Gene expression and trace elements in Greenlandic ringed seals (Pusa hispida).

Marine top predators such as ringed seals biomagnify environmental contaminants; and with the increasing human activities in the Arctic, ringed seals are exposed to biologically significant concentrations of trace elements resulting in reproductive impairment, immunosuppression, and neurological damages. Little is known about the molecular effects of heavy metals on these vulnerable apex predators suffering from a rapidly changing Arctic with significant loss of sea-ice. In the present study, concentrations of cadmium (Cd), mercury (Hg) and selenium (Se) were measured in liver of sixteen Greenlandic ringed seals (nine adults and seven subadults) together with molecular biomarkers involved in bio-transformation, oxidative stress, endocrine disruption and immune activity in blood and blubber. The concentrations of trace elements increased in the following order: Hg > Se > Cd with levels of mercury and selenium being highest in adults. Aryl hydrocarbon receptor nuclear translocator (ARNT), peroxisome proliferator activated receptor alpha (PPARα, estrogen receptor alpha (ESR1), thyroid hormone receptor alpha (TRα) and interleukin - 2 (IL-2) mRNA transcript levels were highest in blubber, while heat shock protein 70 (HSP70) and interleukin - 10 (IL-10) were significantly higher in blood. There were no significant correlations between the concentrations of trace elements and mRNA transcript levels suggesting that stressors other than the trace elements investigated are responsible for the changes in gene expression levels. Since Hg seems to increase in Greenlandic ringed seals, there is a need to re-enforce health monitoring of this ringed seal population.

Mercury in Ringed Seals (Pusa hispida) from the Canadian Arctic in Relation to Time and Climate Parameters.

Mercury is found in Arctic marine mammals that are important in the diet of northern Indigenous peoples. The objectives of the present long-term study, spanning a 45-yr period, were to 1) investigate the temporal trends of total mercury (THg; muscle and liver) and selenium (Se; liver) in ringed seals (Pusa hispida) from different regions of the Canadian Arctic; and 2) examine possible relationships with age, diet, and climate parameters such as air temperature, precipitation, climatic indices, and ice-coverage. Ringed seals were collected by hunters in northern communities in the Beaufort Sea, Central Arctic, Eastern Baffin Island, Hudson Bay, and Ungava/Nunatsiavut regions (Canada) between 1972 and 2017. Mercury levels did not change through time in seal liver, but THg levels in muscle decreased in seals from Hudson Bay (-0.91%/yr) and Ungava/Nunatsiavut (-1.30%/yr). Carbon stable isotope values in seal muscle decreased significantly through time in 4 regions. Selenium-to-THg ratios were found to be >1 for all years and regions. Variation partitioning analyses across regions indicated that THg trends in seals were mostly explained by age (7.3-21.7%), climate parameters (3.5-12.5%), and diet (up to 9%); climate indices (i.e., Arctic and North Atlantic Oscillations, Pacific/North American pattern) explained the majority of the climate portion. The THg levels had a positive relationship with Arctic Oscillation for multiple regions. Associations of THg with air temperature, total precipitation, and sea-ice coverage, as well as with North Atlantic Oscillation and Pacific/North American pattern were found to vary with tissue type and geographical area. Environ Toxicol Chem 2020;39:2462-2474. © 2020 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Fisheries and Oceans Canada.

Rapid changes in Atlantic grey seal milk from birth to weaning - immune factors and indicators of metabolic strain.

True seals have the shortest lactation periods of any group of placental mammal. Most are capital breeders that undergo short, intense lactations, during which they fast while transferring substantial proportions of their body reserves to their pups, which they then abruptly wean. Milk was collected from Atlantic grey seals (Halichoerus grypus) periodically from birth until near weaning. Milk protein profiles matured within 24 hours or less, indicating the most rapid transition from colostrum to mature phase lactation yet observed. There was an unexpected persistence of immunoglobulin G almost until weaning, potentially indicating prolonged trans-intestinal transfer of IgG. Among components of innate immune protection were found fucosyllactose and siallylactose that are thought to impede colonisation by pathogens and encourage an appropriate milk-digestive and protective gut microbiome. These oligosaccharides decreased from early lactation to almost undetectable levels by weaning. Taurine levels were initially high, then fell, possibly indicative of taurine dependency in seals, and progressive depletion of maternal reserves. Metabolites that signal changes in the mother's metabolism of fats, such as nicotinamide and derivatives, rose from virtual absence, and acetylcarnitines fell. It is therefore possible that indicators of maternal metabolic strain exist that signal the imminence of weaning.

