Blubber Layer Research Articles

Blubber biopsy in common bottlenose dolphins using a novel biopsy puncher: Evaluation of the impact on living individuals and possibility of applications in cetacean research.

Biopsy has recently become a preferred protocol for sampling the skin and blubber of many cetacean species, although it is desirable to collect as minimally invasive as possible. Here, the effect of biopsy sampling on the captive common bottlenose dolphins was evaluated by analyzing the process of wound healing and changes in hematological and blood biochemical parameters after biopsy using a puncher developed to collect up to the inner layer of the dolphin blubber. Results showed that the wounds caused by biopsy were closed in as early as 1 day and completely covered with the epidermis within 5-11 days. Blood fibrinogen, which generally increases due to a wound-induced inflammatory response or activation of the coagulation system, was significantly elevated after the biopsy indicating ongoing tissue repair, while other parameters did not exhibit significant differences. Furthermore, histological observation and RNA extraction of samples were performed to investigate the versatility of this method to cetacean research. Histological examination revealed three distinct layers of the blubber in the biopsy samples. Moreover, total RNA extracted from biopsy samples exhibited sufficient quality and quantity for gene expression analyses. Overall, the puncher utilized in our study represents a valuable and minimally invasive tool for investigating various aspects of small cetacean studies.

Cortisol and epinephrine alter the adipose functions and the mobilization of PCBs in adipose tissue slices from elephant seal

Marine mammals are exposed to increasing intensities of anthropogenic stressors such as acoustic disturbance and contaminants. Correlative studies have suggested concerning shifts in behavioral and physiological status of stress-exposed individuals, which could alter the health and survival of marine mammal populations. However, functional studies of the effects of multiple stressors on marine mammals are lacking. To fill in this gap, we recently developed an ex vivo approach of precision-cut adipose tissue slices (PCATS) to study the impact of stressors on the function of an essential endocrine organ: the adipose tissue. In the present study, we investigated the impact of hormones associated with the stress response on adipose tissue from northern elephant seals (Mirounga angustirostris). Blubber biopsies were collected from 17 weaned northern elephant seal pups, separated into inner and outer layers, dissected into PCATS, and cultured for 48 hours. To mimic prolonged and short-term exposure to physiological stress, PCATS were treated with 2 µM cortisol (CORT) for the entire 48 hours or 100 nM epinephrine (EPI) for the last 12 hours of culture, respectively. Hormones were applied individually or in combination. CORT and EPI exhibited an interacting, blubber layer-dependent, effect on adipose tissue biology, as quantified by gene expression in PCATS, and release of glycerol, free fatty acids, leptin and polychlorinated biphenyls into culture media. EPI stimulated a higher rate of lipolysis than CORT in PCATS from both blubber layers. The combination of CORT and EPI upregulated the expression of adipose triglyceride lipase in inner blubber PCATS and downregulated hormone-sensitive lipase in outer blubber-derived PCATS. Expression of the leptin gene and secretion of the leptin adipokine were both decreased by EPI, while addition of CORT attenuated this effect in inner blubber PCATS only. CORT also increased the expression of the antioxidant enzyme glutathione peroxidase 3 in PCATS from both blubber layers. Polychlorinated biphenyls exhibited selective and limited mobilization from PCATS treated with stress hormones, highlighting the lipophilic properties of these toxic compounds. This study showed that physiological stress can impact several essential functions of marine mammal blubber, such as lipolysis and adipokine production.

First report of a common dolphin (Delphinus delphis) death following penetration of a biopsy dart

