Comparison of analytical and predictive methods for water, protein, fat, sugar, and gross energy in marine mammal milk.

Mammalian milks may differ greatly in composition from cow milk, and these differences may affect the performance of analytical methods. High-fat, high-protein milks with a preponderance of oligosaccharides, such as those produced by many marine mammals, present a particular challenge. We compared the performance of several methods against reference procedures using Weddell seal (Leptonychotes weddellii) milk of highly varied composition (by reference methods: 27-63% water, 24-62% fat, 8-12% crude protein, 0.5-1.8% sugar). A microdrying step preparatory to carbon-hydrogen-nitrogen (CHN) gas analysis slightly underestimated water content and had a higher repeatability relative standard deviation (RSDr) than did reference oven drying at 100°C. Compared with a reference macro-Kjeldahl protein procedure, the CHN (or Dumas) combustion method had a somewhat higher RSDr (1.56 vs. 0.60%) but correlation between methods was high (0.992), means were not different (CHN: 17.2±0.46% dry matter basis; Kjeldahl 17.3±0.49% dry matter basis), there were no significant proportional or constant errors, and predictive performance was high. A carbon stoichiometric procedure based on CHN analysis failed to adequately predict fat (reference: Röse-Gottlieb method) or total sugar (reference: phenol-sulfuric acid method). Gross energy content, calculated from energetic factors and results from reference methods for fat, protein, and total sugar, accurately predicted gross energy as measured by bomb calorimetry. We conclude that the CHN (Dumas) combustion method and calculation of gross energy are acceptable analytical approaches for marine mammal milk, but fat and sugar require separate analysis by appropriate analytic methods and cannot be adequately estimated by carbon stoichiometry. Some other alternative methods-low-temperature drying for water determination; Bradford, Lowry, and biuret methods for protein; the Folch and the Bligh and Dyer methods for fat; and enzymatic and reducing sugar methods for total sugar-appear likely to produce substantial error in marine mammal milks. It is important that alternative analytical methods be properly validated against a reference method before being used, especially for mammalian milks that differ greatly from cow milk in analyte characteristics and concentrations.

The evolution of milk secretion and its ancient origins.

Lactation represents an important element of the life history strategies of all mammals, whether monotreme, marsupial, or eutherian. Milk originated as a glandular skin secretion in synapsids (the lineage ancestral to mammals), perhaps as early as the Pennsylvanian period, that is, approximately 310 million years ago (mya). Early synapsids laid eggs with parchment-like shells intolerant of desiccation and apparently dependent on glandular skin secretions for moisture. Mammary glands probably evolved from apocrine-like glands that combined multiple modes of secretion and developed in association with hair follicles. Comparative analyses of the evolutionary origin of milk constituents support a scenario in which these secretions evolved into a nutrient-rich milk long before mammals arose. A variety of antimicrobial and secretory constituents were co-opted into novel roles related to nutrition of the young. Secretory calcium-binding phosphoproteins may originally have had a role in calcium delivery to eggs; however, by evolving into large, complex casein micelles, they took on an important role in transport of amino acids, calcium and phosphorus. Several proteins involved in immunity, including an ancestral butyrophilin and xanthine oxidoreductase, were incorporated into a novel membrane-bound lipid droplet (the milk fat globule) that became a primary mode of energy transfer. An ancestral c-lysozyme lost its lytic functions in favor of a role as α-lactalbumin, which modifies a galactosyltransferase to recognize glucose as an acceptor, leading to the synthesis of novel milk sugars, of which free oligosaccharides may have predated free lactose. An ancestral lipocalin and an ancestral whey acidic protein four-disulphide core protein apparently lost their original transport and antimicrobial functions when they became the whey proteins ß-lactoglobulin and whey acidic protein, which with α-lactalbumin provide limiting sulfur amino acids to the young. By the late Triassic period (ca 210 mya), mammaliaforms (mammalian ancestors) were endothermic (requiring fluid to replace incubatory water losses of eggs), very small in size (making large eggs impossible), and had rapid growth and limited tooth replacement (indicating delayed onset of feeding and reliance on milk). Thus, milk had already supplanted egg yolk as the primary nutrient source, and by the Jurassic period (ca 170 mya) vitellogenin genes were being lost. All primary milk constituents evolved before the appearance of mammals, and some constituents may have origins that predate the split of the synapsids from sauropsids (the lineage leading to 'reptiles' and birds). Thus, the modern dairy industry is built upon a very old foundation, the cornerstones of which were laid even before dinosaurs ruled the earth in the Jurassic and Cretaceous periods.

