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Northern elephant seal platelets: analysis of shape change and response to platelet agonists.

Blood platelets have a vital role in the maintenance of normal mammalian hemostasis. Rapid pressure changes and temperatures lower than 20 degrees C cause activation of human and terrestrial mammal platelets. Elephant seals are routinely subjected to pressures as high as 150 atm, yet do not suffer from the thrombotic effects of platelet activation associated with rapid decompression. We examined the ultrastructure of Northern elephant seal (Mirounga angustirostris) platelets and their functional and morphological response to various platelet agonists. Unstimulated elephant seal platelets are discoid cells, with a microtubule coil, randomly dispersed alpha and dense granules, and glycogen granules. There are well-defined areas of membranous invaginations that indicate the presence of an open canalicular system (OCS). Aggregometry was used to determine the response of elephant seal platelets to various platelet agonists. Dose-dependent curves were generated for thrombin, collagen, and adenosine diphosphate (ADP). Platelet response to thrombin was dose-dependent and was maximal at 2.5 U/ml. Platelets collected in sodium citrate had blunted responses to both ADP and collagen. ADP stimulation caused only reversible, primary activation (shape change) at > or = 5 microM, while platelets did not aggregate in response to any concentration of collagen. Platelets collected in sodium heparin did respond fully to both to ADP and collagen. There was small, reversible shape change in response to ristocetin, but no response to epinephrine. Decreased sensitivity of elephant seal platelets to agonists may be a protective mechanism developed in response to rapid pressure changes and cold temperatures associated with adaptation to an extreme environment.

Adaptation, Physiological↗

Fur seal adaptations to lactation: insights into mammary gland function.

The fur seal (Arctocephalus spp. and Callorhinus spp., members of the pinniped family) is a mammal with the unusual capability to modulate its lactation cycle by turning milk production on and off without the typical mammalian regression and involution of the mammary gland. Lactation has evolved from constraints arising from the spatial and temporal separation of infant nursing and maternal foraging as the mother gives birth and feeds the pup on land while acquisition of nutrients for milk production occurs at sea. The lactation cycle begins with the female fur seal undergoing a perinatal fast of approximately 1 wk, after which time she departs the breeding colony to forage at sea. For the remainder of the long lactation period (116-540 days), the mother alternates between short periods ashore suckling the young with longer periods of up to 4 wk of foraging at sea. Milk production continues while foraging at sea, but at less than 20% the rate of production on land. Fur seals produce one of the richest milk reported, with a very high lipid content contributing up to 85% of total energy. This feature serves as an adaptation to the young's need to produce an insulating blubber layer against heat loss and to serve as an energy store when the mother is away foraging at sea. This atypical pattern of lactation means mothers have long periods with no suckling stimulus and can transfer high-energy milk rapidly while on land to minimize time away from foraging grounds. The absence of suckling stimulus and milk removal during foraging does not result in the onset of involution with associated apoptosis of mammary secretory cells and a subsequent progressive breakdown of the cellular structure of the mammary gland. The mechanisms controlling lactation in the fur seal mammary gland have been investigated using molecular and cellular techniques. These findings have shed light on the processes by which the unique features of lactation in the fur seal are regulated.

Adaptation, Physiological↗

Evidence for a long-lasting single administration contraceptive vaccine in wild grey seals.

A single-administration birth control vaccine based on liposome delivery of porcine zona pellucida antigens reduced pup production in grey seals (Halichoerus grypus) by about 90%. Anti-porcine zona pellucida titers of individual seals with two or more recaptures were variable but without a diminishing trend during the 5 year post-immunization period. Seals that produced at least one or more pups during the 2-5 year post-immunization period when the vaccine is fully effective, had an average anti-porcine zona pellucida titer of 5% of the reference serum. In contrast, the subset of seals that did not reproduce but were recaptured during the breeding season had an average titer of 31% of the reference serum. As measured by antibody titers and pup production, there were no differences in efficacy of the vaccine in 14-, 20- and 21-year-old female grey seals.

Age Factors↗

Contaminant-induced immunosuppression and mass mortalities among harbor seals.