The remote collection of skin and blubber biopsy samples from free-ranging cetaceans is a powerful technique which has been increasingly used by scientists in recent years in a wide range of applications, particularly with respect to genetic and contaminant studies. Biopsy sampling, if carried out responsibly, is known to cause low-level reactions, and is unlikely to produce long-term deleterious effects. However, this technique is not completely devoid of risk for the sampled animals, particularly for smaller odontocetes. This paper reports the death of a common dolphin in the central Mediterranean Sea, following penetration of a biopsy dart and subsequent handling. The dolphin was hit in the dorsal muscle mass below the dorsal fin by a lightweight pneumatic dart fired from a distance of 6m by a variable-power CO2 dart projector. The methods and equipment had been previously successfully used with minimal effect on common dolphins and other species under similar conditions; it was therefore considered to be relatively uninvasive and more likely to reduce disturbance while increasing sample retrieval. However, in the reported event, a dart stuck in the dorsal muscle mass instead of recoiling as expected. Less than 2min after the hit, the dolphin began catatonic head-up sinking, and was recovered by a team member at depth. Basic medical care was given to ensure haemostasis, but the animal died 16min later. Minimal overall bleeding and a small wound in the thick muscle mass were not among the suspected causes of death. This may have been the consequence of either indirect vertebral trauma or stress. Furthermore, the dolphin had a relatively thin (7mm) blubber layer, that may have contributed to the unwanted outcome of the biopsy attempt. The author stresses that scientists should only adopt even mildly intrusive research methods after careful review and risk assessment in the light of the precautionary principle, and that their decisions must be reviewed on a regular basis according to the best available evidence.

Variation in blubber thickness and histology metrics across the body topography of a false killer whale (Pseudorca crassidens).

Blubber is a multifunctional tissue essential to the survival of cetaceans. Histological assessment of blubber may be useful in determining odontocete nutritional state but a greater understanding of specific variation across the body is needed. We report on morphological variation of the blubber according to girth axes and sampling planes in a sub-adult male, bycaught false killer whale (Pseudorca crassidens) using metrics of blubber thickness (BT), adipocyte area (AA), and adipocyte index (AI). 48 full depth blubber samples were taken along 6 girth axes at 5 equidistant sampling points on both sides of the body. At these sampling locations BT was recorded, and AA and AI were determined for three distinct blubber layers. Linear mixed effect models were used to assess variation of the blubber across layers and body topography. BT was somewhat non-uniform across the body but was generally thicker in the dorsal region and thinner laterally. AA was greater cranially and AI was greater caudally. The middle and inner layer blubber showed significant differences dorsoventrally with larger AA and smaller AI in the ventral region of the body. Variation of the blubber metrics across the body are indicative of variable functions of the blubber within an individual. Due to the variability observed, we expect that AI of the dynamic inner layer blubber is most informative of overall body condition and that biopsy samples of the outer and middle blubber may still be useful in determining the nutritional status of live false killer whales.

Stratification of lipids, fatty acids and organochlorine contaminants in blubber of white whales and killer whales

The biopsy 2 via dart, trocar or surgery 2 is becoming the preferred protocol for sampling skin and blubber of many cetacean species, because a small sample from a healthy animal may provide better information than a larger sample collected via necropsy from an ill or emaciated animal. Furthermore, the biopsy is often the only means of obtaining samples (e.g. for threatened or endangered species). Because biopsy darts collect only a small sample of tissue 2 and blubber can be heterogeneous in structure and composition 2 it is essential to compare the results obtained from biopsies to those found by analysing full-thickness blubber samples obtained via necropsy. This manuscript compares blubber stratification in two odontocete species, white whales (Delphinapterus leucas) and killer whales (Orcinus orca). Five parameters (i.e. lipid percent and classes, contaminant concentrations and profiles, fatty acid profiles) were measured by blubber depth. Results of these comparisons strongly suggest that biopsy results must be interpreted with caution and in conjunction with results from species-specific blubber depth profiling. For example, lipid classes measured in biopsy samples of white whales and killer whales were similar to those for equivalent-depth samples obtained by necropsy. In addition, lipid-adjusted contaminant concentrations measured in dart or trocar samples adequately represented those obtained by necropsy of both species. Conversely, the lipid content in biopsy samples was lower than that found in same-depth necropsied samples due to loss of lipid during sampling. Also, because of the high level of fatty acid stratification observed, fatty acid profiles from the outer blubber layer collected via biopsy from both species are less likely than the metabolically active inner layer to be useful in determining the prey species consumed by these odontocetes. This study demonstrates, for white and killer whales, that properly interpreted results from blubber biopsies can provide valuable information about the body condition, health and life history of individual animals.