Mercury and Selenium Balance in Endangered Saimaa Ringed Seal Depend on Age and Sex.

The endangered Saimaa ringed seal (Pusa hispida saimensis) is exposed to relatively high concentrations of mercury (Hg) in freshwaters poor in selenium (Se), a known antagonist of Hg. The impact of age and sex on the bioaccumulation of Hg and Se was studied by analyzing liver, muscle, and hair samples from seals of different age groups. Adult females were found to accumulate significantly more Hg in the liver (with ca. 60% as HgSe), and less Hg in the muscles compared to adult males, which may be explained by accelerated metabolism during gestation and lactation. In adult seals, molar Se:Hg ratios in the muscles fall below one, which is considered a threshold for the emergence of adverse effects. As a result, Saimaa ringed seals may be at risk of developing health and reproductive problems. According to mass balance calculations, the pups are exposed to considerable amounts (µg/d) of mercury during gestation, although lactation is their main exposure route. In lanugo pups, Hg concentrates in the hair, and molting serves as a main detoxification route. For other age groups, demethylation followed by the formation of HgSe is the main detoxification route, and the demethylation capability develops in pups by the time of weaning.

Mercury and selenium concentrations in skeletal muscle, liver, and regions of the heart and kidney in bearded seals from Alaska, USA.

Mean concentrations of total mercury ([THg]) and selenium ([TSe]) (mass and molar-based) were determined for 5 regions of the heart and 2 regions of the kidney of bearded seals (Erignathus barbatus) harvested in Alaska, USA, in 2010 and 2011. Mean [THg] and [TSe] of bearded seal liver and skeletal muscle tissues were used for intertissular comparison. The Se:Hg molar ratios were used to investigate elemental associations and potential antioxidant protection against Hg toxicosis. Age was an important factor in [THg] and Se:Hg molar ratios in heart and kidney. Small but statistically significant differences in mean [THg] occurred among some of the 5 heart regions (p < 0.05). Mean [THg] was highest in liver, 3.057 µg/g, and lowest in heart left ventricle, 0.017 µg/g. Mean [THg] ranked: liver > kidney cortex > kidney medulla > skeletal muscle > heart left ventricle (p < 0.001). Mean [TSe] was highest in liver, 3.848 µg/g, and lowest in heart left ventricle, 0.632 µg/g. Mean [TSe] ranked: liver > kidney cortex > kidney medulla > skeletal muscle > heart left ventricle (p < 0.001). The Se:Hg molar ratios were significantly greater than 1.0 in all tissues (p < 0.001) and represented baselines for normal [TSe] under relatively low [THg]. Mean Se:Hg molar ratios ranked: heart left ventricle > kidney medulla > kidney cortex (p < 0.001).

The effect of seal oil on paclitaxel induced cytotoxicity and apoptosis in breast carcinoma MCF-7 and MDA-MB-231 cell lines.

Some studies have suggested that omega-3 polyunsaturated fatty acids (PUFAs) have an inhibitory effect on the growth of cancer cells and therefore have the potential to increase the efficacy of cancer chemotherapeutic drugs. Considering that omega-3 PUFAs are present abundantly in harp seal oil, we investigated the effect of seal oil on the cytotoxicity and apoptosis induced by paclitaxel in 2 breast cancer cell lines, MCF-7 and MDA-MB-231, respectively. Cytotoxicity evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed that the concentration of paclitaxel that is required for 50% inhibition of cell growth in the presence of seal oil was significantly lower than that of paclitaxel alone. Apoptosis assessment based on morphological changes and DNA fragmentation results indicated that more cells treated with paclitaxel in combination with seal oil underwent apoptosis than with paclitaxel alone. Western blot analysis showed that the expression of B cell lymphoma-2 (Bcl-2) protein, an apoptosis inhibitory protein, in both cell lines was decreased more significant by paclitaxel in combination with seal oil than by paclitaxel alone. In addition, seal oil alone was found to induce apoptosis in both cell lines tested, which appeared to be due to the increased intracellular lipid peroxides produced. It is therefore concluded that paclitaxel in combination with seal oil demonstrated enhanced cytotoxicity and apoptosis in MCF-7 and MDA-MB-231 cells compared to paclitaxel alone, and the use of seal oil may be beneficial in the treatment of breast cancer.

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.

Effect of supplementation with dietary seal oil on selected cardiovascular risk factors and hemostatic variables in healthy male subjects.

