Vitamin E and fatty acids in the grey seal (Halichoerus grypus)

Summary Vitamin E levels in serum, liver and blubber (subcutaneous adipose tissue) were determined for 66 male and female grey seals of varying age in the pupping colony on Sable Island in the Northwest Atlantic by high-performance liquid chromatography (HPLC). Fatty acid concentrations were determined for all blubber specimens. Adult males and pups had significantly higher levels of vitamin E and cholesterol in serum than females and juveniles. A close relationship between vitamin E and cholesterol in serum could be observed. Suckling pups had significantly higher levels of vitamin E in liver (191 mg·kg^–1) than juveniles and adults (21–41 mg·kg^–1). Levels of vitamin E in blubber showed an age-dependent increase, with the highest levels being found in adult males; overall, these levels were much lower than in man. Vitamin E levels in blubber and liver of lactating females were only half that of adult males. This might be due to an intensive transfer of vitamin E from mother to pup during lactation, a process which may also explain the much higher levels of vitamin E in serum and liver of nursing pups. The low levels of vitamin E in blubber of seals might be a result of its high percentage of unsaturated fatty acids (79%). Highest percentage was represented by 18:1, 16:1, 22:6 and 16:0. Pups had lower values of monounsaturated, and a higher percentage of saturated fatty acids compared to mothers.     

Serum Retinol,alpha‐tocopherol,and lipids in four species of adult captive pinnipeds

The sera of adult aquarium‐held pinnipeds from four species (family Phocidae: harbor seals (Phoca vitulina) and gray seals (Halichoerus grypus); family Otariidae: northern fur seals (Callorhinus ursinus) and California sea lions (Zalophus californianus)) were analyzed for vitamin A (retinol), vitamin E (α‐tocopherol), total cholesterol, triglycerides, phospholipids, and fatty acids. Each subject animal was healthy at the time of blood collection, was fasted for at least 12 hr prior to sampling, and was maintained on a constant diet and supplement regime throughout the study. Retinol values for the four species ranged from 0.16 to 0.92 μg/mL, with the lowest concentrations seen in the harbor seals and the highest in the northern fur seals. Vitamin E values ranged from 10.55 to 43.58 μg/mL, with northern fur seals showing the highest and gray seals the lowest levels. Vitamin E/lipid ratios (cholesterol, triglyceride, phospholipid, and total lipids) were also examined. A significant correlation was seen between vitamin E and total lipids (P<0.05) and phospholipid (P<0.01). Statistical analysis of the retinol, tocopherol, triglyceride, and phospholipid levels showed significant differences between phocid and otariid seals. Otariids had significantly lower tocopherol and phospholipid values (19.36 μg/mL, 4.29 mg/mL) and the phocids had significantly lower retinol and triglyceride levels (0.29 μg/mL, 124 mg/dL). There was no significant difference in serum cholesterol. Zoo Biol 22:83–96, 2003. © 2003 Wiley‐Liss, Inc.     

Pregnancy rate and biomarker validations from the blubber of eastern North Pacific blue whales

Hormonal biomarkers are useful indicators of mammalian reproductive and metabolic states. The present study validated and applied the use of progesterone and cortisol blubber assays for studies of blue whales from the Gulf of California, Mexico. In a validation study for pregnancy detection, blubber progesterone concentrations were correlated with pregnancy status for four female blue whales: three resighted with a calf the year following sampling and the fourth stranded with a fetus. The progesterone concentrations were significantly higher than those measured in juvenile whales (n = 3). In the application study, blubber samples from blue whales (51 noncalf females, 2 female calves, 48 noncalf males, and 1 male calf) with known sighting histories were analyzed. Putative pregnant females had elevated progesterone concentrations. Cortisol concentrations did not differ between male and female blue whales, or among females in different reproductive classes. After correcting for uncertain ages, hence maturity status, the pregnancy rate of noncalf females was 33.4% (95% CI 32.2%–34.3%). Although interpretation of hormone biomarkers must consider all physiological states that may influence progesterone concentrations, these results demonstrate the utility of pairing hormone biomarkers with sighting histories to help assess environmental or anthropogenic impacts on reproduction in blue whales.     