A Whale Tale

A Whale Tale Allegra Hyde (bio) The whale was already dead when it washed ashore. A massive sperm whale, its skin leathery and barnacled, its narrow jaw strutted with teeth. The whole town went down to the beach to see it: mothers and firefighters and stay-at-home dads and dead-beat dads and vacuum cleaner salespeople and endometriologists and dowagers and women of the night and diamond miners and fortune tellers and personal trainers. Children were bussed in from school. Prisoners from prison. Altogether, the townspeople encircled the creature, reaching forward to touch its gray-blue skin. People wept. People waved flags. People complained about not being able to barbecue because of health ordinances. More than anything, people wondered how the whale had died. A coroner-for-whales was summoned to conduct an autopsy. She began to cut the whale open right there on the beach, excavating through layers of blubber. Because it had taken her so long to arrive, however, and because it had started drizzling—and also because the work was gruesome—almost everyone went home before the procedure was complete. Everyone except for a few diehard protesters, local news reporters, police officers, orphans, and a man named Anders. Anders did not usually attend public spectacles such as this. He had merely been driving by on his way home from his job at a nonprofit for bicycle safety. He had ventured onto the beach because of the crowds and had stayed because of the whale. Or, more truthfully: Anders had stayed because it meant he could delay going home. And if he delayed going home, then he could delay returning to his longtime lover, Florian. And if he delayed returning to Florian, he could delay doing what he had promised himself he would do—which was end their relationship. The coroner-for-whales went about her work. She used a special saw to cut into the creature’s vast flank. She was aiming for its stomach, she told onlookers. She was getting close. Like everyone in the crowd, [End Page 44] Anders drew an anticipatory breath, though the sight of the whale disturbed him. Fins motionless. Glassy eyes staring at nothing. The bigness of the animal—the deadness of it, too—put a stone in his gut. Hard and round and heavy, the stone gave Anders the sinking sensation of foreboding. Or else: the foreboding came from what he planned to do when he got home. Anders loved Florian—or had loved him, early on in their seven-year relationship. Florian, who was all sparkles and beauty. Florian, with his sandy hair and sea-glass eyes. Florian, who was the son of a local witch/naturopath, who could whistle like the wind singing through trees, and who had at one time taken Anders’s breath away, but now took all of Anders’s money and time and energy, and who refused to get a job. “And we’re in!” The coroner tossed her saw aside and extended a gloved arm into the cavernous opening of the whale’s stomach. Anders felt the stone in his gut grow heavier. He had no business watching this creature be disemboweled. But his relationship with Florian was over—needed to be over—and he couldn’t yet bear to admit this to Florian’s face. So he pushed through the crowd to see what made the others yelp with surprise. The whale’s stomach was bursting with stuff. Human stuff. A wide-screen television. A wooden bed frame. Woolen socks, both matched and unmatched. Plastic fruit. A two-slice toaster. Several doilies—or so Anders guessed. Some items were hard to identify, covered as they were with the whale’s stomach slime. Next came a corduroy-upholstered couch. A set of collectible spoons from Midwest states. A rolltop desk. A pair of roller skates—which made Anders lean closer, since the skates were similar to a pair he’d just purchased. The couch, too, reminded him of the couch he owned, which he’d believed to be one of a kind. People grabbed items, inspected them, expressed their dismay and confusion. A reporter waved aloft a photo frame containing a...

Comprehensive molecular and morphological resolution of blubber stratification in a deep-diving, fasting-adapted seal.

