1. Pure swards of perennial ryegrass (Lolium perenne L. cv. Melle) or white clover (Trifolium repens L. cv. Blanca) were harvested daily as either primary growth (May-June) or mid- (July) and late- (August-September) season 4-week regrowths and offered to Friesian steers at two levels of feed allowance (18 and 24 g dry matter (DM)/kg live weight), to examine the effect of forage species and stage of harvest on nutrient digestion and supply.

2. The early- and mid-season grasses had low nitrogen (23 g/kg DM) and high water-soluble carbohydrate (169 g/kg) contents whilst the late-season grass had a higher N content (28 g/kg). All clover diets had high N (average 45 g/kg) and low water-soluble carbohydrate (89 g/kg) contents, and DM digestibility on all diets ranged from 0.77 to 0.83 (mean of two feeding levels).

3. Mean total rumen volatile fatty acid concentrations were significantly (P < 0.001) higher on the clover diets, whilst on the grass diets molar proportions of propionate showed a slight but not significant decline with advancing season and tended to be higher than those on the clover diets. Mean rumen ammonia concentrations were significantly (P < 0.001) lower on the early- and mid-season grasses (59 mg NH3-N/1) than the late-season grass (242 mg/l) and early-season clover (283 mg/l) which were all significantly (P < 0.01) lower than the mid-and late-season clovers (372 and 590 mg/l) respectively.

4. Rates of organic matter (OM) and N digestion in the rumen were estimated using established nylon-bag techniques and found to be high on all diets, but significant effects due to forage species (clover > grass; P < 0.001) were detected, whilst overall potential degradability in the rumen exceeded 0.89 for both OM and N on all diets.

5. Significantly (P < 0.001) more OM entered the small intestine of calves fed on white clover (10.2 g/kg live weight) than those fed on ryegrass (8.33 g/kg) and similar effects due to level of feeding (g/kg; low 7.9, high 10.6; P < 0.05) and stage of harvest (g/kg; early 8.3 v. mid 10.0, late 9.50; P < 0.05) were also detected. Non-NH3-N (NAN) flow (g/kg) to the small intestine was increased by forage species (grass 0.56, clover 0.69; P < 0.05) and stage of harvest (early 0.59 v. mid 0.65, late 0.64; P > 0.05) whilst NAN flow/N intake ranged from 0.96 to 1.65 g/g (mean 1.25) on the grass diets and from 0.64 to 0.84 g/g (mean 0.75) on the clovers (P < 0001).

6. Microbial N flow to the small intestine averaged 0.72 of duodenal NAN (grass 0.76, clover 0.69). Efficiency of microbial N synthesis was high on all diets, (g/kg OM truly digested in the rumen; grass 33.5, clover 36.3), as was the estimated extent of in vivo feed N degradation (g/g N intake; grass 0.75, clover 0.79).

7. A model is described to simulate the progress curves of the ratio, degraded N:degraded OM in the rumen for the six diets, using indices obtained in the present study. The results are ratified with the in vivo observations of N utilization in the rumen for the grass and clover diets.

8. It is concluded that both forage species and stage of harvest can significantly influence the processes of rumen digestion and nutrient supply, but with the fresh forages examined in the present study, it would appear that the processes of digestion in the rumen greatly outweighed the passage of potentially digestible nutrients from the rumen.

The ability of nuts to improve the blood lipid profile and reduce the risk of CHD is now well established. The interest that health effects of nuts have gained recently has brought the possible benefits of consuming nuts, such as improvement in the conditions of the metabolic syndrome, and their potential to prevent and control diabetes into focus. Results from cohort studies have associated nut consumption with a reduced risk of developing diabetes and CVD. However, few randomised controlled trials have assessed the effect of nuts on diabetes control, and those that have been undertaken have shown improvements in blood lipids but not in the glycaemic control. Diabetes agencies are increasingly recognising the importance of controlling postprandial glycaemia fluctuations. Acute feeding studies indicate that nuts have minimal effects on rising postprandial blood glucose levels when eaten alone, and diminish the postprandial glycaemic response when consumed with high-glycaemic index carbohydrate foods in both normoglycaemic and type 2 diabetic individuals. Nuts have a healthy nutritional profile, high in MUFA and PUFA, are a good source of vegetable protein and are rich in fibre, vitamins and minerals. Incorporation of nuts in the diet may therefore improve the overall nutritional quality of the diet. While more research is required to establish the ability of nuts to improve glycaemic control in the long run, early data indicate that the inclusion of nuts in the diets of individuals with diabetes and the metabolic syndrome is warranted, in view of their potential to reduce CHD risk.

