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Mercury and major essential elements in seals, penguins, and other representative fauna of the Antarctic.

Concentrations of total Hg and major essential elements, Ca, Mg, Na, and K, were measured in the muscle, liver, and kidney of three species of seals, crabeater seal (Lobodon carcinophagus), leopard seal (Hydrurga leptonyx), and Weddell seal (Leptonychotes weddelli) collected in the Antarctic. The muscle and liver of three species of penguins, i.e., gentoo penguin (Pygoscelis papua), Adelie penguin (P. adeliae), chinstrap penguin (P. antarctica), and other representative fauna were also analyzed for the elements. Distinct inter-tissue differences in the metal concentration were observed; liver had the greatest concentrations of Hg, kidney showed maximum concentration of Ca and Na, while muscle was characterized by the greatest content of Mg and K. Inter-specimen differentiation of concentrations among the same species was distinctly visualized for Hg but not for the major essential elements. The Hg concentration in the seals analyzed are in keeping with those reported previously by other authors. Such comparison for Ca, Mg, Na, and K was impossible because of the lack of available literature data for their concentrations in marine seals. Numerous significant correlations were observed between concentrations of the several metals analyzed. There was no correlation between Hg, which is a non-essential element and both Zn and Cd which are essential and non-essential elements, respectively in the liver or kidney of seals. However, there were significant correlations between concentration of Zn and the sum of molar concentrations of Hg+Cd in kidney (r = 0.82) and liver (r = 0.76). The results suggest that several control mechanisms operate to maintain physiologically required levels that decreases any effect of heavy metal toxicants such as Hg and Cd.

Animals↗

The isolation and partial characterization of a highly pathogenic herpesvirus from the harbor seal (Phoca vitulina).

This report describes the first isolation and partial characterization of a herpesvirus from the harbor seal (Phoca vitulina). The virus was isolated during a disease outbreak in a group of young seals nursed in a seal orphanage in The Netherlands. Almost half of the seals died with symptoms of acute pneumonia and focal hepatitis and the virus was isolated of organs of most of the dead animals. Seven out of ten seals of which paired serum samples were obtained showed seroconversion in a virus neutralization test during this outbreak. The virus was tentatively characterized as a herpesvirus (seal herpesvirus: SeHV or phocid herpesvirus 1) on the basis of its characteristic morphology in electron microscopy, buoyant density in sucrose, sensitivity to ether and heat treatment and its antigenic relationship with other probable members of the Alphaherpesvirinae subfamily. The virus caused cytopathic changes within 24 hours after inoculation in seal kidney cells, consisting of a focal rounding of cells and syncytium formation. No cytopathic changes were observed in the cells of nine other mammalian species tested.

Animals↗

Infection studies with canine distemper virus in harbour seals.

Infection studies in harbour seal (Phoca vitulina) were conducted with the Snyder-Hill strain of canine distemper virus (CDV) that is virulent for dog and mink. The inoculated seals showed clinical symptoms which were to some degree similar to those observed in CDV infections of sensitive species of carnivores. Viral replication in lymphoid cells was followed by an extended period of immunosuppression. The results did not provide conclusive evidence for viral replication in surface epithelia of seals, and accordingly no spread of the infection to contact seals and mink was demonstrated. The pathogenicity of the infection did not increase upon a second viral passage in seal. The serological data showed that CDV-infected seals mounted an early virus specific antibody response. Overall, the results indicated that the harbour seal was not especially sensitive to CDV infection. The differences in the in vivo biological properties of CDV and PDV add to the distinction between these viruses at the genomic and antigenic levels.

Animals↗

Elevated concentrations of trace elements in Caspian seals (Phoca caspica) found stranded during the mass mortality events in 2000.

Concentrations of V, Mn, Fe, Cr, Co, Cu, Zn, As, Se, Mo, Ag, Cd, Tl, Hg, Pb, and organic mercury (Org-Hg) were determined in liver, kidney, and muscle of healthy Caspian seals ( Phoca caspica) collected in 1998. These concentrations were compared with those of seals infected with canine distemper virus (CDV) found stranded along the coastal areas in 2000. Concentrations of toxic elements (As, Ag, Cd, Tl, Hg, Pb, and Org-Hg) in Caspian seals stranded in 2000 were comparable or lower than those of samples collected in 1998 and in other pinnipeds. Thus it may be inferred that these elements were not the causative agents in the deaths of the seals. In contrast, concentrations of Zn and Fe were much higher in diseased Caspian seals than those in other pinnipeds. Zinc concentrations in all tissues of Caspian seals also increased during 1993-2000. Furthermore, negative correlations were found between blubber thickness and hepatic and renal Zn concentrations. These results imply the disturbance in homeostatic control and nutritional status of essential elements in Caspian seals stranded in 2000.

