Body and blubber relationships in antarctic pack ice seals: implications for blubber depth patterns.

Morphometrics and blubber depths from all four high Antarctic seals (Weddell, Ross, crabeater, and leopard) were obtained during a midsummer research cruise in the Ross Sea as the physiological ecology component of the U.S. Antarctic Pack Ice Seals project. These data are the only in vivo measurements of all four species from the same location and time of year and focused on variances in morphometrics and blubber depth related to species, sex, and age. By controlling for location and season, this cross-species design provided the means to differentiate how blubber mass might be influenced in these groups. We measured both absolute blubber depth and ratio of blubber depth to body core diameter. We found that adult and younger animals showed differences in blubber depth, but male versus female seals did not show differences within any given species. However, when compared across species, the ratio of blubber ring depth to body core diameter suggests that adult Weddell seals differ in their use of blubber compared with the other three species. We propose that this difference in blubber pattern is most likely related to Weddell nutritional requirements during the breeding season having a greater influence on blubber depth than thermal requirements when compared with the other three species.

Digestive constraints on an aquatic carnivore: effects of feeding frequency and prey composition on harbor seals.

We hypothesized that increased feeding frequency in captive harbor seals would increase nutrient loads and thus reduce retention time and the digestive efficiency of natural prey. We measured daily feed intake and excretion during 6 feeding trials and fed herring (49% lipid), pollock (22% lipid) or an equal mix of each diet over 24 months. Animals were accustomed to feeding at either high or low frequency. Body mass and intake did not vary with season. Although mean retention times were similar between diets and feeding frequencies, solute and particulate digesta markers separated at high feeding frequency. Consistent dry matter digestibility resulted in greater gut fill from pollock than from herring. Digestible energy intakes from pollock were approximately 25% greater than from either herring or the mixed diet. Lipid digestibility of herring declined from 90% to 50% when lipid intake exceeded 60 g kg(-0.75) day(-1). Our hypothesis of a trade-off between intake and digestion was not supported for protein but was supported for lipid. Results of this study imply that a flexible digestive system for harbor seals can compensate for ingesting prey of lower energy density by increasing gut fill and enhancing protein and lipid assimilation, to sustain digestible energy intake.

Adaptations to pressure in the RBC metabolism of diving mammals.

Marine mammals are known to dive up to 2000 m and, therefore, tolerate as much as 200 atm. of hydrostatic pressure. To examine possible metabolic adaptations to these elevated pressures, fresh blood samples from marine and terrestrial mammals were incubated for 2 h at 37 degrees C under 136 atm (2000 psi) of hydrostatic pressure. The consumption of plasma glucose and the production of lactate over the 2-h period were used to assess glycolytic flux in the red cells. The results indicate that glycolytic flux as measured by lactate production under pressure can be significantly depressed in most terrestrial mammals and either not altered or accelerated in marine mammals. The data also suggest that there is a significant shift in the ratio of lactate produced to glucose consumed under pressure. Interestingly, human and dolphin blood do not react to pressure. These combined data imply a metabolic adaptation to pressure in marine mammal RBC that may not be necessary in human or dolphin cells due to their unique patterns of glucose metabolism.

Health status of young Alaska Steller sea lion pups (Eumetopias jubatus) as indicated by blood chemistry and hematology.

Blood chemistry and hematology were examined in 238 Steller sea lion pups (Eumetopias jubatus) to assess the health status of pups <1 month of age. Failure of juvenile recruitment (possibly due to nutritionally or physiologically compromised pups) into breeding populations has been proposed as a cause of recent declines of this endangered species in Alaska. To identify potential correlations with areas of high population decline, blood chemistry data were considered for three areas: eastern Aleutian Islands (low rates of population decline to stable populations), Gulf of Alaska (high rates of decline), and Southeast Alaska (stable to increasing population). Southeast Alaska pups showed elevated ketone body concentrations (beta-hydroxybutyrate,(beta-HBA)) and depressed glucose levels than pups in the Gulf of Alaska. Over 40% of the pups from Southeast Alaska had elevated beta-HBA concentrations suggesting they underwent longer periods of fasting than seen in pups from other areas. Hematocrit (Hct), hemoglobin concentration (Hb) and water content of the blood exhibited typical mammalian relationships. In summary, blood chemistry and hematology data showed no indication that Steller sea lion pups <1 month old from areas of population decline were nutritionally compromised.

