Ungulate management in European national parks: Why a more integrated European policy is needed.

1. Primary objectives of national parks usually include both, the protection of natural processes and species conservation. When these objectives conflict, as occurs because of the cascading effects of large mammals (i.e., ungulates and large carnivores) on lower trophic levels, park managers have to decide upon the appropriate management while considering various local circumstances. 2. To analyse if ungulate management strategies are in accordance with the objectives defined for protected areas, we assessed the current status of ungulate management across European national parks using the naturalness concept and identified the variables that influence the management. 3. We collected data on ungulate management from 209 European national parks in 29 countries by means of a large-scale questionnaire survey. Ungulate management in the parks was compared by creating two naturalness scores. The first score reflects ungulate and large carnivore species compositions, and the second evaluates human intervention on ungulate populations. We then tested whether the two naturalness score categories are influenced by the management objectives, park size, years since establishment, percentage of government-owned land, and human impact on the environment (human influence index) using two generalized additive mixed models. 4. In 67.9% of the national parks, wildlife is regulated by culling (40.2%) or hunting (10.5%) or both (17.2%). Artificial feeding occurred in 81.3% of the national parks and only 28.5% of the national parks had a non-intervention zone covering at least 75% of the area. Furthermore, ungulate management differed greatly among the different countries, likely because of differences in hunting traditions and cultural and political backgrounds. Ungulate management was also influenced by park size, human impact on the landscape, and national park objectives, but after removing these variables from the full model the reduced models only showed a small change in the deviance explained. In areas with higher anthropogenic pressure, wildlife diversity tended to be lower and a higher number of domesticated species tended to be present. Human intervention (culling and artificial feeding) was lower in smaller national parks and when park objectives followed those set by the International Union for the Conservation of Nature (IUCN). 5. Our study shows that many European national parks do not fulfil the aims of protected area management as set by IUCN guidelines. In contrast to the USA and Canada, Europe currently has no common ungulate management policy within national parks. This lack of a common policy together with differences in species composition, hunting traditions, and cultural or political context has led to differences in ungulate management among European countries. To fulfil the aims and objectives of national parks and to develop ungulate management strategies further, we highlight the importance of creating a more integrated European ungulate management policy to meet the aims of national parks.

Hormone, vitamin and contaminant status during the moulting/fasting period in ringed seals (Pusa [Phoca] hispida) from Svalbard.

This study investigates the potential effects of moulting, and the concomitant period of fasting undertaken by ringed seals, on hormone, vitamin and contaminant status in adult animals in a population from Svalbard, Norway, which has relatively low contaminant levels. Concentrations of circulating total and free thyroxine and triiodothyronine, circulating and hepatic vitamin A, hepatic persistent organic pollutants and their circulating hydroxyl metabolites were higher in moulting seals compared to pre-moulting seals. The opposite trend was observed for body condition, circulating calcitriol levels and hepatic mRNA expression of thyroid hormone receptor beta. No differences were observed for circulating or hepatic vitamin E levels or hepatic mRNA expressions for deioidinase 1 or 2, or retinoic acid receptor alpha between the two seal groups. The observed differences are likely the result of increased metabolic rates required during moulting to maintain thermal balance and replace the pelage, in combination with mobilization of lipid soluble compounds from blubber stores during the fasting period that is associated with moulting. The present study shows that contaminant levels and their relationships with physiological or endogenous variables can be highly confounded by moulting/fasting status. Thus, moulting status and body condition should be taken into consideration when using variables related to thyroid, calcium or vitamin A homeostasis as biomarkers for contaminant effects.

Bone-related effects of contaminants in seals may be associated with vitamin D and thyroid hormones.

