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Physiological effects of seawater intake in adult harp seals during phase I of fasting.

Previous studies have shown that harp seals may drink considerable amounts of seawater. The current study was undertaken to study the physiological responses to bolus administration of seawater. Adult harp seals (Phoca groenlandica) were fasted without access to water for 48 h and then given 1000 or 1500 ml of seawater by a stomach tube. Changes in urine and plasma parameters were thereafter monitored for another 12-20 h. Urine production and urine excretion rate of Na+ and Cl- increased soon after administration and reached a maximum 3-4 h later. Urine osmolality was kept rather stable and high ( approximately 1500 mOsm x kg(-1)) following seawater administration, due to a drop in urine concentration of urea that was proportional to the simultaneous increase in urine concentration of NaCl. Plasma osmolality remained at approximately 340 mOsm x kg(-1), while plasma concentration of urea decreased some 20-25% due to increased excretion of urea when seawater was ingested. Despite bolus administrations of seawater of up to approximately 2% of body mass, homeostasis was maintained and no ill effects observed. It is concluded that the concentrating abilities of the kidneys of harp seals are sufficient to prevent net loss of body water following seawater ingestion. Seawater ingestion may, moreover, increase urinary osmotic space and thus serve as a mechanism to excrete additional urea produced during phase I of fasting.

Animals↗

Fasting metabolism in Antarctic fur seal (Arctocephalus gazella) pups.

The metabolism of 52-73-day old Antarctic fur seal pups from Bird Island, South Georgia, was investigated during fasting periods of normal duration while their mothers were at sea foraging. Body mass decreased exponentially with pups losing 3.5-3.8% of body mass per day. Resting metabolic rate also decreased exponentially from 172-197 ml (O2) x min(-1) at the beginning of the fast and scaled to M(b)(0.74) at 2.3 times the level predicted for adult terrestrial mammals of similar size. While there was no significant sex difference in RMR, female pups had significantly higher (F(1,18)=6.614, P<0.019) mass-specific RMR than male pups throughout the fasting period. Fasting FMR was also significantly (t(15)=2.37, P<0.035) greater in females (823 kJ x kg(-1) x d(-1)) than males (686 kJ x kg(-1) x d(-1)). Average protein turnover during the study period was 19.3 g x d(-1) and contributed to 5.4% of total energy expenditure, indicating the adoption of a protein-sparing strategy with a reliance on primarily lipid catabolism for metabolic energy. This is supported by observed decreases in plasma BUN, U/C, glucose and triglyceride concentrations, and an increase in beta-HBA concentration, indicating that Antarctic fur seals pups adopt this strategy within 2-3 days of fasting. Mean RQ also decreased from 0.77 to 0.72 within 3 days of fasting, further supporting a rapid commencement of protein-sparing. However, RQ gradually increased thereafter to 0.77, suggesting a resumption of protein catabolism which was not substantiated by changes in plasma metabolites. Female pups had higher TBL (%) than males for any given mass, which is consistent with previous findings in this and other fur seal species, and suggests sex differences in metabolic fuel use. The observed changes in plasma metabolites and protein turnover, however, do not support this.

3-Hydroxybutyric Acid↗

Fasting and lactation effect fat-soluble vitamin A and E levels in blood and their distribution in tissue of grey seals (Halichoerus grypus).

Grey seals among other phacoids represent a good model to study the mobilisation, transfer and deposition of fat-soluble components such as vitamins in lactating females and suckling pups because during the lactation period mothers may fast completely while secreting large quantities of high fat milks, and pups deposit large amounts of fat as blubber. The level of vitamins A and E in different tissues (liver, adipose tissue, kidney, heart, skeletal muscle, testis) and blood plasma of adult grey seal females and males changed as a result of fasting and lactation; changes were also observed in pups. The most obvious effects were a significant increase of retinol and a decrease of vitamin E levels in plasma of females with the onset of lactation as well as a substantial decrease in liver vitamin E. In suckling pups both retinol and vitamin E levels in plasma increased with the onset of suckling; after weaning no changes in retinol but a significant decrease in plasma vitamin E was observed. While liver vitamin A levels tended to be unaffected by suckling or post-weaning fast, liver vitamin E levels increased with the uptake of milk substantially (P<0.01) and returned at weaning to low levels similar to that in fetuses. Adipose tissue levels of vitamin A and E in both females and pups were only marginally affected by lactation, suckling or post-weaning fast. Results indicate that both plasma and liver levels of vitamin A and E are affected by the mobilisation, absorption and deposition of these components during lactation in seals to a much greater extent than adipose tissue, from which fat-soluble vitamins are mobilized at rates similar to that of lipids.