Plasma angiotensin II, arginine vasopressin and atrial natriuretic peptide in free ranging and captive seals and sea lions.

We used radioimmunoassay methods to quantify arginine vasopressin (AVP), atrial natriuretic peptide (ANP), and angiotensin II (Ang II) in plasma samples from harbor seals (Phoca vitulina richardsii), Weddell seals (Leptonychotes weddellii), northern elephant seals (Mirounga angustirostris), ringed seals (Phoca hispida), California sea lions (Zalophus californianus), and Steller sea lions (Eumetopias jubatus). Plasma concentrations of AVP, ANP, and Ang II in these pinniped species were within the ranges reported for other vertebrates under resting conditions. However, there were species, geographic and developmental variations in these hormones: Levels of AVP in plasma samples from adult Steller sea lions and harbor seals were higher than in pups of the same species; higher levels of plasma ANP were found in wild captured Alaskan Steller sea lions and in hunted ringed seals; differences in plasma levels of all three hormones were found throughout the geographic distribution of harbor seals and Steller sea lions in Alaska. This is the first report on circulating concentrations of vasoactive hormones in pinnipeds, and demonstrates that further studies are needed to ascertain the natural variability in these levels with the impact of molting, fasting, diving and environmental factors in seals and sea lions.

Changes in the plasma levels of vasoactive hormones during apnea in seals.

Prolonged and repetitive breath-hold periods (apnea) during and sleep are a routine component in the ecological physiology of marine mammals. Seals are among the few mammals in which control of heart rate (HR) can be studied independent of respiration, without pharmacochemical manipulation. We hypothesized that the vasoactive hormones angiotensin II (Ang II), arginine vasopressin (AVP), and atrial natriuretic peptide (ANP) were involved in the control of cardiovascular function in seals, and that the relationship was dependent upon input from the respiratory system. Venous plasma samples were collected and electrocardiograms were recorded from Northern elephant seal (Mirounga angustirostris) and Weddell seal (Leptonychotes weddellii) pups during both spontaneous breathing (eupnea) and apnea. Instantaneous HR and simultaneous plasma levels of ANP, AVP, and Ang II from periods of eupnea and apnea were compared. In these seal pups, apnea was associated with bradycardia, increased ANP, and decreased AVP and Ang II. The results support the hypothesis of a complex involvement between the vasoactive hormones and the control of cardiovascular function, and provide evidence for differential levels of control during periods of eupnea and apnea.

Plasma haptoglobin levels in threatened Alaskan pinniped populations.

We evaluated the plasma concentration of the acute phase protein haptoglobin (Hp) from Steller sea lions (Eumetopias jubatus) and harbor seals (Phoca vitulina) in regions of Alaska (USA) where the populations of these pinnipeds were declining and compared the values with concentrations of Hp from the same species in areas where the populations were stable. Samples were collected from 1992 through 1994 at sites in Southeast Alaska, Prince William Sound, the Gulf of Alaska, and the Aleutian Islands. Significantly higher levels of Hp were found in the samples from the areas of decline compared to those from stable populations. Based on these findings, we propose that one may be able to distinguish these compromised pinniped populations using Hp as a biomedical indicator.

Developmental changes in cardiorespiratory patterns of sleep-associated apnea in northern elephant seals.

