The phospholipid and sphingolipid composition of milk is of considerable interest regarding their nutritional and functional properties. The objective of this article was to determine the lipid composition of the milk fat globule membrane (MFGM) of milk from cows fed a diet rich in polyunsaturated fatty acids. The experiments were performed with 2 groups of 6 cows feeding on (i) maize silage ad libitum (+ grassland hay, mixture of cereals, soyabean meal) or (ii) the maize silage-based diet supplemented with extruded linseed (bringing a lipid proportion of 5% of dry matter). The phospholipid and sphingolipid composition of the MFGM was determined using HPLC/ELSD. The fatty acid (FA) composition of total lipids and phospholipids was determined using GC. As expected, the linseed-supplemented diet decreased the saturated FA and increased the unsaturated FA content in milk fat. MFGM in milk from cows fed the diet rich in polyunsaturated FA resulted in (i) a higher amount of phospholipids (+ 18%), which was related to a smaller size of milk fat globules (ii) an increase of 30% (w/w) of the concentration in sphingomyelin, (iii) a higher content in stearic acid (1.7-fold), unsaturated FA (1.36-fold), and C18:1 trans FA: 7.2 +/- 0.5% (3.7-fold). The MFGM contained a higher concentration of unsaturated FA (C18:1, C18:2, and C18:3) and very long-chain FA (C22:0, C23:0, C24:0, EPA, DHA) compared with total lipids extracted from milk. The technological, sensorial, and nutritional consequences of these changes in the lipid composition of the MFGM induced by dietary manipulation remain to be elucidated.
The aim of this work was to characterize UV-absorbing compounds (UAC) in milk in relation to diet. In winter, 4 groups of cows each received a different diet: concentrate rich containing 35% cocksfoot hay (CCH), maize silage (MS), rye grass silage (RS), or rye grass hay (RH). In summer, 2 additional diets were given: mountain grassland hay (GH) and mountain grassland pasture (GP). Polyphenols were analyzed by HPLC and Folin reaction on forages and UAC were extracted from milks and analyzed by HPLC. In forages, the number of polyphenols was lowest in MS (57) and greatest in GP (85). Twenty-four peaks were identified, accounting for 28 to 47% peak area at 280 nm. Caffeoyl compounds and flavonoid glycosides were mainly found in RH, GH, and GP. Hydrolyzed compounds such as hydroxycinnamic acids and aglycones were found in MS and RS. Estimated amounts of polyphenols were lowest for MS (3.7 g/kg), roughly similar for CH, RS, and RH (about 15 g/kg), and greatest for GH and GP (21.6 and 35.3 g/kg, respectively). About 230 different peaks were separated in milks. Milks from RH and GP contained the lowest (87) and the greatest (127) numbers of peaks, respectively. Only 10 peaks were identified, accounting for 21 to 54% of the total spectra area. In addition to the major compound hippuric acid, phenylacetic acid, benzoic acid, 4-hydroxybenzoic acid benzaldehyde, catechol, and small amounts of ferulic acid were found in varying amounts depending on the diet. Flavonoids such as quercetin, luteolin, and apigenin were also present. Hippuric acid was clearly related to the presence of caffeoylquinic compounds in forages. Other identified UAC may originate essentially from forage simple polyphenols or from cell wall aromatics. Some of the several unknown compounds may also originate from the transformation of other nutrients. Estimated amounts of UAC were widely variable within each animal group. They were surprisingly high in CCH and roughly similar in all milks from preserved forages (about 3.6 mg/L), with generally greater values for GH milks, whereas the greatest amount was found in GP milks (13.3 mg/L). Hierarchical clustering clearly discriminated the 6 diets, showing that there were major differences in GP milks. Some UAC were specific to one or a group of diets. Ultraviolet-absorbing compounds are therefore a potential tool to distinguish between milks according to diet. In addition, they may have a bioactive effect on milk component conservation or on human health.
