Blood oxygen transport and depletion in diving emperor penguins.

Oxygen store management underlies dive performance and is dependent on the slow heart rate and peripheral vasoconstriction of the dive response to control tissue blood flow and oxygen uptake. Prior research has revealed two major patterns of muscle myoglobin saturation profiles during dives of emperor penguins. In Type A profiles, myoglobin desaturated rapidly, consistent with minimal muscle blood flow and low tissue oxygen uptake. Type B profiles, with fluctuating and slower declines in myoglobin saturation, were consistent with variable tissue blood flow patterns and tissue oxygen uptake during dives. We examined arterial and venous blood oxygen profiles to evaluate blood oxygen extraction and found two primary patterns of venous hemoglobin desaturation that complemented corresponding myoglobin saturation profiles. Type A venous profiles had a hemoglobin saturation that (a) increased/plateaued for most of a dive's duration, (b) only declined during the latter stages of ascent, and (c) often became arterialized [arterio-venous (a-v) shunting]. In Type B venous profiles, variable but progressive hemoglobin desaturation profiles were interrupted by inflections in the profile that were consistent with fluctuating tissue blood flow and oxygen uptake. End-of-dive saturation of arterial and Type A venous hemoglobin saturation profiles were not significantly different, but did differ from those of Type B venous profiles. These findings provide further support that the dive response of emperor penguins is a spectrum of cardiac and vascular components (including a-v shunting) that are dependent on the nature and demands of a given dive and even of a given segment of a dive.

Diving physiology of marine mammals and birds: the development of biologging techniques.

In the 1940s, Scholander and Irving revealed fundamental physiological responses to forced diving of marine mammals and birds, setting the stage for the study of diving physiology. Since then, diving physiology research has moved from the laboratory to the field. Modern biologging, with the development of microprocessor technology, recorder memory capacity and battery life, has advanced and expanded investigations of the diving physiology of marine mammals and birds. This review describes a brief history of the start of field diving physiology investigations, including the invention of the time depth recorder, and then tracks the use of biologging studies in four key diving physiology topics: heart rate, blood flow, body temperature and oxygen store management. Investigations of diving heart rates in cetaceans and O2 store management in diving emperor penguins are highlighted to emphasize the value of diving physiology biologging research. The review concludes with current challenges, remaining diving physiology questions and what technologies are needed to advance the field. This article is part of the theme issue 'Measuring physiology in free-living animals (Part I)'.

Field Physiology: Studying Organismal Function in the Natural Environment.

Continuous physiological measurements collected in field settings are essential to understand baseline, free-ranging physiology, physiological range and variability, and the physiological responses of organisms to disturbances. This article presents a current summary of the available technologies to continuously measure the direct physiological parameters in the field at high-resolution/instantaneous timescales from freely behaving animals. There is a particular focus on advantages versus disadvantages of available methods as well as emerging technologies "on the horizon" that may have been validated in captive or laboratory-based scenarios but have yet to be applied in the wild. Systems to record physiological variables from free-ranging animals are reviewed, including radio (VHF/UFH) telemetry, acoustic telemetry, and dataloggers. Physiological parameters that have been continuously measured in the field are addressed in seven sections including heart rate and electrocardiography (ECG); electromyography (EMG); electroencephalography (EEG); body temperature; respiratory, blood, and muscle oxygen; gastric pH and motility; and blood pressure and flow. The primary focal sections are heart rate and temperature as these can be, and have been, extensively studied in free-ranging organisms. Predicted aspects of future innovation in physiological monitoring are also discussed. The article concludes with an overview of best practices and points to consider regarding experimental designs, cautions, and effects on animals. © 2021 American Physiological Society. Compr Physiol 11:1979-2015, 2021.

Cervical air sac oxygen profiles in diving emperor penguins: parabronchial ventilation and the respiratory oxygen store.

Some marine birds and mammals can perform dives of extraordinary duration and depth. Such dive performance is dependent on many factors, including total body oxygen (O2) stores. For diving penguins, the respiratory system (air sacs and lungs) constitutes 30-50% of the total body O2 store. To better understand the role and mechanism of parabronchial ventilation and O2 utilization in penguins both on the surface and during the dive, we examined air sac partial pressures of O2 (PO2 ) in emperor penguins (Aptenodytes forsteri) equipped with backpack PO2  recorders. Cervical air sac PO2  values at rest were lower than in other birds, while the cervical air sac to posterior thoracic air sac PO2  difference was larger. Pre-dive cervical air sac PO2  values were often greater than those at rest, but had a wide range and were not significantly different from those at rest. The maximum respiratory O2 store and total body O2 stores calculated with representative anterior and posterior air sac PO2  data did not differ from prior estimates. The mean calculated anterior air sac O2 depletion rate for dives up to 11 min was approximately one-tenth that of the posterior air sacs. Low cervical air sac PO2  values at rest may be secondary to a low ratio of parabronchial ventilation to parabronchial blood O2 extraction. During dives, overlap of simultaneously recorded cervical and posterior thoracic air sac PO2  profiles supported the concept of maintenance of parabronchial ventilation during a dive by air movement through the lungs.