Blubber is a modified subcutaneous adipose tissue in marine mammals that provides energy storage, thermoregulation, hydrodynamic locomotion, and buoyancy. Blubber displays vertical stratification by lipid content, fatty acid composition, and vascularization, leading to the assumption that deeper blubber layers are metabolically active, while superficial layers are mainly structural and thermoregulatory. However, few studies have examined functional stratification of marine mammal blubber directly, especially in pinnipeds. We characterized morphological and transcriptional differences across blubber layers in the northern elephant seal, a deep-diving and fasting-adapted phocid. We collected blubber from seals early in their fasting period and divided blubber cores into three similarly sized portions. We hypothesized that the innermost blubber portion would have higher 1) heterogeneity in adipocyte size, 2) microvascular density, and 3) expression of genes associated with metabolism and hormone signaling than outer blubber. We found that adipocyte area and variance increased from outermost (skin-adjacent) to innermost (muscle-adjacent) blubber layers, suggesting that inner blubber has a higher capacity for lipid storage and turnover than outer blubber. Inner blubber had a higher proportion of CD144+ endothelial cells, suggesting higher microvascular density. In contrast, outer blubber had a higher proportion of CD4+ immune cells than inner blubber, suggesting higher capacity for response to tissue injury. Transcriptome analysis identified 61 genes that were differentially expressed between inner and outer blubber layers, many of which have not been studied previously in marine mammals. Based on known functions of these genes in other mammals, we suggest that inner blubber has potentially higher 1) adipogenic capacity, 2) cellular diversity, and 3) metabolic and neuroendocrine signaling activity, while outer blubber may have higher 1) extracellular matrix synthesis activity and 2) responsiveness to pathogens and cell stressors. We further characterized expression of nine genes of interest identified by transcriptomics and two adipokines with higher precision across blubber layers using targeted assays. Our study provides functional insights into stratification of blubber in marine mammals and a molecular key, including CD144, CD4, HMGCS2, GABRG2, HCAR2, and COL1A2, for distinguishing blubber layers for physiological and functional studies in seals.

Variation of blubber thickness for three marine mammal species in the southern Baltic Sea.

Evaluating populational trends of health condition has become an important topic for marine mammal populations under the Marine Strategy Framework Directive (MSFD). In the Baltic Sea, under the recommendation of Helsinki Commission (HELCOM), efforts have been undertaken to use blubber thickness as an indicator of energy reserves in marine mammals. Current values lack geographical representation from the entire Baltic Sea area and a large dataset is only available for grey seals (Halichoerus grypus) from Sweden and Finland. Knowledge on variation of blubber thickness related to geography throughout the Baltic Sea is important for its usage as an indicator. Such evaluation can provide important information about the energy reserves, and hence, food availability. It is expected that methodological standardization under HELCOM should include relevant datasets with good geographical coverage that can also account for natural variability in the resident marine mammal populations. In this study, seasonal and temporal trends of blubber thickness were evaluated for three marine mammal species-harbor seal (Phoca vitulina), grey seal (Halichoerus grypus) and harbor porpoise (Phocoena phocoena)-resident in the southern Baltic Sea collected and investigated under stranding networks. Additionally, the effects of age, season and sex were analyzed. Seasonal variation of blubber thickness was evident for all species, with harbor seals presenting more pronounced effects in adults and grey seals and harbor porpoises presenting more pronounced effects in juveniles. For harbor seals and porpoises, fluctuations were present over the years included in the analysis. In the seal species, blubber thickness values were generally higher in males. In harbor seals and porpoises, blubber thickness values differed between the age classes: while adult harbor seals displayed thicker blubber layers than juveniles, the opposite was observed for harbor porpoises. Furthermore, while an important initial screening tool, blubber thickness assessment cannot be considered a valid methodology for overall health assessment in marine mammals and should be complemented with data on specific health parameters developed for each species.

Functional Attenuation of UCP1 as the Potential Mechanism for a Thickened Blubber Layer in Cetaceans.

Uncoupling protein 1 (UCP1) is an essential protein in the mitochondrial inner membrane that mediates nonshivering thermogenesis (NST) and plays an important role in thermoregulation and fat deposition. However, the relationship between the evolution of UCP1 and fat deposition in the blubber layer in cetaceans remains unclear. Here, frameshift mutations, premature termination, and relaxed selection pressure (ω = 0.9557, P < 0.05) were detected in UCP1 in cetaceans, suggesting that UCP1 was inactivated during cetacean evolution. By time estimation, it was found that the inactivation of UCP1 in cetaceans occurred between 53.1 and 50.2 Ma. However, combined with findings from immunohistochemical analysis of the blubber layer of the Yangtze finless porpoise and in vitro functional assays, a premature termination of cetacean UCP1 resulted in a reduction of UCP1-mediated NST capacity (about 50%) and lipolytic capacity (about 40%), both of which were beneficial to maintain blubber layer and body temperature without excessive fat consumption. This study provides new insights into the molecular mechanisms of the blubber thickening in cetaceans and highlights the importance of UCP1 attenuation in cetaceans for secondary aquatic adaptation.

Histomorphological stratification of blubber of three dolphin species from subtropical waters.