The high levels of polychlorinated biphenyls (PCBs) and DDT in gray seal (Halichoerus grypus) and ringed seal (Phoca hispida botnica) in the Baltic Sea have been associated with pathological disruptions, including bone lesions and reproductive failures. The underlying environmental and toxicological mechanisms leading to these pathological changes are not yet fully understood. The present study investigated the relationship between the individual contaminant load and bone- and thyroid-related effects in adult gray seals (n=30) and ringed seals (n=46) in the highly contaminated Baltic Sea and in reference areas (Sable Island, Canada, and Svalbard, Norway). In the gray seals, multivariate and correlation analyses revealed a clear relationship between circulating 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D), calcium, phosphate, and thyroid hormone (TH) levels and hepatic PCB and DDT load, which suggests contaminant-mediated disruption of the bone and thyroid homeostasis. Contaminants may depress 1,25(OH)(2)D levels or lead to hyperthyroidism, which may cause bone resorption. In the ringed seals, associations between circulating 1,25(OH)(2)D, THs, and hepatic contaminants were less prominent. These results suggest that bone lesions observed in the Baltic gray seals may be associated with contaminant-mediated vitamin D and thyroid disruption.

Circumpolar pattern of mercury and cadmium in ringed seals.

Mercury (Hg) and cadmium (Cd) concentrations in ringed seal liver and kidney were compared from 11 locations across the Arctic, from Alaska, Canada, Greenland, Svalbard to the White Sea. Adult and sub adult seals were evaluated separately to account for age accumulations of Hg and Cd. Only recently (1995-2001) collected samples were included to minimize influence of changes over time. The Hg and Cd concentrations in ringed seal liver and kidney differed significantly among the studied locations. The Hg concentrations in liver of ringed seals was highest in the western Canadian Arctic locations, while Cd in liver was highest in the eastern Canadian and West Greenland locations. In general, Hg and Cd concentrations in liver and kidney were significantly higher in adult ringed seals than in sub adults and the circumpolar patterns were most pronounced in adult ringed seals. The Hg and Cd concentrations in kidney of ringed seals in general supported the geographical pattern found in livers although the coverage was more limited. The most likely explanation for the observed circumpolar pattern appears mainly to be related to natural geological differences in mineral (Hg and Cd) among regions.

Accumulation of dietary organochlorines and vitamins in Baltic seals.

We studied the accumulation of polychlorinated biphenyls (PCBs), 1,1,1.trichloro-2,2-bis[p-chlorophenyl]ethane (DDT) and its metabolites, and vitamins A and E in the top levels of the Baltic Sea food web. The investigation focused on the transfer of contaminants and vitamins to the ringed seal (Phoca hispida baltica) and the grey seal (Halichoerus grypus) from their main prey species. The trophic level of the seals was investigated using stable isotopes of nitrogen and the results indicated that both species of Baltic seal feed at approximately the same level. PCBs accumulated to a greater extent in the grey seal than in the ringed seal. Biomagnification factors for DDT compounds were similar for both species of Baltic seal (85-140). Differences in observed DDT levels were due to different prey selection by the two species, while differences in PCB levels indicated a species-specific metabolic system. There was a clearly greater accumulation of DDT compounds than of PCBs in both species of seal. The supply of dietary vitamin A was normally above the recommended level in all the seal populations studied. Low levels of hepatic vitamin A in the Baltic seals could therefore indicate the toxic effects of a high level of persistent organic pollutants on vitamin A dynamics, at least in the ringed seal. In the grey seals, low hepatic vitamin A levels could also be explained by lower levels of dietary vitamin A, compared to the reference grey seals, as it is not known if seals can store unlimited amounts of vitamin A. The greater uptake of vitamin E by Baltic seals, compared to seals in the reference areas, demonstrated by elevated levels of vitamin E in the blubber, could be a response to oxidative stress caused by the high contaminant load. These results further support our previous hypothesis that the toxic effects of environmental contaminants could be causing the observed divergence in vitamin levels between the Baltic seals and the reference seal populations.

Current levels of DDT, PCB and trace elements in the Baltic ringed seals (Phoca hispida baltica) and grey seals (Halichoerus grypus).