Effect of lactation stage and concurrent pregnancy on milk composition in the bottlenose dolphin.

ALTHOUGH MANY TOOTHED WHALES (CETACEA: Odontoceti) lactate for 2-3 years or more, it is not known whether milk composition is affected by lactation stage in any odontocete species. We collected 64 pooled milk samples spanning 1-30 months postpartum from three captive bottlenose dolphins Tursiops truncatus. Milks were assayed for water, fat, crude protein (TN × 6.38) and sugar; gross energy was calculated. Ovulation and pregnancy were determined via monitoring of milk progesterone. Based on analysis of changes in milk composition for each individual dolphin, there were significant increases (P<0.05) in fat (in all three dolphins) and crude protein (in two of three), and a decrease (P<0.05) in water (in two of three) over the course of lactation, but the sugar content did not change. In all three animals, the energy content was positively correlated with month of lactation, but the percentage of energy provided by crude protein declined slightly but significantly (P<0.05). At mid-lactation (7-12 months postpartum, n=17), milk averaged 73.0±1.0% water, 12.8±1.0% fat, 8.9±0.5% crude protein, 1.0±0.1% sugar, 1.76±0.09 kcal g(-1) (=7.25 kJ g(-1)) and 30.3±1.3% protein:energy per cent. This protein:energy per cent was surprisingly high compared with other cetaceans and in relation to the growth rates of calves. Milk progesterone indicated that dolphins ovulated and conceived between 413 and 673 days postpartum, following an increase in milk energy density. The significance of these observed compositional changes to calf nutrition will depend on the amounts of milk produced at different stages of lactation, and how milk composition and yield are influenced by sampling procedure, maternal diet and maternal condition, none of which are known.

Interspecific and intraspecific variation in proximate, mineral, and fatty acid composition of milk in Old World fruit bats (Chiroptera: Pteropodidae).

We examine the effect of body mass on milk composition among Old World fruit bats, including Pteropus pumilus (0.175 kg), Pteropus rodricensus (0.265 kg), Pteropus hypomelanus (0.571 kg), and Pteropus vampyrus (1.133 kg). We describe intra- and interspecific differences in the proximate composition of milk among these four species and the minerals and fatty acids in the milk of the latter two species. There were no differences between species in the concentrations of dry matter, fat, or lactose in milk. However, there were significant, although small, differences in the protein content of milk among species, with protein being significantly greater in P. rodricensus than in P. pumilus and P. hypomelanus and protein being significantly less in P. hypomelanus than in P. rodricensus and P. vampyrus. There were no differences in mineral content between P. hypomelanus and P. vampyrus in milk minerals, but minor differences were evident in fatty acids 12:0, 14:0, 18:0, 18:1n11, and 18:2n6. Our findings suggest that milk composition is relatively constant across lactation for most proximate, mineral, and fatty acid components. We found a significant increase in dry matter and energy across lactation in the concentration of dry matter and energy in P. pumilus and fat in P. hypomelanus. In P. hypomelanus, we found a significant increase in the concentration of fatty acids 10:0 and 20:1n9 and a significant decrease in Iso15 and 20:1n7. No other differences associated with day of lactation were found. These findings suggest that milk composition is generally similar within the genus Pteropus, despite a 6.5-fold difference in body mass between species that we evaluated.

Relations among measures of body composition, age, and sex in the common marmoset monkey (Callithrix jacchus).