Virus-associated mass mortalities among seals inhabiting northwestern Europe have generated an interest in immunotoxicology in this species. A morbillivirus has been isolated from victims, but a contribution of immunotoxic contaminants to the severity of the outbreaks could not be ruled out. Fish-eating seals occupy high trophic levels in the aquatic food chain, and accumulate high levels of contaminants including polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). Such chemicals have been found to be immunotoxic at low doses in studies of laboratory animals. We carried out an immunotoxicological study, in which captive harbor seals (Phoca vitulina) were fed herring from either relatively uncontaminated sites of the Atlantic Ocean, or from the highly contaminated Baltic Sea. In this report we summarize the contaminant-related immunosuppression observed in the captive group of seals fed herring from the Baltic Sea. In addition, we describe two parallel studies, in which laboratory rats are exposed as adults or perinatally to the contaminants in the Baltic Sea herring, exhibiting immunotoxicity. On the basis of these studies we conclude that complex mixtures of environmental contaminants including PCBs, PCDFs, and PCDDs may represent a real immunotoxic risk to free-ranging seals.

Animals↗

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.

Angiotensin II↗

Three-dimensional movements and swimming activity of a northern elephant seal.

We attached a video system and data recorder to a northern elephant seal to track its three-dimensional movements and observe propulsive strokes of the hind flippers. During 6 h of recording, the seal made 20 dives and spent 90% of the time submerged. Average dive duration, maximum depth and swimming speed were 14.9 min+/-6.1 S.D., 289 m+/-117 S.D. and 1.1 m s(-1)+/-0.12 S.D., respectively. The distance swum during a dive averaged 925 m+/-339 S.D., and the average descent and ascent angles were 41 degrees +/-18 S.D. and 50 degrees +/-21 S.D., respectively. Dive paths were remarkably straight suggesting that the seal was navigating while submerged. We identified three modes of swimming based on the interval between propulsive strokes: continuous stroking; stroke-and-glide swimming; and prolonged gliding. The seal used continuous stroking from the surface to a mean depth of 20 m followed by stroke-and-glide swimming. Prolonged gliding started at a mean depth of 60 m and continued to the bottom of dives. For dives to depths of 300 m or more, 75% of the descent time was spent in prolonged gliding and 10% in stroke-and-glide swimming, amounting to 5.9-9.6 min of passive descent per dive. Average swimming speed varied little with swimming mode and was not a good indicator of propulsive effort. It appears that the seal can use prolonged gliding to reduce the cost of transport and increase dive duration. Energetically efficient locomotion may help explain the long and deep dives that routinely exceed the theoretical aerobic dive limit in this species.

Animals↗

A gray seal's (Halichoerus grypus) responses to experimenter-given pointing and directional cues.

A gray seal (Halichoerus grypus) was trained to touch a target on its left or right by responding to pointing signals. The authors then tested whether the seal would be able to generalize spontaneously to altered signals. It responded correctly to center pointing and head turning, center upper body turning, and off-center pointing but not to head turning and eye movements alone. The seal also responded correctly to brief ipsilateral and contralateral points from center and lateral positions. Pointing gestures did not cause the seal to select an object placed centrally behind it. Like many animals in similar studies, this gray seal probably did not understand the referential character of these gestures but rather used signal generalization and experience from initial operant conditioning to solve these tasks.

Animals↗

Reproductive failure in common seals feeding on fish from polluted coastal waters.

The population of common seal Phoca vitulina in the westernmost part of the Wadden Sea, The Netherlands, has collapsed during the past few decades. Between 1950 and 1975 the population dropped from more than 3,000 to less than 500 animals. Comparative studies of common seal populations from different parts of the Wadden Sea reveal that pup production has declined sharply only in the western (Dutch) part. A comparative toxicological study on the levels of heavy metals and organochlorines in tissues of seals from the western and northern parts of the Wadden Sea shows that only the polychlorinated biphenyl (PCB) levels differ significantly. This is predominantly a result of PCB pollution from the river Rhine, which mainly affects the western (Dutch) part. PCBs are thought to be responsible for the low rate of reproduction in Dutch common seals on the basis of epidemiological and experimental data on the ability of PCBs to interfere with mammalian reproduction. Here I report that reproductive failure in common seals from the Dutch Wadden Sea is related to feeding on fish from that polluted area. This is the first demonstration of a causal relationship between naturally occurring levels of pollutants and a physiological response in marine mammals.

