Alternative and legacy flame retardants in marine mammals from three northern ocean regions.

Flame retardants are globally distributed contaminants that have been linked to negative health effects in humans and wildlife. As top predators, marine mammals bioaccumulate flame retardants and other contaminants in their tissues which is one of many human-imposed factors threatening population health. While some flame retardants, such as the polybrominated diphenyl ethers (PBDE), have been banned because of known toxicity and environmental persistence, limited data exist on the presence and distribution of current-use alternative flame retardants in marine mammals from many industrialized and remote regions of the world. Therefore, this study measured 44 legacy and alternative flame retardants in nine marine mammal species from three ocean regions: the Northwest Atlantic, the Arctic, and the Baltic allowing for regional, species, age, body condition, temporal, and tissue comparisons to help understand global patterns. PBDE concentrations were 100-1000 times higher than the alternative brominated flame retardants (altBFRs) and Dechloranes. 2,2',4,5,5'-pentabromobiphenyl (BB-101) and hexabromobenzene (HBBZ) were the predominant altBFRs, while Dechlorane-602 was the predominant Dechlorane. This manuscript also reports only the second detection of hexachlorocyclopentadienyl-dibromocyclooctane (HCDBCO) in marine mammals. The NW Atlantic had the highest PBDE concentrations followed by the Baltic and Arctic which reflects greater historical use of PBDEs in North America compared to Europe and greater industrialization of North America and Baltic countries compared to the Arctic. Regional patterns for other compounds were more complicated, and there were significant interactions among species, regions, body condition and age class. Lipid-normalized PBDE concentrations in harbor seal liver and blubber were similar, but HBBZ and many Dechloranes had higher concentrations in liver, indicating factors other than lipid dynamics affect the distribution of these compounds. The health implications of contamination by this mixture of compounds are of concern and require further research.

Exposure to legacy and alternative flame retardants in two harbor seal populations and the association with blubber fatty acid profiles.

Flame retardants (FRs) are ubiquitously present in various environmental compartments due to widespread application. However, there have been few reports on the alternative FRs in harbor seals, and their relationship with fatty acid (FA) profiles have largely been overlooked. Here, we investigated the levels of legacy and alternative FRs and FA profiles in the blubber of harbor seals from the coasts of South Sweden (2009-2016) and Northeastern US (NE US) (1999-2010). We observed different proportions of mono- and poly-unsaturated FAs (MUFAs and PUFAs) between the two populations, which may reflect variations in the diet. Significantly higher concentrations of ΣPBDE were also observed in harbor seals from US compared to those from Sweden, both dominated by BDE 47. By comparison, the levels of alternative FRs, noticeably HBBZ and PBEB were much lower compared to those of PBDEs. Moreover, we found a positive correlation between BDE 99 and Σn-6/Σn-3 PUFA in harbor seals from Sweden. In addition, BDE 153 and BDE 154 were positively correlated with ΣUFA/ΣSFA in seals from Sweden and US, respectively. Our results imply the influence of diet in FA profiles and FR concentrations in top predators, as well as the importance of blubber FA characteristics in indicating FR exposure. Further investigations are required to assess the risk of exposure in these harbor seals, as well as to elucidate the underlying mechanisms associating FA profiles with FR exposure.

Spatiotemporal Trends of Legacy and Alternative Flame Retardants in Harbor Seals from the Coasts of California, the Gulf of Maine, and Sweden.

Temporal trends of polybrominated diphenyl ethers (PBDEs) have been extensively studied in various environmental compartments globally. However, despite the increasing use of alternative flame retardants following PBDE bans, the spatiotemporal trends of these replacements have rarely been studied, and the available results are often inconsistent. In the present study, we retrospectively investigated the spatiotemporal trends of PBDEs and a suite of alternative brominated FRs (aBFRs) and chlorinated FRs (i.e., dechloranes or DECs) in three harbor seal (Phoca vitulina) populations from the coasts of California, the Gulf of Maine, and southern Sweden during 1999-2016. We observed significantly decreasing trends of ΣPBDEs in all the three populations at an annual rate of 9-11%, which were predominantly driven by the declining concentrations of tetra- and penta-BDEs. The levels of ΣaBFRs decreased significantly in seals from California (mainly 1,3,5-tribromobenzene) and Sweden (mainly hexabromobenzene), while no trend was observed for those from Maine. By contrast, DECs (dominated by DEC 602) did not decrease significantly in any population. Compared with the consistent PBDE congener profiles across regions, aBFRs and DECs exhibited varying compositional profiles between regions, likely indicating region-specific sources of these alternative FR mixtures. Spatial analysis also revealed regional differences in the concentrations of PBDEs, aBFRs, and DECs in harbor seals. Our reconstructed spatiotemporal trends suggest the effective regulation of commercial penta-BDE mix in these regions and warrant further monitoring of the higher brominated BDEs and alternative FRs.

