Dynamics of mercury stable isotope compounds in Arctic seals: New insights from a controlled feeding trial on hooded seals Cystophora cristata.

Accurate interpretation of mercury (Hg) isotopic data requires the consideration of several biotic factors such as age, diet, geographical range, and tissue metabolic turnover. A priori knowledge of prey-predator isotopic incorporation rates and Hg biomagnification is essential. This study aims to assess Hg stable isotopes incorporation in an Arctic species of Phocidae, the hooded seal Cystophora cristata, kept in human care for 24 months (2012-2014) and fed on a constant diet of Norwegian Spring Spawning herring Clupea harengus. We measured THg, MMHg and iHg levels, as well as Hg stable isotope composition with both mass dependent (MDF) and mass independent (MIF) fractionation (e.g. δ202Hg and Δ199,200,201,204Hg) in hooded seal kidney, liver, hair and muscle, in addition to herring muscle. We then calculated Hg MDF and MIF isotopic fractionation between hooded seals and their prey. We found a significant shift in δ202Hg between hooded seal hair (+0.80‰) and kidney (-0.78‰), and herring muscle. In hooded seals tissues δ202Hg correlated positively with MMHg percentage. These findings suggest that tissue-specific Hg speciation is the major driver of changes in Hg isotopic fractionation rates in this Arctic predator. Δ199Hg, Δ200Hg, Δ201Hg and Δ204Hg values did not vary between herring and hooded seal tissues, confirming their utility as tracers of Hg marine and atmospheric sources in top predators. To our knowledge, this represents the first attempt to assess complex Hg isotope dynamics in the internal system of Arctic Phocidae, controlling the effects of age, diet, and distribution. Our results confirm the validity of Hg stable isotopes as tracers of environmental Hg sources even in top predators, but emphasize the importance of animal age and tissue selection for inter-study and inter-species comparisons.

The Bacterial Microbiome in the Small Intestine of Hooded Seals (Cystophora cristata).

Arctic hooded seals (Cystophora cristata) are monogastric carnivores that go through extreme fasting and re-feeding in early life. They are born isolated on sea ice; suckle high-fat milk for four days and may then fast for up to one month before they start hunting and feeding on small prey (fish and crustaceans). Previous studies of the gut microbiota in pinnipeds have focused on the large intestine, while little data exist on the small intestinal microbiota. In this study, the bacterial microbiome in the proximal and distal small intestine of four captive two-year old seals (two males and two females) fed herring (Clupea harengus) was sampled post-mortem and characterized using 16S rRNA metabarcoding from the V1-V3 hypervariable region of the 16S ribosomal RNA (rRNA) genes. The seals were originally born in the wild and taken into human care at the end of the suckling period. Molecular-based analysis using Illumina Hiseq resulted in 569,910 16S rRNA sequences from the four seals (both sampling sites together). Taxonomical classification applying a naive Bayesian algorithm gave 412 Operational Taxonomic Units (OTUs). Firmicutes was the major phylum across samples (Proximal (P): 90.5% of total sequences, on average; Distal (D): 94.5%), followed by Actinobacteria (P: 7%; D: 0.3%) and Proteobacteria (P: 1.7%; D: 1.9%). Bacterial spp. belonging to the Clostridium (P: 54.1%; D: 41.6%) and SMB53 (P: 15.3%; D: 21.5%) constituted the major genera in both the proximal and distal small intestine. Furthermore, comparison with hindgut and fecal samples from geographically diverse marine mammals highlighted similarities in the microbiome between our seals and those sharing similar aquatic environments. This study has provided a first reliable glimpse of the bacterial microbiota in the small intestine microbiome of hooded seals.

Energy efficiency of respiration in mature and newborn reindeer.

Reindeer (Rangifer tarandus) have evolved elaborate nasal turbinate structures that are perfused via a complex vascular network. These are subject to thermoregulatory control, shifting between heat conservation and dissipation, according to the animal's needs. The three-dimensional design of the turbinate structures is essential in the sense that they determine the efficiency with which heat and water are transferred between the structure and the respired air. The turbinates have already a relatively large surface area at birth, but the structures have yet not reached the complexity of the mature animal. The aim of this study was to elucidate the structure-function relationship of the heat exchange process. We have used morphometric and physiological data from newborn reindeer calves to construct a thermodynamic model for respiratory heat and water exchange and present novel results for the simulated respiratory energy losses of calves in the cold. While the mature reindeer effectively conserves heat and water through nasal counter-current heat exchange, the nose of the calf has not yet attained a similar efficiency. We speculate that this is probably related to structure-size limitations and more favourable climate conditions during early life. The fully developed structure-function relationship may serve as inspiration for engineering design. Simulations of different extents of mucosal vascularization suggest that the abundance and pattern of perfusion of veins in the reindeer nasal mucosa may contribute to the control of temperature profiles, such that nasal cavity tissue is sufficiently warm, but not excessively so, keeping heat dissipation within limits.