The development of heart rate and respiratory patterns related to sleep-associated apnea were studied in northern elephant seal (Mirounga angustirostris) pups and adult males. Heart rate patterns became more refined in conjunction with an age-related increase in apnea duration in pups. That is, older pups showed significant sinus arrhythmia, while breathing and apneic heart rate were both stable and similar in magnitude to the sinus arrhythmia minimum. By contrast, younger pups showed poor or nonexistent sinus arrhythmia and shorter apnea durations with a variable and/or high heart rate during apnea. Apnea duration was positively correlated with the development of sinus arrhythmia and negatively correlated with both eupneic and apneic heart rate. Adult males showed extremely well-developed patterns of sinus arrhythmia and, in all cases, the minimum heart rate during the sinus arrhythmia was lower than the average heart rate recorded during apnea. These results suggest that seal pups are not born with the cardiac control associated with voluntary long duration apnea, but that apnea tolerance increases with refined cardiorespiratory control.

Patterns of respiration and heart rate during wakefulness and sleep in elephant seal pups.

Although breath holding during diving has been studied extensively in seals, the recent observation that these mammals also exhibit long-duration apnea while apparently sleeping has not been systematically examined. This project examined sleep apnea in northern elephant seal pups (Mirounga angustirostris). The animals exhibited a sequential sleep pattern of wakefulness-slow-wave sleep (SWS)-rapid eye movement (REM) sleep that resembled the normal pattern of mammalian sleep. The typical respiratory pattern during sleep in 4-mo-old pups consisted of short periods of continuous breathing separated by periods of apnea of up to 12 min. Several cycles of apnea and eupnea could occur during a single sleep episode. Breathing during a sleep cycle occurred only in SWS, never during REM sleep. The eupneic heart rate was characterized by significant sinus arrhythmia, and the apneic heart rate was similar to the minimum value during normal sinus arrhythmia. Patterns of change in breathing and heart rate associated with wakefulness and sleep were similar in seals sleeping underwater and on land. When sleeping underwater, the seals raised their heads to the surface to breathe without awakening. The changes in heart rate associated with normal sinus arrhythmia, sleep apnea, and diving apnea appear to be similar, suggesting regulation by a common homeostatic control mechanism.

Muscle temperature and swim velocity profiles during diving in a Weddell seal, Leptonychotes weddellii.

Locomotory muscle temperature and swim velocity profiles of an adult Weddell seal were recorded over a 21 h period. The highest temperatures occurred during a prolonged surface period (mean 37.3 degrees C, S.D. 0.16 degrees C). Muscle temperature averaged 36.8 and 36.6 degrees C (S.D. 0.25 degrees C, 0.19 degrees C) during two dive bouts and showed no consistent fluctuations between dive and interdive surface intervals. Swim velocities were also constant, near 1.3 m s-1. These data indicate that past records of low aortic temperatures (35 degrees C) during and after prolonged dives are not indicative of whole-body temperature changes, and that muscle temperature, even during dives as long as 45 min, remains near 37 degrees C.

Effects of standard anticoagulants and storage procedures on plasma glucose values in seals.

Standard methods for the preservation of vertebrate blood samples for glucose analysis include collecting and storing the blood in evacuated tubes containing sodium fluoride (glycolytic inhibitor) and potassium oxalate (anticoagulant). We found that blood collected from 5 seals by venipuncture and transferred into these tubes had a significantly (P less than 0.05) lower plasma glucose value than blood transferred into tubes containing heparin. In species in which RBC glucose concentration is significantly less than that in the plasma, fluoride and oxalate-induced hemolysis dilutes the plasma with cytoplasm and lowers the measured concentration of glucose in plasma. Therefore, although plasma glucose is used extensively in experimental and clinical analyses, standard techniques for handling the blood may create errors that could confuse comparisons between individuals or species.

The biochemistry of natural fasting at its limits.

There are several groups of animals that are adapted for extremely long duration fasting as part of their reproductive cycle. Penguins, bears and seals routinely fast without food or water for months at time. However, they do not 'starve', as the biochemical implications of starving are very different from those of successful fasting. There are distinct biochemical adaptations in lipid, carbohydrate and especially protein metabolism that allow these animals to survive. It appears, at least for penguins and seals, that the duration of the fast may be limited by changes that occur in biochemical regulation near the ned of the fast. In all of these species, the biochemistry of fasting and the ecological and behavioral demands of their breeding cycles are closely interrelated.

Heart rates and swim speeds of emperor penguins diving under sea ice.