The objective of the present study was to evaluate anti-diabetic effects of chromium picolinate (CrPic) and biotin supplementations in type 2 diabetic rats. The type 2 diabetic rat model was induced by high-fat diet (HFD) and low-dose streptozotocin. The rats were divided into five groups as follows: (1) non-diabetic rats fed a regular diet; (2) diabetic rats fed a HFD; (3) diabetic rats fed a HFD and supplemented with CrPic (80 μg/kg body weight (BW) per d); (4) diabetic rats fed a HFD and supplemented with biotin (300 μg/kg BW per d); (5) diabetic rats fed a HFD and supplemented with both CrPic and biotin. Circulating glucose, cortisol, total cholesterol, TAG, NEFA and malondialdehyde concentrations decreased (P< 0·05), but serum insulin concentrations increased (P< 0·05) in diabetic rats treated with biotin and CrPic, particularly with a combination of the supplements. Feeding a HFD to diabetic rats decreased PPAR-γ expression in adipose tissue and phosphorylated insulin receptor substrate 1 (p-IRS-1) expression of liver, kidney and muscle tissues, while the supplements increased (P< 0·001) PPAR-γ and p-IRS-1 expressions in relevant tissues. Expression of NF-κB in the liver and kidney was greater in diabetic rats fed a HFD, as compared with rats fed a regular diet (P< 0·01). The supplements decreased the expression of NF-κB in diabetic rats (P< 0·05). Results of the present study revealed that supplementing CrPic and biotin alone or in a combination exerts anti-diabetic activities, probably through modulation of PPAR-γ, IRS-1 and NF-κB proteins.

The aim of the present study was to identify the genes differentially expressed in the visceral adipose tissue in a well-characterised mouse model of high-fat diet (HFD)-induced obesity. Male C57BL/6J mice (n 20) were fed either HFD (189 % of energy from fat) or low-fat diet (LFD, 42 % of energy from fat) for 16 weeks. HFD-fed mice exhibited obesity, insulin resistance, dyslipidaemia and adipose collagen accumulation, along with higher levels of plasma leptin, resistin and plasminogen activator inhibitor type 1, although there were no significant differences in plasma cytokine levels. Energy intake was similar in the two diet groups owing to lower food intake in the HFD group; however, energy expenditure was also lower in the HFD group than in the LFD group. Microarray analysis revealed that genes related to lipolysis, fatty acid metabolism, mitochondrial energy transduction, oxidation–reduction, insulin sensitivity and skeletal system development were down-regulated in HFD-fed mice, and genes associated with extracellular matrix (ECM) components, ECM remodelling and inflammation were up-regulated. The top ten up- or down-regulated genes include Acsm3, mt-Nd6, Fam13a, Cyp2e1, Rgs1 and Gpnmb, whose roles in the deterioration of obesity-associated adipose tissue are poorly understood. In conclusion, the genes identified here provide new therapeutic opportunities for prevention and treatment of diet-induced obesity.

The lipid distribution and fatty acid (FA) composition of total lipids, polar lipids and free fatty acids (FFA) were determined in liquid-associated bacteria (LAB) and solid-adherent bacteria (SAB) isolated from the rumen contents of seven dairy cows fitted with rumen fistulas. Two experiments, arranged according to a 4 x 4 and 3 x 3 Latin Square design, were performed using two basal diets consisting of one part hay and one part barley-based concentrate, and five lipid-supplemented diets consisting of the basal diet plus (g/kg dry matter): 53 or 94 rapeseed oil, 98 tallow, 87 soya-bean oil or 94 palmitostearin. For all diets used, total lipids were 1.7–2.2 times higher in SAB than in LAB (P < 005); this probably resulted from a preferential incorporation of dietary FA adsorbed onto food particles. Addition of oil or fat to the diets did not modify the polar lipid content but increased the FFA content of SAB and LAB by 150%. Lipid droplets were observed in the cytoplasm in SAB and LAB using transmission electron microscopy, which suggested that part of the additional FFA was really incorporated into the intracellular FFA rather than associated with the cell envelope by physical adsorption. Linoleic acid was specifically incorporated into the FFA of SAB, which emphasized the specific role of this bacterial compartment in the protection of this acid against rumen biohydrogenation.

