Stratification, composition, and function of marine mammal blubber: the ecology of fatty acids in marine mammals.

Abstract This study of vertical fatty acid profiles, based on analysis of 58 fatty acids sampled at 3-mm intervals throughout the blubber column of a model marine mammal, the ringed seal (Pusa hispida), revealed three chemically distinct layers. The average depths of the outer and inner layers were quite consistent (approximately 1.5 and approximately 1 cm, respectively). Consequently, the middle layer varied greatly in thickness, from being virtually absent in the thinnest animals to 2.5 cm thick in the fattest. The relative consistencies of the thickness and composition of the layers as well as the nature of the fatty acids making up each layer support the generally assumed function of the various layers: (1) the outer layer is primarily structural and thermoregulatory, (2) the inner layer is metabolically active with a fatty acid composition that is strongly affected by recent/ongoing lipid mobilization/deposition, and (3) the middle layer is a storage site that contracts and expands with food availability/consumption. The remarkable dynamics of the middle layer along with the discrete pattern of stratification found in the vertical fatty acid profiles have important implications for methodological sampling design for studies of foraging ecology and toxicology based on analyses of blubber of marine mammals.

Lipid metabolism in the adipose tissues of a carnivore, the raccoon dog, during prolonged fasting.

Previous studies on laboratory rodents, rabbits, and humans have demonstrated that adipose tissue fatty acid (FA) mobilization is selective, and its efficiency is related to the molecular structure of FAs. This study was undertaken to find out whether such preferences of FA mobilization are a general feature of mammalian white adipose tissue (WAT) and are also manifested in carnivores. Fractional mobilization of a wide spectrum of FAs was studied by gas-liquid chromatography from six subcutaneous (scapular, rump, ventral) and intra-abdominal (omental, mesenteric, retroperitoneal) WAT depots of raccoon dogs (Nyctereutes procyonoides) fed or fasted for 2 months. Fasting stimulated the mobilization of shorter-chain saturated, mono-unsaturated (MUFAs), and polyunsaturated FAs (PUFAs). The effects of unsaturation and the position of the first double bond from the methyl end were more inconsistent. The effect of double-bond position may be due to chain shortening of longer-chain MUFAs and preferential utilization of n-3 PUFAs over n-6 PUFAs. Moreover, there were site-specific differences in fractional mobilization, the omental adipose tissue being the most divergent. The in vivo FA mobilization from the regional WAT depots of a carnivore was selective, and the molecular structure of the FA affected its efficiency.

Importance of the kidneys in metabolism of vitamins A1 and A2 and their fatty acyl esters in mink feeding on fish-based diets and exposed to Aroclor 1242.

Two-month-old female mink (Mustela vison) were fed diets based on freshwater smelt (Osmerus eperlanus), Baltic herring (Clupea harengus membras), or North-Atlantic herring (Clupea harengus harengus) for 21 weeks. Half of the smelt-fed mink were exposed to the commercial polychlorinated biphenyl (PCB) preparation Aroclor 1242 (1 mg/day) in the feed. Retinol (vitamin A(1)), 3,4-didehydroretinol (vitamin A(2)), their fatty acyl esters, and vitamin E were studied in the kidneys by reversed-phase high-performance liquid chromatography. Exposure to Aroclor 1242 resulted a significant decrease in the alcoholic and esterified vitamin A(2). Levels of the A(1) analogs did not change due to the PCBs and were the same in mink fed either smelt or Baltic herring. In mink fed very fatty Atlantic herring, the renal levels of vitamin A(1) and E were significantly lower than in the other mink and apparently consumed by lipid peroxidation. The vitamins were located mainly in the cortex, and the fatty acyl esters showed a fatty acid composition that differed from those in liver and plasma. In the kidneys of the smelt-fed mink (control or Aroclor-exposed) the ratio of vitamin A(2) to A(1) was much lower than the ratios in the liver or plasma, suggesting inefficient uptake of A(2) in the kidneys. In the PCB-exposed mink, in which vitamin losses are increased, tissue levels of vitamin A(2) may be more difficult to maintain than levels of vitamin A(1).

Effects of Aroclor 1242 and different fish-based diets on vitamins A1 (retinol) and A2 (3,4-didehydroretinol), and their fatty acyl esters in mink plasma.

The effects of a 21-week exposure to Aroclor 1242 (1mg per day in feed) on plasma concentrations of vitamins A(1) (retinol) and A(2) (3,4-didehydroretinol) and their principal fatty acyl esters (A(1)-16:0, A(2)-16:0 (palmitates), A(1)-18:1n-9; A(2)-18:1n-9 (oleates), and A(1)-18:0; A(2)-18:0 (stearates)) were studied in young female mink (Mustela vison) fed a diet based on freshwater smelt. These vitamin levels were also examined in mink fed diets containing Baltic herring or fatty marine fish. In the Aroclor-exposed smelt-fed mink, the plasma concentrations of A(1) and A(2) esters were significantly lower than the levels in controls fed the uncontaminated smelt diet. In addition, the A(2) esters reacted more sensitively to the polychlorinated biphenyls than did A(1) esters. In contrast, in the plasma of the exposed mink the level of alcoholic A(1) was normal, and transport of thyroxine (T(4)) and nonspecific lipoprotein transport of major lipids were not impaired. Despite the large dietary supply of vitamin A(2) and high levels of plasma A(2) esters, the mink fed freshwater smelt had only trace amounts of alcoholic vitamin A(2) in their plasma. The concentrations of A(1) and A(2) esters in the plasma of all the mink studied correlated with the hepatic total concentrations of the vitamins. Thus, in carnivores that have nonspecific lipoprotein transport of vitamin A esters, determination of plasma levels of the esterified vitamins may be a useful nondestructive way to estimate stores of the vitamin A analogs in the body and to assess the organochlorine-induced decrease in the vitamin stores.