Heart rate during overnight rest and while diving were recorded from five emperor penguins with a microprocessor-controlled submersible recorder. Heart rate, cardiac output and stroke volume were also measured in two resting emperor penguins using standard electrocardiography and thermodilution measurements. Swim velocities from eight birds were obtained with the submersible recorder. The resting average of the mean heart rates was 72 beats min-1. Diving heart rates were about 15% lower than resting rates. Cardiac outputs of 1.9-2.9 ml kg-1 s-1 and stroke volumes of 1.6-2.7 ml kg-1 were similar to values recorded from mammals of the same body mass. Swim velocities averaged 3 m s-1. The swim speeds and heart rates suggest that muscle O2 depletion must occur frequently: therefore, many dives require a significant energy contribution from anaerobic glycolysis.

Metabolic rates of freely diving Weddell seals: correlations with oxygen stores, swim velocity and diving duration.

The metabolic rates of freely diving Weddell seals were measured using modern methods of on-line computer analysis coupled to oxygen consumption instrumentation. Oxygen consumption values were collected during sleep, resting periods while awake and during diving periods with the seals breathing at the surface of the water in an experimental sea-ice hole in Antarctica. Oxygen consumption during diving was not elevated over resting values but was statistically about 1.5 times greater than sleeping values. The metabolic rate of diving declined with increasing dive duration, but there was no significant difference between resting rates and rates in dives lasting up to 82 min. Swimming speed, measured with a microprocessor velocity recorder, was constant in each animal. Calculations of the aerobic dive limit of these seals were made from the oxygen consumption values and demonstrated that most dives were within this theoretical limit. The results indicate that the cost of diving is remarkably low in Weddell seals relative to other diving mammals and birds.

Fuel homeostasis in the harbor seal during submerged swimming.

1. The turnover rates and oxidation rates of plasma glucose, lactate, and free fatty acids (FFA) were measured in three harbor seals (average mass = 40 kg) at rest or during voluntary submerged swimming in a water flume at 35% (1.3 m.s-1) and 50% (2 m.s-1) of maximum oxygen consumption (MO2max). 2. For seals resting in water, the total turnover rates for glucose, lactate, and FFA were 23.2, 26.2, and 7.5 mumols.min-1.kg-1, respectively. Direct oxidation of these metabolites accounted for approximately 7%, 27%, and 33% of their turnover and 3%, 7%, and 18% of the total ATP production, respectively. 3. For swimming seals, MO2max was achieved at a drag load equivalent to a speed of 3 m.s-1 and averaged 1.85 mmol O2.min-1.kg-1, which is 9-fold greater than resting metabolism in water at 18 degrees C. 4. At 35% and 50% MO2max, glucose turnover and oxidation rates did not change from resting levels. Glucose oxidation contributed about 1% of the total ATP production during swimming. 5. At 50% MO2max, lactate turnover and anaerobic ATP production doubled, but the steady state plasma lactate concentration remained low at 1.1 mM. Lactate oxidation increased 63% but still contributed only 4% of the total ATP production. Anaerobic metabolism contributed about 1% of the total ATP production at rest and during swimming. 6. The plasma FFA concentration and turnover rate increased only 24% and 37% over resting levels, respectively, at 50% MO2max. However, the oxidation rate increased almost 3.5-fold and accounted for 85% of the turnover.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationships between plasma ketones and fasting duration in neonatal elephant seals.

Long-duration fasting in mammals can ultimately lead to stage three terminal starvation, which is characterized by depleted fat stores, a metabolic shift away from fat metabolism toward lean tissue catabolism, and a sharp decline in circulating levels of plasma fatty acids and ketone bodies. However, this biochemical shift has never been observed outside of the laboratory in a naturally fasting, nonhibernating mammal. In the current study, plasma levels of the ketone body D-beta-hydroxybutyrate (beta-HBA) were assayed in 10 Northern elephant seal pups during suckling and the postweaning fast and in 12 fasting adult seals. Plasma beta-HBA concentration in the pups was minimal during suckling (0.09 +/- 0.06 mM; n = 10) and began to increase immediately after weaning. The concentration rose until about 55 days into the fast (1.34 +/- 0.36 mM; n = 10) and then declined sharply. Within 10 days of this deflection point, the seal pups left for sea. By contrast, adult elephant seals showed consistently low levels of beta-HBA after several months of fasting (0.06 +/- 0.07 mM; n = 12). The data suggest that the duration of fasting in elephant seal pups may be determined, in part, by biochemical shifts that occur near the end of the fast and that the regulation of ketone concentration is different in fasting neonatal and adult elephant seals.