Hepatic Fatty Acid Profiles Associated with Exposure to Emerging and Legacy Halogenated Contaminants in Two Harbor Seal Populations across the North Atlantic.

Fatty acids (FAs) have been extensively used as indicators of foraging ecology in marine mammals, yet their association with exposure to contaminants has rarely been investigated. The present study provided the first characterization of the relationship between hepatic FA profiles and exposure to a suite of contaminants in a sentinel species─the harbor seal (Phoca vitulina)─from the Gulf of Maine and the south coast of Sweden. FA profiles differed in the two seal populations, and the levels of legacy and alternative brominated flame retardants and polyhalogenated carbazoles were significantly elevated in Maine seals. Correlations between individual FAs and multiple flame retardants (FRs) and poly- and perfluoroalkyl substances (PFASs) were found in seals from both populations. Moreover, several FR and PFAS chemicals were significantly associated with the estimated desaturating enzyme activity inferred from the FA profiles. The ratios of poly to monounsaturated FAs (∑PUFAs/∑MUFAs) and those of unsaturated to saturated FAs (∑UFAs/∑SFAs) were significantly associated with HBBZ, PFHxS, or BDE 47 in seals from Maine and Sweden, whereas ∑n - 6/∑n - 3 PUFAs was significantly associated with BDE 154 and 36-CCZ in Swedish and Maine seals, respectively. Our results suggest the lipid metabolism-disrupting potential of these contaminants in marine mammals and warrant continuous biomonitoring and risk assessment, considering the critical role of PUFAs in vital biological processes.

Life cycle bioenergetics of the gray seal (Halichoerus grypus) in the Baltic Sea: Population response to environmental stress.

Wildlife population dynamics are shaped by multiple natural and anthropogenic factors, including predation, competition, stressful life history events, and external environmental stressors such as diseases and pollution. Marine mammals such as gray seals rely on extensive blubber layers for insulation and energy storage, making this tissue critical for survival and reproduction. This lipid rich blubber layer also accumulates hazardous fat soluble pollutants, such as polychlorinated biphenyls (PCBs), that can directly impact adipose function or be mobilized during periods of negative energy balance or transferred to offspring to exert further impacts on target tissues or vulnerable life stages. To predict how marine mammals will respond to ecological and anthropogenic stressors, it is necessary to use process-based modelling approaches that integrate environmental inputs, full species life history, and stressor impacts with individual dynamics of energy intake, storage, and utilization. The purpose of this study was to develop a full lifecycle dynamic energy budget and individual based model (DEB-IBM) that captured Baltic gray seal physiology and life history, and showcase potential applications of the model to predict population responses to select stressors known to threaten gray seals and other marine mammals around the world. We explore variations of three ecologically important stressors using phenomenological simulations: food limitation, endocrine disrupting chemicals that reduce fertility, and infectious disease. Using our calibrated DEB-IBM for Baltic gray seals, we found that continuous incremental food limitation can be more detrimental to population size than short random events of starvation, and further, that the effect of endocrine disruptors on population growth and structure is delayed due to bioaccumulation, and that communicable diseases significantly decrease population growth even when spillover events are relatively less frequent. One important finding is the delayed effect on population growth rate from some stressors, several years after the exposure period, resulting from a decline in somatic growth, increased age at maturation and decreased fecundity. Such delayed responses are ignored in current models of population viability and can be important in the correct assessment of population extinction risks. The model presented here provides a test bed on which effects of new hazardous substances and different scenarios of future environmental change affecting food availability and/or seal energetic demands can be investigated. Thus, the framework provides a tool for better understanding how diverse environmental stressors affect marine mammal populations and can be used to guide scientifically based management.

Health effects from contaminant exposure in Baltic Sea birds and marine mammals: A review.