Animals↗

Drinking behaviour and water turnover rates of Antarctic fur seal pups: implications for the estimation of milk intake by isotopic dilution.

The estimation of milk consumption in free-ranging seals using tritium dilution techniques makes the key assumption that the animals drink no pre-formed water during the experimental period. However, frequent observations of unweaned Antarctic fur seal pups drinking water at Iles Kerguelen necessitated the testing of this assumption. We estimated water flux rates of 30 pups (10.7+/-0.3 kg) in four experimental groups by isotopic dilution over 4 days. The groups were: (1) pups held in an open air enclosure without access to water to estimate fasting metabolic water production (MWP); (2) free-ranging pups not administered additional water; (3) pups held in an open air enclosure and given a total of 300 ml of fresh water to verify technique accuracy; and (4) free-ranging pups given 200 ml of fresh water. Pups without access to water exhibited water flux rates (20.5+/-0.8 ml kg(-1)d(-1)), which were significantly lower than those observed for the free-ranging group (33.0+/-1.7 ml kg(-1) d(-1)). Mean estimated pre-formed water intake for the free-ranging experimental groups was 12.6 ml kg(-1) d(-1). Thus, MWP, measured as total water intake during fasting, may be significantly over-estimated in free-ranging Antarctic fur seal pups at Iles Kerguelen and at other sites and subsequently milk intake rates may be underestimated.

Animals↗

Chemical characterization of the oligosaccharides in milk of high Arctic harbour seal (Phoca vitulina vitulina).

Carbohydrates were extracted from high Arctic harbour seal milk, Phoca vitulina vitulina (family Phocidae). Free neutral oligosaccharides were separated by gel filtration and preparative thin layer chromatography, while free sialyl oligosaccharides were separated by gel filtration and then purified by ion exchange chromatography, gel filtration and high performance liquid chromatography. Oligosaccharide structures were determined by 1H-NMR spectroscopy. The structures of the neutral oligosaccharides were as follows: lactose, 2'-fucosyllactose, lacto-N-neotetraose, lacto-N-neohexaose, monofucosyl lacto-N-neohexaose and difucosyl lacto-N-neohexaose. Thus, all of the neutral saccharides contained lactose or lacto-N-neotetraose or lacto-N-neohexaose as core units and/or non-reducing alpha(1-2) linked fucose. These oligosaccharides have also been found in hooded seal milk. The structures of the silalyl oligosaccharides were: monosialyl lacto-N-neohexaose, monosialyl monofucosyl lacto-N-neohexaose, monosialyl difucosyl lacto-N-neohexaose and disialyl lacto-N-neohexaose. These oligosaccharides contained lacto-N-neohexaose as core units, and one or two alpha(2-6) linked Neu5Ac, and/or non-reducing alpha(1-2) linked Fuc. The Neu5Ac residues were found to be linked to GlcNAc or penultimate Gal residues. The acidic oligosaccharides are the first to have been characterized in the milk of any species of seal.

Animals↗

Heterogeneous cholinergic reactions of ringed seal coronary arteries.