Cardiac output and stroke volume in swimming harbor seals.

Cardiac output was measured by the thermodilution method in three young harbor seals, at rest and while swimming up to the maximum effort for which they could be trained. Stroke volume was determined by counting heart rate simultaneously with determination of cardiac output. Cardiac outputs varied widely between surface breathing (7.8 ml.kg-1.s-1) and breath-holding while swimming under water (1.8 ml.kg-1.s-1). Stroke volume while at the surface was almost twice the volume while submerged. Surface cardiac output was always near maximal despite work effort, whereas submerged cardiac output gradually increased at higher work efforts. The cardiovascular performance of seals at the maximum MO2 we could induce from them is equivalent to that of the domestic goat.

Circulatory water concentration in suckling and fasting northern elephant seal pups.

This study examined circulatory water concentrations in the neonatal northern elephant seal (Mirounga angustirostris) in order to determine how suckling and fasting would alter the percentage of water in the whole blood, plasma, and red blood cells (RBC). Plasma water concentration dropped by about 3% during suckling and recovered about 1% during the fast (92.38 +/- 0.48% less than 1 week old, 90.15 +/- 0.36% weaning, 91.02 +/- 0.68% end of fast). RBC water values during this time were more variable than plasma values: RBC water increased about 1% during the first 2 weeks of suckling (from 67.80 +/- 0.28% to 68.68 +/- 0.51%) but dropped to slightly below original neonatal values by weaning (67.15 +/- 0.63%). The first several weeks of fasting were marked by wide variability in RBC water, but by the end of the fast RBC water was comparable to that at weaning. These results indicate: 1) Northern elephant seal pups do not exhibit circulatory dehydration during 10 weeks of fasting; 2) Measurements of plasma or RBC metabolites (such as plasma glucose or RBC hemoglobin) may show variations or trends due not to metabolic regulation but rather to changes in circulatory water concentration.

Behavior of freely diving animals.

While diving animals are capable of both long-duration and deep dives, their normal behavior does not routinely approach these limits of time and depth. Studies of the physiologic and biochemical capabilities of diving species have defined the maximum limits of their underwater periods. However, by also studying their diving behavior, we can examine how diving animals most efficiently work within those limits. If we assume for example that the behavioral goal of a foraging seal is to spend as much time underwater as possible, then it must also minimize time at the surface and make many repetitive dives instead of single, longer dives that require long surface recovery periods. To understand the efficiency of bout diving, we must study the physiologic impact of many repetitive dives and how the seal manipulates its behavior to both stay within its physiologic limits and to maximize time underwater.

Influence of hematocrit on whole blood glucose levels: new evidence from marine mammals.

The water content and glucose concentration in the whole blood of marine mammals were found to be correlated to red blood cell concentration. Because hematocrit (Hct) undergoes significant periodic shifts in these mammals during periods of apnea and/or diving, the measured values of whole blood glucose change due to alterations in Hct, independent of shifts in metabolite regulatory pathways. In contrast to humans, where red blood cell and plasma glucose concentrations are equivalent, in most other mammalian species red blood cell glucose concentration is much lower than that in plasma. Therefore, as Hct increases, the whole blood concentration of glucose must decrease in a linear manner, depending on the magnitude of the Hct change and the concentration of glucose in the blood cells. This study demonstrates that water and metabolite distribution must be considered when interpreting whole blood metabolite data. The results suggest that plasma or serum analysis of metabolites would avoid distribution problems. This is especially critical in species where Hct varies during the course of the experiment, as in exercising or diving mammals.