Here we review contaminant exposure and related health effects in six selected Baltic key species. Sentinel species included are common eider, white-tailed eagle, harbour porpoise, harbour seal, ringed seal and grey seal. The review represents the first attempt of summarizing available information and baseline data for these biomonitoring key species exposed to industrial hazardous substances focusing on anthropogenic persistent organic pollutants (POPs). There was only limited information available for white-tailed eagles and common eider while extensive information exist on POP exposure and health effects in the four marine mammal species. Here we report organ-tissue endpoints (pathologies) and multiple biomarkers used to evaluate health and exposure of key species to POPs, respectively, over the past several decades during which episodes of significant population declines have been reported. Our review shows that POP exposure affects the reproductive system and survival through immune suppression and endocrine disruption, which have led to population-level effects on seals and white-tailed eagles in the Baltic. It is notable that many legacy contaminants, which have been banned for decades, still appear to affect Baltic wildlife. With respect to common eiders, changes in food composition, quality and contaminant exposure seem to have population effects which need to be investigated further, especially during the incubation period where the birds fast. Since new industrial contaminants continuously leak into the environment, we recommend continued monitoring of them in sentinel species in the Baltic, identifying possible effects linked to climate change, and modelling of population level effects of contaminants and climate change.

Emergence of the Zoonotic Biliary Trematode Pseudamphistomum truncatum in Grey Seals (Halichoerus grypus) in the Baltic Sea.

The biliary trematode Pseudamphistomum truncatum parasitizes a wide range of fish-eating mammals, including humans. Here we report the emergence of this parasite in grey seals (Halichoerus grypus) in the Baltic Sea. One hundred eighty-three of 1 554 grey seals (11.9%) examined from 2002-2013 had detectable hepatobiliary trematode infection. Parasite identification was confirmed as P. truncatum by sequencing the ITS2 region of a pool of five to 10 trematodes from each of ten seals collected off the coast of seven different Swedish counties. The proportion of seals parasitized by P. truncatum increased significantly over time and with increasing age of seals. Males were 3.1 times more likely to be parasitized than females and animals killed in fishery interactions were less likely to be parasitized than animals found dead or hunted. There was no significant difference in parasitism of seals examined from the Gulf of Bothnia versus those examined from the Baltic Proper. Although the majority of infections were mild, P. truncatum can cause severe hepatobiliary disease and resulted in liver failure in at least one seal. Because cyprinid fish are the second intermediate host for opisthorchiid trematodes, diets of grey seals from the Baltic Sea were analysed regarding presence of cyprinids. The proportion of gastrointestinal tracts containing cyprinid remains was ten times higher in seals examined from 2008 to 2013 (12.2%) than those examined from 2002 to 2007 (1.2%) and coincided with a general increase of trematode parasitism in the host population. The emergence and relatively common occurrence of P. truncatum in grey seals signals the presence of this parasite in the Baltic Sea ecosystem and demonstrates how aquatic mammals can serve as excellent sentinels of marine ecosystem change. Investigation of drivers behind P. truncatum emergence and infection risk for other mammals, including humans, is highly warranted.

Influence of age, season, body condition and geographical area on concentrations of chlorinated and brominated contaminants in wild mink (Neovison vison) in Sweden.

The wild mink has gained acceptance as a sentinel species in environmental monitoring. However, only limited data are available in the literature on factors driving variability in concentrations of organic pollutants in this species. This study characterizes the differences in contaminant concentrations in subcutaneous fat of male mink from four different areas in Sweden and demonstrates how age, season and body condition influence concentrations of polychlorinated biphenyl (PCB) congeners, polybrominated diphenyl ether (PBDE) congeners (including methoxylated forms, MeO-PBDEs), as well as the pesticides dichlorodiphenyldichloroethylene (DDE), chlordane and hexachlorobenzene (HCB). The data were statistically treated using multiple regression and principal component analysis. The ∑PCB concentration and concentrations of PCB congeners 138, 156, 157, 180, 170/190, 189, 194, 206, 209 as well as PBDE 153/154 varied with age. Season had an influence on ∑PCB, PBDE 47 and PBDE 153/154 concentrations, as well as concentrations of most PCB congeners, with the exception of PCB 101, 110, 141 and 182/187. Lean mink had higher concentrations of most PCBs and PBDEs than mink with larger fat depots. The analyzed pesticides (DDE, oxychlordane, HCB) showed no systematic variation with season, age or body condition. The concentrations of MeO-PBDEs were generally low and 6MeO-PBDE 47 was the most commonly detected MeO-PBDE in mink from marine, brackish and freshwater areas. The results indicate that age, season and body condition are factors that may influence the concentrations of PCBs and PBDEs, and it is thus recommended to take these factors into account when analyzing mink exposure data.

Improved reproductive success in otters (Lutra lutra), grey seals (Halichoerus grypus) and sea eagles (Haliaeetus albicilla) from Sweden in relation to concentrations of organochlorine contaminants.

