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See detailXenobiotic and Immune-Relevant Molecular Biomarkers in Harbor Seals as Proxies for Pollutant Burden and Effects?
Lehnert, Kristina; Ronnenberg, Katrin; Weijs, Liesbeth et al

in Archives of Environmental Contamination & Toxicology (2016), 70(1), 106-120

Harbor seals are exposed to increasing pressure caused by anthropogenic activities in their marine environment. Persistent organic pollutants (POPs) and trace elements are hazardous contaminants which ... [more ▼]

Harbor seals are exposed to increasing pressure caused by anthropogenic activities in their marine environment. Persistent organic pollutants (POPs) and trace elements are hazardous contaminants which accumulate in tissues of harbor seals. POPs and trace elements can negatively affect the immune-system and have been reported e.g. to increase susceptibility to viral infections in seals. Biomarkers of the xenobiotic metabolism, cytokines and heat-shock-protein as cell mediators of the immune-system were established to evaluate the impact of environmental stressors on harbor seals. Harbor seals (n=54) were captured on sandbanks in the North Sea during 2009 to 2012. Medicals including hematology were performed, RNAlater blood samples were taken and analyzed using quantitative Polymerase Chain Reaction. Normalized transcript copy numbers were correlated to hematology and POP concentration in blood, and trace metals in blood and fur. [less ▲]

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See detailEffects of Methylmercury on Harbour Seal Peripheral Blood Leucocytes In Vitro Studied by Electron Microscopy
Dupont, Aurélie; De Pauw-Gillet, Marie-Claire ULg; Schnitzler, Joseph ULg et al

in Archives of Environmental Contamination & Toxicology (2016), 70(1), 133-142

Methylmercury MeHg is highly immunotoxic and can alter the health status of the harbour seal, Phoca vitulina, from the North Sea. To investigate the mechanism of MeHg-induced toxicity in harbour seal ... [more ▼]

Methylmercury MeHg is highly immunotoxic and can alter the health status of the harbour seal, Phoca vitulina, from the North Sea. To investigate the mechanism of MeHg-induced toxicity in harbour seal lymphocytes, ConA-stimulated peripheral blood leucocytes were exposed in vitro to sub-lethal concentrations of MeHgCl (0.2, 1 and 2 µM) for 72h, and then analysed for their viability and ultrastructure. After 72h of incubation, cells were counted with a propidium iodide staining technique, a metabolic MTS assay was performed, and cells exposed to 1 µM MeHgCl were observed by transmission electron microscopy (TEM). Alive cell numbers decreased with increased MeHgCl concentrations. In presence of ConA and 1µM of MeHgCl, TEM images revealed a higher frequency of apoptotic cells. Exposed cells displayed condensation of the chromatin at the nuclear membrane and mitochondrial damages. The results suggest that in vitro MeHgCl induced apoptosis in harbour seal lymphocytes through a mitochondrial pathway. [less ▲]

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See detailAbsence of selenium protection against methylmercury toxicity in harbour seal leucocytes in vitro
Das, Krishna ULg; Dupont, Aurélie; De Pauw-Gillet, Marie-Claire ULg et al

in Marine Pollution Bulletin (2016), 108

Previous studies described high concentrations of mercury (Hg) and selenium (Se) in the blood of harbour seals, Phoca vitulina from the North Sea. In the present study, we evaluated the in vitro potential ... [more ▼]

Previous studies described high concentrations of mercury (Hg) and selenium (Se) in the blood of harbour seals, Phoca vitulina from the North Sea. In the present study, we evaluated the in vitro potential protective effects of sodium selenite (Na2SeO3) and selenomethionine (SeMet) on cell proliferation of harbour seal lymphocytes exposed to MeHgCl 0.75 µM. In vitro exposure of ConA-stimulated T lymphocytes resulted in severe inhibition of DNA synthesis, likely linked to severe loss of mitochondrial membrane potential at 0.75 µM. Neither selenite nor SeMet showed a protective effect against MeHg toxicity expressed at the T lymphocyte proliferation level for harbour seals. Selenite and SeMet did not show negative effects regarding lymphocyte proliferation and mitochondrial membrane potential. To conclude, our results clearly demonstrated that MeHg affected in vitro immune cells exposure with no protective effects of selenium at a molar ratio Hg:Se of 1:10 in harbour seals from the North Sea. [less ▲]

