Brazilian Kefir-Fermented Sheep’s Milk,a Source of Antimicrobial and Antioxidant Peptides

Fermented milks are a source of bioactive peptides and may be considered as functional foods. Among these, sheep’s milk fermented with kefir has not been widely studied and its most relevant properties need to be more thoroughly characterized. This research study is set out to investigate and evaluate the antioxidant and antimicrobial properties of peptides from fermented sheep’s milk in Brazil when produced by using kefir. For this, the chemical and microbiological composition of the sheep’s milk before and after the fermentation was evaluated. The changes in the fermented milk and the peptides extracted before the fermentation and in the fermented milk during its shelf life were verified. The antimicrobial and antioxidant activities of the peptides from the fermented milk were evaluated and identified according to the literature. The physicochemical properties and mineral profile of the fermented milk were like those of fresh milk. The peptide extract presented antimicrobial activity and it was detected that 13 of the 46 peptides were able to inhibit the growth of pathogenic microorganisms. A high antioxidant activity was observed in the peptides extracted from fermented milk (3.125 mg/mL) on the 28th day of storage. Two fractions displayed efficient radical scavenging properties by DPPH and ABTS methods. At least 11 peptides distributed in the different fractions were identified by tandem mass spectrometry. This sheep’s milk fermented by Brazilian kefir grains, which has antioxidant and antimicrobial activities and probiotic microorganisms, is a good candidate for further investigation as a source for bioactive peptides. The fermentation process was thus a means by which to produce potential bioactive peptides.     

Kefir micro-organisms: their role in grain assembly and health properties of fermented milk

Kefir is a homemade viscous and slightly effervescent beverage obtained by milk fermentation with kefir grains, which are built up by a complex community of lactic acid and acetic acid bacteria and yeasts confined in a matrix of proteins and polysaccharides. The present review summarizes the role of kefir micro-organisms in grain assembly and in the beneficial properties attributed to kefir. The use of both culture-dependent and independent methods has made possible to determine the micro-organisms that constitute this ecosystem. Kefir consumption has been associated with a wide range of functional and probiotic properties that could be attributed to the micro-organisms present in kefir and/or to the metabolites synthetized by them during milk fermentation. In this context, the role of micro-organisms in kefir health promoting properties is discussed with particular attention to the contribution of yeast as well as bioactive metabolites such as lactic and acetic acid, exopolysaccharides and bioactive peptides. Even though many advances on the knowledge of this ancient fermented milk have been made, further studies are necessary to elucidate the complex nature of the kefir ecosystem.     

Kefir: a symbiotic yeasts-bacteria community with alleged healthy capabilities

Kefir is a fermented milk beverage. The milk fermentation is achieved by the of kefir grains, a cluster of microorganisms held together by a polysaccharide matrix named kefiran. Kefir grains are an example of symbiosis between yeast and bacteria. They have been used over years to produce kefir, a fermented beverage that is consumed all over the world, although its origin is Caucasian. A vast variety of different species of organisms forming the kefir grains, comprising yeast and bacteria, have been isolated and identified. Kefir is a probiotic food. Probiotics have shown to be beneficial to health, being presently of great interest to the food industry. Kefir has been accredited with antibacterial, antifungal and antitumoural activities among other beneficial attributes. This review includes a critical revision of the microbiological composition of kefir along with its beneficial properties to human health.     

Fatty acid composition and biogenic amines in acidified and fermented fish silage: a comparison study

In the presented study, ensiling of discard fish by acidification or fermentation was evaluated. Klunzinger’s ponyfish which is a discard fish was used for the production of fish silage by acidification (3% formic acid for Method FA; 1.5% formic and 1.5% sulphuric acid for Method FASA) and fermentation (Lactobacillus plantarum for Method LP and Streptococcus thermophilus for Method ST). The chemical, microbiological and nutritional properties of the differently preserved fish silages were estimated during a storage period of 60 d at ambient temperature. Compared to the raw material, a slight increase in saturated fatty acids and a slight decrease in polyunsaturated fatty acids were observed in all silages. At the end of the storage period, the aerobic bacteria counts after applying Methods FA, FASA, LP and ST amounted to 2.35, 2.39, 5.77 and 5.43 log cfu/g, respectively. The analysis of thiobarbituric acid revealed that acidification of silages accelerated the lipid oxidation. Nine biogenic amines were found in raw fish and different silages. The initial histamine concentration in raw fish was 0.17 mg/100 g and in all silages it remained at low levels during the storage period. The initial tyramine content was found to be 1.56 mg/100 g in raw fish and increased significantly in all silages. The increase of the tyramine content in fermented silages was considerably higher than in acidified silages (23–48 mg/100 g and 5–10 mg/100 g, respectively). It can be concluded that acidified or fermented fish silage should be considered as potential feed component for animals because of its high nutritional value and appropriate microbiological and chemical quality.     