Animals↗

Vaccination of harbour seals (Phoca vitulina) against phocid distemper with two different inactivated canine distemper virus (CDV) vaccines.

Two inactivated canine distemper virus (CDV) vaccines--an adjuvanted whole inactivated virus and a subunit ISCOM preparation--were tested for their ability to induce protective immunity in harbour seals (Phoca vitulina) against phocid distemper, a disease that recently killed greater than 17,000 harbour seals in the North and Baltic seas, and was shown to be caused by infection with a newly discovered morbillivirus, which is antigenically closely related to CDV. Four CDV seronegative harbour seals were vaccinated three times with the whole-virus vaccine, two with the ISCOM subunit vaccine and two were sham-vaccinated with an antigen-free preparation. Ten days after the last vaccination, when all six vaccinated animals had developed CDV neutralizing antibody titres ranging from 300 to 3000, all eight animals were challenged by the oculonasal and the peritoneal routes, with an organ suspension from dead seals. None of the six vaccinated animals developed clinical signs. The two sham-vaccinated seals died on days 14 and 18, respectively, after having shown a body temperature rise, respiratory symptoms and weight loss. In organs from both dead animals morbillivirus antigen was demonstrated with an enzyme-linked immunosorbent assay and an immunofluorescence assay. One of these two animals had developed a low titre of CDV-specific antibodies just before death. These data clearly indicate that seals can be protected from fatal challenge with the phocid distemper virus (PDV), by vaccination with certain inactivated CDV vaccines. They also reconfirm that infection with PDV should be considered the primary cause of the recent epizootic in seals.

Animals↗

Pathological findings indicative of distemper in European seals.

The first recorded cases of the recent epizootic were harbour seals observed at the Danish island of Anholt, 12 April 1988. The disease then spread throughout the sea waters of north-western Europe. The total mortality in Europe up to November 1988, was estimated to be at least 17,000 seals. The mortality rate in Danish-Swedish waters was about 60%. Autopsies including sampling for histology of most organs were performed on 37 harbour seals and 12 grey seals, collected mainly at the Swedish west coast and in the southern Baltic. In most of the harbour seals and in three of the grey seals we found histological changes in the upper and lower respiratory tracts, in the lower urinary tract and in the lymphatic system consistent with those diagnostic of distemper viral infection in the canine. These diagnostic criteria were: presence of intracytoplasmic eosinophilic inclusion bodies of epithelial cells of the trachea and the urinary bladder, interstitial pneumonia, and atrophy of lymphatic organs due to depletion of lymphocytes. Our findings in pathology of a canine distemper-like disease in the seals were presented in late August 1988, together with the Dutch findings in virology by Dr. Osterhaus and collaborators.

Animals↗

A 2,000-year record of mercury and ancient civilizations in seal hairs from King George Island, West Antarctica.

The concentrations of total mercury (Hg(T)) and three bio-essential elements (phosphor, potassium, sodium) were analyzed in Antarctic seal hairs from a lake core spanning the past 2,000 years and collected from King George Island (63 degrees 23'S, 57 degrees 00'W), West Antarctica. The Hg(T) concentration shows a significant fluctuation while the levels of the three bio-essential elements remain almost constant. The rise and fall of the Hg(T) concentration in the seal hairs are found to be closely coincided with ancient activities of gold and silver mining using Hg-amalgamation process around the world, especially in the Southern Hemisphere. Briefly, Hg(T) levels are high during five episodes of extensive gold and silver mining activities--Rome Empire and China Han Dynasty (approximately 18-300 A.D.), Maya period and China Tang (750-900 A.D.), Incas civilization and Christian Kingdom (1200-1500 A.D.), New world (1650-1800 A.D.), and modern industry period (1840 A.D.-present); they are low during four time periods of reduced gold and silver mining activities--the China Han and Rome fall (since 300 A.D.), Maya fall and Wartime period in China (1050-1250 A.D.), Pizarro coming (ca. 1532 A.D.) and Independence War of South America (1800-1830 A.D.). Two profiles of Hg(T) in other two lake cores, one affected by seal excrements and the other by penguin droppings, from the same region are similar to the one in seal hairs. The Hg concentration profile in the seal hairs is significantly correlated with the one in a peat bog of Southern Chile near King George Island. Since Hg is existent mainly at the form of methyl-mercury in seal hairs, this correlation supports a relationship and link between atmospheric mercury concentration and methyl-mercury production. Comparing with samples from American and European continents, the Antarctic seal hairs provide an archive of total mercury concentration in surface seawater of the South Ocean less affected by regional human activities, and this archive may provide a good reference for assessing the global Hg emissions, depositions and recycling in the past thousand years.