Blubber is a highly specialized and dynamic tissue unique to marine mammals and presents a reflection of the individuals' nutrition, environment, and life history traits. Few studies have investigated the histomorphology of cetacean blubber in subtropical environments. The aim of this study was to investigate the blubber histomorphology of three different dolphin species off the subtropical KwaZulu-Natal coast, South Africa, using adipocyte cell size, number, and density. Blubber tissue samples from the saddle area of 43 incidentally bycaught animals (fourSousa plumbea, 36 Tursiops aduncus, and threeDelphinus delphis) were used to compare cell parameters between blubber layers. Samples were divided into the upper third (corresponding to the superficial layer closest to the epidermis), middle third, and lower third (corresponding to the deep layer). For T. aduncus, factors potentially affecting blubber histomorphology, such as sex, age class, and season, were also assessed. Our results showed that no stratification was present in S. plumbea, which could be ascribed to the species' warmer inshore habitat, large body size, and apparent lower mobility. For T. aduncus and D. capensis, however, blubber stratification was determined, characterized by a gradual transition of cell size, number, and density between layers rather than clearly defined layers. Significant differences in adipocyte cell number and density were found for different sexes and age classes of T. aduncus. However, there were no significant differences between seasons, which was attributed to the small temperature differences between seasons. This study represents the first investigation of odontocete blubber histomorphology in subtropical waters. It is recommended that future studies investigate blubber lipid content, while also taking into consideration the reproductive status of the females and the temperature range of their study area. It is hoped that our results, in conjunction with histopathology and other health indicators, could assist in assessing health and body condition.

Age group differences in blubber fatty acid profiles in northern elephant seals (Mirounga angustirostris)

Northern elephant seals (Mirounga angustirostris), like many marine mammals, rely on internal lipid stores, specifically fatty acids (FAs) stored in the blubber layer, to meet metabolic needs. The energetic demands of northern elephant seals vary with ontogeny, as each life-history stage experiences different metabolic requirements due to development, growth, and breeding. To date, no comprehensive study has reported on changes in blubber FA profiles across northern elephant seal age groups or sex. Therefore, our objective was to determine how blubber FAs differ across ontogeny and sex in northern elephant seals. As a sexually dimorphic mammal, northern elephant seals go through sex-specific ontogenetic changes in morphology and physiology; we hypothesized that these changes would be reflected in their FAs profiles. To determine this, FAs profiles were compared from full blubber cores collected from 79 northern elephant seals across four age groups. We provide the first evidence of blubber fatty acids differing across ontogeny as NES transition from young, developing seals to mature fully developed adults. However, we did not find differences in blubber FAs profiles between the sexes. Monounsaturated fatty acids (MUFAs) are found in the highest proportions across all NES age classes and sexes, followed by SFAs and PUFAs; this highlights the important role MUFAs play in maintaining fluidity of the blubber layer and in thermoregulation. The individual FAs with the highest concentrations (C16:1, C18:1n9 and C16:0) in northern elephant seal blubber are similar to those in other marine mammals. Weaned pup FAs profiles were significantly different from all other age classes; adults and juveniles also showed age-specific differences. Specifically, weaned pups had the highest proportions of SFAs and the lowest proportions of PUFA, suggesting use of PUFAs to aid development. Each life history stage of NES is interconnected to previous and future stages, making FA accumulation, mobilization, and storage an important process throughout an individual’s life. Further, any changes to this process can have cascading consequences throughout ontogeny in this species. Future monitoring of the FA composition of blubber across age classes could potentially indicate the costs of different environmental changes on blubber storage in NES.

Stratification, sex and ontogenetic effects on the lipid and fatty acid profiles in the blubber of sperm whales from Tasmanian waters.

