Influence of season, age, sex, and time of day on the endocrine profile of the common bottlenose dolphin (Tursiops truncatus).

Understanding baseline hormone levels, the magnitude of intra-individual variability, and their variation as a function of life history is difficult in toothed whales (e.g. dolphins and porpoises) because of the effects of capture stress. To determine the endocrine profile of the common bottlenose dolphin (Tursiops truncatus) as a function of season, time of day (TOD), age, sex, and reproductive status, blood corticosteroids, thyroid hormones, and catecholamines were repeatedly measured in a managed-care population exposed to ambient light and water temperatures of San Diego Bay. Additionally, fecal hormone metabolites were assessed for cortisol, aldosterone, and triiodothyronine. Samples were collected at two to four-week intervals over a period of two years, and multiple times within a day at monthly intervals over a year. Samples were collected through the voluntary participation of the dolphins in the blood draws and fecal collections in order to avoid the effects of handling stress. All serum hormones except aldosterone significantly varied with season and all serum hormones except total thyroxine significantly varied as a function of TOD. Fecal glucocorticoid metabolites significantly correlated with circulating cortisol levels, and there was a significant seasonal effect on triiodothyronine fecal metabolites. Strong seasonal effects demonstrated complex interactions with age and sex suggesting that contextual information is critical to interpreting differences in endocrine profiles. Strong circadian patterns further suggest that sampling design is important to the interpretation of blood or fecal collections, particularly since diurnal changes in some serum hormone levels are similar to the magnitude of seasonal differences. Despite potential impacts of feeding schedules on diurnal patterns, managed care populations can provide important insights into seasonal and age-related endocrine changes in toothed whales.

Variation in adrenal and thyroid hormones with life-history stage in juvenile northern elephant seals (Mirounga angustirostris).

The classical approach to quantifying the impact of stressors on wildlife is through characterization of hormones associated with the generalized stress response. However, interpretation of hormone data can be difficult due to the range of natural variation within a species and potential confounds of individual and life-history variables. Blood adrenal and thyroid hormones were measured in 144 chemically immobilized yearling northern elephant seals (Mirounga angustirostris) to characterize variation between sexes and across semiannual haul-outs. There was no relationship between hormone concentrations and time needed for collecting blood nor evidence of diel patterns, suggesting that collection of samples for baseline values can be accomplished without bias due to handling artifacts or time of day. Serum cortisol concentrations did not vary with gender or across haul-out fasts but increased dramatically during molting. Cortisol was correlated with aldosterone across all measured life-history stages. Thyroid hormone levels were lower in females and decreased with fasting in both sexes during the fall haul-out. Cortisol concentrations were inversely associated with total triiodothyronine (T3) and positively associated with reverse T3 concentrations across all measured life-history stages suggesting an important impact of cortisol on deiodinase enzymes and thyroid function. Epinephrine concentrations increased across fasts and norepinephrine concentrations were higher in males than in females. Significant variation in stress hormone concentrations with gender and life-history stage emphasizes the importance of contextual variables when interpreting serum hormone concentrations.

Deadly diving? Physiological and behavioural management of decompression stress in diving mammals.

Decompression sickness (DCS; 'the bends') is a disease associated with gas uptake at pressure. The basic pathology and cause are relatively well known to human divers. Breath-hold diving marine mammals were thought to be relatively immune to DCS owing to multiple anatomical, physiological and behavioural adaptations that reduce nitrogen gas (N(2)) loading during dives. However, recent observations have shown that gas bubbles may form and tissue injury may occur in marine mammals under certain circumstances. Gas kinetic models based on measured time-depth profiles further suggest the potential occurrence of high blood and tissue N(2) tensions. We review evidence for gas-bubble incidence in marine mammal tissues and discuss the theory behind gas loading and bubble formation. We suggest that diving mammals vary their physiological responses according to multiple stressors, and that the perspective on marine mammal diving physiology should change from simply minimizing N(2) loading to management of the N(2) load. This suggests several avenues for further study, ranging from the effects of gas bubbles at molecular, cellular and organ function levels, to comparative studies relating the presence/absence of gas bubbles to diving behaviour. Technological advances in imaging and remote instrumentation are likely to advance this field in coming years.