Effects of gender, diet, exogenous melatonin and subchronic PCB exposure on plasma immunoglobulin G in mink.

Effects of different fish-based diets (freshwater smelt, Baltic herring, marine herring/cod offal or their mixtures), gender, beta-glucan supplement, exogenous melatonin, and PCB exposure (Aroclor 1242((R)), 1 mg per animal per day in feed) on plasma immunoglobulin G (IgG) in the mink (Mustela vison) were studied. The aims of the study were to find out whether plasma IgG of the mink is affected by the subchronic PCB exposure, and whether biological, nutritional and hormonal effects are large enough to mask the possible IgG response. The concentration of IgG was determined using enzyme-linked immunosorbent assay (ELISA). Sexual dimorphism was detected, the males having higher levels of plasma IgG. In addition, melatonin tended to decrease IgG in females but not males. Diet also affected the humoral immune arm; the mixed-fish diets caused an unfavorable ratio of the oxidation products of lipids vs. vitamin E in liver, and resulted in low IgG concentration in plasma. In males fed Baltic herring, the beta-glucan supplement also lowered IgG levels. The PCBs failed to affect the plasma IgG of the smelt-fed female mink, and IgG concentration was not correlated with increased hepatic EROD activity or with the decreased total retinol in the liver of exposed mink. It is concluded that hormonal/seasonal and dietary factors affect the plasma IgG levels to such an extent that possible change in plasma IgG level due to PCBs in wild populations of mink is difficult to detect without a large amount of reference data.

Analysis of hepatic vitamins A1, A2, their fatty acyl esters, and vitamin E for biomonitoring mammals feeding on freshwater fish.

In tissues of freshwater fish-feeding mammals, 3,4-didehydroretinol (A2) is a major form of vitamin A. In mink liver, with organochlorine exposure, this analog has been found to decrease more than retinol (A1) and thus has potential as a sensitive freshwater biomarker. The presence of the analogs A1 and A2 as alcohol and different fatty acyl esters, which react to polychlorinated biphenyls differently, necessitates detailed analyses achieved by using direct extraction of tissue homogenate. In direct hexane extraction, compared to total levels of the vitamins obtained in the saponification procedure, a large proportion of the vitamins was released only after repeated and long-time vortex mixing with the extraction solvent. Thus, in tissue extraction, the use of internal standardization alone can lead to a rough underestimation of the levels of these fat-soluble vitamins. For analyses of vitamins A1 and A2 in liver, we applied the argentation high-performance liquid chromatography, which provided good separation of individual A1 and A2 fatty acyl esters. We report retention times for numerous esters of A1 and A2 and, to aid identification, the change in their retention properties after adding AgNO3 to the mobile phase. The argentation did not affect the recoveries of any forms of the retinoids studied but destroyed half the vitamin E. Despite selective acylation of fatty acids into the vitamin A esters, the fatty acids of the esters were the same as those found to be the major fatty acids in the gas-liquid chromatography of total lipids. The goal of this work was to create a methodology that is suitable for biomonitoring alcoholic and esterified vitamins A1 and A2 in tissues of freshwater fish-feeding mammals.

Vitamins A1 and A2 in hepatic tissue and subcellular fractions in mink feeding on fish-based diets and exposed to Aroclor 1242.

Two-month-old female mink were fed diets based on either Baltic herring (Clupea harengus membras) or freshwater smelt (Osmerus eperlanus) for 21 weeks. A portion of the smelt-fed mink were exposed orally to polychlorinated biphenyls (PCBs), Aroclor 1242 (1 mg/d). Retinol (vitamin A1), 3,4-didehydroretinol (vitamin A2), and their different fatty acyl esters were studied in hepatic tissue, microsomes, and cytosol by argentated reversed-phase high-performance liquid chromatography. As a result of Aroclor exposure, concentrations of the fatty acyl esters of vitamins A1 and A2 were about one-tenth and those of unesterified A2 one-fourth those of the control levels. In the fatty acyl esters, percentages of stearates (A1--18:0 and A2--18:0) increased at the expense of the other fatty acyl esters. The Aroclor exposure decreased concentrations of alcoholic and esterified forms of the A2 analog more than those of the corresponding A1 analog. In microsomes. Aroclor decreased the alcoholic and esterified vitamin analogs to the same extent (to 9-17%). In the cytosol compared to the control, the concentrations of the vitamin esters fell below 10%, but the alcoholic analogs remained at 30 to 40%. Despite equal dietary supply, in mink fed on Baltic herring, the hepatic levels of vitamin A, were only about one-third of the values found in the smelt-fed mink. The organochlorines also altered hepatic lipid composition and impaired breeding and kit growth. In the kits of the females fed on Baltic herring, blood hemoglobin was decreased.