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See detailSeasonal Variation of Harbor Seal's Diet from the Wadden Sea in Relation to Prey Availability
de la Vega, Camille; Lebreton, Benoit; Siebert, Ursula et al

in PLoS ONE (2016)

The Wadden Sea has an important role for marine mammals in terms of resting, nursing and foraging. Harbor seal is the most abundant marine mammal species in this area. The use of the food resources of the ... [more ▼]

The Wadden Sea has an important role for marine mammals in terms of resting, nursing and foraging. Harbor seal is the most abundant marine mammal species in this area. The use of the food resources of the Wadden Sea by seals is not clear, and previous studies showed that this species can travel kilometers away from their haul-outs to forage in the North Sea. In this study, we analyzed the stable isotopes of vibrissae from 23 dead harbor seals found on the island of Sylt to investigate their diet. The predator´s carbon and nitrogen isotope compositions were compared to the compositions of different potential prey items from the Sylt-Rømø Bight and from the North Sea in order to study seasonal pattern in the diet and in the foraging location. In parallel, seasonal variation of abundance and biomass of the potential prey items from the Sylt-Rømø Bight were studied and compare to their contribution to the seal´s diet. The results revealed a change in the seal´s diet from pelagic sources in spring to a benthic based diet in summer, and an increasing use of the North Sea resources in fall and winter in accordance with the seasonal variation of the availability of prey in the Sylt-Rømø Bight. [less ▲]

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See detailImpacts of underwater noise on marine vertebrates: Project introduction and first results
Liebschner, Alexander; Seibel, Henrike; Teilmann, Jonas et al

in Advances in Experimental Medicine and Biology (2015), 875

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See detailRelationships between in vitro lymphoproliferative responses and levels of contaminants in blood of free-ranging adult harbour seals (Phoca vitulina) from the North Sea
Dupont, Aurélie ULg; Siebert, Ursula; Covaci, Adrian et al

in Aquatic Toxicology (2013), 142-143

tIn vitro culture of peripheral blood leucocytes (PBLs) is currently used in toxicological studies of marinemammals. However, blood cells of wild individuals are exposed in vivo to environmental ... [more ▼]

tIn vitro culture of peripheral blood leucocytes (PBLs) is currently used in toxicological studies of marinemammals. However, blood cells of wild individuals are exposed in vivo to environmental contaminantsbefore being isolated and exposed to contaminants in vitro. The aim of this study was to highlightpotential relationships between blood contaminant levels and in vitro peripheral blood lymphocyteproliferation in free-ranging adult harbour seals (Phoca vitulina) from the North Sea. Blood samplesof 18 individuals were analyzed for trace elements (Fe, Zn, Se, Cu, Hg, Pb, Cd) and persistent organiccontaminants and metabolites ( PCBs, HO-PCBs, PBDEs, 2-MeO-BDE68 and 6-MeO-BDE47, DDXs,hexachlorobenzene, oxychlordane, trans-nonachlor, pentachlorophenol and tribromoanisole). The samesamples were used to determine the haematology profiles, cell numbers and viability, as well as thein vitro ConA-induced lymphocyte proliferation expressed as a stimulation index (SI). Correlation tests(Bravais-Pearson) and Principal Component Analysis with multiple regression revealed no statisticallysignificant relationship between the lymphocyte SI and the contaminants studied. However, the numberof lymphocytes per millilitre of whole blood appeared to be negatively correlated to pentachlorophenol(r = −0.63, p = 0.005). In adult harbour seals, the interindividual variations of in vitro lymphocyte pro-liferation did not appear to be directly linked to pollutant levels present in the blood, and it is likelythat other factors such as age, life history, or physiological parameters have an influence. In a generalmanner, experiments with in vitro immune cell cultures of wild marine mammals should be designed soas to minimize confounding factors in which case they remain a valuable tool to study pollutant effectsin vitro. [less ▲]