Distribution and development of the highly specialized lipids in the sound reception systems of dolphins

Fat bodies in the heads of toothed whales, which serve to transmit and receive sound, represent extraordinary examples of physiological specialization in adipose tissues among mammals, yet we know surprisingly little about their biochemical composition. We describe the spatial distributions and development of unusual endogenous lipids (branched-chain [“iso”] molecules and wax esters) in the mandibular fat bodies of bottlenose dolphins (Tursiops truncatus) using an ontogenetic series (fetus to adult; n = 10). Although concentrations of iso-acids, iso-alcohols and waxes were lower in younger dolphins than in adults, the same relative spatial arrangement was present in all age classes, implying a set “pattern” of acoustic lipid distribution that is established very early in life. In all age classes, a small region of blubber overlying the lateral region contained unusually high concentrations of iso-acids, exhibiting a tenfold increase over “normal” adjacent blubber. Being chemically more similar to the acoustic fat bodies, this region may serve as an entry point for sound into the head. Developmental accumulations of some iso-acids and iso-alcohols occurred more rapidly than others, implying that not only are the spatial distributions of branched-chain molecules under extremely fine-scale control, but the regulatory mechanisms controlling acoustic lipid synthesis are also highly complex.     

Flow injection methods for the determination of retinol and α‐tocopherol using lucigenin‐enhanced chemiluminescence

Flow injection (FI) methods are reported to determine retinol and α‐tocopherol based on its enhancement affect of lucigenin chemiluminescence (CL) in alkaline medium. Surfactants including Brij‐35, Triton X‐100, cetyltrimethyl ammonium bromide (CTAB) and sodium dodecyl sulfate have been reported for the first time to enhance lucigenin CL intensity in the presence of retinol and α‐tocopherol. With Brij‐35, the CL intensity was enhanced by 67% for retinol and 58% for α‐tocopherol. CTAB was found to enhance the CL intensity by 16% for retinol whereas for α‐tocopherol, the CL intensity was quenched up to 95%. Retinol could be determined specifically in the presence of α‐tocopherol using CTAB. The calibration graphs were found to be linear up to 1.43 mg/L (R² = 0.9985, n = 8) with a detection limit (3s) of 1.43 × 10^?3 mg/L for retinol and 2.15 mg/L (R² = 0.9989; n = 8) with a detection limit (3s) of 4.31 × 10^?4 mg/L for α‐tocopherol. An injection throughput of 120/h, and relative standard deviations of 0.9–2.8% (n = 4) were achieved in the concentration range studied. The influence of common ions, excipients in pharmaceutical formulations and related organic compounds on the determination of retinol and α‐tocopherol individually was studied. The proposed methods were applied to determine retinol and α‐tocopherol in pharmaceutical formulations and human blood serum. The results did not differ significantly from the CL method and HPLC reference method at 95% confidence level. Copyright © 2010 John Wiley & Sons, Ltd.     

Serum α- and γ-tocopherols, retinol, retinyl palmitate, and carotenoid concentrations in captive and free-ranging bottlenose dolphins (Tursiops truncatus)

Concentrations of retinol, retinyl palmitate, β-carotene, α-carotene, cryptoxanthin, lutein, lycopene, α-tocopherol, and γ-tocopherol were measured in blood samples collected from 15 captive and 55 free-ranging bottlenose dolphins (Tursiops truncatus). From June 1991 to June 1994, blood samples were collected from captive animals residing at two locations; at Seven Seas (Brookfield Zoo, Brookfield, IL) and Hawk’s Cay (Marathon Key, FL). Blood samples were collected from free-ranging animals from June 1991 to June 1996. Retinol levels were not significantly different between captive dolphin groups. However, Seven Seas animals had higher (P<0.01) serum retinol concentrations compared to free-ranging animals (0.061 vs 0.041 μg/ml). Retinyl palmitate was not detected in the serum of captive or free-ranging dolphins. Alpha-tocopherol levels were significantly (P<0.05) higher for Seven Seas dolphins (16.4 μg/ml) than for Hawk’s Cay (13.0 μg/ml) and free-ranging dolphins (12.5 μg/ml). Gamma-tocopherol concentrations were similar among captive and free-ranging dolphins. Free-ranging dolphins showed levels of circulating carotenoids (lutein and β-carotene) while the captive animals did not. Additional carotenoids (lycopene, α-carotene and cryptoxanthin) were analyzed but not detected in any samples. Serum vitamin differences between captive and free-ranging dolphins may reflect the natural diet or indicate some potential biological or nutritional status significance.     