The average daily consumption of seal oil by the Inuit people is approximately 8-9 g, yet there is very little information on the effect of seal oil consumption on cardiovascular disease risk factors. In this study, 19 healthy, normocholesterolemic subjects consumed 20 g of encapsulated seal oil containing eicosapentaenoic acid (EPA; 20:5n-3), docosahexaenoic acid (DHA; 22:6n-3), and docosapentaenoic acid (DPA; 22:5n-3) or 20 g of vegetable oil (control) per day for 42 days. Levels of selected cardiovascular and thrombotic risk factors as well as fatty acid profiles of serum phospholipid and nonesterified fatty acid (NEFA) were determined. EPA levels in serum phospholipid and NEFA increased by 4.3- and 2.7-fold, respectively, in the seal oil supplemented group. DHA levels rose 1.5- and 2.1-fold, respectively, and DPA levels rose 0.5- and 0.7-fold, respectively. Arachidonic acid (AA) levels dropped by 26% in both serum phospholipid and serum NEFA. There was a significant decrease in the ratio of n-6 to n-3 fatty acids in serum phospholipid from 7.2 to 2.1 and a significant increase in the ratio of EPA/AA in NEFA. Ingestion of seal oil raised the coagulant inhibitor, protein C, values by 7% and decreased plasma fibrinogen by 18%. No alterations in other hemostatic variables, including plasma activity of Factors VII, VIII, IX, and X and antithrombin, or in the concentrations of von Willebrand Factor, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglyceride, glucose, Apo A-1, or lipoprotein(a) were observed in either group. Other risk factors for cardiovascular disease, including hematocrit, white blood cell count, plasma viscosity, systolic and diastolic blood pressures, heart rate, and platelet aggregation after stimulation with ADP or collagen did not change. Our results indicate that seal oil supplementation in healthy, normocholesterolemic subjects decreased the n-6/n-3 ratio and increased EPA, DHA, and DPA and the ratio of EPA/AA and DHA/AA in the serum phospholipid and NEFA, while exhibiting a modest beneficial effect on fibrinogen and protein C levels.

Composition of polyunsaturated fatty acids in the liver of freshwater and marine ringed seals (Phoca hispida ssp.) differs largely due to the diet of the seals.

Fatty acid composition in the liver of four ringed-seal (Phoca hispida ssp.) populations, from ocean (Spitsbergen), brackish water (the Baltic Sea) and freshwater (Lake Saimaa and Lake Ladoga) were determined by gas liquid chromatography. The fatty acid compositions in liver were compared with those of the blubber of the seals, which largely reflect the fatty acid supply from marine or freshwater fish. When the ratios of 20:4n-6/20:5n-3 (arachidonic acid/eicosapentaenoic acid, AA/EPA, from 0.06 to 0.55) and n-6 PUFA/n-3 PUFA (n-6/n-3, from 0.09 to 0.35) increased from the marine to freshwater blubber, the corresponding increase in these ratios in the total lipids of the liver was 10-fold or more (AA/EPA: from 1.2 to 13.5 and n-6/n-3: from 0.9 to 3.2). Thus, when available in the diet, AA is preferentially incorporated into the lipids of pinniped liver.

Methylmercury and total mercury in tissues of arctic marine mammals.

Concentrations of methylmercury, total mercury and selenium in marine mammal tissues were determined in liver, muscle, skin (muktuk) and blubber of belugas, ringed seals and narwhal, using atomic absorption and capillary gas chromatography with ECD detection. Mean MeHg levels in the types of tissues analysed, except blubber, generally exceeded the Canadian Federal Consumption Guideline for mercury in fish (0.5 micrograms/g wet wt.). A spatial trend of higher MeHg levels in western compared to eastern Arctic belugas and ringed seals was found which followed a similar trend observed for total mercury. Factors which could explain this trend are discussed. Robust linear regression of MeHg on total Hg and MeHg on age of animals was performed and a strong correlation between the two variables was found in each case. The ratio of MeHg to total mercury as indicated by the regression coefficients was close to one for muscle and skin (muktuk) while for liver it was < 1. The mean percentage of MeHg in the liver of marine mammals was 3-12% of the total Hg in this tissue depending on species and location. It is postulated that the formation and deposition of mercuric selenide in the liver is part of the demethylation process in this tissue. This is based on the relatively low fraction of MeHg in the liver not withstanding the fact that the predominant form of mercury taken up via food is MeHg. The long half-life for total mercury and the relatively short half-life for MeHg in this organ are in accord with this postulate as is the 1:1 stoichiometric relationship between mercury and selenium in the liver.