Animal Feed↗

Genetic variation in Phoca vitulina (the harbour seal) revealed by DNA fingerprinting and RAPDs.

Genetic variation in two harbour seal (Phoca vitulina) populations from the Dutch Wadden Sea and Scotland was examined by RAPD analysis and DNA fingerprinting. For comparison a population of grey seals (Halichoerus grypus) was studied. The RAPD method revealed a very low number of polymorphic bands. The multilocus DNA fingerprinting technique showed very low levels of variation both within and between populations of the harbour seals. For one probe a significant difference in variation between populations was demonstrated. The estimated average heterozygosity, however, diverged more strongly between the two harbour seal populations because of more frequent occurrence of rare alleles in the Scottish population. Low levels of genetic variation seem to be characteristic for the subspecies Phoca vitulina vitulina, but not for all marine mammals in the North Sea, as the grey seal population showed significantly higher variation.

Animals↗

Low reproductive success in territorial male Antarctic fur seals (Arctocephalus gazella) suggests the existence of alternative mating strategies.

Microsatellites were used to conduct an analysis of paternity of Antarctic fur seals (Arctocephalus gazella) from Bird Island, South Georgia. At most, only 28% of pups at our study site could be assigned a father, even though the majority (approximately 90%) of candidate males within this colony were sampled. The behavioural and genetic evidence from this study suggests that a number of alternative mating strategies may exist within this fur seal population. Holding a land-based territory conferred an advantage to male reproductive success. However, this advantage was much smaller than expected from behavioural observations. At least 70% of fur seal pups born at our study site in a given year are not fathered by males who held a territory or were observed copulating with females in the previous year, implying that there exists a pool of males that seldom venture ashore at this site. To explain this discrepancy we suggest that female choice is an integral component of the Antarctic fur seal mating system and that aquatic mating may play a much larger role in the Antarctic fur seal than previously thought.

Animals↗

Bioaccumulation of polychlorinated biphenyls (PCBs) and dichlorodiphenylethane (DDE) methyl sulfones in tissues of seal and dolphin morbillivirus epizootic victims.

Polychlorinated biphenyl (PCB) and dichlorodiphenylethane (DDE) methyl sulfone (MSF) metabolites possess high affinities for binding two homologous 16,000 Da homodimeric receptor proteins in the lung (Clara cell secretory protein, CCSP) and the uterus (uteroglobin, UG), leading to selective bioaccumulation of MSFs in these tissues. As marine mammals are highly exposed to organochlorines, concentrations of PCBs, PCB MSFs, DDT, and DDE MSF were analyzed in blubber, lung, and uterus samples from harbor seal (Phoca vitulina) and striped dolphin (Stenella coeruleoalba) morbillivirus epizootic victims to investigate uterine and lung MSF accumulation. Mean uterus concentrations of PCB MSFs and DDE MSF in harbor seals were 0.61 and 0.04 microg/g lipid weight and in striped dolphins 0.05 and 0.01 microg/g lipid weight. Mean lung concentrations of PCB MSFs and DDE MSF in harbor seals were 0.96 and 0.02 microg/g lipid weight and in striped dolphins 0.16 and 0.01 microg/g lipid weight. To ascertain whether uterine and lung bioaccumulation of MSFs is possible due to the presence of CCSP and UG in seals, CCSP and UG proteins in uterine flushings and in uterine and lung and epithelial tissue from Baltic gray and ringed seals were characterized using gel electrophoresis and Western blotting techniques. UG- and CCSP-like proteins with molecular weights of 16,000 Da were resolved in all samples. This is the first demonstration of this protein in any marine mammalian species. The toxicological implications of MSF binding with UG and CCSP in marine mammals are discussed.

Adipose Tissue↗

Age, sex, and reproductive state influence free amino acid concentrations in the fasting elephant seal.