Fatty acid composition of the blubber in white whales (Delphinapterus leucas)

Fatty acid (FA) composition of the blubber in free-ranging white whales (Delphinapterus leucas) from Svalbard's waters was determined and compared with the fatty acid composition of potential prey species in an attempt to assess diet. This methodology is based on the common assumption that unique arrays of FAs found within groups of organisms are transferred, largely unaltered, up marine food chains and thus may be useful for assessment of diet composition. Complete-column blubber biopsies were sampled from white whales (n=7) during the summers of 1996 and 1997. All captured animals were adult males. FAs were extracted from 2–4 replicates taken from an area about 10 cm in front of the mid-dorsal ridge. FA data from a total of 12 potential prey species from the Svalbard area were compared to the white-whale blubber samples. Twenty-two FAs were consistently found in relative amounts >0.5% of the total FA composition in white whales. These FAs accounted for 94–96% of the total FAs present. The blubber was composed almost entirely of triacylglycerols. The major saturated FAs were 14:0 and 16:0; 16:1(n-7), 18:1(n-9) and 20:1(n-9) were the major monounsaturated FAs and 20:5(n-3) and 22:6(n-3) were the major polyunsaturated FAs. Sixteen of the 22 FAs consistently found in the white-whale blubber were also found in considerable amounts (>0.5% of total FAs) in most of the potential species. Principal Component Analysis run on these 16 FAs suggests that polar cod (Boreogadus saida) had the most similar FA composition to the white-whale blubber, followed by capelin (Mallotus villosus), the copepod Calanus hyperboreus and the shrimp Pandalus borealis. Accepted: 27 November 1999     

Serum cytokines and muscle strength after anterior cruciate ligament surgery are not modulated by high-doses of vitamins E (α- and γ-tocopherol’s) and C

The purpose of this investigation was to identify if supplemental vitamin E (consisting of α- and γ-tocopherol’s) and C modulate serum cytokine and muscle strength following an ACL injury and surgery. Subjects were randomly assigned to one of two groups: (1) placebo (n = 14) or (2) vitamins E (α-[600 mg RRR-α-tocopherol, αT] and γ-[600 mg of RRR-γT]) and C (1000 mg ascorbic acid, AA) (EC; n = 15). Supplements were taken daily starting ∼2-wk prior to and concluding 16-wk after surgery. Fasting blood samples were obtained and single-leg peak isometric force measurements were performed at baseline (prior to supplementation), before surgery (∼120-min – blood draw only), and 8-wk, 12-wk, and 16-wk after surgery. αT, γT, AA, and cytokines were measured in each blood sample, and peak isometric force was measured on the injured and non-injured legs separately at each testing session. An exercise protocol consisting of repetitive knee and hip extension and flexion contractions to exhaustion was performed on the injured limb at 16-wk. Vitamin E and C supplementation significantly (all p < 0.05) increased plasma αT (∼40%), γT (∼160%), and AA (∼50%) concentrations. Serum cytokine concentrations, peak isometric force, and time to exhaustion during the exercise protocol were not significantly different between groups. Based on these findings, we conclude that vitamin E and C supplementation increases their endogenous levels without minimizing muscular weakness or modulating serum cytokine concentrations after ACL surgery.     