Few studies of body composition have been done in New World primates. In the study reported here, four methods of assessing body composition (body weight, anthropometry, labeled-water dilution, and total body electroconductivity) were compared in 20 marmosets, aged 0.96 to 7.97 years. Males and females did not differ in any measure (P > 0.05). Body weight ranged from 272 to 466 g, and body fat estimates varied from 1.6 to 19.5%. Strong positive correlations were observed between total body water and total body electroconductivity (R2 = 0.77), body weight and fat-free mass (males R2 = 0.95; females R2 = 0.91), and body weight and fat mass (males R2 = 0.86; females R2 = 0.85; P < 0.01). Male and female slopes were equivalent (P > 0.05) for the regressions of fat and fat-free mass against body weight. Positive correlations also were observed between girth measures and fat-free mass (R2 = 0.48 to 0.78) and fat mass (R2 = 0.60 to 0.74; P < 0.01). A good second- order polynomial relationship was observed between age and fat-free mass for the combined sample (R2 = 0.64). Results indicated that: subjects were lean; there was no sexual dimorphism relative to measures; body weight provided a reliable estimate of fat and fat-free mass; and within-subject body weight changes reflected a similar relationship between body weight and fat-free mass as did that across subjects.

Comparison of oligosaccharides in milk specimens from humans and twelve other species.

Human milk contains large amounts of many oligosaccharides, most of which are fucosylated; several inhibit pathogenic bacteria, viruses, and toxins that cause disease in humans. Although bovine milk is known to have much less and many fewer types of oligosaccharides, no studies heretofore have indicated whether the amount or complexity of human milk oligosaccharides is unique to our species. Toward this end, a comparison was made of the major individual oligosaccharides in milk specimens from a variety of species, including the great apes. The neutral compounds, which represent the bulk of oligosaccharides in human milk, were isolated, perbenzoylated, resolved by high performance liquid chromatography (HPLC), and detected at 229nm. Ambiguous structures were determined by mass spectrometry. All milk specimens contained lactose, although levels were quite low in bear and kangaroo milk. The types of oligosaccharides in milk specimens from the primates resembled those of human milk, but the amounts, especially of the larger molecules, were markedly lower. The relative amounts of oligosaccharides in the bonobo changed over the course of lactation, as they do in humans. Marine mammals generally had few oligosaccharides in their milk other than 2'-fucosyllactose. Grizzly and black bear milk specimens contained a wide range of oligosaccharides, many of which had novel, fucosylated structures. Milk specimens from humans, bears, and marsupials had the greatest quantity of, and the most complex, neutral oligosaccharides. Although human milk contained more oligosaccharide than did milk specimens from the other species studied, the presence of appreciable amounts of complex oligosaccharides was not unique to humans. This finding suggests that in animal milk specimens, as in human milk, neutral fucosylated oligosaccharides potentially offer protection from pathogens to offspring with immature immune systems.

Use of maternal reserves as a lactation strategy in large mammals.

The substrate demands of lactation must be met by increased dietary intake or by mobilization of nutrients from tissues. The capacity of animals to rely on stored nutrients depends to a large extent on body size; large animals have greater stores, relative to the demands of lactation, than do small animals. The substrate demands of lactation depend on the composition and amount of milk produced. Animals that fast or feed little during lactation are expected to produce milks low in sugar but high in fat, in order to minimize needs for gluconeogenesis while sustaining energy transfers to the young. The patterns of nutrient transfer are reviewed for four taxonomic groups that fast during part of or throughout lactation: sea lions and fur seals (Carnivora: Otariidae), bears (Carnivora: Ursidae), true seals (Carnivora: Phocidae) and baleen whales (Cetacea: Mysticeti). All these groups produce low-sugar high-fat milks, although the length of lactation, rate of milk production and growth of the young are variable. Milk protein concentrations also tend to be low, if considered in relation to milk energy content. Maternal reserves are heavily exploited for milk production in these taxa. The amounts of lipid transferred to the young represent about one-fifth to one-third of maternal lipid stores; the relative amount of the gross energy of the body transferred in the milk is similar. Some seals and bears also transfer up to 16-18 % of the maternal body protein via milk. Reliance on maternal reserves has allowed some large mammals to give birth and lactate at sites and times far removed from food resources.

