization is not affected by the rate of transition from rest to exercise. Consequently, the O2 deficit at the onset of exercise is not due to a delay in O2 transport, but may be due to a limited peripheral O2 utilization as a result of metabolic adjustments at the cellular level. Increases in ADP and Pi are suggested to be primary metabolic regulators which activate both aerobic and anaerobic energy production resulting in the O2 deficit.

Operation of the purine nucleotide cycle in animal tissues.

Summary1. The operation of the purine nucleotide cycle, consisting of the enzymes adenylate deaminase (E.C. 3.5.4.6), adenylosuccinate synthetase (E.C. 6.3.4.4) and adenylosuccinate lyase (E.C. 4.3.2.2), has been reviewed with reference to its metabolic function in animal tissues.2. Abundant evidence, both from in vitro and in vivo studies, suggests that the purine nucleotide cycle serves to stabilize the adenylate ‘energy charge’ (or ‘phosphorylation potential’) in the cytoplasm of vertebrate cells during a temporary imbalance between ATP‐consumption and ATP‐production. This stabilization, however, is absent or much less efficient in tissues of invertebrates.3. The hypothesis that AMP‐deaminase is involved in the regulation of glycolysis is not supported by recent work. In a variety of cell types, including skeletal muscle and blood platelets, blocking of AMP‐deaminase activity (due to a genetic defect or to pharmacological inhibition) is without effect on the glycolytic rate. Detailed kinetic and histochemical analysis of energy metabolism shows lack of correlation between AMP‐deaminase activity and glycolysis in skeletal muscle during exercise.4. The purine nucleotide cycle appears to control the level of citric acid cycle intermediates in skeletal muscle. Pharmacological inhibition of adenylosuccinate lyase or adenylosuccinate synthetase leads to a reduced availability of four‐carbon ‘sparker’ molecules to the Krebs cycle with a concomitant impairment of aerobic energy production during muscular work.5. The cycle appears to be a major pathway for amino acid deamination in skeletal muscle and brain of vertebrates, but not in kidney or liver.

Free amino acids in developing eggs and yolk-sac larvae of the cod,Gadus morhuaL.

Abstract The cod egg contains about 200 nmoles of free amino acids (FAA) at spawning, decreasing to about 100 nmoles at hatching and about 25 nmoles at the end of the yolk-sac stage. The decrease occurs without a corresponding buildup of body proteins but is correlated with ammonia production. The measured oxygen uptake of the developing cod eggs and yolk-sac larvae matches with the oxygen demand necessary to catabolize the FAA lost during this period. The conclusion drawn is that FAA, mainly alanine, leucine, and serine, are the substrate for aerobic energy production of cod eggs and yolk-sac larvae. The implication for the preparation of a favourable first-feed diet of cod larvae, is stressed.

Mechanical work and efficiency in ergometer bicycling at aerobic and anaerobic thresholds.

Internal and external mechanical work, energy consumption and mechanical efficiency were studied in constant-load ergometer bicycling at five different power outputs below, equal to, and above the aerobic (AerT) and anaerobic (AnT) thresholds. The gross, net and true efficiencies of the whole body in five male subjects were calculated. The work against the external load was defined as the external mechanical work. The internal mechanical work was calculated as the sum of the increments of kinetic and potential energy in all body segments by using methods of film analysis. Total energy consumption was measured by combining aerobic and anaerobic energy production. When the power output of the bicycle ergometer was increased from 146 +/- 15 to 283 +/- 17 W, oxygen consumption increased from 2.20 +/- 0.98 to 4.22 +/- 0.20 l min-1 (P less than 0.001), while the oxygen consumption at rest was 0.30 +/- 0.03 l min-1. The concentration of blood lactate increased from 2.2 +/- 0.4 at the lowest work load to 8.6 +/- 1.2 mmol l-1 at the highest work load (P less than 0.001). The amount of external work done per revolution increased from 139 +/- 20 to 277 +/- 29 J (P less than 0.001), while the amount of internal work per revolution remained almost constant (56 +/- 12 J). The gross efficiency in the present study was 17-20%, net efficiency 18-22% and true efficiency 21-30%, respectively. The highest gross and net efficiencies were reached at the AerT. The lowest efficiencies were obtained at highest work load.(ABSTRACT TRUNCATED AT 250 WORDS)

Free amino acids as energy substrate in developing eggs and larvae of the cod Gadus morhua

The content of free amino acids (FAA) in the cod (Gadus morhua L.) egg is about 200 nmol at spawning, decreasing by about 100 nmol/egg during the egg stage and about 75 nmol/larva during the yolksac larval stage. Together, alanine, leucine, serine, isoleucine, lysine, and valine account for about 75% of the decrease. Ammonium accumulates gradually during the egg stage and is quickly excreted after hatching. The body protein content is maintained during the egg and yolksac larval stages. The measured oxygen uptake of the cod embryo during the egg and yolksac larval stages accounts for about 85% of the oxygen necessary to catabolize the FAA disappearing during this period. Ammonia excretion of the cod embryo, as taken from literature data, is similar to the expected ammonia production from catabolism of the FAA. Our data suggest that FAA are a major substrate for aerobic energy production in cod eggs and yolksac larvae. The implication of this finding for the production of a favourable first-feed for cod and other cultivated marine fish larvae, and for the selection of high quality eggs of marine fishes, is stressed.