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See detailThe stranding anomaly as population indicator: the case of harbour porpoise Phocoena phocoena in North-Western Europe.
Peltier, Helene; Baagoe, Hans J.; Camphuysen, Kees C. J. et al

in PloS one (2013), 8(4), 62180

Ecological indicators for monitoring strategies are expected to combine three major characteristics: ecological significance, statistical credibility, and cost-effectiveness. Strategies based on stranding ... [more ▼]

Ecological indicators for monitoring strategies are expected to combine three major characteristics: ecological significance, statistical credibility, and cost-effectiveness. Strategies based on stranding networks rank highly in cost-effectiveness, but their ecological significance and statistical credibility are disputed. Our present goal is to improve the value of stranding data as population indicator as part of monitoring strategies by constructing the spatial and temporal null hypothesis for strandings. The null hypothesis is defined as: small cetacean distribution and mortality are uniform in space and constant in time. We used a drift model to map stranding probabilities and predict stranding patterns of cetacean carcasses under H0 across the North Sea, the Channel and the Bay of Biscay, for the period 1990-2009. As the most common cetacean occurring in this area, we chose the harbour porpoise Phocoena phocoena for our modelling. The difference between these strandings expected under H0 and observed strandings is defined as the stranding anomaly. It constituted the stranding data series corrected for drift conditions. Seasonal decomposition of stranding anomaly suggested that drift conditions did not explain observed seasonal variations of porpoise strandings. Long-term stranding anomalies increased first in the southern North Sea, the Channel and Bay of Biscay coasts, and finally the eastern North Sea. The hypothesis of changes in porpoise distribution was consistent with local visual surveys, mostly SCANS surveys (1994 and 2005). This new indicator could be applied to cetacean populations across the world and more widely to marine megafauna. [less ▲]

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See detailMercury in blood of free-ranging seals Phoca vitulina from the North Sea: Time-trend and association with environmental factors
Das, Krishna ULg; Brochoire, Charlène ULg; Chambosse, Mélanie et al

Conference (2012, March 27)

The harbour seal (Phoca vitulina) population from the North Sea has experienced various fluctuations these last decades due to habitat loss, prey fluctuation and pollution of the marine environment ... [more ▼]

The harbour seal (Phoca vitulina) population from the North Sea has experienced various fluctuations these last decades due to habitat loss, prey fluctuation and pollution of the marine environment. Recently, development of monitoring programs and non-invasive sampling techniques, including seal catches allowed blood sampling on a regular basis together with measurements of blubber thickness, body mass, sex and body length. Concentrations of total mercury (T-Hg) and other trace elements (Cd, Pb, Zn, Cu, Fe) as well as ∂13C and ∂15N values were determined by mass spectrometry in blood of 75 wild harbour seals caught in the German Wadden Sea between 1997 and 2011. ∂13C and ∂15N mean values (-17.5‰ and 18.1 ‰ respectively) were strongly similar to that measured previously in the muscle of stranded harbour seals from the Wadden Sea. In contrast, ∂15N mean value was strikingly higher than that recorded in grey seals (Halichoerus grypus) from Scotland (14.1 ‰ respectively; Habran et al. submitted.) while ∂13C values remained similar between the two seal species. These values confirmed the high trophic position of the harbour seal in the North Sea. In contrast to Cd and Pb, T-Hg in blood harbour seals reached concentrations as high as 2.1 μg.g-1 dry weight (10 times higher than the 0.21 μg.g-1 dry weight recorded for grey seals from Scotland) but depended on several factors including ∂15N values, age group and the body mass. T-Hg was detected in juveniles confirming maternal transfer to offspring and time-trend revealed no decrease of T-Hg in blood of harbour seals these last 15 years. [less ▲]