Oscars, Astronotus ocellatus, have a dietary requirement for vitamin C.

We found that vitamin C is an essential nutrient for an Amazonian ornamental fish, the oscar (Astronotus ocellatus). This was demonstrated by the absence of L-gulonolactone oxidase activity, the enzyme responsible for the biosynthesis of vitamin C, in liver or kidney of oscars and by a feeding trial in which oscars without vitamin C dietary supplementation developed clinical deficiency signs. Fish weighing 29.2 +/- 1.9 g were divided into four groups, and each group was fed a casein-based semipurified diet containing 0, 25, 75 or 200 mg ascorbic acid equivalent (AA)/kg diet for 26 wk. Vitamin C was supplemented in the diets as L-ascorbyl-2-polyphosphate, a mixture of phosphate esters of ascorbate, which is more stable to oxidation than AA. At the end of 26 wk, fish fed no AA had significantly lower weight gain than fish fed the AA-supplemented diets (P < 0.05). Oscars without dietary AA supplementation gained only 37% of their initial weight, compared with 112, 102 and 91% gained by fish fed 25, 75 and 200 mg AA/kg diet, respectively. After 25 wk without dietary supplementation of AA, fish began to develop clinical deficiency signs, including deformed opercula and jaws, hemorrhage in the eyes and fins, and lordosis. Histology indicated that fish without AA supplementation had deformed gill filament support cartilage and atrophied muscle fibers. Collagen content of the vertebral column was significantly lower in fish devoid of dietary AA (P < 0.05). Liver AA concentration varied in proportion to dietary concentration of AA. The minimum dietary AA concentration tested in this study, 25 mg AA/kg diet, was sufficient to prevent growth reduction and AA deficiency signs in oscars.

Dietary fiber influences nutrient utilization, growth and dry matter intake of green iguanas (Iguana iguana).

Herbivory is an uncommon feeding strategy in lizards. Appropriate diet formulations for captive lizards should be based on performance measures, yet few data are available on the effect of plant fiber on food intake, nutrient utilization and growth of captive herbivorous lizards. This study was conducted to determine the effect of three levels of dietary fiber on dry matter intake, nutrient and energy metabolizability and growth rate of the green iguana (Iguana iguana). Twenty-one captive iguanas were fed nutritionally complete diets containing three levels of dietary fiber: 19, 24, and 27% neutral detergent fiber. The iguanas were fed each diet for at least 12 wk, and total excreta were collected for 11.3 +/- 4.0 d (means +/- , range of 7 to 25 d). Diets and excreta were analyzed for dry matter, organic matter, gross energy, neutral detergent fiber, acid detergent fiber, and acid detergent lignin. The study was designed as a Latin square crossover. Across all diets, dry matter intake was proportional to body mass1.0 (BM). Growth rate was greater (P < 0. 05) when iguanas were fed the low and medium fiber diets (2.2 and 2. 4 g/d, respectively) than when fed the high fiber diet (1.4 g/d). However, mean daily dry matter intake of the three diets [7.2 g/(d. kg BM)] was not different. In general, digestibility of fiber fractions and the metabolizability of dietary energy decreased (P < 0.05) as the level of dietary fiber increased. These data suggest that a diet containing less than 27% neutral detergent fiber should be fed if rapid growth is to be sustained during intensive captive production of green iguanas.

Milk composition and lactational output in the greater spear-nosed bat, phyllostomus hastatus.