We examined the differential deposition of lipids according to layer, sex and ontogeny in the blubber of 31 adult sperm whales (n = 22 females, 9 males) and two calves that stranded off the Tasmanian coast from 2002 to 2004. Total lipid (TL) content varied widely across the blubber layers of adults (27-77%). Overall, females had higher TL content than males possibly representing higher energy needs due to reproduction. Higher TL content in the middle layer of adults (69%) suggests this layer may act as an energy reserve. Wax esters (WE) dominated the blubber and were highest in the outer layer of adults and calves, likely providing insulative qualities for this deep-diving odontocete. Triacyclglycerols, an easily mobilized energy source, were highest in the inner layer of females (37.3 ± 13.5%) and calves (32.1 ± 1.8%) compared to males (17.1 ± 8.2%). Monounsaturated fatty acids (MUFA) also dominated the blubber. An increasing gradient from the inner to outer layer reflected an increasing source of endogenously synthesized lipids, whereas an increasing gradient of saturated fatty acids and polyunsaturated fatty acids (PUFA) toward the inner layer reflected an increasing source of dietary lipids. Although body site did not affect lipid profiles, stratification between the outer and more metabolically active inner layers suggests that only using the outer layer may result in an incomplete lipid profile for sperm whales.

Validation of quantitative fatty acid signature analysis for estimating the diet composition of free-ranging killer whales

Accurate diet estimates are necessary to assess trophic interactions and food web dynamics in ecosystems, particularly for apex predators like cetaceans, which can regulate entire food webs. Quantitative fatty acid analysis (QFASA) has been used to estimate the diets of marine predators in the last decade but has yet to be implemented on free-ranging cetaceans, from which typically only biopsy samples containing outer blubber are available, due to a lack of empirically determined calibration coefficients (CCs) that account for fatty acid (FA) metabolism. Here, we develop and validate QFASA for killer whales using full blubber from managed-care and free-ranging individuals. First, we compute full, inner, and outer blubber CCs from the FA signatures across the blubber layers of managed-care killer whales and their long-term diet items. We then run cross-validating simulations on the managed-care individuals to evaluate the accuracy of diet estimates by comparing full-depth and depth-specific estimates to true diets. Finally, we apply these approaches to subsistence-harvested killer whales from Greenland to test the utility of the method for free-ranging killer whales, particularly for the outer blubber. Accurate diet estimates for the managed-care killer whales were only achieved using killer whale-specific and blubber-layer-specific CCs. Modeled diets for the Greenlandic killer whales largely consisted of seals (75.9 ± 4.7%) and/or fish (20.4 ± 2.4%), mainly mackerel, which was consistent with stomach content data and limited literature on this population. Given the remote habitats and below surface feeding of most cetaceans, this newly developed cetacean-specific QFASA method, which can be applied to outer-layer biopsies, offers promise to provide a significant new understanding of diet dynamics of free-ranging odontocetes and perhaps other cetacean species throughout the world’s oceans.

Variability of Lipids and Fatty Acids in Pacific Walrus Blubber

The variability of lipid content and fatty acid (FA) composition across blubber depth and body sites are important considerations for condition and diet studies of marine mammals. We investigated lipid and FA variability among inner and outer blubber layers, three body sites, four study years, and lactation status of adult female Pacific walruses (Odobenus rosmarus divergens) using blubber samples collected from subsistence-harvested walruses in spring 2007–2010. Percent lipid content did not differ between the inner and outer blubber layers at the rump, flank, or sternum of walruses. Although FA composition differed between the inner and outer blubber layers, the difference was consistent across body sites, and differences between layers within individual FAs were small (&amp;lt;2%). Lipid content at the sternum of lactating females was 6% higher than non-lactating females, consistent with known variation in body condition among these reproductive classes. Across study years, lipid content varied 18% and individual FAs varied 6%, likely reflecting population-level interannual variability in energy budgets and small differences in diet among years. Consistency in blubber lipid content across blubber depth and body sites and detectable variation in blubber lipid content among reproductive classes and years suggests the potential for lipid content to be a useful indicator of walrus body condition. In addition to information on condition, FA composition of blubber samples could potentially provide insights into changes in walrus diet that may be expected to occur from changes in their access to prey resources resulting from continued sea ice loss.

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.

Thermoregulatory costs in molting Antarctic Weddell seals: impacts of physiological and environmental conditions: Themed Issue Article: Conservation of Southern Hemisphere Mammals in a Changing World.