Vitamin A concentrations in serum and liver from Florida panthers

Many of the anomalies and clinical signs afflicting the Florida panther (Felis concolor coryi) are suggestive of vitamin A deficiency. Our objectives in this study were to determine if a vitamin A deficiency exists in the free-ranging panther population and to determine if there are differences in vitamin A levels among various subgroups of free-ranging panthers. Retinol concentrations were used as an index to Vitamin A concentrations and were determined in serum and liver from free-ranging (serum, n = 45; liver, n = 22) and captive (serum, n = 9; liver, n = 2) juvenile and adult Florida panthers from southern peninsular Florida (USA), and in liver from free-ranging cougars (F. concolor subspp.) from Washington (USA) and Texas (USA) between November 1984 and March 1994. Combined juvenile (6- to 24-mo-old) and adult (> 24-mo-old) free-ranging Florida panthers had mean +/- SD serum retinol concentrations of 772.5 +/- 229 pmol/ml. Adult free-ranging Florida panthers had mean liver retinol concentrations of 4794.5 +/- 3747 nmol/g. Free-ranging nursing Florida panther kittens (age < 1 mo) had mean serum retinol concentrations of 397.9 +/- 69 pmol/ml. Among subgroups of free-ranging Florida panthers, females had higher corrected mean serum retinol concentrations than males and adult free-ranging Florida panthers had higher mean liver retinol concentrations than juveniles. Retinol concentrations in free-ranging Florida panthers did not differ significantly from those in captive panthers (liver and serum) or other free-ranging cougars (liver). Based on limited published values and our controls, a vitamin A deficiency could not be demonstrated in the Florida panther population nor were any subgroups or individuals considered deficient.     

Effects of Vitamin A and Its Analogs on Nonenzymatic Lipid Peroxidation in Rat Brain Mitochondria

Vitamin A (retinol) and some of its analogs exhibited varying degrees of inhibition on induced iron and ascorbic acid lipid peroxidation of rat brain mitochondria. Malonyldialdehyde production was used as an index of the extent of in vitro lipid peroxidation. The fat-soluble vitamins retinol, retinol acetate, retinoic acid, retinol palmitate, and retinal at concentrations between 0.1 and 10.0 mmol/L inhibited brain lipid peroxidation. Retinol and retinol acetate were the most effective inhibitors. It is concluded from this study that retinol and its analogs can be considered as potential antioxidant factors, more potent than some of the well-known antioxidants such as alpha-tocopherol and butylated hydroxytoluene.     

Blood serum retinol levels in Asinara white donkeys reflect albinism‐induced metabolic adaptation to photoperiod at Mediterranean latitudes

Previous works on albinism form of Asinara white donkeys (Equus asinus) identified the mutation leading to the peculiar phenotype spread to all specimens of the breed. Inbreeding naturally occurred under geographic isolation, on Asinara Island, in the Mediterranean Sea. Albino individuals can be more susceptible to develop health problems when exposed to natural sun radiation. Alternative metabolic pathways involved in photoprotection were explored in this trial. Nutrition‐related metabolites are believed to contribute to the conservation of Asinara donkeys, in which melanin, guaranteeing photoprotection, is lacking. Biochemical profiles with particular focus on blood serum β‐carotene and retinol levels were monitored. Identical natural grazing conditions for both Asinara (albino) and Sardo (pigmented) donkey breeds were assured on same natural pastures throughout the experimental period. A comparative metabolic screening, with emphasis on circulating retinol and nutrient‐related metabolites between the two breeds, was carried out over one year. Potential intra‐ and interspecimen fluctuations of metabolites involved in photoprotection were monitored, both during negative and positive photoperiods. Differences (p = .064) between blood serum concentrations of retinol from Asinara versus Sardo breed donkeys (0.630 vs. 0.490 μg/ml, respectively) were found. Retinol levels of blood serum turned out to be similar in the two groups (0.523 vs. 0.493 μg/ml, respectively, p = .051) during the negative photoperiod, but markedly differed during the positive one (0.738 vs. 0.486, respectively, p = .016). Blood serum β‐carotene levels displayed to be constantly around the limit of sensitivity in all animals of both breeds. Variations in blood serum concentrations of retinol in Asinara white donkeys can reflect the need to cope with seasonal exposure to daylight at Mediterranean latitudes, as an alternative to the lack of melanin. These results may suggest that a pulsed mobilization of retinol from body stores occurs to increase circulating levels during positive photoperiod.     