Phenolics contribute to the micronutrient composition of forages, which in turn may affect animal product composition. To assess the importance of these compounds, the polyphenolic and botanical composition of a permanent mountain pasture in the Massif Central (France) were studied at three stages of growth. Phenolic fractions (acids, flavonoids and total) were analysed using HPLC and specific colorimetric methods over the whole pasture and in nine main species. On a botanical level, 43 species including 31 dicotyledons composed a mountain variant of Cynosurion grouping. Over time, the species growing on the field changed, the main plants being Festuca nigrescens and eight dicotyledons. In whole pasture, about 170 different compounds were separated by HPLC, of which only 30 were common to all stages. The total polyphenolic content was estimated as 31, 32 and 19 g kg −1 DM for the three stages. They were mainly composed of dihydroxycinnamic derivatives (3,5-di-O-caffeoylquinic, chlorogenic, 1,5-di-O-caffeoylquinic acids), and of flavonoids in lesser proportions. Some polyphenols were peculiar to a few species and others were ubiquitous. The two major dicaffeoylquinic acids were identified for the first time in Achillea millefolium and Knautia arvernensis. Colorimetric measurements of phenolic fractions were in good agreement with HPLC analysis (P < 0.01) for all species studied. Thus, on the mountain pasture studied, a cow would be expected to ingest up to 500 g of polyphenols daily, which could affect the quality of animal products.
To improve the digestibility of the forage crop alfalfa (Medicago sativa L.), cinnamyl alcohol dehydrogenase (CAD), which catalyses the last step in the biosynthesis of the lignin monomers, was down-regulated by using an antisense approach. A subset of six transgenic lines with reduced CAD activity and control lines were analysed when grown in the greenhouse and in the field. The down-regulation of the CAD enzyme was associated with a red coloration of the stem. The lignin quantity remained unchanged, but the lignin composition, as determined by thioacidolysis, was altered. The highest reduction of CAD activity was associated with a lower syringyl/guaiacyl (S/G) ratio and a lower S+G yield, mainly because of a decreased amount of S units. An increase in in situ disappearance of dry matter and of cell wall residue was detected in one of the transgenic lines grown in the greenhouse, and for two of the lines grown in the field the rate of disappearance of dry matter slightly improved. Furthermore, these two lines had a higher solubility in alkali as shown by the lower yield of saponified residue. This study opens perspectives for improving forage crop digestibility by the modulation of enzymes involved in lignin biosynthesis.
We studied the nutritional behaviour of hinds foraging on a mixed-forest edge by direct observation of their choices at each season and by measuring nutrient concentration in the plants. We compared nutrient concentrations in the observed diets with those in the total available vegetation, and with those of 1,000 randomly simulated diets in which we included only those plants that were actually eaten by the animal. Whether the available or the consumed feeds were used as the basis for comparisons had important consequences due to the presence of conifers and ferns, which were high in soluble sugars but were never eaten by the animals (potentially due to their toxicity). The selected diets were lower in sugars than the total available vegetation in summer, but were actually higher in sugars than the random diets generated from consumed forage species only. Hind diets contained more soluble sugars but not more protein than simulated diets in all seasons. Contrary to our prediction, anti-nutritional compounds (ADL and tannins) were avoided only in winter. Compared to simulated diets, hinds consumed more tannins in spring and summer and more ADL in summer and autumn. We suggest that this was a consequence of selection for soluble sugars, because the preferred plant species, which had high soluble sugar concentrations, also contained a large proportion of the anti-nutritional compounds eaten. In winter, the grass-dominated diets contained more fibre (NDF) and less ADL than the simulated diets, indicating that hinds orient their feeding towards digestible fibres. The switch from a browser to a grazer diet was related to a change in the availability of the nutrients, mainly soluble sugars. In our study, grasses contained more soluble sugars and proteins than deciduous browse during winter. This calls into question the dichotomy usually assumed in the literature between grass and browse in terms of nutrient content.
