Cassondra L. Williams

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.

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

SUMMARY 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) ≤24°C (9.5±5.7%) increased with increasing mean maximum dive depths (22.6±7.1 m; all P≤0.001). The FMRs increased with longer mean dive durations, bottom times and surface intervals and increased time spent in Tw≤24°C (all r2≥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.

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

Summary 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 O2 stores that underlie the ADL. Therefore, in order to assess strategies of O2 store management, we review (a) the magnitude of O2 stores, (b) past studies of O2 store depletion and (c) our recent investigations of O2 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 O2 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.

Heart rate regulation and extreme bradycardia in diving emperor penguins

SUMMARY To investigate the diving heart rate (fH) 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 (fH significantly<fH at rest) occurred during diving [dive fH (total beats/duration)=57±2 beats min–1, fH at rest=73±2 beats min–1 (mean ± s.e.m.)]. For dives less than the aerobic dive limit (ADL; duration beyond which [blood lactate] increases above resting levels), dive fH=85±3 beats min–1, whereas fH in dives greater than the ADL was significantly lower (41±1 beats min–1). In dives greater than the ADL, fH reached extremely low values: fH during the last 5 mins of an 18 min dive was 6 beats min–1. Dive fH and minimum instantaneous fH during dives declined significantly with increasing dive duration. Dive fH was independent of swim stroke frequency. This suggests that progressive bradycardia and peripheral vasoconstriction (including isolation of muscle) are primary determinants of blood oxygen depletion in diving emperor penguins. Maximum instantaneous surface interval fH in this study is the highest ever recorded for emperor penguins (256 beats min–1), equivalent to fH at V̇O2 max., presumably facilitating oxygen loading and post-dive metabolism. The classic Scholander–Irving dive response in these emperor penguins contrasts with the absence of true bradycardia in diving ducks, cormorants, and other penguin species.

Returning on empty: extreme blood O2 depletion underlies dive capacity of emperor penguins.

SUMMARY Blood gas analyses from emperor penguins (Aptenodytes forsteri) at rest, and intravascular PO2 profiles from free-diving birds were obtained in order to examine hypoxemic tolerance and utilization of the blood O2 store during dives. Analysis of blood samples from penguins at rest revealed arterial PO2s and O2 contents of 68±7 mmHg (1 mmHg= 133.3 Pa) and 22.5±1.3 ml O2 dl–1 (N=3) and venous values of 41±10 mmHg and 17.4±2.9 ml O2 dl–1 (N=9). Corresponding arterial and venous Hb saturations for a hemoglobin (Hb) concentration of 18 g dl–1 were >91% and 70%, respectively. Analysis of PO2 profiles obtained from birds equipped with intravascular PO2 electrodes and backpack recorders during dives revealed that (1) the decline of the final blood PO2 of a dive in relation to dive duration was variable, (2) final venous PO2 values spanned a 40-mmHg range at the previously measured aerobic dive limit (ADL; dive duration associated with onset of post-dive blood lactate accumulation), (3) final arterial, venous and previously measured air sac PO2 values were indistinguishable in longer dives, and (4) final venous PO2 values of longer dives were as low as 1–6 mmHg during dives. Although blood O2 is not depleted at the ADL, nearly complete depletion of the blood O2 store occurs in longer dives. This extreme hypoxemic tolerance, which would be catastrophic in many birds and mammals, necessitates biochemical and molecular adaptations, including a shift in the O2–Hb dissociation curve of the emperor penguin in comparison to those of most birds. A relatively higher-affinity Hb is consistent with blood PO2 values and O2 contents of penguins at rest.

O2 store management in diving emperor penguins

SUMMARY In order to further define O2 store utilization during dives and understand the physiological basis of the aerobic dive limit (ADL, dive duration associated with the onset of post-dive blood lactate accumulation), emperor penguins (Aptenodytes forsteri) were equipped with either a blood partial pressure of oxygen (PO2) recorder or a blood sampler while they were diving at an isolated dive hole in the sea ice of McMurdo Sound, Antarctica. Arterial PO2 profiles (57 dives) revealed that (a) pre-dive PO2 was greater than that at rest, (b) PO2 transiently increased during descent and (c) post-dive PO2 reached that at rest in 1.92±1.89 min (N=53). Venous PO2 profiles (130 dives) revealed that (a) pre-dive venous PO2 was greater than that at rest prior to 61% of dives, (b) in 90% of dives venous PO2 transiently increased with a mean maximum PO2 of 53±18 mmHg and a mean increase in PO2 of 11±12 mmHg, (c) in 78% of dives, this peak venous PO2 occurred within the first 3 min, and (d) post-dive venous PO2 reached that at rest within 2.23±2.64 min (N=84). Arterial and venous PO2 values in blood samples collected 1–3 min into dives were greater than or near to the respective values at rest. Blood lactate concentration was less than 2 mmol l–1 as far as 10.5 min into dives, well beyond the known ADL of 5.6 min. Mean arterial and venous PN2 of samples collected at 20–37 m depth were 2.5 times those at the surface, both being 2.1±0.7 atmospheres absolute (ATA; N=3 each), and were not significantly different. These findings are consistent with the maintenance of gas exchange during dives (elevated arterial and venous PO2 and PN2 during dives), muscle ischemia during dives (elevated venous PO2, lack of lactate washout into blood during dives), and arterio-venous shunting of blood both during the surface period (venous PO2 greater than that at rest) and during dives (arterialized venous PO2 values during descent, equivalent arterial and venous PN2 values during dives). These three physiological processes contribute to the transfer of the large respiratory O2 store to the blood during the dive, isolation of muscle metabolism from the circulation during the dive, a decreased rate of blood O2 depletion during dives, and optimized loading of O2 stores both before and after dives. The lack of blood O2 depletion and blood lactate elevation during dives beyond the ADL suggests that active locomotory muscle is the site of tissue lactate accumulation that results in post-dive blood lactate elevation in dives beyond the ADL.

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

SUMMARY 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 (O2) store. A dual wavelength near-infrared spectrophotometer was developed and used to measure myoglobin (Mb) O2 saturation levels in the locomotory muscle during dives of emperor penguins (Aptenodytes forsteri). Two distinct patterns of muscle O2 depletion were observed. Type A dives had a monotonic decline, and, in dives near the ADL, the muscle O2 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 O2 depletion during dives. The mean type A Mb desaturation rate allowed for calculation of a mean muscle O2 consumption of 12.4 ml O2 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 O2 store depletion.

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 (O2) 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 O2 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 O2 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.

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

ABSTRACT 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. Summary: Review of the literature and heart rate profiles of diving California sea lions emphasizes the dominance of the parasympathetic nervous system in heart rate regulation and the unlikelihood of morbid arrhythmias.