The relationship between micronuclei in human lymphocytes and selected micronutrients in vegetarians and non-vegetarians

A vegetarian diet results in higher intake of vitamins and micronutrients, which – although providing antioxidant defence – may lead to deficiency in other micronutrients involved in DNA metabolism and stability (such as vitamins belonging to the B group). The principal difference among various vegetarian diets is the extent to which animal products are avoided. We have performed a pilot study to determine the relationship between the micronucleus frequency in lymphocytes and diet, and we compared the levels of Vitamins C and E, β-carotene, B₁₂, folic acid, homocysteine and total antioxidant capacity in healthy vegetarians and non-vegetarians. The vegetarian group, consisting of 24 volunteers (13 women and 11 men), were matched for age and sex with 24 volunteers (12 women and 12 men) with a traditional dietary habit. Among the vegetarians were 13 lacto-ovo-vegetarians with average duration of vegetarian diet 10.8 years (ranging from 5 to 26 years) and 11 lacto-vegetarians with average duration of vegetarian diet 8.2 years (ranging from 3 to 15 years). Homocysteine, Vitamins C and E and β-carotene levels in plasma were assayed by HPLC, and serum folate and Vitamin B₁₂ were determined with Elecsys Immunoassay tests. The total antioxidant capacity of plasma was estimated by measuring the ferric-reducing activity in a spectrophotometric assay. Micronuclei were measured in cytokinesis-blocked lymphocytes. Vegetarians had significantly higher levels of Vitamin C and β-carotene (but not Vitamin E) in plasma compared with non-vegetarians (P < 0.001). There were no significant differences in serum levels of folic acid and Vitamin B₁₂ between the monitored groups. Levels of folic acid in vegetarians correlated with length of vegetarianism (r = 0.62, P = 0.001, N = 24). Vegetarians had elevated levels of homocysteine compared with non-vegetarians (P = 0.007), as did vegetarian women compared with non-vegetarian women (P = 0.031). We did not find any differences in total antioxidant capacity or in micronucleus frequency between the groups. Micronuclei correlated with age (r = 0.62, P < 0.001, N = 48), women having higher frequencies than men. Multifactorial regression analysis showed significant effects of age, sex and total antioxidant capacity on micronucleus frequency (N = 48, P < 0.001).     

Evaluating morphometric and metabolic markers of body condition in a small cetacean,the harbor porpoise (Phocoena phocoena)

Mammalian body condition is an important individual fitness metric as it affects both survival and reproductive success. The ability to accurately measure condition has key implications for predicting individual and population health, and therefore monitoring the population‐level effects of changing environments. No consensus currently exists on the best measure to quantitatively estimate body condition in many species, including cetaceans. Here, two measures of body condition were investigated in the harbor porpoise (Phocoena phocoena). First, the most informative morphometric body condition index was identified. The mass/length² ratio was the most appropriate morphometric index of 10 indices tested, explaining 50% of the variation in condition in stranded, male porpoises with different causes of death and across age classes (n = 291). Mass/length² was then used to evaluate a second measure, blubber cortisol concentration, as a metabolic condition marker. Cortisol is the main glucocorticoid hormone involved in the regulation of lipolysis and overall energy balance in mammals, and concentrations could provide information on physiological state. Blubber cortisol concentrations did not significantly vary around the girth (n = 20), but there was significant vertical stratification through the blubber depth with highest concentrations in the innermost layer. Concentrations in the dorsal, outermost layer were representative of concentrations through the full blubber depth, showed variation by sex and age class, and were negatively correlated with mass/length². Using this species as a model for live cetaceans from which standard morphometric measurements cannot be taken, but from which blubber biopsy samples are routinely collected, cortisol concentrations in the dorsal, outermost blubber layer could potentially be used as a biomarker of condition in free‐ranging animals.     