Microbial Species Diversity,Community Dynamics,and Metabolite Kinetics of Water Kefir Fermentation

Water kefir is a sour, alcoholic, and fruity fermented beverage of which the fermentation is started with water kefir grains. These water kefir grains consist of polysaccharide and contain the microorganisms responsible for the water kefir fermentation. In this work, a water kefir fermentation process was followed as a function of time during 192 h to unravel the community dynamics, the species diversity, and the kinetics of substrate consumption and metabolite production. The majority of the water kefir ecosystem was found to be present on the water kefir grains. The most important microbial species present were Lactobacillus casei/paracasei, Lactobacillus harbinensis, Lactobacillus hilgardii, Bifidobacterium psychraerophilum/crudilactis, Saccharomyces cerevisiae, and Dekkera bruxellensis. The microbial species diversities in the water kefir liquor and on the water kefir grains were similar and remained stable during the whole fermentation process. The major substrate, sucrose, was completely converted after 24 h of fermentation, which coincided with the production of the major part of the water kefir grain polysaccharide. The main metabolites of the fermentation were ethanol and lactic acid. Glycerol, acetic acid, and mannitol were produced in low concentrations. The major part of these metabolites was produced during the first 72 h of fermentation, during which the pH decreased from 4.26 to 3.45. The most prevalent volatile aroma compounds were ethyl acetate, isoamyl acetate, ethyl hexanoate, ethyl octanoate, and ethyl decanoate, which might be of significance with respect to the aroma of the end product.     

Survival of Escherichia coli O157:H7, Listeria monocytogenes 4b and Yersinia enterocolitica O3 in different yogurt and kefir combinations as prefermentation contaminant

AIMS: To compare microbiological safety of yogurt, kefir and different combinations of yogurt and kefir samples by using three foodborne pathogenic strains (Escherichia coli O157:H7, Listeria monocytogenes 4b and Yersinia enterocolitica O3) as indicators. METHODS AND RESULTS: Fresh yogurt and kefir drinks were added to pasteurized milk at a 5% rate either separately or together, and then incubated at different temperatures (43 degrees C for yogurt and 30 degrees C for kefir), depending on appropriate growth temperature of their starter microflora. While traditional yogurt was found to be the least suppressive on the three pathogenic micro-organisms, samples obtained from two subsequent fermentation process (samples fermented at 43 degrees C for 3 h and at 30 degrees C for 21 h) were more suppressive than that of traditional kefir. There was no significant survival difference between E. coli O157:H7 and L. monocytogenes 4b in samples tested (P > 0.05), but Y. enterocolitica O3 was more susceptible than other two test strains (P < 0.05). CONCLUSIONS: The microbiological safety of the dairy product fermented at two consecutive periods was superior than that of traditional yogurt or kefir alone. SIGNIFICANCE AND IMPACT OF THE STUDY: These experiments may mimic what happens when yogurt and kefir starter micro-organisms are combined in a milk fermentation process with different time and temperature periods.     

Influence of pumpkin seed cake and extruded linseed on milk production and milk fatty acid profile in Alpine goats

The aim was to determine the effect of substituting pumpkin seed cake (PSC) or extruded linseed (ELS) for soya bean meal in goats’ diets on milk yield, milk composition and fatty acids profile of milk fat. In total, 28 dairy goats were divided into three groups. They were fed with concentrate mixtures containing soya bean meal (Control; n=9), ELS (n=10) or PSC (n=9) as main protein sources in the trial lasting 75 days. Addition of ELS or PSC did not influence milk yield and milk gross composition in contrast to fatty acid profile compared with Control. Supplementation of ELS resulted in greater branched-chain fatty acids (BCFA) and total n-3 fatty acids compared with Control and PSC (P<0.05). Total n-3 fatty acids were accompanied by increased α-linolenic acid (ALA, C18:3n-3; 0.56 g/100 g fatty acids) and EPA (C20:5n-3; 0.12 g/100 g fatty acids) proportions in milk of the ELS group. In contrast, ELS and PSC resulted in lower linoleic acid (LA, C18:2n-6; 2.10 and 2.28 g/100 g fatty acids, respectively) proportions compared with Control (2.80 g/100 g fatty acids; P<0.05). Abovementioned resulted in lower LA/ALA ratio (3.81 v. 7.44 or 6.92, respectively; P<0.05) with supplementation of ELS compared with Control or PSC. The PSC diet decreased total n-6 fatty acids compared with the Control (2.96 v. 3.54 g/100 g fatty acids, P<0.05). Oleic acid (c9-C18:1), CLA (c9,t11-18:2) and t10-,t11-C18:1 did not differ between treatments (P⩾0.08), although stearic acid (C18:0) increased in ELS diets compared with Control (12.7 v. 10.2 g/100 g fatty acids, P<0.05). Partially substituted soya bean meal with ELS in hay-based diets may increase beneficial n-3 fatty acids and BCFA accompanied by lowering LA/ALA ratio and increased C18:0. Pumpkin seed cake completely substituted soya bean meal in the diet of dairy goats without any decrease in milk production or sharp changes in fatty acid profile that may have a commercial or a human health relevancy.     