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See detailPotential effects of blood contaminants on immune responses in harbour seals (Phoca vitulina)
Dupont, Aurélie ULg; Weijs, Liesbeth; Siebert, Ursula et al

Poster (2011, August 23)

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See detailSTUDY OF SELENITE AND SELENOMETHIONINE EFFECT ON METHYLMERCURY IN VITRO TOXICITY
Dupont, Aurélie ULg; Siebert, Ursula; Rosenberger, Tanja et al

Conference (2011, May 16)

Methylmercury (MeHg) and selenium (Se) can be found at elevated concentrations in blood of marine mammals and both display modulatory effects on the immune system. Whereas mercury (Hg)-Se antagonism in ... [more ▼]

Methylmercury (MeHg) and selenium (Se) can be found at elevated concentrations in blood of marine mammals and both display modulatory effects on the immune system. Whereas mercury (Hg)-Se antagonism in liver of marine mammals is well known, the protective role of Se against Hg immunotoxicity in marine mammals has been poorly described. We propose here an in vitro approach using combined Hg and Se in vitro exposure of peripheral blood mononuclear cells (PBMCs) of the harbor seal (Phoca vitulina). PBMCs were isolated from the blood of 10 harbor seals and exposed to environmental concentrations of MeHg (1µM) and selenite or selenomethionine (5µM), respectively inorganic and organic forms of Se. MeHg leaded to a decrease of lymphocyte proliferation, to an increase of cells with compromised mitochondrial membrane potentials and cell death. Preliminary results evidenced that none of the two Se forms had a protective effect against MeHg toxicity, although cells were slightly stimulated by Se alone. Therefore MeHg expresses its toxicity among blood circulating lymphocytes in presence or absence of selenite or selenomethionine. [less ▲]

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See detailThe harbor seal and the harbor porpoise from the North Sea: review of their ecotoxicological status based on stranded and free-ranging individuals and potential threaths to the population
Das, Krishna ULg; Weijs, Liesbeth; Habran, Sarah ULg et al

Conference (2011, May)

The harbor seal (Phoca vitulina) and the harbor porpoise (Phocoena phocoena) from the North Sea have experienced major fluctuations these last decades due to habitat loss, prey fluctuation and pollution ... [more ▼]

The harbor seal (Phoca vitulina) and the harbor porpoise (Phocoena phocoena) from the North Sea have experienced major fluctuations these last decades due to habitat loss, prey fluctuation and pollution of the marine environment. Recently, development of monitoring programs and non-invasive sampling techniques, including seal catches in Germany allowed blood sampling together with measurements of blubber thickness, body mass, sex and body length. This approach is complementary to the study of stranded and by-caught individuals sampled during necropsies. Essential (Se, Zn, Cu, Fe) and non-essential elements (T-Hg, MeHg, Cd, Pb), perfluorinated organochemicals (PFCs) polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) were measured in blubber and in blood (for selected compounds) of stranded, by-caught and free-ranging individuals from Belgian and German coasts. In contrast to Cd and Pb, hepatic T-Hg can reach concentrations as high as 2.1 μg.g-1 dry weight but depended on several factors including position in the trophic chain (inferred from δ13C and δ15N values), age group and the body mass. This level has been shown to generate in vitro toxicity on harbor seal lymphocytes. Organic compound analysis (PFCs, PCBs, PBDEs) revealed widespread dispersion of contaminants in the marine environment with higher concentrations in seals compared to porpoises in agreement with the higher trophic position of the harbor seal. However, metabolization of these compounds differed between the two species, as revealed by the higher contribution of the persistent BDE-47 and lower chlorinated and non-persistent congeners (e.g. CB 52, CB95) in tissues of harbor porpoises. T-Hg, PFCs, PCBs and PBDEs were detected in calves confirming maternal transfer to offspring. These pollutants are strongly suspected to affect the immune and endocrine systems as well as vitamin A metabolism and this raises concern about exposure-related health effects, especially in younger individuals. [less ▲]