Growth rates of mammalian young are closely linked to the ability of the mother to provide nutrients; thus, milk composition and yield provide a direct measure of maternal investment during lactation in many mammals. We studied changes in milk composition and output throughout lactation in a free-ranging population of the omnivorous bat, Phyllostomus hastatus. Fat and dry matter of milk increased from 9 to 21% and from 21 to 35% of wet mass, respectively, throughout lactation. Energy increased from 6 to 9 kJ.g-1 wet mass, primarily due to the increase in fat concentration. Total sugar levels decreased slightly but non-significantly. Mean sugar level was 4.0% of wet mass. Protein concentration increased from 6 to 11% of wet mass at peak lactation and then decreased as pups approached weaning age. Total milk energy output until pups began to forage was 3609 kJ. Milk levels of Mg, Fe, Ca, K, and Na averaged 0.55 %/- 0.26, 0.23 +/- 0.2, 8.75 +/- 4.17, 5.42 +/- 2.11, and 9.87 +/- 4.3 mg.g-1 dry matter, respectively. Of the minerals studied, calcium appears to be most limiting in this species. The high degree of variability in foraging time, milk composition and milk yield between individuals at the same stage of lactation could potentially yield high variance in reproductive success among females of this polygynous species.

Lactation in whales and dolphins: evidence of divergence between baleen- and toothed-species.

Although it has been more than one hundred years since the first publication on the milks of whales and dolphins (Order Cetacea), information on lactation in these species is scattered and fragmentary. Yet the immense size of some cetaceans, and the recent evidence that another group of marine mammals, the true seals, have remarkable rates of secretion of milk fat and energy, make this group of great comparative interest. In this paper information on lactation patterns, milk composition and lactation performance is reviewed. Two very different patterns are evident. Many of the baleen whales (Suborder Mysticeti) have relatively brief lactations (5-7 months) during which they fast or eat relatively little. At mid-lactation they produce milks relatively low in water (40-53%), high in fat (30-50%), and moderately high in protein (9-15%) and ash (1.2-2.1%). From mammary gland weights and postnatal growth rates, it is predicted that their energy outputs in milk are exceptional, reaching on the order of 4000 MJ/ d in the blue whale. This is possible because pregnant females migrate to feeding grounds where they can ingest and deposit great amounts of energy, building up blubber stores prior to parturition. On the other hand, the toothed whales and dolphins (Suborder Odontoceti) have much more extensive lactations typically lasting 1-3 years, during which the mothers feed. At mid-lactation their milks appear to be higher in water (60-77%) and lower in fat (10-30%) and ash (0.6-1.1%), with similar levels of protein (8-11%). At least some odontocetes resemble primates in terms of low predicted rates of energy output and a long period of dependency of the young. However, these hypotheses are based on small numbers of samples for a relatively small number of species. Much of the available data on milk composition is of rather poor quality; for example, it is not possible to determine if milk composition changes over the course of lactation among odontocetes. Additional research on cetacean mammary glands and their secretions is needed to understand the reproductive strategies of these fascinating animals.

Milk composition reflects pattern of maternal care in prosimian primates.

Hypothesized relationships between milk composition and life history traits were examined by analyzing mid-lactation milks of seven lemurs (Eulemur fulvus, E. macaco, E. rubriventer, E. mongoz, Varecia variegata, Hapalemur griseus, Lemur catta), three bushbabies (Otolemur crassicaudatus, O. garnettii, Galago moholi), and two lorises (Nycticebus coucang, Loris tardigradus); partial data were also obtained for the lemuroid Cheirogaleus medius. There were no significant differences in milk composition among species within either Eulemur or Otolemur, but the four genera for which multiple samples were available (Eulemur, Varecia, Otolemur, and Nycticebus) exhibited large composition differences. Eulemur milk was, on average, very dilute (9.9% dry matter) and low in energy (0.49 kcal/g). These milks contained 0.9% fat, 1.2% protein, and 8.4% carbohydrate on a fresh weight basis. Protein energy comprised only about 15% of total milk energy. Varecia had significantly higher dry matter (13.5%), fat (3.2%), protein (4.2%), gross energy (0.80 kcal/g), and protein energy:total energy ratio (28%) than Eulemur. Milks of the lorisoid genera Otolemur and Nycticebus were very similar, and both had significantly higher dry matter (18.3, 16.3%), fat (7.6, 7.0%), and gross energy concentration (1.27, 1.13 kcal/g) than either lemuroid genus. Otolemur milk was higher in protein than Nycticebus milk. We conclude that lorises, bushbabies, and perhaps cheirogaleids produce relatively rich, energy-dense milks in comparison with anthropoid primates. However, dilute milks appear to be uniformly found among species of Eulemur and perhaps in Lemur catta. The milk of Varecia (and perhaps Hapalemur) is intermediate in composition. Differences in milk composition among prosimians may be related to differences in maternal care: prosimians that carry their young during lactation produce more dilute milks than do species which leave their young unattended for prolonged periods. When looking at primates as a whole, however, the picture somewhat less clear, since the milks of some "parkers" like Varecia do closely resemble those of large anthropoid primates who carry their young.