Temporal variability in organochlorine contamination in blood and blubber of captive Steller sea lions (Eumetopias jubatus)

Three adult captive Steller sea lions (SSL; two females, one male) housed in Alaska were longitudinally sampled for organochlorine contaminant (OC) analysis in both blubber (n= 19) and blood (n= 69) over a 2 yr period (March 2001 to March 2003). Blood OC concentrations were similar between individuals, and exhibited significant increases in summer months (July through September) relative to winter (January through March). Additionally, paired blood and blubber sample (n= 18) OC were significantly related for all animals. The relationship of blubber OC concentrations to lipid content was significant in all animals. Although limited to a small number of animals, our study results indicate that in SSLs, blood OC were both consistent among all animals and likely changed in association with physiologically driven metabolism of blubber.     

Organochlorine contaminant levels in Eskimo harvested bowhead whales of arctic Alaska

Organochlorine (OC) levels in liver and blubber of 20 bowhead whales (Balaena mysticetus) collected during the Eskimo subsistence harvest at Barrow (Alaska, USA) in 1992 and 1993 are presented. Liver sum DDT (lipid weight) was significantly greater in male whales than in females. Most of the organochlorines measured were at higher levels in longer (older) than in shorter (younger) males. For female bowhead whales, hexachlorobenzene and lipid levels decreased and other OC levels did not change significantly with increasing length. Most organochlorine contaminants have low concentrations in tissues of the bowhead whale compared to concentrations in tissues of other cetaceans, especially Odontocetes. Based on allowable daily intakes (ADI) levels established by the Canadian Northern Contaminants Program (Ottawa, Ontario, Canada) "safe" levels of blubber to consume were calculated. Chlordane levels in bowhead whale blubber results in the most restrictive consumption amount (50 g blubber/day). We expect no adverse effects related to these organochlorine contaminants to occur in bowhead whales or in consumers of their tissues. However, investigation of low level chronic exposure effects and a more rigorous assessment of histopathology, biomarkers, and immune status in the bowhead whale would be required to conclude "no effect" with more certainty.     

Effects of Vitamin E and Selenium on Serum Trace and Major Elements in Horses

The combined effects of vitamin E and selenium were studied in native Anatolian horses subject to strenuous exercise. The concentrations of copper, zinc, iron, calcium, potassium, and magnesium were determined in serum by atomic absorption spectrometry in two study groups (n = 25 each), one of which served as untreated controls. After exercising the horses by running 1,500 m in about 7 min, only the copper level and the copper/zinc ratio significantly increased (p < 0.05), but the concentrations of calcium, potassium, iron, and magnesium remained unchanged. In horses treated with vitamin E and selenium, the calcium and potassium levels decreased to levels lower than those of untreated controls before and after exercise. The iron levels were not changed by exercise or treatment alone but increased when the horses had been supplemented and exercised. The copper level and the copper/zinc ration increased as a result of exercise in both treated and untreated horses. These changes suggest that supplementation with vitamin E and selenium had an important effect on the serum concentrations of calcium, potassium, copper, iron, and the copper/zinc ratio.     

Retinol and α-tocopherol concentrations in whole fish commonly fed in zoos and aquariums

Retinol (n = 17 spp.) and α-tocopherol (n = 9 spp.) concentrations in whole fish utilized for captive animal feeding programs were determined by high-performance liquid chromatography (HPLC) following routine storage and preparation after commercial purchase by two zoological institutions. Vitamin A activity was calculated from retinol values and ranged from 55 IU/100 g (immature herring) to >2,000 IU/100 g (salmon) on an as-fed basis. α-Tocopherol values, a measure of vitamin E activity, ranged from 0.9 IU/100 g (butterfish) to 12.3 IU/100 g (tilapia) on a wet basis. Vitamin levels in whole fish were intermediate to values previously quantified for muscle or liver tissues alone. Vitamin concentrations in fish livers were quantified separately in seven of these species; liver contributed 35–63% of total retinol measured and 8–34% of total α-tocopherol. Based on these analyses, whole fish commonly fed in zoos, aquariums, and marine zoological parks would appear to meet vitamin A requirements established for most species without additional supplementation, whereas levels of vitamin E quantified indicate a need for supplementation of diets for piscivores.     