Animals↗

Persistent organochlorine pollutants in ringed seals and polar bears collected from northern Alaska.

Blubber samples from ringed seal (Phoca hispida; n = 8) and polar bear subcutaneous fat (Ursus maritimus; n = 5) were collected near Barrow, Alaska in 1996 as part of the Alaska Marine Mammal Tissue Archival Project (AMMTAP) and retained in the National Biomonitoring Specimen Bank at the National Institute of Standards and Technology in Gaithersburg, Maryland (USA). The samples were analyzed for a variety of persistent organochlorine pollutants (POPs) including polychlorinated biphenyls (PCBs), hexachlorocyclohexanes (HCHs), chlordane and metabolites, hexachlorobenzene (HCB) and DDTs and metabolites. The geometric mean, on a wet mass basis, of sigmaPCBs (sum of 29 congeners and congener groups) were 732+/-282 ng/g (1 S.D.) in seals and 3395+/-1442 ng/g in polar bears. The geometric mean of sigmaDDTs, sigmaHCHs (alpha-, beta- and gamma- HCH) and HCB concentrations (wet mass basis) in seals and bears were 562+/-261 ng/g vs. 74.8+/-39 ng/g, 380+/-213 ng/g vs. 515 ng/g, and 17.4+/-10.1 ng/g vs. 183+/-153 ng/g, respectively. The geometric mean sum of chlordane (sigmachlordane, sum of cis- and trans-chlordane, cis- and trans-nonachlor, oxychlordane and heptachlor epoxide) and dieldrin concentrations in ringed seals and polar bears were 753+/-617 ng/g vs. 720+/-315 ng/g and 38.6+/-22.8 ng/g vs. 130+/-65 ng/g, respectively. Apparent bioaccumulation factors (polar bear/ringed seal POP concentrations) were lower in the animals sampled near Barrow, Alaska than in those from locations in the Canadian Arctic. This suggests that polar bears are also preying on marine mammals from lower trophic levels than the ringed seals with correspondingly lower organochlorine levels, such as bowhead whale carcasses. PCB congener patterns in the samples demonstrated the metabolism of certain PCB congeners in the polar bear relative to the ringed seal in agreement with previous studies. Regional comparisons of animals collected in Alaska and Arctic Canada are presented.

Adipose Tissue↗

Seal gammaherpesviruses: identification, characterisation and epidemiology.

Phocid herpesvirus type 2 (PhHV-2), tentatively classified as a gammaherpesvirus, has been isolated from European and American harbour seals (Phoca vitulina). Here we describe the isolation and the molecular as well as biological characterisation of different PhHV-2 isolates from harbour seals and grey seals (Halichoerus grypus). Of 522 harbour seals and 231 grey seals that had been admitted to the seal research and rehabilitation centre in Pieterburen, The Netherlands, between 1992 and 2000, 38 and 18%, respectively, proved to have PhHV-2 neutralising antibodies. PhHV-2 was isolated from peripheral blood mononuclear cells (PBMCs) of 12 and 28% of these seropositive animals, respectively, and 26 and 56% of these cell samples, respectively, were positive by PCR analysis. Analysis of amino acid sequences of PCR products and of the growth characteristics of different PhHV-2 isolates indicated that harbour and grey seals are infected with distinct gamma-herpesviruses, which however, may co-circulate between the two species.

Amino Acid Sequence↗

Mercury, cadmium, lead and selenium in ringed seals (Phoca hispida) from the Baltic Sea and from Svalbard.

The concentrations of mercury (Hg), cadmium (Cd), lead (Pb) and selenium (Se) were determined in liver, kidney and muscle samples from 20 Baltic ringed seals (Phoca hispida botnica) (3-32 years), and from 17 ringed seals (Phoca hispida) (0-20 years) from Svalbard, in the Arctic. The concentrations of Hg and Se were considerably higher in the Baltic ringed seals, but the Cd concentrations lower than in the Svalbard ringed seals. There was no big geographical difference with respect to Pb concentrations. Se and Hg concentrations showed a significant positive correlation in both regions. By comparison with earlier studies on Baltic seals, the metal concentrations have remained at the same level since the 1980s. Of the metals we studied, only the level of Hg in Baltic ringed seals can be considered high (mean 53 mg/kg, range 6.5-124 mg/kg wet wt. for liver), but probably not high enough to cause metal intoxication. No pathological changes associated with metal intoxication were observed in the seals.

