Thea Ø. Bechshøft

Cortisol levels in hair of East Greenland polar bears.

To demonstrate the ability to assess long-term hypothalamic-pituitary-adrenocortical (HPA) axis activity in polar bears (Ursus maritimus), a pilot study was conducted in which cortisol concentrations was analyzed in hair from 7 female (3-19 years) and 10 male (6-19 years) East Greenland polar bears sampled in 1994-2006. The hair was chosen as matrix as it is non-invasive, seasonally harmonized, and has been validated as an index of long-term changes in cortisol levels. The samples were categorized according to contamination: eight were clean (2 females, 6 males), 5 had been contaminated with bear blood (2 F, 3 M), and 4 with bear fat (3 F, 1 M). There was no significant difference in cortisol concentration between the three categories after external contamination was removed. However, contaminated hair samples should be cleaned before cortisol determination. Average hair cortisol concentration was 8.90 pg/mg (range: 5.5 to 16.4 pg/mg). There was no significant correlation between cortisol concentration and age (p=0.81) or sampling year (p=0.11). However, females had higher mean cortisol concentration than males (females mean: 11.0 pg/mg, males: 7.3 pg/mg; p=0.01). The study showed that polar bear hair contains measurable amounts of cortisol and that cortisol in hair may be used in studies of long-term stress in polar bears.

Three decades (1983-2010) of contaminant trends in East Greenland polar bears (Ursus maritimus). Part 2: Brominated flame retardants

Brominated flame retardants were determined in adipose tissues from 294 polar bears (Ursus maritimus) sampled in East Greenland in 23 of the 28years between 1983 and 2010. Significant linear increases were found for sum polybrominated diphenyl ether (ΣPBDE), BDE100, BDE153, and hexabromocyclododecane (HBCD). Average increases of 5.0% per year (range: 2.9-7.6%/year) were found for the subadult polar bears. BDE47 and BDE99 concentrations did not show a significant linear trend over time, but rather a significant non-linear trend peaking between 2000 and 2004. The average ΣPBDE concentrations increased 2.3 fold from 25.0ng/g lw (95% C.I.: 15.3-34.7ng/g lw) in 1983-1986 to 58.5ng/g lw (95% C.I.: 43.6-73.4ng/g lw) in 2006-2010. Similar but fewer statistically significant trends were found for adult females and adult males likely due to smaller sample size and years. Analyses of δ(15)N and δ(13)C stable isotopes in hair revealed no clear linear temporal trends in trophic level or carbon source, respectively, and non-linear trends differed among sex and age groups. These increasing concentrations of organobromine contaminants contribute to complex organohalogen mixture, already causing health effects to the East Greenland polar bears.

Two decades of biomonitoring polar bear health in Greenland: a review

SummaryWe present an overview of studies of anthropogenic pollutants in East Greenland polar bears over the period of 1999-2011. East Greenland polar bears are among the most polluted species, not just in the Arctic but globally, and represent an excellent biomonitoring species for levels and effects of global pollution in an apex predator. Therefore, an international multidisciplinary team joined to monitor and assess the patterns and concentrations of contaminants and their potential negative impact on polar bears. The review showed that East Greenland polar bears are exposed to a mix of chlorinated, brominated and fluorinated organic compounds as well as mercury which are all known to have endocrine, immune and organ-system toxic properties. For example, the concentrations of PCBs (polychlorinated biphenyls) in blubber ranged approximately 800-21,000 ng/g lw while mercury concentrations in liver and kidney ranged 0.1-50 μg/g ww. Regarding health endpoints, bone density seemed to decrease as a function of time and OHC (organohalogen compound) concentrations and further T-score for adult males indicated risk for osteoporosis. .The size of sexual organs decreased with increasing OHC concentrations. In the lower brain stem, mercury-associated decreases in NMDA-receptor levels and DNA-methylation was found The present review indicated that age was one of the major drivers for liver and renal lesions, although contaminants and infectious diseases may also play a role. Lesions in thyroid glands were most likely a result of infectious and genetic factors and probably, together with endocrine disrupting chemical (EDCs), the reason for disturbances/fluctuations in blood plasma thyroid hormone concentrations. Except for bone density reductions and neurological measures, all findings were supported by case-control studies of Greenland sledge dogs exposed long-term orally to similar combinations of contaminant concentrations. The studies of sledge dogs also indicated that the mixture of contaminants and fatty acids in the blubber of prey similar to that of polar bears induces cellular as well as humoral immune toxic changes. These controlled studies using model species for polar bears indicate that the correlative findings between health endpoint and contaminants in polar bears could be a cause-and-effect relationship. Physiologically based pharmacokinetic (PBPK) modelling showed that the risk quotients were ≥1 for ΣPCB, dieldrin and PFOS, which indicate an increased risk of prenatally reproductive pathology. In conclusion polar bears are susceptible to long-range transported chemicals that may have various adverse effects on multiple organ systems such as the reproductive and immune system.