Seroprevalence for Brucella spp. in Baltic ringed seals (Phoca hispida) and East Greenland harp (Pagophilus groenlandicus) and hooded (Cystophora cristata) seals.

Zoonotic infections transmitted from marine mammals to humans in the Baltic and European Arctic are of unknown significance, despite given considerable potential for transmission due to local hunt. Here we present results of an initial screening for Brucella spp. in Arctic and Baltic seal species. Baltic ringed seals (Pusa hispida, n = 12) sampled in October 2015 and Greenland Sea harp seals (Pagophilus groenlandicus, n = 6) and hooded seals (Cystophora cristata, n = 3) sampled in March 2015 were serologically analysed for antibodies against Brucella spp. The serological analyses were performed using the Rose Bengal Test (RBT) followed by a confirmatory testing of RBT-positive samples by a competitive-enzyme linked immunosorbent assay (C-ELISA). Two of the Baltic ringed seals (a juvenile male and a juvenile female) were seropositive thus indicating previous exposure to a Brucella spp. The findings indicate that ringed seals in the Baltic ecosystem may be exposed to and possibly infected by Brucella spp. No seropositive individuals were detected among the Greenland harp and hooded seals. Although our initial screening shows a zoonotic hazard to Baltic locals, a more in-depth epidemiological investigation is needed in order to determine the human risk associated with this.

Effects of Polar Bear and Killer Whale Derived Contaminant Cocktails on Marine Mammal Immunity.

Most controlled toxicity studies use single chemical exposures that do not represent the real world situation of complex mixtures of known and unknown natural and anthropogenic substances. In the present study, complex contaminant cocktails derived from the blubber of polar bears (PB; Ursus maritimus) and killer whales (KW; Orcinus orca) were used for in vitro concentration-response experiments with PB, cetacean and seal spp. immune cells to evaluate the effect of realistic contaminant mixtures on various immune functions. Cytotoxic effects of the PB cocktail occurred at lower concentrations than the KW cocktail (1 vs 16 µg/mL), likely due to differences in contaminant profiles in the mixtures derived from the adipose of each species. Similarly, significant reduction of lymphocyte proliferation occurred at much lower exposures in the PB cocktail (EC50: 0.94 vs 6.06 µg/mL; P < 0.01), whereas the KW cocktail caused a much faster decline in proliferation (slope: 2.9 vs 1.7; P = 0.04). Only the KW cocktail modulated natural killer (NK) cell activity and neutrophil and monocyte phagocytosis in a concentration- and species-dependent manner. No clear sensitivity differences emerged when comparing cetaceans, seals and PB. Our results showing lower effect levels for complex mixtures relative to single compounds suggest that previous risk assessments underestimate the effects of real world contaminant exposure on immunity. Our results using blubber-derived contaminant cocktails add realism to in vitro exposure experiments and confirm the immunotoxic risk marine mammals face from exposure to complex mixtures of environmental contaminants.

Carbon, nitrogen and sulphur isotopic fractionation in captive juvenile hooded seal (Cystophora cristata): Application for diet analysis.

RATIONALE: Intrinsic biogeochemical markers, such as stable isotope ratios of carbon, nitrogen and sulphur, are increasingly used to trace the trophic ecology of marine top predators. However, insufficient knowledge of fractionation processes in tissues continues to hamper the use of these markers. METHODS: We performed a controlled feeding experiment with eight juvenile hooded seals (Cystophora cristata) that were held on a herring-based diet (Clupea harengus) for two years. Stable isotope ratios were measured via isotope ratio mass spectrometry in three of their tissues and related to values of these markers in their diet. RESULTS: Diet-tissue isotope enrichment (trophic enrichment factor, TEF) values between dietary herring and seal tissues for carbon (Δ13 C) were +0.7 ‰ for red blood cells, +1.9 ‰ for hair and +1.1 ‰ for muscle. The TEFs for nitrogen trophic (Δ15 N) were +3.3 ‰ for red blood cells, +3.6 ‰ for hair and +4.3 ‰ for muscle. For sulphur, the Δ34 S values were +1.1 ‰ for red blood cells, +1.0 ‰ for hair and +0.9 ‰ for muscle. CONCLUSIONS: These enrichment values were greater than those previously measured in adult seals. This increase may be related to the higher rate of protein synthesis and catabolism in growing animals. This study is the first report on sulphur isotope enrichment values for a marine mammal species.