For polar marine mammals, the energetic cost of thermoregulation depends on ambient conditions in the highly variable surrounding environment. Heat conservation strategies used by pinnipeds to reduce total heat loss include small surface area to volume ratios, the ability to limit perfusion and thick subcutaneous blubber layers. There are limits to how cool the skin surface may remain without compromising function, especially during the annual pelage molt, when hair and skin are replaced. To determine if actively molting seals incur higher thermoregulatory costs, surface temperature (ST) and heat flux (HF) were measured in 93 adult female Weddell seals (Leptonychotes weddellii) both prior to and during the active molting period using direct sensors and infrared imaging. Linear mixed-effect models revealed that ST increased significantly with increased ambient temperature and decreased wind speed (contributing 44.6 and 41.7% of the attributed variance, respectively). Seal STs were not impacted by molt status, but were maintained at 11.2 ± 0.3°C warmer than the ambient temperature. Infrared imaging results averaged 15.1 ± 1.4°C warmer than direct ST measurements. In contrast, HF was significantly higher in seals in early molting stages compared to the pre-molt season ( P < 0.001) and molt status accounted for 66.5% of the variance in HF. Thermoregulatory costs calculated from estimated basal metabolic rate and measured HF were more than double for molting seals as compared to those in pre-molt. This suggests that perfusion is increased during molt to support follicle development, despite the increased energetic costs associated with higher HF rates. Because ST, HF and thermoregulatory costs are strongly influenced by ambient conditions, molt timing is likely under selective pressure to occur during the warmest period of the year. Shifts in environmental conditions that delay molt phenology or increase HF rates could negatively impact seal populations by further increasing thermoregulatory costs.

Histological Variation in Blubber Morphology of the Endangered East Asian Finless Porpoise (Neophocaena asiaeorientalis sunameri) with Ontogeny and Reproductive States.

The highly lipid-rich blubber in the hypodermis is a specialized structure that functions in thermoregulation, energy storage, buoyancy control, locomotion, and streamlining the body shape in marine mammals. The key objective of this study was to investigate blubber development in the East Asian Finless Porpoise (Neophocaena asiaeorientalis sunameri) across the ontogenetic (fetuses, calves, and adults) and reproductive states (adults, pregnant, and lactating). Blubber samples were collected from East Asian Finless Porpoises (EAFP) that were accidentally caught in the fishing nets in the Bohai/Yellow Sea from late April to mid May of 2015. The mean blubber depth was significantly thinner in fetuses across the ontogenetic groups and significantly thicker in pregnant and lactating vs. adult females across the reproductive states. Across the four regions in each group, we did not find significant variations in blubber depth. However, the correlations between body length and weight vs. blubber depth was significant. Histological observation identified three layers of stratified blubber with a significantly smaller adipocyte cell size in fetuses and a significantly higher area ratio of structural fiber in the middle and inner layers across the ontogenetic groups. Across the reproductive states, we did not observe a statistically significant difference in the adipocyte cell size or area ratio of the structural fiber. Our results suggest that prenatal blubber growth is characterized by an increase in the adipocyte cell count, while postnatal growth is the result of an increase in cell size. They also indicate that ontogeny can affect blubber depths and cellular measurements in the EAFP.

Are vitamins A and E associated with persistent organic pollutants and fatty acids in the blubber of highly contaminated killer whales (Orcinus orca) from Greenland?

We quantified blubber concentrations of vitamins A (retinol) and E (α-tocopherol) and evaluated associations with persistent organic pollutants (ΣPOPs) in 14 highly-contaminated killer whales (Orcinus orca) sampled in Greenland from 2012 to 2014. We considered the influence of blubber depth, sex/age class and diet (based on biomass % of major fatty acids) in these relationships. Blubber concentrations of vitamin A averaged 34.1 ± 4.7 μg g−1 wet weight (ww) and vitamin E averaged 35.6 ± 4.4 μg g−1 ww. Although overall vitamin A concentrations did not vary between inner (closer to the muscle) and outer (closer to the skin) blubber layer or between sub-adults and adult females, concentrations in the outer layer of sub-adults were lower compared to the outer layer of adult females (p = 0.03). Outer layer may therefore reflect age accumulation of vitamin A, while in the more active inner layer, age effects might be masked by metabolic needs such as lactation. Neither diet nor ΣPOPs affected vitamin A variation, suggesting this vitamin is highly regulated in the body. Given the high exposures in these killer whales, vitamin A might not be a sensitive biomarker for POPs adverse effects. Vitamin E concentrations were significantly higher in inner compared to outer layer (p < 0.001), likely associated with blubber composition, suggesting that biopsies may not fully represent vitamin E concentrations in blubber. Age-accumulation of vitamin E also occurred with higher concentrations in adult females compared to sub-adults, independent of blubber depth (p < 0.01). Diet, ΣPOPs, and an interaction between these two variables significantly affected vitamin E variation in inner blubber, explaining 91% of this variation. The negative relationship between ΣPOPs (especially Σdichlorodiphenyltrichloroethane (DDT) and Σchlordanes in outer layers) and vitamin E was observed only in killer whales with a diet poorer in polyunsaturated fatty acids, suggested that killer whales feeding more consistently on marine mammals in Arctic environments over a fish-based diet, may be at higher risk of POP-induced disruption in vitamin E homeostasis. Considering diet is therefore important to understand the potential effects of elevated contaminant exposures on levels of certain essential nutrients, i.e., vitamin E, in killer whales.