The aerobic dive limit: After 40 years, still rarely measured but commonly used.

The aerobic dive limit (ADL) and the hypothesis that most dives are aerobic in nature have become fundamental to the understanding of diving physiology and to the interpretation of diving behavior and foraging ecology of marine mammals and seabirds. An ADL, the dive duration associated with the onset of post-dive blood lactate accumulation, has only been documented with blood lactate analyses in five species. Applications to other species have involved behavioral estimates or use of an oxygen store / metabolic rate formula. Both approaches have limitations, but have proved useful to the evaluation of the dive behavior and ecology of many species.

Activity, not submergence, explains diving heart rates of captive loggerhead sea turtles.

Marine turtles spend their life at sea and can rest on the seafloor for hours. As air-breathers, the breath-hold capacity  of marine turtles is a function of oxygen (O2) stores, O2 consumption during dives and hypoxia tolerance. However, some physiological adaptations to diving observed in mammals are absent in marine turtles. This study examined cardiovascular responses in loggerhead sea turtles, which have even fewer adaptations to diving than other marine turtles, but can dive for extended durations. Heart rates (fH) of eight undisturbed loggerhead turtles in shallow tanks were measured using self-contained ECG data loggers under five conditions: spontaneous dives, resting motionless on the tank bottom, resting in shallow water with their head out of water, feeding on squid, and swimming at the surface between dives. There was no significant difference between resting fH while resting on the bottom of the tank, diving or resting in shallow water with their head out of water. fH rose as soon as turtles began to move and was highest between dives when turtles were swimming at the surface. These results suggest cardiovascular responses in captive loggerhead turtles are driven by activity and apneic fH is not reduced by submergence under these conditions.

Heart rate regulation in diving sea lions: the vagus nerve rules.

Recent publications have emphasized the potential generation of morbid cardiac arrhythmias secondary to autonomic conflict in diving marine mammals. Such conflict, as typified by cardiovascular responses to cold water immersion in humans, has been proposed to result from exercise-related activation of cardiac sympathetic fibers to increase heart rate, combined with depth-related changes in parasympathetic tone to decrease heart rate. After reviewing the marine mammal literature and evaluating heart rate profiles of diving California sea lions (Zalophus californianus), we present an alternative interpretation of heart rate regulation that de-emphasizes the concept of autonomic conflict and the risk of morbid arrhythmias in marine mammals. We hypothesize that: (1) both the sympathetic cardiac accelerator fibers and the peripheral sympathetic vasomotor fibers are activated during dives even without exercise, and their activities are elevated at the lowest heart rates in a dive when vasoconstriction is maximal, (2) in diving animals, parasympathetic cardiac tone via the vagus nerve dominates over sympathetic cardiac tone during all phases of the dive, thus producing the bradycardia, (3) adjustment in vagal activity, which may be affected by many inputs, including exercise, is the primary regulator of heart rate and heart rate fluctuations during diving, and (4) heart beat fluctuations (benign arrhythmias) are common in marine mammals. Consistent with the literature and with these hypotheses, we believe that the generation of morbid arrhythmias because of exercise or stress during dives is unlikely in marine mammals.

Continuous arterial PO2 profiles in unrestrained, undisturbed aquatic turtles during routine behaviors.

Mammals and birds maintain high arterial partial pressure of oxygen (PO2 ) values in order to preserve near-complete hemoglobin (Hb) oxygen (O2) saturation. In diving mammals and birds, arterial O2 follows a primarily monotonic decline and then recovers quickly after dives. In laboratory studies of submerged freshwater turtles, arterial O2 depletion typically follows a similar pattern. However, in these studies, turtles were disturbed, frequently tethered to external equipment and confined either to small tanks or breathing holes. Aquatic turtles can alter cardiac shunting patterns, which will affect arterial PO2  values. Consequently, little is known about arterial O2 regulation and use in undisturbed turtles. We conducted the first study to continuously measure arterial PO2  using implanted microelectrodes and a backpack logger in undisturbed red-eared sliders during routine activities. Arterial PO2  profiles during submergences varied dramatically, with no consistent patterns. Arterial PO2  was also lower than previously reported during all activities, with values rarely above 50 mmHg (85% Hb saturation). There was no difference in mean PO2  between five different activities: submerged resting, swimming, basking, resting at the surface and when a person was present. These results suggest significant cardiac shunting occurs during routine activities as well as submergences. However, the lack of relationship between PO2  and any activity suggests that cardiac shunts are not regulated to maintain high arterial PO2  values. These data support the idea that cardiac shunting is the passive by-product of regulation of vascular resistances by the autonomic nervous system.

