Clare Andvik

Steroid hormones can give an indication of stress, physiological health and reproductive status in whales; however, analysis is often limited by sample mass. We developed a sensitive analytical method using solid phase extraction–liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS) for the identification and quantification of eight steroid hormones (cortisol, cortisone, testosterone, androstenedione, progesterone, 11-deoxycortisol, 11-deoxycorticosterone and 17-hydroxyprogesterone) in small amounts of whale blubber (≤50 mg). Blubber biopsies were collected from stranded killer whales (Orcinus orca; n = 7), harbour porpoises (Phocoena phocoena; n = 3) and sperm whales (Physeter macrocephalus; n = 4), harvested common minke whales (n = 10; Balaenoptera acutorostrata) and free-living killer whales (n = 2) in Norway and the Barents Sea. Steroid hormones were separated using pentafluorophenyl propyl stationary phase and an optimized gradient programme with a total 20-min run. All samples (n = 26) were analysed at 50-mg sample mass, and a subset (n = 11) at 25 mg or lower. Low limit of detection and limit of quantification values resulted in high detection rates of hormones, and we found similar hormone concentrations between parallel samples and at the lowest tested sample mass (14 mg). Cortisol concentrations in sperm whales (4.3 ± 1.6 ng/g) were four times higher than stranded killer whales (1.1 ± 0.96 ng/g), which in turn were up to four times higher than free-living killer whales (0.23 ± 0.12 ng/g), harbour porpoise (0.64 ± 0.38 ng/g) and harvested minke whales (0.22 ± 0.13), possibly due to fatality type and duration of stress prior to death. We provide the first quantification of blubber steroid hormones in killer whale, minke whale, sperm whale and harbour porpoise, and in less than 50-mg sample mass. By minimizing sample mass without compromising precision or sensitivity, our method allows for steroid hormone analysis even on samples with limited availability, strengthening our ability to monitor and conserve marine mammal populations.

Short-, medium-, and long-chain chlorinated paraffins (CPs) (SCCPs, MCCPs, and LCCPs) and dechloranes are chemicals of emerging concern; however, little is known of their bioaccumulative potential compared to legacy contaminants in marine mammals. Here, we analyzed SCCPs, MCCPs, LCCPs, 7 dechloranes, 4 emerging brominated flame retardants, and 64 legacy contaminants, including polychlorinated biphenyls (PCBs), in the blubber of 46 individual marine mammals, representing 10 species, from Norway. Dietary niche was modeled based on stable isotopes of nitrogen and carbon in the skin/muscle to assess the contaminant accumulation in relation to diet. SCCPs and dechlorane-602 were strongly positively correlated with legacy contaminants and highest in killer (Orcinus orca) and sperm (Physeter macrocephalus) whales (median SCCPs: 160 ng/g lw; 230 ng/g lw and median dechlorane-602: 3.8 ng/g lw; 2.0 ng/g lw, respectively). In contrast, MCCPs and LCCPs were only weakly correlated to recalcitrant legacy contaminants and were highest in common minke whales (Balaenoptera acutorostrata; median MCCPs: 480 ng/g lw and LCCPs: 240 ng/g lw). The total contaminant load in all species was dominated by PCBs and legacy chlorinated pesticides (63-98%), and MCCPs dominated the total CP load (42-68%, except 11% in the long-finned pilot whale Globicephala melas). Surprisingly, we found no relation between contaminant concentrations and dietary niche, suggesting that other large species differences may be masking effects of diet such as lifespan or biotransformation and elimination capacities. CP and dechlorane concentrations were higher than in other marine mammals from the (sub)Arctic, and they were present in a killer whale neonate, indicating bioaccumulative properties and a potential for maternal transfer in these predominantly unregulated chemicals.

Killer whales (Orcinus orca) are at risk from high levels of biomagnifying pollutants, such as polychlorinated biphenyls (PCBs) and mercury (Hg). Previous toxicological risk assessments for the Norwegian killer whale population have assumed fish as the primary prey source, and assessed the population as below established effect thresholds. However, some individuals have recently been identified to also feed on seals. This study is the first to quantify levels of pollutants in seal-eating killer whales from northern Norway, and to measure Hg levels in the skin of killer whales worldwide. We found higher levels of all pollutants in seal-eating than fish-eating killer whales, including the emerging brominated flame retardants pentabromoethylbenzene (PBEB), pentabromotoluene (PBT) and hexabromobenzene (HBB). Sum polychlorinated biphenyls (ΣPCBs) in the blubber of seal-eaters (n = 7, geometric mean = 46 µg/g l.w.) were four times higher than fish-eaters (n = 24, geometric mean = 11 µg/g l.w.), which pushed all seal-eating individuals above multiple thresholds for health effects. Total Hg levels in skin of seal-eaters (n = 10, arithmetic mean = 3.7 µg/g d.w.) were twice as high as in fish-eaters (n = 28, arithmetic mean = 1.8 µg/g d.w.). Our results indicate that by feeding on higher trophic prey, the Norwegian killer whale population is at higher risk of health effects from pollution than previously assumed.

Ecological diversity has been reported for killer whales (Orcinus orca) throughout the North Atlantic but patterns of prey specialization have remained poorly understood. We quantify interindividual dietary variations in killer whales (n = 38) sampled throughout the year in 2017–2018 in northern Norway using stable isotopic nitrogen (δ¹⁵N: ¹⁵N/¹⁴N) and carbon (δ¹³C: ¹³C/¹²C) ratios. A Gaussian mixture model assigned sampled individuals to three differentiated clusters, characterized by disparate nonoverlapping isotopic niches, that were consistent with predatory field observations: seal‐eaters, herring‐eaters, and lumpfish‐eaters. Seal‐eaters showed higher δ¹⁵N values (mean ± SD: 12.6 ± 0.3‰, range = 12.3–13.2‰, n = 10) compared to herring‐eaters (mean ± SD: 11.7 ± 0.2‰, range = 11.4–11.9‰, n = 19) and lumpfish‐eaters (mean ± SD: 11.6 ± 0.2‰, range = 11.3–11.9, n = 9). Elevated δ¹⁵N values for seal‐eaters, regardless of sampling season, confirmed feeding at high trophic levels throughout the year. However, a wide isotopic niche and low measured δ¹⁵N values in the seal‐eaters, compared to that of whales that would eat solely seals (δ_N‐measured = 12.6 vs. δ_N‐expected = 15.5), indicated a diverse diet that includes both fish and mammal prey. A narrow niche for killer whales sampled at herring and lumpfish seasonal grounds supported seasonal prey specialization reflective of local peaks in prey abundance for the two fish‐eating groups. Our results, thus, show differences in prey specialization within this killer whale population in Norway and that the episodic observations of killer whales feeding on prey other than fish are a consistent behavior, as reflected in different isotopic niches between seal and fish‐eating individuals.