Environment and activity affect skin temperature in breeding adult male elephant seals (Mirounga angustirostris).

The large body size and high rates of metabolic heat production associated with male mating success in polygynous systems creates potential thermoregulatory challenges for species breeding in warm climates. This is especially true for marine predators carrying large blubber reserves intended for thermoregulation in cold water and fuel provision during extended fasts. Thermographic images were used to measure changes in skin temperature (T(S)) in adult male northern elephant seals (Mirounga angustirostris) over the breeding season. Environmental variables, primarily ambient temperature and solar radiation, were the principal determinants of mean and maximum T(S). When controlled for environmental variables, dominance rank significantly impacted mean T(S), being highest in alpha males. Behavioral activity significantly influenced T(S) but in a counter-intuitive way, with inactive males exhibiting the highest T(S). This was likely due to strong impacts of environmental variables on the kinds of behavior exhibited, with males being less active on warm, humid days at peak solar radiation. We classified thermal windows as areas in which T(S) was one standard deviation greater than mean T(S) for the individual seal within a thermograph. Thermal features suggest active physiological thermoregulation during and after combat and significant circulatory adaptations for heat dumping, as evidenced by recurring locations of thermal windows representing widely varying T(S) values. Frequent observations of localized T(S) above 37°C, particularly after combat, suggest the production of thermoregulatory stress during breeding behavior. Our findings demonstrate the importance of environmental drivers in shaping activity patterns during breeding and provide evidence for thermoregulatory costs of successful breeding in large polygynous males.

Investigation of the potential for vascular bubble formation in a repetitively diving dolphin.

The production of venous gas emboli (VGE) resulting from altered dive behavior is postulated as contributing to the stranding of beaked whales exposed to mid-frequency active sonar. To test whether nitrogen gas uptake during repetitive breath-hold diving is sufficient for asymptomatic VGE formation in odontocetes, a bottlenose dolphin (Tursiops truncatus Montagu) was trained to perform 10-12 serial dives with 60 s surface intervals to depths of 30, 50, 70 or 100 m. The dolphin remained at the bottom depth for 90 s on each dive. Doppler and/or two-dimensional imaging ultrasound did not detect VGE in the portal and brachiocephalic veins following a dive series. Van Slyke analyses of serial, post-dive blood samples drawn from the fluke yielded blood nitrogen partial pressure (P(N(2))) values that were negligibly different from control samples. Mean heart rate (HR; +/-1 s.d.) recorded during diving was 50+/-3 beats min(-1) and was not significantly different between the 50, 70 and 100 m dive sessions. The absence of VGE and elevated blood P(N(2)) during post-dive periods do not support the hypothesis that N(2) supersaturation during repetitive dives contributes to VGE formation in the dolphin. The diving HR pattern and the presumed rapid N(2) washout during the surface-interval tachycardia probably minimized N(2) accumulation in the blood during dive sessions.

Renal function in suckling and fasting pups of the northern elephant seal.

Elephant seals fast for prolonged periods without access to water. This is made possible, in part, by reductions in urine production. However, the mechanisms involved in reducing urine production are not understood. In this study, glomerular filtration rate (GFR) was measured in five northern elephant seal pups (Mirounga angustirostris) via the inulin clearance technique. Measurements were made during day 9 and day 18-22 of nursing and the second and eighth week of the postweaning fast. Plasma aldosterone and cortisol concentrations, quantified by radioimmunoassay, were measured in eight other weanlings during the second and eighth week of the fast. Mean GFR was 79.3+/-29.3 ml/min during the early suckling period and 78.2+/-17.1, 89.8+/-52.7, and 80.4+/-12.2 ml/min during the late suckling, early fasting and late fasting periods, respectively. Differences between nursing and fasting were insignificant, possibly because reduced protein oxidation during suckling and rapid recruitment of protein for tissue synthesis obviated the need for postprandial hyperfiltration. Alternatively, maintenance of GFR during fasting may facilitate urea concentration by compensating for reductions in the fractional excretion of urea. It is further hypothesized that aldosterone is primarily responsible for mediating renal water reabsorption in this system.

Can diving-induced tissue nitrogen supersaturation increase the chance of acoustically driven bubble growth in marine mammals?