Coronary blood flow of some seal species is unusual in that it is highly variable in both non-diving and diving conditions and shows intermittent fluctuations, especially during dives when it frequently ceases for brief periods. We sought regulatory mechanisms governing these reactions by studying isometric tension recordings of isolated left circumflex (LC) and left anterior descending (LAD) coronary arteries of ringed seals, Phoca hispida, during reactions to a variety of agents for stimulating or blocking autonomic responses of the vascular smooth muscle. Micromolar acetylcholine (ACh) produced constriction of the small diameter segments of the LAD, but relaxation of the LC and larger segments of LAD. Both constrictions and dilations were prevented by atropine. Small vessel constriction by ACh was prevented by micromolar indomethacin and by a thromboxane receptor antagonist. Large vessel ACh dilations were prevented or reduced by rubbing off the endothelium and by the L-arginine analog, L-NG-nitro-arginine, an inhibitor of nitric oxide synthesis. We conclude that cholinergic, muscarinic, dilation of ringed seal large coronary arteries is mediated by endothelial-derived relaxing factor (EDRF), whereas ACh constriction of small arteries is mediated by a prostaglandin.

Acetylcholine↗

Thyroid hormones in grey seal pups (Halichoerus grypus).

The aim of the present study was to describe the changes in thyroid hormone status in grey seal (Halichoerus grypus) pups from birth to weaning and moulting. Plasma concentrations of total thyroxine (tT4) were highest the first two days after birth, thereafter dropping to a lower, but stable level. This pattern may reflect a high transfer rate of maternal thyroxine prepartum, prior to parturition, or postpartum via colostrum, or it may be caused by active secretory thyrocytes in late foetal stage. Total triiodothyronine (tT3) concentrations were lowest in neonatal pups, and increased as a function of age, indicating that deiodination of T4 to T3 increases as a function of age. Plasma concentrations of free thyroxine (fT4) did not vary as a function of age. All hormone concentrations were higher than previously reported in adults, probably reflecting the important role of these hormones in regulating their high rates of metabolism and tissue synthesis and the growth of their juvenile pelage. Since polychlorinated biphenyls (PCBs) have been reported to affect plasma concentrations of thyroid hormones in seals, the information on thyroid hormone concentrations and dynamics reported in grey seal pups from a pristine Norwegian coastal environment provide valuable reference material for future studies on pups from more polluted areas.

Animals↗

Nutrient, fatty acid, amino acid and mineral analysis of natural prey of the Hawaiian monk seal, Monachus schauinslandi.

Proximate nutrients, gross energy content, mineral, amino acid and fatty acid composition were determined for teleost, cephalopod and crustacean prey of the Hawaiian monk seal. Crude protein was highest in the octopus, Octopus cyanea (80.0%), crude fat was highest in the Muraenid teleost, Gymnothorax eurostus (14.1%), whereas crude ash was highest in the lobster, Panulirus marginatus (11.6%). Gross energies ranged from 4.0 +/- 0.01 kcal g-1 in the Labrid teleost Bodianus bilulunatus to 6.0 +/- 0.12 kcal g-1 in the moray eel, Gymnothorax undulatus. Essential amino acids occurred in lower concentrations as a percentage of the total amino acids (35.8 +/- 2.6%) than non-essential amino acids (64.2 +/- 2.6%), but the ratio of individual amino acids to total amino acid concentrations were similar to those required by some monogastric terrestrial species and fingerling salmon. The fatty acid concentrations varied widely among species (range = 1.2-16.5 mg 100 mg-1); however, the teleosts had higher total fatty acids than the non-teleosts. This study indicates that, from a nutritional standpoint, some prey may be more beneficial to the Hawaiian monk seal; however, these prey are not necessarily the most abundant or available to some populations of the monk seal.

Amino Acids↗

Salmonella in sub-Antarctica: low heterogeneity in Salmonella serotypes in South Georgian seals and birds.

The number of human visitors to Antarctica is increasing rapidly, and with it a risk of introducing infectious organisms to native animals. To study the occurrence of salmonella serotypes in sub-Antarctic wildlife, faecal samples were collected from gentoo penguins, macaroni penguins, gray-headed albatrosses, black-browed albatrosses and Antarctic fur seals on Bird Island in the South Georgian archipelago during the austral summer of 1996 and 1998. In 1996, S. havana, S. typhimurium and S. enteritidis were isolated from 7% of gentoo penguins and 4% of fur seals. In 1998, however, 22% of fur seals were found to be infected with S. havana, S. enteritidis and S. newport. All isolates, except one, showed identical pulsed-field gel electrophoresis-patterns within each serotype, irrespective of sampling year and animal reservoir. No significant antibiotic resistance was found. The very low heterogeneity in the salmonella isolates found could either indicate a high genetic adaptation of the bacteria to the environment or a recent introduction of salmonella into the area.