We studied indices of reproductive outcome in three aquatic species in relation to organochlorine concentrations during four decades. In female otters, the frequency of signs of reproduction increased after 1990. In grey seals, pregnancy rate increased 1990-2010 and uterine obstructions ceased after 1993. The frequency of uterine tumours was highest 1980-2000. The number of sea eagle nestlings per checked nest increased 1985-2000, while the frequency of desiccated eggs decreased. Organochlorine concentrations decreased at annual rates between 3.5 and 10.2%. The estimated mean concentration (mg/kg lw) for total-PCB decreased from 70 to 8 (otters), from 110 to 15 (seals) and from 955 to 275 (eagles). The corresponding concentrations for ΣDDT decreased from 3.4 to 0.2 (otters), from 192 to 2.8 (seals) and from 865 to 65 (eagles). This study adds evidence to support the hypothesis that PCBs and DDTs have had strong negative effects on the reproduction and population levels of these species.

Reprint of: Temporal trends of polyfluoroalkyl compounds (PFCs) in liver tissue of grey seals (Halichoerus grypus) from the Baltic Sea, 1974-2008.

Temporal trends of polyfluoroalkyl compounds (PFCs) were examined in grey seal (Halichoerus grypus) liver from the Baltic Sea over a period of 35 years (1974-2008). In total, 17 of 43 PFCs were found, including the perfluoroalkyl sulfonates (C(4)-C(10) PFSAs), perfluorooctanesulfinate (PFOSi), long chain perfluoroalkyl carboxylates (C(7)-C(14) PFCAs), and perfluoroalkyl sulfonamides (i.e., perfluorooctane sulfonamide (FOSA) and N-ethyl perfluorooctane sulfonamide (EtFOSA)), whereas saturated and unsaturated fluorotelomer carboxylates, shorter chain PFCAs and perfluoroalkyl phosphonic acids were not detected. Perfluorooctane sulfonate (PFOS) was the predominant compound (9.57-1444 ng g(-1) wet weight (ww)), followed by perfluorononanoate (PFNA, 0.47-109 ng g(-1) ww). C(6)-C(8) PFSAs, PFOSi and C(7)-C(13) PFCAs showed statistically significant increasing concentrations between 1974 and 1997, with a peak in 1997 and then decreased or levelled off (except for C(12) and C(13) PFCAs). FOSA had a different temporal trend with a maximum in 1989 followed by significant decreasing concentrations until 2008. Toxicological implications for grey seals are limited, but the maximal PFOS concentration found in this study was about 40 times lower than the predicted lowest observed effect concentrations (LOEC). The statistically significant decreasing concentrations or levelling off for several PFCs in the relative closed marine ecosystem of the Baltic Sea indicate a rapidly responding to reduced emissions to the marine environment. However, the high concentrations of PFOS and continuing increasing concentrations of the longer chain PFCAs (C(12)-C(14)) shows that further work on the reduction of environmental emissions of PFCs are necessary.

Temporal trends of polyfluoroalkyl compounds (PFCs) in liver tissue of grey seals (Halichoerus grypus) from the Baltic Sea, 1974-2008.

Temporal trends of polyfluoroalkyl compounds (PFCs) were examined in grey seal (Halichoerus grypus) liver from the Baltic Sea over a period of 35 years (1974-2008). In total, 17 of 43 PFCs were found, including the perfluoroalkyl sulfonates (C(4)-C(10) PFSAs), perfluorooctanesulfinate (PFOSi), long chain perfluoroalkyl carboxylates (C(7)-C(14) PFCAs), and perfluoroalkyl sulfonamides (i.e., perfluorooctane sulfonamide (FOSA) and N-ethyl perfluorooctane sulfonamide (EtFOSA)), whereas saturated and unsaturated fluorotelomer carboxylates, shorter chain PFCAs and perfluoroalkyl phosphonic acids were not detected. Perfluorooctane sulfonate (PFOS) was the predominant compound (9.57-1,444 ng g(-1) wet weight (ww)), followed by perfluorononanoate (PFNA, 0.47-109 ng g(-1) ww). C(6)-C(8) PFSAs, PFOSi and C(7)-C(13) PFCAs showed statistically significant increasing concentrations between 1974 and 1997, with a peak in 1997 and then decreased or levelled off (except for C(12) and C(13) PFCAs). FOSA had a different temporal trend with a maximum in 1989 followed by significant decreasing concentrations until 2008. Toxicological implications for grey seals are limited, but the maximal PFOS concentration found in this study was about 40 times lower than the predicted lowest observed effect concentrations (LOEC). The statistically significant decreasing concentrations or levelling off for several PFCs in the relative closed marine ecosystem of the Baltic Sea indicate a rapidly responding to reduced emissions to the marine environment. However, the high concentrations of PFOS and continuing increasing concentrations of the longer chain PFCAs (C(12)-C(14)) shows that further work on the reduction of environmental emissions of PFCs are necessary.