Animals↗

Organochlorine and organotin compounds in Caspian seals (Phoca caspica) collected during an unusual mortality event in the Caspian Sea in 2000.

Polychlorinated biphenyls (PCBs), organochlorine pesticides and organotin compounds were determined in the blubber and liver of Caspian seals (Phoca caspica) found stranded on the coast of the Caspian Sea during an outbreak of canine distemper virus (CDV) in 2000. Among organochlorines analyzed, DDTs were the most dominant contaminants with concentrations ranging from 6.3 to 470 microg/g on a lipid-weight basis. Caspian seals collected in 2000 during the epizootic had higher concentrations of organochlorines than healthy individuals sampled in 1998. However, the blubber layer was generally thinner in the seals collected in 2000 than those in the previous surveys. Although compositions of organochlorine pesticides in seals suggested that the contamination status in the Caspian Sea is improving, the levels found in Caspian seals in 2000 were comparable to those in other marine mammals that have suffered from epizootics. This implies that the present status of contamination found in Caspian seals poses a risk of immunosuppression. Concentrations of butyltin compounds in livers of seals ranged from 0.49 to 17 ng/g on a wet-weight basis and octyltin compounds were below limit of detection in all the samples analyzed, suggesting less contamination by organotin compounds in the Caspian Sea.

Algorithms↗

Seal and whale meat: two newly recognized food allergies.

BACKGROUND: Alaska's marine mammals compose a large portion of the diet of indigenous coastal Alaskan people. Bowhead whales (Balaena mysticetus) and bearded seals (Erignathus barbatus), inhabitants of the Bering and Beaufort seas along Alaska's western and northern coasts, are 2 of the most important subsistence species, serving as major food sources to the native population. OBJECTIVE: To describe an Inupiaq boy with symptoms consistent with an IgE-mediated food allergy after ingestion of bowhead whale and bearded seal meat. METHODS: Extracts of cooked bowhead whale and bearded seal were prepared, lyophilized, and evaluated for protein content. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed for each extract, followed by transfer to nitrocellulose and IgE immunoblots. Skin prick testing was conducted using reconstituted extracts of 1:10 wt/vol dilution. RESULTS: Immunoblots revealed serum specific IgE binding with the extracts of bowhead whale and bearded seal meat. Protein bands of approximately 25, 40, 50, and 90 kDa were found in the seal meat. Protein bands of 55 and 90 kDa were found in the whale meat. Skin prick test results were positive to whale and seal extracts with appropriate positive and negative controls. Ten control subjects had negative reactions to both extracts. CONCLUSION: A patient with moderate anaphylaxis to bowhead whale and bearded seal meat demonstrated serum specific IgE by means of immunoblot and positive skin prick test results. This is the first known reported case of specific IgE to these species.

Alaska↗

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.

Animals↗

Molecular scatology: the use of molecular genetic analysis to assign species, sex and individual identity to seal faeces.

Seals and commercial fisheries are potential competitors for fish and cephalopods. Research into the diet of British seal species has been based on conventional dietary analyses, but these methods often do not allow assignment of species identity to scat samples. We present a protocol for obtaining DNA from seal scat (faecal) samples which can be used in polymerase chain reactions to amplify both nuclear and mitochondrial DNA. This can provide a method of identifying the species, sex and individual identity of the seal, from a particular scat sample. Combined with conventional dietary analyses these techniques will allow us to assess sources of variation in seal diet composition. Scat samples have been collected from intertidal haul-out sites around the inner Moray Firth, north-east Scotland. We have assessed methods to extract and purify faecal DNA, a combination of DNA from the individual seal, prey items, and gut bacteria, for use in PCR. Controls using faecal and blood samples from the same individual have enabled microsatellite primer sets from four pinniped species to be tested. Approximately 200 scat samples have been examined for species identity and individual matches. This study will provide essential information for the assessment of interactions between seals and commercial or recreational fisheries.

Animals↗

Metabolic limits on dive duration and swimming speed in the southern elephant seal Mirounga leonina.