Kefir production in Iran

Kefir grains were prepared in a goat-hide bag using pasteurized milk inoculated with sheep intestinal flora, followed by culture of the surface layer in milk. From the grain, 11 strains of lactic acid bacteria, non-lactic acid bacteria and yeasts were isolated and identified. Six samples of kefir were prepared by fermenting pasteurized milk for different lengths of time. Sensory evaluation identified the sample prepared by 24 h fermentation as the best product.     

Use of isolated kefir starter cultures in kefir production

Kefir is a beverage produced by lactic-alcoholic fermentation of milk using kefir grain. For the first time in Iran, the microbial flora of kefir grain was isolated and identified (Motaghi et al. 1997). In this paper various ratios of starter cultures of kefir grains were investigated. Various ratios of lactic acid bacteria, yeasts and acetic acid bacteria were tested and the quality (colour, smell, flavour, acidity, effervescence and viscosity) of the product was assessed. At constant incubation time and temperature (24 h, 25 °C using homogenised milk with 2.5% fat), samples with various ratios of starter culture (3–5% w/v) were examined and analysed for protein, fat, sugar, alcohol, carbon dioxide, acidity, density, and riboflavin content. Samples produced with 3% (v/v) bacterial mixed culture and 2% (v/v) yeast (K3 procedure) culture were considered as best with respect to quality and organoleptic quality. The comparison of the results with the organoleptic tests of previous studies showed that the kefir produced with kefir grain is more desirable as compared with kefir produced with starter cultures. This revised version was published online in July 2006 with corrections to the Cover Date.     

Kefir ameliorates hypertension via gut–brain mechanisms in spontaneously hypertensive rats

Hypertension is associated with gut dysbiosis and dysregulation of the gut–brain axis. Previous work has shown that probiotic treatments exert beneficial cardiovascular effects in humans and animal models of hypertension. Coupled with the evidence of elevated sympathetic outflow and chronic inflammation in hypertension, we hypothesized that both peripherally and centrally mediated mechanisms underlie the antihypertensive effects of kefir, a probiotic obtained from the fermentation of milk by kefir grains. Eight-week-old spontaneously hypertensive rats (SHRs) were treated by oral gavage with either vehicle or kefir (0.3 ml/100 g/day; 9 weeks; SHR-Kefir), and age-matched with vehicle-treated Wistar Kyoto rats (WKY). Long-term kefir treatment attenuated mean arterial pressure elevations in SHR-Kefir relative to vehicle-treated SHRs. Peripherally, SHRs exhibited differences in the wall of the jejunum (fewer Paneth cells per crypt of Lieberkϋhn and increased tunica muscularis thickness) and higher serum lipopolysaccharide levels compared to WKY, alterations which were reversed in SHR-Kefir. Centrally, kefir treatment reduced IL-6 and TNF-α protein densities, and abolished the microglial activation observed in the hypothalamic paraventricular nucleus and rostral ventrolateral medulla of SHRs. Taken together, our findings indicate that the antihypertensive effects of long-term kefir treatment occur, at least in part, through improved structural and functional integrity of the intestinal wall and protection against neuroinflammation within cardioregulatory nuclei.     