Phenolic constituents (lignins and phenolic acids) and carbohydrates are assembled in a tight architecture which differs according to the plant species. During cell wall digestion, the hydrolysis kinetics differ between carbohydrates and seem to depend chiefly on the content and organisation of tissue phenolics. Among the phenylpropanoids, ferulic acid is released more quickly than pcoumaric acid. Lignins remain largely in the cell walls. They also undergo transformations, chiefly solubilisation as lignin-carbohydrate complexes. The limiting effect of lignins on cell wall degradation increases with increasing content. However, their effect on degradation might also depend on qualitative factors such as lignin structure and polymer organisation in walls and tissues. When various grasses (normal and selected genotypes), or grasses and legumes are compared, correlations between certain factors such as lignin uncondensed fraction, syringyl units or phenolic acids contents and cell wall degradation emerge but not clear causal relationship has been shown. Nonetheless, other structural characteristics, related to the alkali reactivity of lignins, seem to have a stronger influence on cell wall degradation. Phenylpropanoids seem to act mainly as a physical and (bio)chemical barrier to the action of the microbial enzymes. In addition, their reactivity as phenolic compounds and their hydrophobicity seem to play a role. Digestion is not limited only by phenolics. The factors that limit glycanolysis-the accessibility, crystallinity and capillary structure of cellulose and the branching of hemicelluloses-seem to have little or no effect on cell wall degradation in oivo. In contrast, other antiquality substances (tannins, cutin and silica), plant antomy, environmental factors, factors modulating microbial growth and animal physiology influence cell wall utilisation. Future research in this field should focus on the effects of phenolic structure and of cell wall and tissue organisation on carbohydrate degradation.
Phenolic compounds contribute to the micronutrient composition of pasture, which in turn may affect animal product composition. To assess the importance and variations in content of these compounds, the polyphenolic and botanical compositions of 24 permanent pastures located in one lowland and two upland regions were studied at equivalent stages of growth. Phenolic fractions were characterized and quantified using HPLC-PDA-ESI-QToF, and the total content was determined by colorimetry over each whole pasture. A rise in altitude was accompanied by a marked increase in total phenolic content, linked to changes in botanical composition, but did not have any influence on the distribution according to molecular class. For all of the pastures, significantly different patterns due to qualitative and quantitative differences among the 92 separate peaks were observed with 31 compounds identified. The involvement of certain plants in the variations of content and composition in phenolic compounds of pastures was statistically evaluated.
The energy value of NSP has been expressed as their metabolizable energy (ME) content. The aim of the present study was to determine whether differences in ME and net energy (NE) contents were similar for insoluble and soluble NSP. Nine healthy young men were offered three diets according to a Latin-square design (3 x 3) with three repetitions: diet C (control), diet B (control + 50 g sugarbeet fibre/d) and diet I (control + 50 g commercial inulin/d). After a 16 d adaptation period to NSP isolate, food intake was controlled (duplicate meal method) and faeces and urine were collected for 8 d. A period of 60 h was devoted to measurement of energy expenditure (EE) by whole-body indirect calorimetry. NSP-isolate ingestion induced significant increases in the number of defecations and stool weight resulting from increases in water, DM and microbial mass excretion. After deduction of microbial N, differences in faecal N excretion between diets were not significantly different. Urinary N excretion was slightly decreased by sugarbeet fibre or commercial inulin ingestion but the N balances for the diets were not significantly different. Diet energy, N and lipid apparent digestibilities decreased by only 1-2%. Commercial inulin was entirely fermented and fermentability of sugarbeet fibre averaged 0.886 (SD 0.117). Sugarbeet fibre and commercial inulin ME values averaged 10.7 (SD 1.2) and 13.0 (SD 2.3) kJ/g DM respectively. NSP-isolate ingestion caused significant (sugarbeet) and nonsignificant (inulin) increases in daily EE. The maintenance NE contents of sugarbeet fibre and inulin averaged 5.0 (SD 5.0) and 11.9 (SD 1.3) kJ/g DM respectively. Differences in maintenance NE contents of NSP isolates were much greater than differences in ME values.
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