Adaptation, Physiological↗

Selective habituation shapes acoustic predator recognition in harbour seals.

Predation is a major force in shaping the behaviour of animals, so that precise identification of predators will confer substantial selective advantages on animals that serve as food to others. Because experience with a predator can be lethal, early researchers studying birds suggested that predator recognition does not require learning. However, a predator image that can be modified by learning and experience will be advantageous in situations where cues associated with the predator are highly variable or change over time. In this study, we investigated the response of harbour seals (Phoca vitulina) to the underwater calls of different populations of killer whales (Orcinus orca). We found that the seals responded strongly to the calls of mammal-eating killer whales and unfamiliar fish-eating killer whales but not to the familiar calls of the local fish-eating population. This demonstrates that wild harbour seals are capable of complex acoustic discrimination and that they modify their predator image by selectively habituating to the calls of harmless killer whales. Fear in these animals is therefore focused on local threats by learning and experience.

Acoustic Stimulation↗

Male mating success in an aquatically mating pinniped, the harbour seal (Phoca vitulina), assessed by microsatellite DNA markers.

Similar to many other pinniped species, harbour seals (Phoca vitulina) mate exclusively at sea. Here we present the first attempt to measure male mating success in an aquatically mating pinniped. Male mating success was estimated by paternity analysis in two cohorts of pups born at Sable Island, Nova Scotia, Canada, using microsatellite DNA markers. The genotypes of 275 pups born in 1994 and 1995 were compared to those of 90 candidate males at six microsatellite loci using a likelihood approach to resolve paternity. Paternity could be assigned for two, 22, 40 and 85 pups at confidence levels of 95, 80, 65 and 50%, respectively. Most successful males were assigned the paternity of a single offspring, suggesting a low variance in male mating success relative to most pinniped species. The proportion of paternal half sibs within cohorts and between maternally related sibs estimated by maximum likelihood were not significantly different from zero. It is thus unlikely that most offspring were sired by a small number of highly successful unsampled males, and that female harbour seals do not usually exhibit fidelity to the same male in sequential breeding seasons. A low level of polygyny in Sable Island harbour seals is consistent with predictions based on their breeding ecology, as females are highly mobile and widely dispersed in the aquatic mating environment at Sable Island.

Animals↗

Clostridium perfringens toxin types in hooded seals in the Greenland Sea, determined by PCR and ELISA.

Very little is known about the occurrence of Clostridium perfringens and of diseases caused by this anaerobic bacterium in marine mammals, especially those that are free-living. During a scientific expedition to the Greenland Sea (West Ice) in spring 1999, faeces samples from 70 hooded seals (Cystophora cristata) were taken to isolate C. perfiringens. Subsequently, PCR analysis of the isolates was performed with oligonucleotide primers of the genes encoding the four major lethal toxins (alpha, beta, epsilon and iota) for classification of toxin type and of the genes encoding C. perfringens beta2-toxin and enterotoxin for further subclassification. In addition, a commercial ELISA kit for detection of C. perfringens alpha, beta- and epsilon-toxin was used. C. perfingens was isolated in samples from 38 (54.3%) hooded seals. All isolates were C. perfringens toxin type A (alpha-toxin positive). This is the first report on the occurrence of C. perfringens in this arctic marine mammal species. Myositis and enterotoxemia caused by C. perfrigens were described in other marine mammals and it may be assumed that the pathogenesis of an outbreak of disease is similar to that encountered in terrestrial animals. Although there is some controversy surrounding the enteropathogenicity and virulence of alpha-toxin (concerning enterotoxemia), this study suggests that a possible outbreak of enterotoxemia caused by C. perfringens type A in hooded seals may, however, not be excluded.

Animals↗

Gross and microscopic visceral anatomy of the male Cape fur seal, Arctocephalus pusillus pusillus (Pinnipedia: Otariidae), with reference to organ size and growth.