Hidden keys to survival: the type, density, pattern and functional role of emperor penguin body feathers.

Antarctic penguins survive some of the harshest conditions on the planet. Emperor penguins breed on the sea ice where temperatures drop below -40°C and forage in -1.8°C waters. Their ability to maintain 38°C body temperature in these conditions is due in large part to their feathered coat. Penguins have been reported to have the highest contour feather density of any bird, and both filoplumes and plumules (downy feathers) are reported absent in penguins. In studies modelling the heat transfer properties and the potential biomimetic applications of penguin plumage design, the insulative properties of penguin plumage have been attributed to the single afterfeather attached to contour feathers. This attribution of the afterfeather as the sole insulation component has been repeated in subsequent studies. Our results demonstrate the presence of both plumules and filoplumes in the penguin body plumage. The downy plumules are four times denser than afterfeathers and play a key, previously overlooked role in penguin survival. Our study also does not support the report that emperor penguins have the highest contour feather density.

Muscle energy stores and stroke rates of emperor penguins: implications for muscle metabolism and dive performance.

In diving birds and mammals, bradycardia and peripheral vasoconstriction potentially isolate muscle from the circulation. During complete ischemia, ATP production is dependent on the size of the myoglobin oxygen (O(2)) store and the concentrations of phosphocreatine (PCr) and glycogen (Gly). Therefore, we measured PCr and Gly concentrations in the primary underwater locomotory muscle of emperor penguin and modeled the depletion of muscle O(2) and those energy stores under conditions of complete ischemia and a previously determined muscle metabolic rate. We also analyzed stroke rate to assess muscle workload variation during dives and evaluate potential limitations on the model. Measured PCr and Gly concentrations, 20.8 and 54.6 mmol kg(-1), respectively, were similar to published values for nondiving animals. The model demonstrated that PCr and Gly provide a large anaerobic energy store, even for dives longer than 20 min. Stroke rate varied throughout the dive profile, indicating muscle workload was not constant during dives as was assumed in the model. The stroke rate during the first 30 s of dives increased with increased dive depth. In extremely long dives, lower overall stroke rates were observed. Although O(2) consumption and energy store depletion may vary during dives, the model demonstrated that PCr and Gly, even at concentrations typical of terrestrial birds and mammals, are a significant anaerobic energy store and can play an important role in the emperor penguin's ability to perform long dives.

In pursuit of Irving and Scholander: a review of oxygen store management in seals and penguins.

Since the introduction of the aerobic dive limit (ADL) 30 years ago, the concept that most dives of marine mammals and sea birds are aerobic in nature has dominated the interpretation of their diving behavior and foraging ecology. Although there have been many measurements of body oxygen stores, there have been few investigations of the actual depletion of those stores during dives. Yet, it is the pattern, rate and magnitude of depletion of O(2) stores that underlie the ADL. Therefore, in order to assess strategies of O(2) store management, we review (a) the magnitude of O(2) stores, (b) past studies of O(2) store depletion and (c) our recent investigations of O(2) store utilization during sleep apnea and dives of elephant seals (Mirounga angustirostris) and during dives of emperor penguins (Aptenodytes forsteri). We conclude with the implications of these findings for (a) the physiological responses underlying O(2) store utilization, (b) the physiological basis of the ADL and (c) the value of extreme hypoxemic tolerance and the significance of the avoidance of re-perfusion injury in these animals.

What triggers the aerobic dive limit? Patterns of muscle oxygen depletion during dives of emperor penguins.