Twenty years of trace metal analyses of marine mammals in Alaska: evaluation and summation.

The compilation of existing data on contaminants in the marine food chain is essential in addressing concerns regarding the magnitude of potential human exposures and in the evaluation of subsistence food safety. This paper presents a summary of studies on trace metals in tissues of Alaska marine mammals from the 1970s to the present, along with derived mean tissue trace metal concentrations. The derived mean can serve as a norm against which future monitoring results may be compared, and may be used to estimate human exposure to trace metals through the consumption of marine mammals. Additionally, the variation among studies in the reported mean tissue concentrations has been described through a derived standard deviation. Sufficient analytical and methodological details were available to derive means and standard deviations for tissues in bearded seal, bowhead whale, beluga whale, fur seal, harbor seal, Pacific walrus, and ringed seal. A high concordance between trace metal values reported in tissues (i.e., liver, kidney, muscle) was observed despite significant differences in reported sampling and analytical methodologies. Consistent with other reviews of trace metal concentrations in marine species, the standard deviation of tissue metal concentrations was generally < or = 100% of the reported mean. Significant gaps in available information remain, particularly for muscle tissues and for methylmercury, despite the considerable efforts to monitor marine mammal species in Alaska.

Overview and regional and temporal differences of heavy metals in Arctic whales and ringed seals in the Canadian Arctic.

Concentrations of mercury, cadmium, and other heavy metals in tissues of belugas (Delphinapterus leucas), narwhal (Monodon monoceros) and ringed seals (Phoca hispida) from across the Canadian Arctic are reported. Published and new information is used to provide an overview of metals in tissues of these animals, to delineate the existence of a spatial trend of mercury and cadmium in belugas and ringed seals, and to show a temporal trend is superimposed on the geochemical trend. Mercury concentrations in tissues of Arctic whales and ringed seals were high relative to the Canadian guideline of 0.5 micrograms/g wet wt., for mercury in fish, except in the skin of belugas (0.59-0.78 micrograms/g wet wt.) and flesh of ringed seals (0.39-0.41 micrograms/g wet wt.). In the flesh of belugas (0.94-1.34 micrograms/g wet wt.), and in the liver of ringed seals, tissues that are also consumed by Native people in the Arctic (8.34-27.5 micrograms/g wet wt.), the guideline value was significantly exceeded. Mean lead concentrations in tissues of belugas, narwhal and ringed seals were generally low (0.002-0.028 micrograms/g wet wt.), except in tissues of belugas in the St. Lawrence River (0.10-0.15 micrograms/g wet wt.). The concentration of zinc in the skin of whales was two to three times higher than in other tissues. The concentration of cadmium in organs was highest in narwhal. There was a positive correlation between mercury and selenium in the liver of all three species. The concentration of mercury in tissues of belugas and ringed seals was higher in the western than the eastern Arctic. This was attributed to different natural background concentrations in the western and eastern Arctic of Canada dictated by different geological formations in the two regions. Cadmium concentrations in tissues of belugas and ringed seals were higher in the eastern than the western Arctic. Zinc and copper in some tissues of belugas and ringed seals were also higher in the eastern than the western Arctic. Mercury in the liver of belugas was found to have increased in the western and eastern Arctic over 10-12 years. Mercury in the liver of ringed seals in the western Arctic and narwhal in the eastern Arctic showed similar increases. In recently collected belugas, the rate of accumulation of mercury in the liver was approximately twice that in belugas collected 10-12 years ago. In ringed seals, the rate was three times higher in recent samples compared to 15-20 years ago. There was no temporal change in cadmium levels in tissues of belugas, ringed seals or narwhal.

Bioaccumulation of vanadium and other trace metals in livers of Alaskan cetaceans and pinnipeds.