The gross and microscopic anatomy of the Cape fur seal heart, lung, liver, spleen, stomach, intestine and kidneys (n = 31 seals) is described. Absolute and relative size of organs from 30 male seals are presented, with histological examination conducted on 7 animals. The relationship between log body weight, log organ weight and age was investigated using linear regression. Twenty five animals were of known age, while 6 were aged from counts of incremental lines observed in the dentine of tooth sections. For the range of ages represented in this study, body weight changes were accurately described by the exponential growth equation, weight = w(o)r(t), with body weight increasing by 23 % per annum until at least 9-10 y of age. Organ weight increased at a rate of between 25 % and 33 % per annum until at least 9-10 y of age, with the exception of the intestines, where exponential increase appeared to have ceased by about 7 y. The relationship between body weight and organ weight was investigated using logarithmic transformations of the allometric equation, y = ax(b), where the exponent b is 1 if organ weight is proportional to body weight. Most organs increased in proportion to the body. However, the heart, liver and spleen had exponents b > 1, suggesting that these organs increased at a faster rate than the body. The basic anatomical features of the viscera were similar to those of other pinnipeds, with some exceptions, including the arrangement of the multilobed lung and liver. Apart from the large liver and kidneys, relative size of the organs did not differ greatly from similar sized terrestrial carnivores. The histological features of the organs were generally consistent with those previously described for this species and other otariids. The heart, as in other pinnipeds, was unlike that of cetacea in not having unusually thick endocardium or prominent Purkinje cells. Notable histological features of the lungs included prominent fibrous septa, prominent smooth muscle bundles, cartilage extending to the level of the alveolar sacs and ample lymphoid tissue. The spleen had a thick capsule, well developed trabeculae and plentiful plasma cells. Abundant parietal cells were present in the fundic glands and lymphoid follicles were present in the gastric lamina propria, particularly in the pyloric region. Small intestinal villi were very long but this could have resulted from underlying chronic inflammation. Lymphoid follicles were prominent in the colon. The kidney reniculi each had a complete cortex, medulla and calyx, but a sportaperi medullaris musculosa was not identified.

Animals↗

The occurrence of Corynosoma strumosum in the grey seal, Halichoerus grypus, caught off the Atlantic coast of Ireland.

The occurrence, location and sex ratio of Corynosoma strumosum (Acanthoceophala: Polymorphidae) from 26 juvenile grey seals, Halichoerus grypus, by-caught from around the Inishkea Islands, Co. Mayo, from March to June 1997 were recorded. The location of C. strumosum within the seal did not vary, with all worms being found in the small intestine. The prevalence of infection was 100%, with no other acanthocephalan species being recorded. The mean abundance was 416 (range 80-846) C. strumosum per seal. There was no significant change in abundance by month. The sex ratio of mature female to mature male parasites was 1.2:1. The lengths of female and male worms did not differ significantly.

Acanthocephala↗

Cloning and sequencing of protein kinase cDNA from harbor seal (Phoca vitulina) lymphocytes.

Protein kinases (PKs) play critical roles in signal transduction and activation of lymphocytes. The identification of PK genes provides a tool for understanding mechanisms of immunotoxic xenobiotics. As part of a larger study investigating persistent organic pollutants in the harbor seal and their possible immunomodulatory actions, we sequenced harbor seal cDNA fragments encoding PKs. The procedure, using degenerate primers based on conserved motifs of human protein tyrosine kinases (PTKs), successfully amplified nine phocid PK gene fragments with high homology to human and rodent orthologs. We identified eight PTKs and one dual (serine/threonine and tyrosine) kinase. Among these were several PKs important in early signaling events through the B- and T-cell receptors (FYN, LYN, ITK and SYK) and a MAP kinase involved in downstream signal transduction. V-FGR, RET and DDR2 were also expressed. Sequential activation of protein kinases ultimately induces gene transcription leading to the proliferation and differentiation of lymphocytes critical to adaptive immunity. PKs are potential targets of bioactive xenobiotics, including persistent organic pollutants of the marine environment; characterization of these molecules in the harbor seal provides a foundation for further research illuminating mechanisms of action of contaminants speculated to contribute to large-scale die-offs of marine mammals via immunosuppression.