Changes in milk composition during lactation in three species of insectivorous bats.

Changes in milk composition are described for three species of free-ranging insectivorous bats (Myotis lucifugus, M. velifer, and Tadarida brasiliensis) from early to mid (peak) lactation. Dry matter and energy concentrations in milk increased from early to mid-lactation. In M. lucifugus and T. brasiliensis, but not M. velifer, these increases were due largely to a rise in fat concentration, since protein and carbohydrate remained relatively constant. Energy content of milk (kJ.g-1) for each species from early through mid-lactation was related to dry matter (DM) as follows: M. lucifugus (y = 0.31 DM-0.32, r2 = 0.68), M. velifer (y = 0.48 DM-5.08, r2 = 0.99), and T. brasiliensis (y = 0.37 DM-1.51, r2 = 0.61). Comparison of the effect of sampling method on milk composition of T. brasiliensis indicated that fat, dry matter, and energy concentrations increased significantly from pre-dawn to pre-noon samples. Relatively high fat and low water levels in T. brasiliensis milk may reflect the limited access that lactating females have to free water, as well as need to minimize mass of stored milk during long foraging trips. Conversely, lower fat concentrations and higher water levels in milk in M. lucifugus and M. velifer may relate to the propensity for colonies of these two species to roost and forage near bodies of water. In addition, differences in milk fat concentrations observed among the three species may correlate to daily suckling schedules.(ABSTRACT TRUNCATED AT 250 WORDS)

Prenatal and postnatal transfer of fatty acids from mother to pup in the hooded seal.

Unlike most mammals, hooded seal (Cystophora cristata) pups are born with a substantial layer of adipose tissue. Subsequently, during the brief lactation period of only 4 days, fasting mothers mobilize enormous amounts of lipid from blubber and secrete milk (60% fat) at rates of 10 kg.day-1. Pups gain 7 kg.day-1 due primarily to the deposition of fat in blubber. We measured blubber content and fatty acid composition of blubber and milk in hooded seal mother-pup pairs at birth and over the 4-day lactation period to examine the nature and source of fetal lipids, the incorporation of maternal blubber fatty acids into milk lipid, and patterns of fatty acid deposition in suckling young. The fatty acid composition of the blubber of the newborn was notably different from that of its mother. Fetal deposition was likely due to a combination of both fetal synthesis and direct placental transfer of maternal circulating fatty acids. The blubber of the newborn was characterized by high levels (> 90% of total fatty acids) of saturated and monounsaturated fatty acids of primarily endogenous origin. In particular, the fetus appeared to have high delta-9 desaturase activity as evidenced by the large amounts of 14:1n-5 (4.2%) and 16:1n-7 (37.0%) in newborn blubber compared to maternal blubber (0.2% and 14.1%, respectively). Nevertheless, essential and long-chain polyunsaturated fatty acids of the n-3 and n-6 families, which could only have originated by direct transfer from the mother, comprised > 7% of pup blubber fatty acids and indicated greater rates of placental transfer than found in humans. In hooded seal mothers, rapid lipid transfer during the brief lactation period appeared to be facilitated by direct incorporation of mobilized fatty acids into milk. Although some differences in proportions of specific fatty acids were found between milk and maternal blubber, most of these differences declined over the course of lactation. However, selective mobilization of 20:5n-3 from maternal blubber into milk was apparent throughout lactation and resulted in elevated levels in pup blubber at weaning compared to maternal blubber. Ingested fatty acids were deposited directly and without modification into the blubber of pups, and by 4 days the fatty acid composition of pup blubber was virtually identical to that of the milk consumed.