Residue levels of polychlorinated biphenyls (PCBs) and 1,1,1-trichloro-2,2-bis[p-chlorophenyl]ethane (DDT) were determined from liver samples of ringed seals (Phoca lispida) from the Baltic Sea and Svalbard, and of grey seals (Halichoerus grypus) from the Baltic Sea and Sable Island in Canada. Both Baltic seal populations showed clearly higher average sum PCB (SPCB) and sum DDT (SDDT) levels than the reference seal populations. Among the Baltic seals, SPCB levels were twice as high as SDDT levels, and both contaminants were higher in ringed seals than in grey seals. A difference in gender was observed only in the Sable Island grey seal population, in which males showed a higher level of contamination than females. A reduction of the SDDT levels, and to a lower extent of the SPCB levels can be observed in the Baltic seals since the peak contaminant levels during the 1970s. The decrease has been more rapid in the grey seals than in the ringed seals. The SPCB levels in the Baltic ringed seals are still high enough to cause a threat to their well being. Residue levels of the trace elements mercury, cadmium, lead and selenium were determined from liver, kidney and muscle samples of grey seals from the Baltic Sea and Sable Island. Only cadmium showed a geographic difference being higher in the seals from Sable Island than from the Baltic Sea. No clear reduction of the metal burden can be observed in the Baltic grey seal population since the 1970s.

Hepatic microsomal cytochrome P450 enzyme activity in relation to in vitro metabolism/inhibition of polychlorinated biphenyls and testosterone in Baltic grey seal (Halichoerus grypus).

Among other factors, cytochrome P450 (CYP) enzyme activity determines polychlorinated biphenyl (PCB) bioaccumulation, biotransformation, and toxicity in exposed species. We measured the oxidative metabolism in vitro of 12 PCB congeners, representing structural groups based on the number and position of the chlorine atoms, by the hepatic microsomes of one Baltic grey seal (Halichoerus grypus). Microsomal metabolism was observed for several PCBs with vicinal H atoms exclusively in the ortho and meta positions and without any ortho-Cl substituents (CB-15 [4,4'-Cl2] and CB-77 [3,3',4,4'-Cl4]), vicinal meta and para-H atoms (CB-52 [2,2',5,5'-Cl4], and -101 [2,2',4,5,5'-Cl5]) or with both characteristics in combination with either only one ortho-Cl (CB-26 [2,3',5-Cl3], CB-31 [2,4',5-Cl3]) or two ortho-Cl substituents (CB-44 [2,2',3,5'-Cl4]). To allocate PCB biotransformation to specific CYPs, the inhibitive effect of compounds with known CYP-specific inhibition properties was assessed on in vitro PCB metabolism and on regio- and stereospecific testosterone hydroxylase activities. Metabolic inhibition was considered relevant at concentrations < or = 1.0 microM because these inhibitors became decreasingly selective at higher concentrations. At < 1.0 microM, ellipticine (CYPIAI/2 inhibitor) selectively inhibited CB-15, -26, -31, and -77 metabolism, with no significant inhibition of CB-44, -52, and -101 metabolism. Inhibition of CB-52 and -101 metabolism by chloramphenicol (CYP2B inhibitor) started at 1.0 microM and maximized at about 100% at 10 microM. Ketoconazole (CYP3A inhibitor) appeared to selectively inhibit CB-26, -31, and -44 metabolism relative to CB-15, -77, and -52 at concentrations < or = 1.0 microM. Major testosterone metabolites formed in vitro were 2beta-(CYP3A), 6beta- (CYP3A, CYPIA), and 16beta- (CYP2B) hydroxytestosterone and androstenedione (CYP2B, CYP2C11). The CYP forms indicated are associated with the specific metabolism of testosterone in laboratory animals. Inhibition of 2beta- and 6beta-hydroxytestosterone formation at ellipticine and ketoconazole concentrations < or = 1.0 microM suggested that both inhibitors were good substrates of CYP3A-like enzymes in grey seal. Chloramphenicol (model for CYP2B) is apparently not a good inhibitor of CYPI A and CYP3A activities in grey seal because the chemical did not inhibit any metabolic route of testosterone at concentrations from 0.1 to 10 microM. Our findings demonstrated that at least CYP1A- and CYP3A-like enzymes in the liver of grey seals are capable of metabolizing PCBs with ortho-meta and/or meta-para vicinal hydrogens. A CYP2B form might also be involved, but this could not be proven by the results of our experiments. Defining the profiles of CYP enzymes that are responsible for PCB biotransformation is necessary to fully understand the bioaccumulation, toxicokinetics, and risk of PCB exposure in seals and other free-ranging marine mammals.