Amino Acid Sequence↗

Effect of a low-Fat diet on body composition and blubber fatty acids of captive juvenile harp seals (Phoca groenlandica).

We investigated the effects of a change from a high-fat diet to a low-fat diet of differing fatty acid (FA) composition on the body composition and blubber FA of five captive juvenile harp seals. Seals that had been maintained for 1 yr on a diet of Atlantic herring (>/=9% fat) were switched to a diet of Atlantic pollock (1. 7% fat) for 30 d. On days 0, 14, and 30, mass and body composition (using isotope dilution) were measured, and blubber biopsies (5 cmx6 mm) were taken for FA analysis. Fat accounted for 38%-49% of body mass at the start of the experiment. When switched to the pollock diet, and despite food intakes averaging 6.5 kg/d (32.3 MJ/d), body fat declined by an average of 6.4 kg or by 32% over the 30-d experiment. In contrast, body protein increased in direct relation to protein intake (r2=0.836, P=0.030). Despite substantial loss of body fat, blubber FA signature changed significantly to reflect the changes in dietary intake of FA, and the deposition of FA was quantifiably predictable. Our results suggest that young growing phocids are unable to maintain body fat stores on low-fat diets even when protein intakes are high. This may have significant implications for juvenile pinniped survival in the wild. In addition, turnover and deposition of dietary FA in blubber takes place in nonfattening seals.

Adipose Tissue↗

Thermoregulation of weaned northern elephant seal (Mirounga angustirostris) pups in air and water.

Fasting weaned northern elephant seal pups (Mirounga angustirostris) experience diverse environmental conditions on land and in water on a daily basis. Each environment undoubtedly induces distinct energetic costs that may vary for pups of differing body condition. To determine the energetic costs associated with different environmental conditions and whether costs vary between individuals, body mass, surface area, volume, body composition, resting metabolic rate, and core body temperature were determined for 17 weaned northern elephant seal pups from Año Nuevo, California. Metabolic rate and body temperature were measured for pups resting in air (20.9 degrees +/-0.8 degrees C), cold water (3.8 degrees+/-0.4 degrees ;C), and warm water (14.5 degrees+/-0.2 degrees C). Resting metabolic rate increased with body mass (range: 62.0-108.0 kg) and was also correlated with lean mass and lipid mass. Metabolic rates ranged from 293.6 to 512.7 mL O(2) min(-1) and were lowest for pups resting in cold water. Thermal conductance, calculated from metabolic rate and core body temperature, ranged from 3.1 to 15.2 W degrees C(-1), with the highest values in air and the lowest values in cold water. Metabolic responses to the three environmental conditions did not differ with individual variation in body condition. For all elephant seal pups, a consequence of high lipid content is that thermoregulatory costs are greatest on land and lowest in cold water, a pattern that contrasts markedly with terrestrial mammals.

Aging↗

Juvenile southern elephant seals exhibit seasonal differences in energetic requirements and use of lipids and protein stores.

Growing juvenile animals undergo many morphological, physiological, and behavioural changes that influence their energetic requirements, patterns of energy use, and ultimately, their survival and reproductive success. We examined changes in mass loss and body composition of juvenile southern elephant seals (1- and 2-yr-olds) during their two annual haul-outs. At the start and end of the midyear and molt haul-outs, we caught, weighed, and measured 41 and 14 seals, respectively. We measured blubber depth using ultrasound to estimate body composition (lean and adipose tissue mass). Using energy densities of the adipose and lean tissue, we calculated total, lean, and adipose mass changes and energy expenditure. While molting, juvenile seals used more energy than during the midyear, which is related to the increased use of lean tissue for hair and skin regeneration. The amount of energy used increases with mass as individuals mature. We found sexual differences in energy use where females retained greater fat reserves than males by utilizing more lean tissue. These differences are most likely related to haul-out function and behavior, growth, and earlier development of females toward sexual maturity.

Adipose Tissue↗