Changes in lipid, fatty acids and phospholipids composition of whole rice bran after solid-state fungal fermentation

The aim of this study was to evaluate fermented rice bran phospholipids, lipids and fatty acid content in a fermentation solid system with Rhizopus oryzae fungus. For this, aliquots were withdrawn every 24h over 120 h. The content of phospholipids was determined by colorimetric method. Esterified fatty acids were separated by gas chromatography, then identified and quantified. The total lipids from fermented rice bran (FB) decreased from 20.4% to 11.2% in the range between 0 h and 120 h of fermentation while phospholipid contents were increased up to 2.4 mg P g(lipid)(-1). In fermented bran, oleic, palmitic and linoleic acids prevailed, with a decrease in saturated fatty acids (20%) and increase in the unsaturated ones (5%). This study showed that rice bran fermentation with R. oryzae can be applied to the production of phospholipids altering the saturated to unsaturated fatty acid ratio.     

配糟对清香型白酒发酵中环境因子及微生物群落的影响

【背景】环境因子是影响微生物生长代谢的重要因素,解析半开放条件下酿造过程中环境因子对微生物群落演替的作用对于清香型白酒生产调控具有重要意义。配糟在白酒发酵过程中起着调节发酵速度的作用,其对微生物群落组成变化的影响尚不明确。【目的】揭示使用不同发酵周期配糟对清香型白酒发酵过程中环境因子及微生物群落演替的影响。【方法】采用PacBio测序平台和多元统计分析比较使用2种配糟酒醅中微生物群落结构组成,结合蒙特卡洛置换检验明确环境因子对微生物群落的影响。【结果】与使用正常发酵周期配糟酒醅相比,使用延长发酵周期配糟酒醅水分较低,而酸度、氨基酸态氮、总游离氨基酸、还原糖和残余淀粉较高;微生物多样性和丰富度分析发现,使用延长发酵周期配糟酒醅中细菌α多样性极显著高于使用正常发酵周期配糟酒醅(P<0.001),而真菌α多样性显著/极显著低于使用正常发酵周期配糟酒醅(P<0.05, P<0.001);通过组间差异性分析发现,细菌群落共产生28个差异指示种,而真菌群落共产生15个差异指示种;水分、酸度、氨基酸态氮、还原糖、残余淀粉和总游离氨基酸对微生物群落结构的影响显著(P<0.05)...     

Transfer of fatty acids from female seal blubber via milk to pup blubber

Fatty acids were determined by gas chromatography followed by principal component analysis (PCA) and soft independent modelling of class analogy (SIMCA) in the blubber of 18 female grey seals, Halichoerus grypus, in their milk and in the blubber of their 1-week-old nursing pups. Large individual differences were observed in both blubber and milk content of fatty acids. The fatty acid composition of the milk was systematically different from the composition in maternal blubber, with higher relative amounts of the saturated acids, the monounsaturated with 20 carbon atoms and the n3 polyunsaturated, except 18:3n3. The composition of the fatty acids in the blubber of the pups was different from that of the milk. The same fatty acids that were enriched in the milk were depleted in the blubber of the pups. Therefore the fatty acid composition in the blubber of the pups was similar to that in the adults, although not identical. The results from this investigation imply that the composition of the fatty acids in the blubber of female seals and in the blubber of their pups cannot be determined directly by analysis of fatty acid composition of milk. Accepted: 16 February 2000     

Nutritional status and intermediate chain-length fatty acids influence the conservation of essential fatty acids in the milk of northern Nigerian women

The milk of 89 women in northern Nigeria was analyzed for the fatty acid composition of the total milk lipids, and assessed for the effect nutritional status has on the conservation of essential and non-essential fatty acids when the proportions of C(10)-C(14)fatty acids are increased. The women were stratified on the basis of their body mass index, and calculations were made to estimate the effects of a 3.3-fold increase in the proportion of C(10)-C(14)fatty acids on the proportion of alpha-linolenic acid, docosahexaenoic acid, linoleic acid and arachidonic in total milk lipids. In the well-nourished group (group III, body mass index >23 kg/m(2)), the critical n-3 and n-6 fatty acids were not conserved, while in poorly nourished women (group I, body mass index <19 kg/m(2)), marked conservation of alpha-linolenic acid, docosahexaenoic acid, arachidonic acid, and palmitic acid was seen. Poor nutritional status of the mother appears to promote selective retention of critical essential and non-essential fatty acids in the milk lipid fraction.     