Concentrations for 38 elements are routinely measured in the marine mammal liver tissues archived in the National Biomonitoring Specimen Bank (NBSB). Results show that hepatic concentrations of vanadium, selenium, silver, cadmium, and mercury are positively correlated with age for beluga whales (Delphinapterus leucas) and of vanadium, selenium, cadmium, and mercury with length for ringed seals (Phoca hispada). Many researchers have reported linear correlations of hepatic selenium, cadmium, and mercury with marine mammal age; however, there is only one other report of a linear correlation of hepatic vanadium with marine mammal age. Vanadium levels are at or below detection limits (< or = 0.01 micrograms/g) in liver tissues of U.S. east coast marine mammals from the NBSB but are present at levels ranging from 0.02 to 1.2 micrograms/g of wet weight in the tissues of Alaskan marine mammals. Although only three bearded seal (Eriganthus barbatus) and three bow-head whale (Balaena mysticetus) liver samples have been analyzed, hepatic vanadium levels also increased with animal size for these species. The presence of relatively high levels of vanadium in the livers of these Alaskan animals may reflect a unique dietary source of vanadium, a unique geochemical source of vanadium, or anthropogenic input to the Alaskan marine environment.

Effects of dietary triacylglycerol structure on triacylglycerols of resultant chylomicrons from fish oil- and seal oil-fed rats.

We investigated the influence of the intramolecular fatty acid distribution of dietary triacyl-sn-glycerols (TAG) rich in n-3 polyunsaturated fatty acids (PUFA) on the structure of chylomicron TAG. Fish oil and seal oil, comparable in fatty acid compositions but with different contents of major n-3 PUFA esterified at the sn-2 position (20:5n-3, 46.6%, and 5.3%; 22:6n-3, 75.5%, and 3.8%, respectively), were fed to rats. Mesenteric lymph was collected and the chylomicrons were isolated by ultracentrifugation. The fatty acid composition of chylomicrons largely reflected the fatty acid composition of the oils administered. The intramolecular fatty acid distributions of the TAG fed were reflected in the chylomicron TAG as the fraction of the total contents observed in the sn-2 position of 20:5n-3 were 23.6 and 13.3%, and of 22:6n-3 were 30.6 and 5.4% for resultant chylomicrons following fish oil and seal oil administration, respectively. Thus, after seal oil administration, significant higher load of n-3 PUFA was esterified in the sn-1,3 positions of chylomicron TAG compared with fish oil administration (P < 0.05).

[PCB methyl sulphone: comparison of tissue levels in Baltic grey seals and a Yusho patient].

Methyl sulphone metabolites of PCB and DDE were isolated from different tissues of a Baltic grey seal (Halichoerus grypus). Main components in the seal blubber were identified as 3-MeSO2-2,2',4',5,5'-pentaCB, 4-MeSO2-2,2',4',5,5'-pentaCB, 3-MeSO2-p,p'-DDE, 4-MeSO2-2,2',3',4',5-pentaCB and 4-MeSO2-2,2',3,4',5',6-hexaCB. Liver and lung in the seal contained different MeSO2-PCB pattern compared to all other tissues. These levels in the both tissues were estimated to be 28 and 15 ppm (lipid basis) which corresponded to the same level as the PCB. Concentrations of MeSO2-PCB in any tissues of a Yusho patient were low compared to those in the seal.

Characterization and measurement of immunoglobulins in the grey seal (Halichoerus grypus).

Immunoglobulin concentrations were measured in the serum and colostrum of adult and pup Grey seals from North Rona in the outer Hebrides and the Isle of May in the Firth of Forth. IgG was shown to be the main immunoglobulin and two subclasses were identified. Serum immunoglobulin content was low in the week-old pups and increased up to 5 weeks, although it remained substantially lower than in the adults. Colostral immunoglobulins were high and cannot explain the low values in the pup serum. It is considered that the increased opportunistic infections seen in Grey seal pups may be related to this low immunoglobulin status.

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.

Pollutants in harp seals (Phoca groenlandica). II. Heavy metals and selenium.

Between 1976 and 1978, 249 harp seals were sampled from five locations in the Northwest Atlantic and Arctic for heavy metal and selenium residue analyses in tissue. Significant loading was apparent only in blood, brain, kidney, liver and muscle. Samples were analysed for mercury (Hg), selenium (Se), copper (Cu), cadmium (Cd) and lead (Pb). The seals carried higher levels of Cd than of the other metals. Residue levels of Cd were highest in kidney tissue; levels of other metals were highest in liver. Both males and females bioaccumulated Cd, Hg and Se. Cd, Hg, Se and Cu residues were detected in tissue from neonatal seals, indicating that transplacental and transmammary transfer of these elements had occurred. Despite the passage of residues from mother to pup, females bore significantly higher levels of Hg and Cd than males. Conversely, levels of Cu, Se or Pb did not appear to differ significantly between sexes. There was considerable individual variation in residue levels. This, coupled with the extensive annual migration undergone by these animals, made it difficult to arrive at definite conclusions regarding geographic accumulation patterns.