The ability of air-breathing marine predators to forage successfully depends on their ability to remain submerged. This is in turn related to their total O(2) stores and the rate at which these stores are used up while submerged. Body size was positively related to dive duration in a sample of 34 adult female southern elephant seals from Macquarie Island. However, there was no relationship between body size and dive depth. This indicates that smaller seals, with smaller total O(2) stores, make shorter dives than larger individuals but operate at similar depths, resulting in less time being spent at depth. Nine adult female elephant seals were also equipped with velocity time depth recorders. In eight of these seals, a plot of swimming speed against dive duration revealed a cloud of points with a clear upper boundary. This boundary could be described using regression analysis and gave a significant negative relationship in most cases. These results indicate that metabolic rate varies with activity levels, as indicated by swimming speed, and that there are quantifiable limits to the distance that a seal can travel on a dive of a given swimming speed. However, the seals rarely dive to these physiological limits, and the majority of their dives are well within their aerobic capacity. Elephant seals therefore appear to dive in a way that ensures that they have a reserve of O(2) available.

Aerobiosis↗

Vitamin D metabolism in the hooded seal (Cystophora cristata).

Fish-eating mammals, such as seals, appear to ingest levels of vitamin D that are toxic to most mammals. To determine how seals cope with high vitamin D intakes, the metabolism of tritiated cholecalciferol ([3H]D3) was investigated in hooded seal (Cystophora cristata) pups during their postweaning fast and pups and adults consuming herring alone or supplemented with 400,000 iu D3 daily. [3H]D3 was metabolized to 25-[3H]OHD3 and 24,25-[3H](OH)2D3. 1,25-[3H](OH)2D3 was not detected, but plasma levels of 1,25-(OH)2D were similar to those in other mammals and were not affected by vitamin D intake. Plasma vitamin D, 25-OHD and 24,25-(OH)2D increased with vitamin D intake, but 25-OHD did not increase to the extent seen in other mammals. The supplemented seals showed no evidence of toxicity. Levels of 24,25-(OH)2D were higher in the unsupplemented seals (4 to 33 ng/mL) than reported in other mammals with similar 25-OHD levels and did not decrease with 25-OHD. High levels of 24,25-(OH)2D relative to 25-OHD have also been found in hooded seals in the wild. The half-lives of vitamin D, 25-OHD and 24,25-(OH)2D were shorter than those reported for most other mammals. Increased conversion of 25-OHD to 24,25-(OH)2D and a high capacity for vitamin D storage in their large blubber mass appeared to be factors in the resistance of seals to vitamin D toxicity.

Animals↗

Elephant seal genetic variation and the use of simulation models to investigate historical population bottlenecks.

Because the northern elephant seal (Mirounga angustirostrus) was heavily exploited during the 19th century, it experienced an extreme population bottleneck. Since then, under legislative protection in the United States and Mexico, northern elephant seals have recovered dramatically in number, although their genomic diversity was greatly reduced, apparently as a consequence of the bottleneck. In this study we investigated DNA sequence diversity in two mtDNA regions (the control region and 16S RNA) and found low genetic variation in the northern elephant seal: there were only two control region haplotypes (sequence difference = 1%), which was consistent with an extreme founder event in the recent history of the northern species. We also reaffirmed the lack of allozyme diversity in this species. In contrast, the southern elephant seal (M. leonina), which though similarly exploited never fell below 1,000 animals, had 23 control region mtDNA haplotypes (average sequence difference = 2.3%). To investigate the extent of the founder event in the northern elephant seal we devised a simulation model based on extensive demographic data. This allowed a statistical analysis of the likely outcome of bottlenecks of different size and duration. Given these historical data, our results indicate (within 95% confidence) a bottleneck of less than 30 seals and 20-year duration, or, if hunting was the primary pressure on the population, a single-year bottleneck of less than 20 seals.

Animals↗

Morbillivirus infection of seals (Phoca vitulina) during the 1988 epidemic in the Bay of Heligoland. I. Mode, frequency and significance of cultural virus isolation and neutralizing antibody detection.

From 16 (14%) out of 112 dead or euthanized seals originating from wildlife and seal orphanages phocine morbillivirus was isolated. The majority of viral isolates in cell cultures was obtained from lung homogenates of 15 out of 71 free-ranging seals (21%). The virus was isolated by longterm cultivation in roller cultures of seal kidney cells. The phocine morbillivirus was detected by typical cytopathogenic alteration and by peroxidase-linked antibody (PLA) assay, respectively. A neutralization test based on PLA was used for antibody detection in seals using a canine distemper virus (CDV) strain and in parallel one of the phocine morbillivirus isolates. All sera tested were proven to contain neutralizing antibodies of higher titres against the latter virus than against the CDV strain. Several seals furnished morbillivirus isolates and at the same time exhibited neutralizing antibodies of low to medium titres. No viral isolates were obtained from the majority of sick animals with moderate to high neutralizing titres (greater than 1/1,000). The significance of these findings is discussed in relation to the cause of the mass mortality amongst seals observed in 1988 in the Bay of Heligoland.

Animals↗