Anthropogenic flank attack on polar bears: interacting consequences of climate warming and pollutant exposure

Polar bears (Ursus maritimus) are subjected to several anthropogenic threats, climate warming and exposure to pollutants being two of these. For polar bears, one of the main effects of climate warming is limited access to prey, due to loss of their sea ice habitat. This will result in prolonged fasting periods and emaciation and condition related negative effects on survival and reproduction success. Prolonged fasting will result in increases of the tissue concentrations of persistent organic pollutants (POPs) in polar bears, and thus increase the probability for POP levels to exceed threshold levels for effects on health, and thus on reproductive success and survival. There are clear potentials for interactions between impacts of climate warming and impacts of pollutant exposure on polar bears. It is likely that that fasting-induced increases of POPs will add to mortality rates and decrease reproductive success beyond effects caused by loss of habitat alone. However, there is a lack of studies that have addressed this. Thus, there is a need to focus on population effects of POP exposure in polar bears, and to consider such effects in relation to the effects of climate induced habitat loss.

Fluctuating Asymmetry in Metric Traits; a Practical Example of Calculating Asymmetry, Measurement Error, and Repeatability

We present an example of background statistics in studies of fluctuating asymmetry (FA); calculations of asymmetry, measurement error (ME), and repeatability. Nine bilateral metric traits in skulls and lower jaws of 691 East Greenland and Svalbard polar bears (Ursus maritimus) were measured twice, and examined for asymmetry. The skulls were collected in the period 1892–2004. In this study, 2.0% of the FA data were identified and treated as outliers, which is less than in comparable studies. FA for each trait amounted to 0.1%–3% of the average size of the corresponding trait. The magnitude of FA generally increased with trait size. For every trait measured, ME was found to be smaller than FA. The repeatability of the traits was inversely proportional to ME. Five of the nine traits had a repeatability of 90% or more, which is similar to what has been reported in other studies.

Temporal and Spatial Variation in Metric Asymmetry in Skulls of Polar Bears (Ursus maritimus) from East Greenland and Svalbard

Fluctuating asymmetry (FA) as a measure of environmental stress was studied in polar bear (Ursus maritimus) skulls from East Greenland (n = 300, collected 1892–2004) and Svalbard (n = 391, collected 1950–2004). Nine metric traits in skull and lower jaw were measured. FA levels of each trait were compared between sex/age groups (subadults, adult females, adult males), periods (≤ 1960, ≥ 1960), and localities (East Greenland, Svalbard). The period ≤ 1960 was chosen to represent a period prior to the appearance of organohalogen pollution in the Arctic. Results indicated that Svalbard bears generally had a higher level of FA than those of East Greenland. Overall, no substantial evidence of a linkage between FA and organohalogens was found. Instead, indications were of subpopulations with declining levels of FA over time, suggesting the existence of positive population level effects powerful enough to overrule the influence of stress caused by global warming, pollution, and overharvesting.

Developing a new research tool for use in free-ranging cetaceans: recovering cortisol from harbour porpoise skin

In this paper we present a new analytical method that allows the extraction of the stress hormone cortisol from naturally sloughed cetacean skin cells. Skin cortisol is unaffected by short-term cortisol fluctuations and could thus be used to assess chronic stress levels in cetaceans.

Temporal monitoring of liver and kidney lesions in contaminated East Greenland polar bears (Ursus maritimus) during 1999–2010