Contaminant exposure and effects in Baltic ringed and grey seals as assessed by biomarkers.

The Baltic Sea ecosystem has suffered from a heavy pollutant load for more than three decades. Persistent organic pollutants (POPs) and heavy metals have been of most concern due to their persistence and toxic properties. Ringed seals (Phoca hispida baltica) and grey seals (Halichoerus grypus) living in the Baltic Sea have been suffering from pathological impairments, including reproductive disturbances, which have resulted in a depressed reproductive capacity. We investigated several biochemical parameters as potential biomarkers for exposure to and effects of the contaminant load in the Baltic seals. Seals from less polluted areas were used as reference material in terms of the pollution load. In both Baltic seal populations, the levels of some biochemical parameters diverged from those in the reference seals, and some of these showed a clear correlation with the individual contaminant load. Of the potential bioindicators, we propose cytochrome P4501A activity and vitamin E levels, in blubber or plasma, as exposure biomarkers for polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT) in both species. The arylhydrocarbon receptor-mediated chemical-activated luciferase gene expression (CALUX) response reflects the whole PCB and DDT burden in ringed seals. Retinyl palmitate (vitamin A) levels showed a negative correlation with the individual POP load, and is proposed as potential effect biomarkers for the depletion of the vitamin A stores. As the nutritional levels of both vitamin A and E have an impact on the vitamin levels in the seals, more information on the dietary vitamin levels is needed before any conclusions can be drawn. As the relationship between biochemical parameters and contaminants varied between the two species, species-specific characteristics has to be considered when monitoring the health status and possible toxic effects of the contaminant load in ringed and grey seals.

Antioxidative defense and oxidative stress in ringed seals (Pusa hispida) from differently polluted areas.

High contaminant levels detected in Baltic seals have been associated with various health effects. In this study several parameters related to antioxidative defense and oxidative stress (concentrations of reduced and oxidised glutathione, lipid hydroperoxide and vitamin E, activities of glutathione reductase, peroxidase and S-transferase, glucose-6-phosphate dehydrogenase, catalase, and superoxidedismutase) were measured in the livers of ringed seals from the Baltic Sea and from a less contaminated reference area, Svalbard, Norway. Seals were caught during two different time periods 1996-1997 and 2002-2007, which represent different levels of contamination. No signs of oxidative damage were found in the Baltic seals. However, glutathione metabolism was enhanced in the ringed seals from the Baltic Sea compared to the seals from Svalbard. The adaptation to dive where repetitive ischemia/reperfusion occurs naturally may contribute to the resistance of oxidative stress and to the capacity to increase enzymatic antioxidant defense in phocid seals. This could explain the similarities in oxidative stress levels despite the differences in antioxidant responses between the ringed seals from the Baltic Sea and Svalbard.

Comparative endocrine disruptive effects of contaminants in ringed seals (Phoca hispida) from Svalbard and the Baltic Sea.

We investigated variables related to thyroid, vitamin A and calcitriol homeostasis, immune function and tumour development in ringed seals (Phoca hispida) from the polluted Baltic Sea and a less polluted reference location at Svalbard, Norway. We also examined the relationships between the biological variables and the concentrations of persistent organic pollutants (POPs) and their hydroxylated (OH) metabolites. Our data show higher plasma concentrations of free triiodothyronine (T3), and ratios of free and total T3 in Baltic seals as compared to Svalbard seals. Baltic seals had also higher hepatic mRNA expressions of deiodinase-I, thyroid hormone receptor beta, retinoic acid receptor alpha, growth hormone receptor and interleukin-1beta compared to Svalbard seals. Levels of plasma retinol were lower in the Baltic seals as compared to Svalbard seals. No geographical difference was observed for other thyroid hormone levels and hepatic retinoid levels. Ratios of free and total T3 were positively correlated to OH-POPs in plasma. The results of the present study suggest that endocrine homeostasis may be affected by contaminant and metabolite exposure in the Baltic ringed seals with respect to circulating hormones and retinol and hepatic mRNA expressions. In addition, OH-POPs may putatively produce the disruption of thyroid hormone transport in plasma.