A cross-species comparison of neutral lipid composition of milk fat of prosimian primates.

The fatty acid composition of milk fat is known to be affected by dietary and genetic differences, while the milk triacylglycerol structure is believed to be attuned to the needs of the subsequent lipolysis during gastrointestinal passage. The availability of milk samples from eight species of prosimian primates, whose milk triacylglycerol structure had not been analyzed, offered an opportunity to further assess these ideas. The milk samples were collected by manual expression and the lipids extracted with chloroform/methanol (2:1, vol/vol). The lipid classes were resolved by thin-layer chromatography, and the neutral lipids subjected to detailed analyses by capillary gas-liquid chromatography of fatty acids and molecular species of triacylglycerols using nonpolar and polarizable liquid phases. The milk samples were found to differ greatly in total fat content (4-73%) and in the composition of the neutral lipid classes and molecular species. The concentration of triacylglycerols ranged from 88-95%, free fatty acids from 0.5-10%, alkyldiacylglycerols from 0.5-5.0%, and diacylglycerols, monoacylglycerols and free and esterified cholesterol made up the remainder. The fatty acid chain length ranged from C8-C24, with palmitic (16-31%) and oleic (13-40%) acids being the major components in most of the species. In all instances, the molecular association of the fatty acids differed from random distribution by a higher proportion of the monoacid (trioleoyl) and diacid (dipalmitoyloleoyl) glycerols. The phylogenetic influences on neutral milk lipid composition, however, remained unclear, as some of the differences between closely related species were greater than those between more distantly related ones.

The adaptation of milk secretion to the constraints of fasting in bears, seals, and baleen whales.

Although lactation is accompanied by increased nutrient demands for milk synthesis, many species of bears, true seals, and baleen whales fast for much or all of lactation. Large body mass in these species confers the advantage of greater stores of fat and protein relative to rates of milk production. Given the constraints on substrate availability during fasting, the milks of fasting mammals are predicted to be low in carbohydrate, protein, and water and to be high in fat. The milks of bears, true seals, and baleen whales conform to this prediction. Mammals that lactate while fasting may lose up to 40% of initial BW. The production of milk entails the export of up to one-third of body fat and 15% of body protein in the dormant black bear and in several seal species, which greatly depletes maternal resources and may represent a physiological threshold, because higher protein and fat outputs have only been measured in species that start feeding. The low K:Na ratio of seal and whale milks and the low Ca:casein and inverse Ca:P ratios in seal milks are unusual and warrant further study.

Nutrition and growth of suckling black bears (Ursus americanus) during their mothers' winter fast.

In black bears the last 6-8 weeks of gestation and the first 10-12 weeks of lactation occur in winter while the mother is in a dormant state, and reportedly does not eat, drink, urinate or defaecate. Measurements were made of the body composition and organ weights of cubs, of the composition of milk, and of milk intake (by dilution of 2H2O), in the first 3 months after birth. Additional milk samples were collected until 10 months postpartum. Bear cubs were small at birth, only 3.7 g/kg maternal weight, and chemically immature, as indicated by the high concentration of water (840 g/kg) in their bodies. Organ weights at birth were similar to those of puppies. In the first month after birth cubs gained 22 g/d or 0.23 g/g milk consumed; the milk was high in fat (220 g/kg) and low in water (670 g/kg). About 30% of the ingested energy and 51% of the ingested N were retained in the body. Over the entire 12-week period bear cubs required about 11 kg milk, containing (kg) water 7, fat 2.5, protein 0.8 and total sugar 0.25, to achieve a 2.5 kg weight gain. The birth of immature young and the production of high-fat, low-carbohydrate milk seem to be maternal adaptations to limit the utilization of glucogenic substrates during a long fast. Isotope recycling indicates that mothers may also recover most of the water (and perhaps much of the N) exported in milk by ingesting the excreta of the cubs. Lactation represents another aspect of the profound metabolic economy of the fasting bear in its winter den.