The potential for acoustically mediated causes of stranding in cetaceans (whales and dolphins) is of increasing concern given recent stranding events associated with anthropogenic acoustic activity. We examine a potentially debilitating non-auditory mechanism called rectified diffusion. Rectified diffusion causes gas bubble growth, which in an insonified animal may produce emboli, tissue separation and high, localized pressure in nervous tissue. Using the results of a dolphin dive study and a model of rectified diffusion for low-frequency exposure, we demonstrate that the diving behavior of cetaceans prior to an intense acoustic exposure may increase the chance of rectified diffusion. Specifically, deep diving and slow ascent/descent speed contributes to increased gas-tissue saturation, a condition that amplifies the likelihood of rectified diffusion. The depth of lung collapse limits nitrogen uptake per dive and the surface interval duration influences the amount of nitrogen washout from tissues between dives. Model results suggest that low-frequency rectified diffusion models need to be advanced, that the diving behavior of marine mammals of concern needs to be investigated to identify at-risk animals, and that more intensive studies of gas dynamics within diving marine mammals should be undertaken.

Protein catabolism in suckling and fasting northern elephant seal pups (Mirounga anglstirostris).

Nursing elephant seal pups are hypothesized to be preadapted to the postweaning fast, yet no comparison of lipid or protein use for meeting metabolic costs has been made between these contrasting nutritional periods. To address this, protein catabolism was estimated in five elephant seal pups from measurements of urea turnover made twice during nursing and twice during the postweaning fast. Changes in body composition were measured in ten separate weaned pups via tritiated water dilution and matched to fasting urea turnover measurements in order to assess errors in protein catabolism derived from urea turnover rates. Estimates of lean mass loss based upon urea turnover and tritiated water dilution were in general agreement, supporting estimates of protein catabolism derived from urea turnover measurements. Protein catabolism was estimated to contribute less than 4% to the average metabolic rate of suckling and fasting pups implying strict protein conservation during both periods and supporting the shypothesis that suckling pups are pre-adapted to fasting. It is proposed that strict protein conservation across suckling and fasting compensates for relative reductions in maternal investment associated with the abbreviated lactation period of the elephant seal.

Classification of dolphin echolocation clicks by energy and frequency distributions.

Dolphins demonstrate an adaptive control over echolocation click production, but little is known of the manner or degree with which control is exercised. Echolocation clicks (N approximately 30,000) were collected from an Atlantic bottlenose dolphin (Tursiops truncatus) performing object discrimination tasks in order to investigate differential click production. Seven categories of clicks were identified using the spectral conformation and relative position of -3 and -10 dB peaks. A counterpropagation network utilizing 16 inputs, 50 hidden units, and 8 output units was trained to classify clicks using the same spectral variables. The network classified novel clicks with 92% success. Additional echolocation clicks (N > 24,000) from two other dolphins were submitted to the network for classification. Classified echolocation clicks were analyzed for animal specific differences, changes in predominant click type within click trains, and task-related specificity. Differences in animal and task performance may influence click type and click train length.

Evidence that a 550,000-dalton cartilage matrix glycoprotein is a chondrocyte membrane-associated protein closely related to ceruloplasmin.

Cartilage matrix glycoprotein (CMGP) is a disulfide-bonded 550,000-dalton protein that is synthesized by chondrocytes and ciliary epithelial cells. We have purified the protein from bovine and porcine articular cartilage and have sequenced two peptides, which both have significant homology with human ceruloplasmin, a copper-binding oxidase. Immunolocation analysis indicates that a commercial polyclonal antiserum to human ceruloplasmin reacts with bovine cartilage CMGP. Chelating columns made with copper bind CMGP from bovine cartilage extracts. CMGP is present in bovine chondrocyte membrane preparations purified from sucrose density gradients. Oligonucleotide probes have been synthesized based on the published sequence of the 3'-untranslated region and a portion of the C terminus of human ceruloplasmin and have been used to amplify a cDNA fragment from bovine cartilage and human liver libraries. CMGP demonstrates oxidase activity towards p-phenylenediamine similar to that of ceruloplasmin. These studies suggest that CMGP is closely related to, if not identical with, ceruloplasmin. It is possible that CMGP may be involved in metal transport into and/or within the chondrocyte.