Dietary influence of kefir on microbial activities in the mouse bowel

AIMS: In this work the microflora present in kefir, a fermented milk product, was studied together with the effect of kefir administration on different groups of indigenous bacteria of mouse bowel. METHODS AND RESULTS: Kefir microflora was composed of lactic acid bacteria, acetic acid bacteria and yeasts. Yeast population was composed of Saccharomyces cerevisiae, S. unisporus, Candida kefir, Kluyveromyces marxianus and K. lactis. The streptococci levels in kefir treated mice increased by 10-fold and the levels of sulfite-reducing clostridia decreased by 100-fold. The number of lactic acid bacteria increased significantly. CONCLUSIONS: The administration of kefir significantly increased the lactic acid bacteria counts in the mucosa of the bowel. Ingestion of kefir specifically lowered microbial populations of Enterobacteriaceae and clostridia. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first long-term study about the effects of the kefir administration on the intestinal microflora of mice.     

Effect of replacing calcium salts of palm oil distillate with rapeseed oil, milled or whole rapeseeds on milk fatty-acid composition in cows fed maize silage-based diets

Inclusion of rapeseed feeds in dairy cow diets has the potential to reduce milk fat saturated fatty acid (SFA) and increase cis-monounsaturated fatty acid (cis-MUFA) content, but effectiveness may depend on the form in which the rapeseed is presented. Four mid-lactation Holstein dairy cows were allocated to four maize silage-based dietary treatments according to a 4 × 4 Latin Square design, with 28-day experimental periods. Treatments consisted of a control diet (C) containing 49 g/kg dry matter (DM) of calcium salts of palm oil distillate (CPO), or 49 g/kg DM of oil supplied as whole rapeseeds (WR), rapeseeds milled with wheat (MR) or rapeseed oil (RO). Replacing CPO with rapeseed feeds had no effect (P > 0.05) on milk fat and protein content, while milk yields were higher (P < 0.05) for RO and MR compared with WR (37.1, 38.1 and 34.3 kg/day, respectively). Substituting CPO with RO or MR reduced (P < 0.05) milk fat total SFA content (69.6, 55.6, 71.7 and 61.5 g/100 g fatty acids for C, RO, WR and MR, respectively) and enhanced (P < 0.05) milk cis-9 18:1 MUFA concentrations (corresponding values 18.6, 24.3, 17.0 and 23.0 g/100 g fatty acids) compared with C and WR. Treatments RO and MR also increased (P < 0.05) milk trans-MUFA content (4.4, 6.8, 10.5 g/100 g fatty acids, C, MR and RO, respectively). A lack of significant changes in milk fat composition when replacing CPO with WR suggests limited bioavailability of fatty acids in intact rapeseeds. In conclusion, replacing a commercial palm oil-based fat supplement in the diet with milled rapeseeds or rapeseed oil represented an effective strategy to alter milk fatty acid composition with the potential to improve human health. Inclusion of processed rapeseeds offered a good compromise for reducing milk SFA and increasing cis-MUFA, whilst minimising milk trans-MUFA and negative effects on animal performance.     

Effect of frozen storage on fatty acid composition of the different tissues of four scombrid and one dussumeriid species

In the current study, the effect of frozen storage at ?18°C was evaluated on fatty acid composition of different body parts (liver, muscle tissue, and viscera) of narrow‐barred Spanish mackerel (Scomberomorus commerson, Lacépède, 1800), longtail tuna (Thunnus tonggol, Bleeker, 1851), kawakawa (Euthynnus affinis, Cantor, 1849), king mackerel (Scomberomorus guttatus, Bloch & Schneider, 1801), and rainbow sardine (Dussumieria acuta, Valenciennes, 1847) caught in the Persian Gulf. Changes in saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid plus docosahexaenoic acid/palmitic acid (EPA+DHA/C16), ω3 PUFA/ω6 PUFA (ω3/ω6), and polyunsaturated fatty acids/saturated fatty acids (PUFA/SFA) were investigated during a 6‐month period. A decrease in unsaturated fatty acids, particularly PUFAs (60–100%) as well as ω3/ω6, EPA+DHA/C16 (polyene index) and PUFA/SFA ratios, indicated a decrease in the nutritional values of the samples.     

Sequencing-Based Analysis of the Bacterial and Fungal Composition of Kefir Grains and Milks from Multiple Sources