Study of the relation between the incidence of uterine leiomyomas and the concentrations of PCB and DDT in Baltic gray seals.

Exposure to environmental contaminants is believed to be associated with the previously described decrease in the reproduction rate of Baltic gray seals. In the present study the prevalence of uterine leiomyomas was investigated in 257 Baltic gray seal females examined during 1973-2007, in relation to the levels of polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT) in Baltic biota, using an estimated exposure index. Additionally, the proliferative activity in leiomyomas, occurrence of corpora lutea, and blubber concentrations of PCB and DDT were investigated in a subset of females. Leiomyomas were only found in females 22-41 years old, at a prevalence of 65%. No differences in blubber concentrations of PCB or DDT were detected between the subset of leiomyoma-bearing females and reference females, but the estimated exposure index indicated that the PCB level in Baltic biota might be related to the leiomyoma prevalence in Baltic gray seal females.

Effects of chlorinated biphenyls and metabolites on human uterine myocyte proliferation.

Uterine myometrial cells are responsive to sex steroids, which could make them susceptible to actions of endocrine disrupting environmental contaminants such as some PCBs. The aim of this investigation was to identify possible effects of some chlorinated biphenyls (CBs) and their metabolites on myometrial cell proliferation. Myometrial cells obtained from women in both phases of the menstrual cycle and from pregnant women were grown in vitro and exposed to CB 101, CB 118, 3' -MeSO2-CB 101, 4'-MeSO2-CB 101, 4-OH-CB 107, 17 beta-estradiol, progesterone, ethinylestradiol or levonorgestrel. The proliferative activity was studied by a BrdU assay. Myometrial cell cultures originating from pregnant women exhibited decreased proliferation in response to 3'-MeSO2-CB 101, 4'-MeSO2-CB 101 and 4-OH-CB 107. Estradiol, a combination of 1 nM 17beta-estradiol and 10 nM progesterone, ethinylestradiol and levonorgestrel also reduced the proliferation of the myometrial cells, regardless of whether the cells were collected from either of the menstrual cycle phases or from pregnant women. To our knowledge this study is the first to demonstrate that some CBs affect the proliferative activity of human uterine myocytes.

Effects of some endocrine disruptors on the proliferation and viability of human endometrial endothelial cells in vitro.

Endocrine disrupting chemicals (EDCs) pose a potential threat to human reproductive health. We studied the proliferation and viability of human endometrial endothelial cells (HEECs) in vitro after exposure to 2,2-bis(o,p-chlorophenyl)-1,1,1-trichloroethane (o,p'-DDT), 3,3',4,4'-tetrachlorobiphenyl (CB 77), 3,3',4,4',5-pentachlorobiphenyl (CB 126), di-n-butyl phthalate (DBP), bisphenol A (BPA), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and 17beta-oestradiol, progesterone, 17alpha-ethynyl oestradiol and levonorgestrel. Cell proliferation was studied using immunocytochemistry for PCNA expression and a 5-bromo-2'-deoxyuridine assay. Cell viability was studied by vital staining with propidium iodide and Hoechst 33258. HEECs in primary culture responded with increased proliferation to oestradiol and with decreased proliferation to levonorgestrel and the EDCs. Some EDCs also affected cell viability and increased the proportion of necrotic cells. However, the decrease in proliferation in response to DBP and TCDD cannot be explained by cell death. In light of these results, it is possible that the EDCs could have effects in vivo as well as in vitro, and influence processes involving for example endometrial angiogenesis.

Proliferative effects of estradiol, progesterone, and two CB congeners and their metabolites on gray seal (Halichoerus grypus) uterine myocytes in vitro.

Gray seal females living in the Baltic Sea have been found to exhibit a high prevalence of uterine leiomyomas. These animals are also known to accumulate lipid-soluble PCBs in their blubber. PCBs have documented endocrine-disrupting effects; to investigate whether the PCBs could be part of the genesis of uterine smooth muscle tumors in this species, gray seal myometrial cell cultures were exposed to two CBs and their metabolites, as well as to estradiol and progesterone, after which the effects were analyzed in terms of proliferative activity by measurements of BrdU absorbance and protein content. Progesterone was found to have an inhibitory effect, whereas one CB acted as a stimulant on the myometrial cell proliferation. One of the CB metabolites also seemed to have an inhibitory effect, although this could not be statistically verified. These results suggest that some CBs have effects on uterine myometrial cell proliferation in gray seals and, thus, may also take part in the growth regulation of uterine leiomyomas.