Organohalogen contaminants bioaccumulate to high concentrations in tissues of polar bears (Ursus maritimus). The exposure levels are in the order to be toxic to inner organs like liver and kidney. We therefore investigated the temporal development of lesions in liver (n=115) and kidney (n=122) samples from East Greenland polar bears taken over the 12 year period from 1999 to 2010. Seven liver and seven kidney lesions were observed of which six were age-related. Controlling for this, the analyses showed that hepatic steatosis and renal cell infiltrations, glomerular sclerosis and tubular hyperplasia decreased over the investigated time period (all p<0.05). Similarly, hypertrophy of hepatic Ito cells, renal glomerular capillary wall thickening and interstitial fibrosis increased over the study period (all p<0.05). Regarding contaminant, concentrations of polybrominated diphenyl ethers in adipose tissue were negatively associated with hepatic mononuclear cell infiltrations (p=0.05) and a similar trend was found for Hg (p=0.09). Hexachlorobenzene was positively associated with portal cell infiltrations and hepatic lipid granulomas, while polychlorinated biphenyls were negatively associated with the prevalence of steatosis (both p<0.05) and a similar trend was found for hexachlorocyclohexanes (p=0.08). Mercury was positively correlated with the frequencies of hypertrophic Ito cells (p=0.002) and a similar trend was found for perfluorooctanesulfonic acid (p=0.07). In renal tissue, hexachlorocyclohexanes were positively associated with medullar hyaline casts (p=0.03) and a similar trend was found for cell infiltrations (p=0.08). There was a trend of trans-nonachlor being positively associated with glomerular sclerosis (p=0.06) while dichlorodiphenyltrichloroethanes were negatively associated with tubular hyperplasia (p=0.02). These results suggest that specific liver and renal lesions have decreased or increased over time and that long-range transported organohalogen contaminants and mercury may be among the co-factors responsible for these observations. These relationships are important to take into account when biomonitoring health and pollution in wildlife species such as polar bears.

East Greenland and Barents Sea polar bears (Ursus maritimus): adaptive variation between two populations using skull morphometrics as an indicator of environmental and genetic differences

A morphometric study was conducted on four skull traits of 37 male and 18 female adult East Greenland polar bears (Ursus maritimus) collected 1892-1968, and on 54 male and 44 female adult Barents Sea polar bears collected 1950-1969. The aim was to compare differences in size and shape of the bear skulls using a multivariate approach, characterizing the variation between the two populations using morphometric traits as an indicator of environmental and genetic differences. Mixture analysis testing for geographic differentiation within each population revealed three clusters for Barents Sea males and three clusters for Barents Sea females. East Greenland consisted of one female and one male cluster. A principal component analysis (PCA) conducted on the clusters defined by the mixture analysis, showed that East Greenland and Barents Sea polar bear populations overlapped to a large degree, especially with regards to females. Multivariate analyses of variance (MANOVA) showed no significant differences in morphometric means between the two populations, but differences were detected between clusters from each respective geographic locality. To estimate the importance of genetics and environment in the morphometric differences between the bears, a PCA was performed on the covariance matrix derived from the skull measurements. Skull trait size (PC1) explained approx. 80% of the morphometric variation, whereas shape (PC2) defined approx. 15%, indicating some genetic differentiation. Hence, both environmental and genetic factors seem to have contributed to the observed skull differences between the two populations. Overall, results indicate that many Barents Sea polar bears are morphometrically similar to the East Greenland ones, suggesting an exchange of individuals between the two populations. Furthermore, a subpopulation structure in the Barents Sea population was also indicated from the present analyses, which should be considered with regards to future management decisions.

Vitamins A and E in liver, kidney, and whole blood of East Greenland polar bears sampled 1994–2008: reference values and temporal trends

Vitamins A (retinol) and E (α-tocopherol) are dietary vitamins, essential for, e.g., growth and development, reproduction, and immune function. Persistent organic pollutants (POPs) have been found to be related to vitamin A and E metabolism. However, few investigations have been published on this health issue in polar bears (Ursus maritimus). The aim of this study was thus to provide reference values for concentrations of vitamin A in liver, kidney cortex, and whole blood and vitamin E in kidney cortex and whole blood from 166 East Greenland polar bears, as well as to assess the relationship between POPs and vitamin concentrations. In addition, vitamin concentrations were analyzed for temporal trends (1994–2008). Results showed vitamin A in liver to be higher in adult bears and the concentrations of vitamin E in kidney and blood to likewise be generally higher in adult bears. In addition, all analyzed contaminant groups were correlated with at least one of the vitamin parameters, predominantly in a negative way. Finally, vitamin A liver concentrations as well as concentration of vitamin E in kidney and blood showed a temporal increase. Together, these results add to the weight of evidence that POPs could be disrupting polar bear vitamin status. However, while the observed temporal increases in vitamin concentrations were likely POP related, the question remains as to whether they stem from influence of contaminants only or also, e.g., changes in prey species. Further studies are needed to tease apart the causes underlying these changes in vitamin concentrations.