The physiological basis of the aerobic dive limit (ADL), the dive duration associated with the onset of post-dive blood lactate elevation, is hypothesized to be depletion of the muscle oxygen (O(2)) store. A dual wavelength near-infrared spectrophotometer was developed and used to measure myoglobin (Mb) O(2) saturation levels in the locomotory muscle during dives of emperor penguins (Aptenodytes forsteri). Two distinct patterns of muscle O(2) depletion were observed. Type A dives had a monotonic decline, and, in dives near the ADL, the muscle O(2) store was almost completely depleted. This pattern of Mb desaturation was consistent with lack of muscle blood flow and supports the hypothesis that the onset of post-dive blood lactate accumulation is secondary to muscle O(2) depletion during dives. The mean type A Mb desaturation rate allowed for calculation of a mean muscle O(2) consumption of 12.4 ml O(2) kg(-1) muscle min(-1), based on a Mb concentration of 6.4 g 100 g(-1) muscle. Type B desaturation patterns demonstrated a more gradual decline, often reaching a mid-dive plateau in Mb desaturation. This mid-dive plateau suggests maintenance of some muscle perfusion during these dives. At the end of type B dives, Mb desaturation rate increased and, in dives beyond the ADL, Mb saturation often reached near 0%. Thus, although different physiological strategies may be used during emperor penguin diving, both Mb desaturation patterns support the hypothesis that the onset of post-dive lactate accumulation is secondary to muscle O(2) store depletion.

Extreme hypoxemic tolerance and blood oxygen depletion in diving elephant seals.

Species that maintain aerobic metabolism when the oxygen (O(2)) supply is limited represent ideal models to examine the mechanisms underlying tolerance to hypoxia. The repetitive, long dives of northern elephant seals (Mirounga angustirostris) have remained a physiological enigma as O(2) stores appear inadequate to maintain aerobic metabolism. We evaluated hypoxemic tolerance and blood O(2) depletion by 1) measuring arterial and venous O(2) partial pressure (Po(2)) during dives with a Po(2)/temperature recorder on elephant seals, 2) characterizing the O(2)-hemoglobin (O(2)-Hb) dissociation curve of this species, 3) applying the dissociation curve to Po(2) profiles to obtain %Hb saturation (So(2)), and 4) calculating blood O(2) store depletion during diving. Optimization of O(2) stores was achieved by high venous O(2) loading and almost complete depletion of blood O(2) stores during dives, with net O(2) content depletion values up to 91% (arterial) and 100% (venous). In routine dives (>10 min) Pv(O(2)) and Pa(O(2)) values reached 2-10 and 12-23 mmHg, respectively. This corresponds to So(2) of 1-26% and O(2) contents of 0.3 (venous) and 2.7 ml O(2)/dl blood (arterial), demonstrating remarkable hypoxemic tolerance as Pa(O(2)) is nearly equivalent to the arterial hypoxemic threshold of seals. The contribution of the blood O(2) store alone to metabolic rate was nearly equivalent to resting metabolic rate, and mean temperature remained near 37 degrees C. These data suggest that elephant seals routinely tolerate extreme hypoxemia during dives to completely utilize the blood O(2) store and maximize aerobic dive duration.

Heart rate regulation and extreme bradycardia in diving emperor penguins.

To investigate the diving heart rate (f(H)) response of the emperor penguin (Aptenodytes forsteri), the consummate avian diver, birds diving at an isolated dive hole in McMurdo Sound, Antarctica were outfitted with digital electrocardiogram recorders, two-axis accelerometers and time depth recorders (TDRs). In contrast to any other freely diving bird, a true bradycardia (f(H) significantly

Bioenergetics and diving activity of internesting leatherback turtles Dermochelys coriacea at Parque Nacional Marino Las Baulas, Costa Rica.

Physiology, environment and life history demands interact to influence marine turtle bioenergetics and activity. However, metabolism and diving behavior of free-swimming marine turtles have not been measured simultaneously. Using doubly labeled water, we obtained the first field metabolic rates (FMRs; 0.20-0.74 W kg(-1)) and water fluxes (16-30% TBW day(-1), where TBW=total body water) for free-ranging marine turtles and combined these data with dive information from electronic archival tags to investigate the bioenergetics and diving activity of reproductive adult female leatherback turtles Dermochelys coriacea. Mean dive durations (7.8+/-2.4 min (+/-1 s.d.), bottom times (2.7+/-0.8 min), and percentage of time spent in water temperatures (Tw) < or =24 degrees C (9.5+/-5.7%) increased with increasing mean maximum dive depths (22.6+/-7.1 m; all P< or =0.001). The FMRs increased with longer mean dive durations, bottom times and surface intervals and increased time spent in Tw< or =24 degrees C (all r2> or =0.99). This suggests that low FMRs and activity levels, combined with shuttling between different water temperatures, could allow leatherbacks to avoid overheating while in warm tropical waters. Additionally, internesting leatherback dive durations were consistently shorter than aerobic dive limits calculated from our FMRs (11.7-44.3 min). Our results indicate that internesting female leatherbacks maintained low FMRs and activity levels, thereby spending relatively little energy while active at sea. Future studies should incorporate data on metabolic rate, dive patterns, water temperatures, and body temperatures to develop further the relationship between physiological and life history demands and marine turtle bioenergetics and activity.