Mercury and selenium concentrations in lanugo of free-ranging California sea lions in the southern Gulf of California, Mexico.

Total mercury ([THg]) and selenium ([TSe]) concentrations were determined in California sea lion (Zalophus californianus) lanugo from the Gulf of California in 2021 and 2022. Relationships with sex, morphometrics, and year were evaluated. Following toxicological thresholds of concern for piscivorous mammals, most pups had a [THg] < 10 ppm, one pup (2021) had a [THg] > 20 ppm, no pups had a [THg] > 30 ppm. Females had significantly higher [TSe] than males; sex did not influence [THg]. [THg] and [TSe] in 2022 were significantly higher in the general population and male cohorts compared to 2021. Significant negative correlations were observed between [THg], [TSe], and morphometrics (2021). These results indicate that, compared to other pinniped species, regional California sea lions may have a decreased likelihood of experiencing Hg-related adverse health effects. Year-related changes in element concentrations suggest continued monitoring of this population to assess pinniped, environmental, and potentially, human health.

Indicators of oxidative stress in leukocytes isolated from bottlenose dolphins (Tursiops truncatus) in response to a proinflammatory challenge.

Few studies have analyzed the indicators of oxidative stress in marine mammals following exposure to lipopolysaccharides (LPS); sex and maturity-related differences have not been explored. The objective of this study was to compare the indicators of oxidative stress following exposure to LPS for 24 and 48 h in isolated Pacific bottlenose dolphin (Tursiops truncatus; N = 12) leukocytes in relation to sex and maturity stage, using spectrophotometry. Following 48 h under experimental conditions (10 µg LPS mL-1), the leukocytes from males (n = 5) produced significantly more superoxide radical (O2•-; F (1, 8) = 13.965, p = 0.006) and displayed significantly greater activities of catalase (CAT; F (1, 8) = 9.465, p = 0.015) and glutathione S-transferase (GST; p = 0.028) compared to the leukocytes from females (n = 7). Following 48 h under experimental conditions, maturity-stage did not significantly influence the indicators of oxidative stress. Mature bottlenose dolphins (n = 7) had a significantly higher average daily dietary intake compared to immature bottlenose dolphins (n = 5; F (1, 10) = 5.825, p = 0.036). These results suggest that sex-related strategies for coping with a proinflammatory challenge may be present within the leukocytes from bottlenose dolphins, while potential maturity stage-related strategies require further investigation.

The effect of predator exposure and reproduction on oxidative stress parameters in the Catarina scallop Argopecten ventricosus.

Predation is known to impact growth and reproduction, and the physiological state of the prey, including its susceptibility to oxidative stress. In this study, we investigated how prolonged exposure to predators modulates tissue specific antioxidant defense and oxidative damage in the short-lived epibenthic scallop Argopecten ventricosus (2years maximum lifespan). Scallops that were experimentally exposed to predators had not only lower antioxidant capacities (superoxide dismutase and catalase), but also lower oxidative damage (protein carbonyls and TBARS=thiobarbituric acid reactive substances including lipid peroxides) in gills and mantle compared to individuals not exposed to predators. In contrast, oxidative damage in the swimming muscle was higher in predator-exposed scallops. When predator-exposed scallops were on the verge of spawning, levels of oxidative damage increased in gills and mantle in spite of a parallel increase in antioxidant defense in both tissues. Levels of oxidative damage increased also in the swimming muscle whereas muscle antioxidant capacities decreased. Interestingly, post-spawned scallops restored antioxidant capacities and oxidative damage to immature levels, suggesting they can recover from spawning-related oxidative stress. Our results show that predator exposure and gametogenesis modulate oxidative damage in a tissue specific manner and that high antioxidant capacities do not necessarily coincide with low oxidative damage.

Changes in oxidative stress parameters in relation to age, growth and reproduction in the short-lived catarina scallop Argopecten ventricosus reared in its natural environment.

Increase in oxidative damage and decrease in cellular maintenance is often associated with aging, but, in marine ectotherms, both processes are also strongly influenced by somatic growth, maturation and reproduction. In this study, we used a single cohort of the short-lived catarina scallop Argopecten ventricosus, to investigate the effects of somatic growth, reproduction and aging on oxidative damage parameters (protein carbonyls, TBARS and lipofuscin) and cellular maintenance mechanisms (antioxidant activity and apoptosis) in scallops, caged in their natural environment. The concentrations of protein carbonyls and TBARS increased steeply during the early period of fast growth and during reproduction in one-year-old scallops. However, oxidative damage was transient, and apoptotic cell death played a pivotal role in eliminating damage in gill, mantle and muscle tissues of young scallops. Animals were able to reproduce again in the second year, but the reduced intensity of apoptosis impaired subsequent removal of damaged cells. In late survivors low antioxidant capacity and apoptotic activity together with a fast accumulation of the age pigment lipofuscin was observed. Rates of oxygen consumption and oxidative stress markers were strongly dependent on somatic growth and reproductive state but not on temperature. Compared to longer-lived bivalves, A. ventricosus seems more susceptible to oxidative stress with higher tissue-specific protein carbonyl levels and fast accumulation of lipofuscin in animals surviving the second spawning. Superoxide dismutase activity and apoptotic cell death intensity were however higher in this short-lived scallop than in longer-lived bivalves. The life strategy of this short-lived and intensely predated scallop supports rapid somatic growth and fitness as well as early maturation at young age at the cost of fast cellular degradation in second year scallops.