Long-term fasting is a component of northern elephant seal (Mirounga angustirostris) life history requiring physiological adaptations to nitrogen conservation. Plasma free amino acids (FAAs) were determined for five elephant seal pups during the second and eighth weeks of the postweaning fast, six lactating female seals at 4-6 and 25 d postpartum, and seven sexually competitive adult male seals taken midway through the breeding season. Total FAAs declined in lactating females (11%) and pups (30%) with time fasting, but cystine concentration more than doubled in pups while decreasing by approximately 43% in lactating females. Methionine concentration significantly increased (approximately 68%) across lactation in adult females but was low for all classes of seal. Alanine was the most abundant FAA in adult males, and glycine became the dominant FAA in adult females late in lactation. Glutamine dominated the FAAs of weaned pups across the fast. Reductions in total FAAs of weanlings mirrored reductions in protein catabolism, but reductions in total FAAs also occurred in lactating females concomitant with an increase in protein catabolism. Observed variation in FAA concentrations may reflect ontogenetic requirements for certain amino acids in fasting weanlings. Similarly, increases in specific FAA concentrations across lactation may reflect variations in FAA flux resulting from the nutrient demands of lactogenesis.

Adaptation, Physiological↗

Angiotensin II and aldosterone increase with fasting in breeding adult male northern elephant seals (Mirounga angustirostris).

The renin-angiotensin-aldosterone system (RAAS) appears to contribute significantly to osmoregulation of fasting northern elephant seal (Mirounga angustirostris) pups; however, RAAS has not been characterized in fasting adult seals. Therefore, this study examined the contribution of RAAS to water turnover rates in fasting adult male northern elephant seals. Blood samples were obtained twice during their breeding fast at an interval of 6.5 wk, and water efflux rate was estimated by isotopic dilution during the same period. Serum electrolytes (Na+, K+, Cl-) and osmolality were unaltered between the two sampling periods, indicating ionic and osmotic homeostasis during the fast. Despite the lack of an increase in vasopressin, serum angiotensin II and aldosterone were increased and were significantly and positively correlated. Changes in aldosterone concentration and water efflux rate were significantly and negatively correlated, suggesting that the greater the increase in aldosterone, the smaller the loss of water. Adult male seals maintain ionic and osmotic homeostasis similar to that of fasting weaned pups, and this homeostasis appears to be mediated, at least in part, by RAAS, which probably contributes to increased water retention as well. The hormonal mechanisms by which northern elephant seals maintain water and electrolyte balance during fasting conditions appear to be similar regardless of age.

Aldosterone↗

Genetic consequences of a severe population bottleneck in the Guadalupe fur seal (Arctocephalus townsendi).

Population bottlenecks may lead to diminished genetic variability and correlative effects on fitness. The Guadalupe fur seal was nearly exterminated by commercial sealers during the late 18th and early 19th centuries. To determine the genetic consequences of this population bottleneck, we compared the variation at a 181 bp section of the mitochondrial DNA (mtDNA) control region from the bones of 26 prebottleneck fur seals versus variation in the extant population. We found 25 different mtDNA genotypes in the prebottleneck fur seals and only 7 genotypes among 32 extant fur seals, including only one of the ancient genotypes. These data demonstrate a substantial loss of genetic variability correlating with the recent population bottleneck. We also found from several genetic measures that the prehistoric population of Guadalupe fur seals was robust and that it had been increasing at some time during the late prehistoric period. Continued recovery of this species may, however, owe more to more immediate nongenetic factors, such as poaching and local availability of food resources during the breeding season and consequent effects on pup survival, than on the reduced genetic variability.

Animals↗

Genetic variation of southern hemisphere fur seals (Arctocephalus spp.): investigation of population structure and species identity.

We have examined phylogenetic and geographic patterns of variation in the mitochondrial cytochrome b gene of Southern Hemisphere fur seals (Arctocephalus spp.). Our survey of 106 individuals from four putative species reveals three distinct patterns of variation reflecting ancient, recent historic, and contemporary gene flow. For the combined samples of Subantarctic (Arctocephalus tropicalis) and Antarctic (Arctocephalus gazella) fur seals, we find low levels of sequence diversity and reciprocal paraphyly of hapiotypes (where representative haplotypes of a species are found to occur infrequently in another species and vice versa). For the Australian and Cape fur seal subspecies (Arctocephalus pusillus doriferus and A. p. pusillus, respectively), we find low levels of sequence diversity but significant differences in the regional distribution of haplotypes that are consistent with, but not conclusive of, the current subspecies definition based on nonmolecular data. For the New Zealand fur seal (Arctocephalus forsteri), we find high levels of average sequence diversity because of the survival of two divergent lineages of mitochondrial hapiotypes with differences approaching that found in interspecific comparisons of other mammals. The two divergent clades are distributed sympatrically in some regions, but the overall geographic structure of the variation is significant across the range of this species. These new molecular data are inconsistent with current taxonomic definitions of species within the Southern Hemisphere fur seals and argue for reevaluation of these "species" definitions. For management purposes, the definition of evolutionarily significant units (Ryder 1986) and genetic management units (Moritz 1994) in relation to these species may also be evaluated in light of this molecular genetic information.