Effects of hydrostatic pressure per se (101 ATA) on energetic processes in fish

1. 1. Nucleotides concentrations (ATP, ADP, AMP) have been measured in brain and muscle of eels exposed to 101 ATA of hydrostatic pressure (HP) for 3 hr. Survival times (ST) and oxygen arterial content (CaO 2) have been measured in trouts exposed to HP= 101 ATA. 2. 2. The results show that at HP = 101 ATA, AMP increases ( P < 0.05) and ATP decreases (−12%; NS) in muscle but are not modified in brain; ST values are similar in normoxic and hyperoxic conditions, and CaO 2 are similar at 1 ATA and 101 ATA of HP. 3. 3. It is concluded that HP tends to decrease aerobic production of energy. This phenomenon is not due to a failure in O 2 transport from ambient medium to the cell but to a possible perturbation of the aerobic cellular processes leading to energy production (Krebs cycle and/or respiratory chain coupled to oxydative phosphorylation).

Importance of purine nucleotide cycle to energy production in skeletal muscle.

The purpose of this study was to determine the role of the purine nucleotide cycle in aerobic energy production. Rats received either saline or 5-amino-4-imidazolecarboxamide riboside (AICAriboside), a precursor to an inhibitor of adenylosuccinate lyase (AICAR). Muscle tension was quantified during gastrocnemius stimulation, and muscle metabolite content was measured to obtain an estimate of the activity of the enzymes of the cycle. AICAriboside prevented the increase in synthetase and lyase activities observed in control animals during moderate (aerobic) stimulation, and was accompanied by marked muscle dysfunction. Although glycolytic energy production was not impaired in the AICAriboside-treated animals (lactate production occurred), total energy production did not meet energy demand. These results suggest that disruption of the purine nucleotide cycle impairs aerobic energy metabolism. Tetanic (anaerobic) stimulation produced more rapid fatigue in the AICAriboside-treated group. Total energy production was again impaired in the AICAriboside-treated animals, but lactate production was similar in both groups. These findings suggest the loss of the initial aerobic component of energy generation in tetanically stimulated muscle of AICAriboside-treated animals. The results of this study indicate that disruption of the purine nucleotide cycle at the level of the synthetase and lyase reactions is associated with skeletal muscle dysfunction, and suggest that the cycle plays an anapleurotic role in providing citric acid cycle intermediates that enhance aerobic energy production in contracting skeletal muscle.

The anaerobic threshold: physiological and clinical significance. International Symposium on the Clinical Value of the Anaerobic Threshold in Heart and Lung Diseases. Veruno, November 22-23, 1985. Proceedings.

During exercise, the oxygen consumption above which aerobic energy production is supplemented by anaerobic mechanisms, causing a sustained increase in lactate and metabolic acidosis, is termed the anaerobic threshold (AT). The oxygen consumption at the AT depends on factors that affect oxygen delivery to the tissues. It is increased when oxygen flow is enhanced and decreased when oxygen flow is diminished. Its value is quite low in patients with heart disease. The AT is an important functional demarcation since the physiological responses to exercise are different above the AT compared to below the AT. Above the AT, in addition to the development of metabolic acidosis, exercise endurance is reduced, VO2 kinetics are slowed so that a steady state is delayed, and VE increases disproportionately to the metabolic requirement and a progressive tachypnea develops. The AT can be measured directly from the lactate concentration with precise threshold detection from a log-log transformation of lactate and VO2. This threshold also defines the VO2 above which the lactate/pyruvate ratio increases. As bicarbonate changes reciprocally with lactate, its measurement can also be used to estimate the lactate threshold. But most convenient are gas exchange measurements made during exercise testing which can be used to noninvasively detect the lactate or anaerobic threshold. These methods are based on the physical-chemical event of buffering lactic acid with bicarbonate, and the increased CO2 output which occurs in association with the acute development of a metabolic acidosis.

Mitochondrial diseases

The last two decades have witnessed the discovery of a novel group of inborn metabolic errors which directly impinge upon the pathways of aerobic energy production in the mitochondrial inner membrane and matrix space. The spectrum of reported biochemical abnormalities includes defects which impair the entry of high energy substrates into the mitochondria or the capacity to generate reducing potential from these substrates, as well as those that block the oxidative phosphorylation pathway itself. Clinical expression varies according to the nature, severity and tissue distribution of the metabolic block. Aerobically active organs with a high demand for ATP, such as the brain, retina, cardiac and skeletal muscle, are particularly vulnerable to defects of this type and may be affected either singly or in different combinations. The molecular and genetic mechanisms underlying these disorders have not yet been clearly identified but there are strong theoretical reasons for believing that some of them may be due to mutations affecting the mitochondrial genetic system.

Energetics of the Exercising Wharf Crab Sesarma cinereum

Crabs (1.9 g) were exercised for up to 60 min on a miniature treadmill respirometer. O₂ consumption ($\dot{V}O_{2}$) at all velocities was low, a maximum of only 1.5 times the resting rate. $\dot{V}O_{2}$ was not related to the velocity of running. Five weeks of treadmill training produced a minor rise in $\dot{V}O_{2}$ at the maximum sustainable velocity, but the highest $\dot{V}O_{2}$ was still only two times the resting $\dot{V}O_{2}$. Crabs exercising in 100% O₂ significantly increased their endurance capacity, indicating that this crab's O₂ conductance system is severely limited. Anaerobic metabolism with lactate as an end product supplemented aerobic energy production to a minor extent. Wholebody lactate (WBL) increased at a constant but small rate (0.10 mg·g⁻¹·min⁻¹) when the crab ran at the highest velocity (0.14 km·h⁻¹). Wharf crabs fatigued rapidly when run in a 100% N₂ atmosphere, indicating they do not have unusual anaerobic capacities or unusually high levels of O⠂ stores or phosphagen reserv...