DETERMINATION OF PREGNANCY STATUS FROM BLUBBER SAMPLES IN MINKE WHALES (BALAENOPTERA ACUTOROSTRATA)

Measurement of progesterone concentration in blubber was developed as a method to detect pregnancy in minke whales. Progesterone was extracted and quantified from blubber samples of minke whale carcasses by radioimmunoassay. Results showed a highly significant difference (almost 60-fold) between blubber progesterone concentrations of anatomically determined pregnant females versus non-pregnant female or male carcasses. The results of the study suggest that the blubber progesterone concentrations might be used to determine pregnancy status in free-ranging whales.     

Blue whale population structure along the eastern South Pacific Ocean: evidence of more than one population

Blue whales (Balaenoptera musculus) were among the most intensively exploited species of whales in the world. As a consequence of this intense exploitation, blue whale sightings off the coast of Chile were uncommon by the end of the 20th century. In 2004, a feeding and nursing ground was reported in southern Chile (SCh). With the aim to investigate the genetic identity and relationship of these Chilean blue whales to those in other Southern Hemisphere areas, 60 biopsy samples were collected from blue whales in SCh between 2003 and 2009. These samples were genotyped at seven microsatellite loci and the mitochondrial control region was sequenced, allowing us to identify 52 individuals. To investigate the genetic identity of this suspected remnant population, we compared these 52 individuals to blue whales from Antarctica (ANT, n = 96), Northern Chile (NCh, n = 19) and the eastern tropical Pacific (ETP, n = 31). No significant differentiation in haplotype frequencies (mtDNA) or among genotypes (nDNA) was found between SCh, NCh and ETP, while significant differences were found between those three areas and Antarctica for both the mitochondrial and microsatellite analyses. Our results suggest at least two breeding population units or subspecies exist, which is also supported by other lines of evidence such as morphometrics and acoustics. The lack of differences detected between SCh/NCh/ETP areas supports the hypothesis that eastern South Pacific blue whales are using the ETP area as a possible breeding area. Considering the small population sizes previously reported for the SCh area, additional conservation measures and monitoring of this population should be developed and prioritized.     

Stable carbon and nitrogen isotope ratios in muscle and epidermis of arctic whales

Collection of minimally invasive biopsy samples has become an important method to establish normal stable isotopes reference ranges in various wildlife species. Baseline data enhance the understanding of feeding ecology, habitat use, and potential food limitation in apparently healthy, free‐ranging cetaceans. Epidermis and muscle were collected from subsistence‐hunted northern Alaskan bowhead (n= 133 epidermis/134 muscle) and beluga whales (n= 42/49) and subsistence‐hunted Russian gray whales (n= 25/17). Additional samples were obtained from gray whales stranded in California (n= 18/11) during mortality events (1999, 2000). Both δ¹⁵N and δ¹³C are trophic position and benthic/pelagic feeding indicators, respectively, in muscle and epidermis. Epidermis is generally enriched in ¹⁵N over muscle, while epidermal ¹³C is more depleted. Lipid extraction does not alter δ¹⁵N in either tissue, but affects epidermal δ¹³C. Nitrogen‐15 is enriched in muscle, but not epidermis of stranded compared to subsistence‐hunted gray whales, indicating probable protein catabolism and nutritional stress in stranded whales. Similarly, epidermal δ¹³C of harvested whales is lower than in stranded whales, suggesting depleted lipid stores and/or food limitation in stranded animals. Epidermal isotope signatures are similar in both present‐day bowheads and in an ancient sample from the Northern Bering Sea region. Although only one specimen, this suggests trophic level of the ancient whale compares to modern bowheads after a millennium.