Animals↗

Sex-specific foraging strategies and resource partitioning in the southern elephant seal (Mirounga leonina).

The evolution of resource specializations is poorly understood, especially in marine systems. The southern elephant seal (Mirounga leonina) is the largest of the phocid seals, sexually dimorphic, and thought to prey predominantly on fish and squid. We collected vibrissae from male and female southern elephant seals, and assessed stable C and N isotope ratios along the length of the vibrissae. Given that whiskers grow slowly, this sampling strategy reflects any variation in feeding behaviour over a period of time. We found that isotopic variation among females was relatively small, and that the apparent prey choice and trophic level of females was different from that for males. Further, males showed a very broad range of trophic/prey choice positions, grouped into several clusters, and this included isotopic values too low to match a broad range of potential fish and cephalopod prey tested. One of these clusters overlapped with data for South American sea lions (Otaria flavescens), which were measured for comparison. Both male southern elephant seals and southern sea lions forage over the continental shelf, providing the potential for competition. We discuss the possibility that individual southern elephant seals are pursuing specialist foraging strategies to avoid competition, both with one another, and with the South American sea lions that breed nearby.

Animals↗

Impaired cellular immune response in harbour seals (Phoca vitulina) feeding on environmentally contaminated herring.

In a 2.5-year immunotoxicological study, two groups of captive harbour seals (Phoca vitulina) were fed herring from the heavily polluted Baltic Sea or from the relatively uncontaminated Atlantic Ocean. Blood samples were collected at regular intervals, and functional immunological parameters were monitored. T cell mitogen and mixed lymphocyte-induced proliferative responses of peripheral blood mononuclear cells (PBMC) obtained from seals fed Baltic herring were significantly reduced over the course of experiment. Upon immunization with rabies virus antigen (RV) and tetanus toxoid (TT), specific proliferative responses of PBMC from the seals fed Baltic herring were also significantly reduced. Impairment of T cell-mediated immune responses became especially apparent during the second year on the respective diets, and correlated significantly to 2,3,7,8-tetrachloro-dibenzo-p-dioxin toxic equivalent levels in blubber biopsies taken from the seals after 2 years on the respective diets. Humoral immune responses, including lipopolysaccharide (LPS)-induced lymphoproliferative responses, in vitro immunoglobulin production by PBMC, as well as RV-, TT-and poliovirus-specific serum antibody responses following immunization, remained largely unaffected. We conclude that suppression of the cellular immune response in the seals fed Baltic herring was induced by the chronic exposure to immunotoxic environmental contaminants accumulated through the food chain. Since cellular immune responses are known to be of crucial importance in the clearance of morbillivirus infections, these results suggest that environmental pollution-related immunosuppression may have contributed to the severity and extent of recent morbillivirus-related mass mortalities among marine mammals.

Animals↗

Paternity exclusion by DNA fingerprinting, and mate guarding in the hooded seal Cystophora cristata.

Hooded seal Cystophora cristata trios consist of an adult female, her pup, and an attending adult male. Using DNA fingerprinting, we excluded the possibility that the attending males within hooded seal trios were the fathers of the pups, proving that these hooded seals did not remain paired from one breeding season to the next. Behavioural observations of the trios after capture and release revealed that male hooded seals displace one another in attending nursing females. Mate guarding appears to be the preferred mating strategy available to male hooded seals given intense competition for females, a very brief nursing period, and oestrus occurring soon after weaning, but its effectiveness remains unclear.

Animals↗