Hypoxemic and ischemic tolerance in emperor penguins.

Oxygen store depletion and a diving bradycardia in emperor penguins (Aptenodytes forsteri) expose tissues to critical levels of hypoxemia and ischemia. To assess the prevention of re-perfusion injury and reactive oxygen species (ROS) damage in emperor penguins, superoxide radical production, lipid peroxidation (thiobarbituric acid reactive substances (TBARS)), and antioxidant enzyme activity profiles in biopsy samples from muscle and liver were determined and compared to those in the chicken and 8 species of flighted marine birds (non-divers and plunge divers). In muscle of emperor penguins, superoxide production and TBARS levels were not distinctly different from those in the other species; among the antioxidant enzymes, catalase (CAT) and glutathione-S-transferase (GST) activities were significantly elevated above all species. In the liver of emperor penguins, TBARS levels were not significantly different from other species; only CAT activity was significantly elevated, although GST and glutathione peroxidase (GPX) activities were 2-3 times higher than those in other species. The potential for ROS formation and lipid peroxidation is not reduced in the pectoral muscle or liver of the emperor penguin. Scavenging of hydrogen peroxide by CAT and the conjugation of glutathione with reactive intermediates and peroxides by GST and GPX appear to be important in the prevention of ROS damage and re-perfusion injury in these birds.

Diving seals: are they a model for coping with oxidative stress?

The diving lifestyle of seals depends upon cardiovascular adjustments that result in frequent vasoconstriction of numerous organs. With the first post-dive breath, reperfusion allows for eliminating accumulated carbon dioxide (CO(2)) and reloading oxygen (O(2)) stores. Reintroduction of oxygenated blood raises the potential for production of reactive oxygen species (ROS) and the possibility that they may overwhelm the antioxidant defenses. This study addresses the question of possible adaptive responses that allow ringed seal (Phoca hispida) tissues to tolerate repeated cycles of ischemia and reperfusion, and thus protect them from oxidative insult. We obtained samples of ringed seal heart, muscle and kidney through the cooperation of native subsistence hunters at Barrow, Alaska. Samples were subjected to oxidative stress by addition of xanthine oxidase. Production of superoxide radical (O(2)(.-)), lipid peroxidation (as determined by the presence of thiobarbituric acid reactive substances, TBARS) and antioxidant capacity (AOX) were quantified by spectrophotometric analysis. Similarly treated pig tissues were anticipated to be more susceptible to oxidative stress. Contrary to expectations, pig tissues revealed less O(2)(.-) and TBARS compared with ringed seal tissues. These results show that ringed seal muscle, heart and kidney can be induced in vitro to generate ROS, and suggest that the living seal's protective defenses may depend upon O(2)(.-) production, similar to the protective effect of experimental preconditioning, or on enhanced intermediate scavenging, as evidenced by the larger AOX found in ringed seal tissues.

Effects of arginine vasopressin in the heart are mediated by specific intravascular endothelial receptors.

Arginine vasopressin induces vascular, inotropic and arrhythmogenic effects in the heart. Existing evidence, obtained indirectly, suggests that these effects occur through paracrine endothelial mechanisms. To demonstrate this, vasopressin was confined to the intravascular space by covalent coupling to high molecular weight (2x10(6) Da, vasopresin-dextran) dextran. Isolated guinea pig hearts were infused with equivalent concentrations of vasopressin and vasopressin-dextran. The negative inotropic and coronary vasopressor effects of vasopressin-dextran were similar to those evoked by vasopressin; in both cases effects were reversible. Free dextran had no effect on vascular resistance nor in ventricular developed pressure. The inotropic and vascular effects of both vasopressin and vasopressin-dextran were blocked by the vasopressin receptor antagonist [Adamantaneacetyl(1), o-Et-D-Tyr(2), Val(4), Aminobutyryl(6), Arg(8,9)]vasopressin (Adam-vasopressin), indicating that the effects of the two agonists were vasopressin receptor-mediated. To elucidate possible endothelial intermediaries of these effects, isolated guinea pig hearts were infused simultaneously with vasopressin or vasopressin-dextran and several inhibitors either of synthesis or blockers of receptors of possible endothelial mediators. Only reactive blue 2, a P(2y) purinoceptor antagonist, and suramin, a P(2y) and a P(2x) purinoceptor antagonist, caused a total reversal of vascular and inotropic effects of vasopressin and vasopressin-dextran. Pyridoxalphosphate-6-Azophenyl-2'-4'disulphonic acid, a P(2x) purinoceptor antagonist, was without effect. Our results provide direct evidence that the short-term cardiac effects of vasopressin are due to selective activation of intravascular purinoceptors and suggest that an intermediary of these effects is ATP.

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.