Positional specificity of gastric hydrolysis of long-chain n-3 polyunsaturated fatty acids of seal milk triglycerides.

Long-chain n-3 polyunsaturated fatty acids (n-3 PUFA) of marine oils are important dietary components for both infants and adults, and are incorporated into milks following maternal dietary intake. However, little is known about the hydrolysis of these PUFA from milk triglycerides (TG) by lipases in suckling young. Seals, like humans, possess gastric lipase; however, the milk lipids of seals and sea lions are almost devoid of the readily hydrolyzable medium-chain fatty acids, and are characterized by a large percentage (10-30%) of n-3 PUFA. Gastric hydrolysis of milk lipids was studied in vivo in suckling pups of three species (the California sea lion, the harp seal and the hooded seal) in order to elucidate the actions and specificity of gastric lipases on milk TG in relation to fatty acid composition and TG structure. Regardless of milk fat content (31-61% fat) or extent of gastric hydrolysis (10-56%), the same fatty acids were preferentially released in all three species, as determined by their relative enrichment in the free fatty acid (FFA) fraction. In addition to 16:1 and 18:0, these were the PUFA of 18 carbons and longer, except for 22:6n-3. Levels of 20:5n-3 were most notably enriched in FFA, at up to five times that found in the TG. Although 22:6n-3 was apparently also released from the TG (reduced in the diglyceride), it was also notably reduced in FFA.(ABSTRACT TRUNCATED AT 250 WORDS)

Fatty acid composition of black bear (Ursus americanus) milk during and after the period of winter dormancy.

Black bears give birth and lactate during the 2-3-mon fast of winter dormancy. Thereafter the female emerges from the den with her cubs and begins to feed. We investigated fatty acid patterns of milk from native Pennsylvania black bears during the period of winter dormancy, as well as after den emergence. Throughout winter dormancy, milk fatty acid composition remained relatively constant. The principal fatty acids at all times were 14:0, 16:0, 16:1, 18:0, 18:1, 18:2n-6, 18:3n-3 and 20:4n-6. After den emergence, large changes occurred in almost all the fatty acids, particularly in 18:2n-6 and 18:3n-3. Large variability among the active free-ranging animals likely reflected differences in diet. In a carnivore, with apparently limited de novo synthesis of fatty acids, milk fatty acid composition may be affected by factors such as transition from reliance on stored lipids to feeding, and by temporal changes in dietary intake.

Fatty-acyl esters of retinol (vitamin A) in the liver of the harp seal (Phoca groenlandica), hooded seal (Cystophora cristata), and California sea lion (Zalophus californianus).

The fatty-acid composition of retinyl esters in the livers of two species of phocid seal, the harp seal (Phoca groenlandica, n = 20) and the hooded seal (Cystophora cristata, n = 15), and one species of otariid seal, the California sea lion (Zalophus californianus, n = 6), was determined. Vitamin A ranged in concentration from 4 to 1024 nmol retinol/g liver for the phocids and from 381 to 979 nmol/g liver for the otariids. In most of the livers, retinyl palmitate was not the principal ester, and the palmitate + stearate + oleate trio of retinyl esters represented less than 50% of the total. In all samples, the retinyl esters contained 20:1, 20:4, 20:5, and 22:6 in unusually large amounts. Retinyl esters tended to be richer than whole-liver lipids in 20:5 + 22:6, whereas whole-liver lipids were richer in 18:0 and 18:2. Therefore, the pool of acyl donors used for the esterification of retinol may be distinct from that used for other lipids. Birth-to-weaning changes were seen only in the harp seals. In the pups, the hepatic vitamin-A concentration increased 454%, while the proportion of 18:0 and 20:1 in the retinyl esters rose and that of 14:0 + 16:1 and 20:4 fell. Concomitantly, in their mothers, the proportion of 20:4 increased but that of 16:0 and 18:0 decreased.