High Levels of Food Intake in Harbor Porpoises (Phocoena phocoena): Insight into Recovery from Disturbance

If harbor porpoises (Phocoena phocoena) are impaired in their foraging ability because they need to move away from anthropogenic sound sources, their fitness may be reduced. Understanding how much harbor porpoises can eat after a disturbance, and how quickly they can replenish their energy reserves, is important for assessing the significance of disturbances. After fasting for various time periods (2 to 24 h), four captive harbor porpoises, housed in water and air temperatures similar to those encountered by wild conspecifics, were fed a structured diet of meals larger than usual (each normal meal was 20% of the daily food mass requirement). A few times they were fed ad libitum, but this led to severe constipation, so this feeding method was abandoned for welfare and health reasons. The food ingested over a period of one hour following fasting for 2 to 24 hours was quantified (i.e., mass, volume, and as a percentage of normal daily food mass intake in that period). The results show that, in contrast to established belief, harbor porpoises can eat a large percentage (up to ~98%) of their normal daily food mass intake in a single feeding bout without showing physical problems. Adult animals of around 155 cm in body length can eat up to ~3 kg (~2,700 ml) in one feed. If food is abundantly available after a period of fasting due to a disturbance, wild harbor porpoises could eat a large percentage of their daily energetic requirement in one feeding bout to compensate for the period of fasting. However, if food availability is limited in terms of prey numbers, size, or species, or if the fish are widely dispersed (so that more time is required to find and capture them), this may limit or reduce the speed of the recovery of body mass and blubber layer.

Factors affecting the degree of vertical stratification of fatty acids in grey seal blubber

The biochemistry of marine mammal blubber differs vertically from skin to muscle, which forms a challenge for using fatty acids (FAs) from differently sampled blubber as a proxy for dietary studies required for ecosystem-based management of coastal resources. In the blubber of some phocid seal individuals, the vertical stratification of several FAs is pronounced whereas in others the FAs distribute almost evenly through the blubber column. Using gas chromatography, we analysed the blubber vertical FA profiles of 30 adult male grey seals from the Baltic Sea, and examined which factors induced the largest vertical change of FA composition detected at the depth of 15–18 mm (outer and middle blubber boundaries). It was revealed that the degree of this compositional shift did not depend on the blubber thickness. Seal age only affected the vertical distribution of the FAs 16:0 and 16:1n-7. However, the outer blubber ratio of ∆9-desaturated monounsaturated FAs (MUFAs) to their saturated FA (SFA) precursors was not increased by grey seal age, contrasting earlier findings for ringed seals. A major determinant of the degree of FA stratification between the outer and middle blubber was the mismatch between the individually varying FA composition of the innermost blubber, regarded to reflect the dietary FA supply the most, and the uniform FA composition of endogenously regulated MUFA-rich outer blubber. Thus, discarding a fixed-depth layer of the grey seal outermost blubber, which we here show to span 0–18 mm from skin and which to a lesser extent reflects the diet of the individual, may in the case of small pinnipeds improve the sensitivity of the FA analysis in assessing spatial, temporal and individual dietary differences. When studying the outer blubber samples using only the diet-derived PUFA variables (SFAs and MUFAs omitted), the sensitivity of the analysis was better than when using this sample type with all main FA variables included.