Kefir is a fermented milk-based beverage to which a number of health-promoting properties have been attributed. The microbes responsible for the fermentation of milk to produce kefir consist of a complex association of bacteria and yeasts, bound within a polysaccharide matrix, known as the kefir grain. The consistency of this microbial population, and that present in the resultant beverage, has been the subject of a number of previous, almost exclusively culture-based, studies which have indicated differences depending on geographical location and culture conditions. However, culture-based identification studies are limited by virtue of only detecting species with the ability to grow on the specific medium used and thus culture-independent, molecular-based techniques offer the potential for a more comprehensive analysis of such communities. Here we describe a detailed investigation of the microbial population, both bacterial and fungal, of kefir, using high-throughput sequencing to analyse 25 kefir milks and associated grains sourced from 8 geographically distinct regions. This is the first occasion that this technology has been employed to investigate the fungal component of these populations or to reveal the microbial composition of such an extensive number of kefir grains or milks. As a result several genera and species not previously identified in kefir were revealed. Our analysis shows that the bacterial populations in kefir are dominated by 2 phyla, the Firmicutes and the Proteobacteria. It was also established that the fungal populations of kefir were dominated by the genera Kazachstania, Kluyveromyces and Naumovozyma, but that a variable sub-dominant population also exists.     

Influence of roughage/concentrate ratio and linseed oil on the concentration of trans-fatty acids and conjugated linoleic acid in duodenal chyme and milk fat of late lactating cows

The objective of the study was to investigate the influence of two roughage-to-concentrate ratios, with or without linseed oil supplementation, on the flow of fatty acids in the intestinal chyme and the secretion in milk fat in late lactating cows. Seven late lactating cows fitted with cannulae in the dorsal rumen and simple T-shaped cannulae in the proximal duodenum were randomly assigned to four experimental periods applying an incomplete replicated 2 x 2 Latin square design. The rations consisted of meadow hay and a concentrate mixture given in a ratio of 70:30 or 30:70 on dry matter basis. The basal rations were fed without or with 200 g linseed oil daily. After three weeks of adaptation, samples from the duodenal chyme were taken to study the flow of fatty acids. Additionally, milk samples were analysed for their milk fat composition. Decreasing roughage/concentrate ratio and linseed oil supplementation significantly increased the flow of monounsaturated fatty acids (MUFA), trans-fatty acids (tFA) and conjugated linoleic acids (CLA) in the duodenum. Furthermore, linseed oil increased the flow of saturated fatty acids (SFA) in the duodenum. Higher concentrate portion (H 30) and linseed oil supplementation significantly decreased the milk fat content. SFA were lower (p < 0.05) and MUFA were higher (p < 0.05) in milk fat after linseed oil supplementation; H 30 resulted in more polyunsaturated fatty acids (PUFA, p < 0.05) in the milk. Linseed oil supplementation significantly increased tFA and CLA in milk fat. The higher CLA content in milk fat as compared to that in the digesta suggests that a substantial endogenous synthesis of CLA in the mammary gland tissue through A9-desaturase took place. Between 21% and 48% of duodenal t11-C(18:1) were converted into c9, t11-CLA in milk fat.     

Milk composition in free-ranging polar bears (Ursus maritimus) as a model for captive rearing milk formula

The goals of this study were to have an improved understanding of milk composition and to help create a suitable milk formula for cubs raised in captivity. Milk samples were evaluated for fat, fatty acids, carbohydrate, vitamin D(3), 25(OH)D(3), vitamin A (retinol), vitamin E (α-tocopherol), protein, and amino acids. Total lipids in milk did not differ for cubs (mean ± SEM = 26.60 ± 1.88 g/100 ml vs. yearlings 27.80 ± 2.20 g/100 ml). Milk lipids were of 23.6% saturated fatty acid for cubs and 22.4% for yearlings. Milk consumed by cubs and yearlings contained 43.8 and 42.0% mono-unsaturated fatty acids and 23.4 and 21.9% polyunsaturated fatty acids, respectively. Carbohydrate content was higher in milk for cubs (4.60 ± 0.64 g/100 ml) than for yearlings (2.60 ± 0.40 g/100 ml). Vitamin D(3) concentration of milk was 18.40 ± 5.00 ng/ml in early lactation compared with 7.60 ± 2.00 ng/ml for mid-lactation. 25(OH)D(3) was lower in milk consumed by cubs (162.00 ± 6.70 pg/ml) than in milk consumed by yearlings (205.00 ± 45.70 pg/ml). Vitamin A concentrations were 0.06 ± 0.01 and 0.03 ± 0.01 μg/ml for cubs and yearlings, respectively. Vitamin E was higher in milk consumed by cubs (20.16 ± 4.46 μg/ml) than by yearlings (7.30 ± 1.50 μg/ml). Protein content did not differ in milk available to cubs (11.40 ± 0.80 g/100 ml compared with milk for yearlings 11.80 ± 0.40 g/100 ml). Taurine was the most abundant free amino acid at 3,165.90 ± 192.90 nmol/ml (0.04% as fed basis).