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See detailNew insights in the toxicology and health status of marine marine mammals: Use of free-ranging harbour seals from the Wadden Sea
Das, Krishna ULg; Seibel, Henrike; Hasseilmeier, Ilka et al

Conference (2011, March 20)

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See detailEvaluation of the ecotoxicological effects of POPs and heavy metals, reflecting pathological, microbiological and genetic analyses, on the Mekong River population of Irrawaddy dolphins (Orcaella brevirostris)
Siebert, Ursula; Das, Krishna ULg

Report (2011)

For this report, samples of Irrawaddy River dolphins found dead along the Mekong River between 2006 and 2010 were analysed in different laboratories in Europe, Canada and Cambodia. The aim of the ... [more ▼]

For this report, samples of Irrawaddy River dolphins found dead along the Mekong River between 2006 and 2010 were analysed in different laboratories in Europe, Canada and Cambodia. The aim of the investigation was to gain additional knowledge on histological and toxicological findings as well as on the genetic variation of the Mekong population. The report summarizes those results in relation to previous investigations conducted in Cambodia and the United States of America. [less ▲]

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See detailPOTENTIAL EFFECTS OF BLOOD CONTAMINANTS ON IMMUNE RESPONSES IN HARBOUR SEALS (PHOCA VITULINA)
Dupont, Aurélie ULg; Weijs, Liesbeth; Siebert, Ursula et al

in Organohalogen Compounds (2011), 73

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See detailMethylmercury and selenium in vitro effects on harbor seal (Phoca vitulina) lymphocytes : a multidisciplinary approach
Dupont, Aurélie ULg; Bouquegneau, Jean-Marie ULg; Das, Krishna ULg et al

Poster (2010, May 26)

Methylmercury (MeHg) bioaccumulates along the food web, leading to the highest levels in tissues of predatory species. It constitutes the predominant form present in the blood of marine mammals. The blood ... [more ▼]

Methylmercury (MeHg) bioaccumulates along the food web, leading to the highest levels in tissues of predatory species. It constitutes the predominant form present in the blood of marine mammals. The blood cells, including the immune cells, are therefore exposed to the toxic properties of that chemical. Nevertheless, selenium (Se) is an essential element absorbed concomitantly to MeHg which seems to modulate this toxicity. The goal of this study is to evaluate the immunotoxicity of MeHg on the harbor seal (Phoca vitulina) T lymphocytes, highly important in the adaptive immune response, and to investigate the modulating effect of Se on that toxicity. In parallel, the concentrations of MeHg, total mercury (T-Hg) and Se are determined in free-ranging harbour seal blood in order to follow their contamination levels. The T lymphocytes were isolated from the whole blood, exposed to various MeHg and Se concentrations and the exposure effects were estimated by functional tests including the evaluation of viability, proliferation, metabolic activity, DNA and protein synthesis, and by morphological analysis by transmission electron microscopy. The mean T-Hg concentration was 172 ± 143 µg/l of whole blood. The T lymphocytes cultures in vitro displayed a decreasing number of viable cells with increasing concentrations of MeHg, and numerous ultrastructural defects. The cells exposed to MeHg notably displayed distortion of the plasmic membrane, nucleus fragmentations, chromatin compaction, swelling mitochondrias and cytoplasmic vacuolisations. Those results highlighted various immunotoxic effects of MeHg, both at the functional and ultrastructural levels. The antagonistic role of Se on MeHg immunotoxicity is discussed. [less ▲]

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See detailAspects of population biology: Epizootics in harbour seals (Phoca vitulina): clinical aspects
Siebert, Ursula; Gulland, Frances; Harder, Timm et al

in Desportes, G.; Bjorge, A.; Rosing-Avid, A. (Eds.) et al Harbour seals in the North Atlantic and the Baltic (2010)

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See detailMethylmercury in vitro exposure of harbor seal (Phoca vitulina) lymphocytes: a multidisciplinary approach
Dupont, Aurélie ULg; Siebert, Ursula; Rosenberger, Tanja et al

Poster (2009, June 08)

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