We have previously demonstrated that maternal protein restriction during pregnancy leads to raised blood pressure and endothelial dysfunction in the offspring (F1). Here we show that these characteristics are transmitted to the F2 offspring through the maternal line, in the absence of any additional challenges to the F1. Female Wistar rats were fed either a control (18 % casein) or protein-restricted diet (PR; 9 % casein) throughout pregnancy. Female F1 offspring, maintained on standard chow postpartum, were mated with breeding males to produce F2 progeny. Systolic blood pressure (SBP) in male F2 offspring was assessed by tail-cuff plethysmography at age 100 d and vascular function of small mesenteric arteries by wire myography at age 80 and 200 d. SBP was raised in PR F2 offspring compared with controls (control 122·1 (sem 2·3) mmHg, n 7; PR 134·7 (sem 3·2) mmHg, n 6; P < 0·01) and endothelial function, assessed by vasodilatation to acetylcholine, was impaired at both age 80 d (% maximal response: control 89·7 (sem 2·6), n 14; PR 72·7 (sem 4·4), n 15; P < 0·01) and 200 d (effective concentration equal to 50 % of maximum (pEC50): control 7·67 (sem 0·10), n 10; PR 7·33 (sem 0·07), n 8; P < 0·05). The present study demonstrates that both raised blood pressure and endothelial dysfunction are passed via the maternal line to grand-offspring in the absence of any additional dietary challenges to their F1 mothers. Risk factors for chronic disease may therefore be heritable by non-genomic processes.

The aim of the present study was to examine the effect of consumption of a high-fruit and vegetable diet, or a spray-dried extract of selected fruits and vegetables of high antioxidant content, on indices of antioxidant status of individuals consuming a background diet with minimal antioxidant intake. Plasma antioxidant concentrations were determined in twenty-five men following a 2-week depletion period during which they consumed self-selected low-antioxidant diets (less than three servings of fruit and vegetables with no tea, coffee, red wine or fruit juice). Following this period the volunteers consumed either a self-selected diet containing five to seven servings of fruit and vegetables/d, or 30 g of a spray-dried supplement designed to provide the equivalent antioxidant activity of five to seven servings of fruit and vegetables for 2 weeks in a crossover trial. Following consumption of a high-antioxidant diet for 2 weeks, plasma concentrations of ascorbic acid, α- and β-carotene and lutein+zeaxanthin were all significantly increased (P<0.05) over the depletion period. However, concentrations of lycopene, retinol and tocopherol were not affected. Consumption of the supplement also raised the concentrations of these same antioxidants in plasma. Despite the increases in the concentrations of measured antioxidant nutrients, the 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-equivalent antioxidant capacity of plasma, as estimated by inhibition of metmyoglobin activity, was not significantly affected by any of the dietary treatments.

1.Fifteen female farmers (aged 18–47 years) from two villages of the Mossi Plateau in Upper-Volta participated in a survey in which their daily activity pattern and their energy expenditure were assessed. Eight of the subjects were investigated twice, in March (dry season) when there is no agricultural activity, and in July–August (rainy season) when heavy physical work is performed: mostly hoeing, weeding and replanting sorghum (Sorghum vulgare) and millet (Pennisetum typhoïdes).

2. The mean height was 1.57 m and the mean weight 50.6 kg. The average percentage of body fat, calculated from skinfold thickness, was comparable to that of European females but the triceps skinfold was more than 60% below the standard value (Jelliffe, 1969). The type of activities and the period of time spent on each activity changed significantly with the season. The mean energy output rose from 9.7 MJ (2320 kcal) in March to 12.1 MJ (2890 kcal) in July–August for a 55 kg standard weight.

3. In this paper, the extent of both the dally activity pattern of women living in a subsistence agriculture and their energy output is estimated. The results suggest that during the rainy season, the energy requirements of female farmers are much higher than usually estimated.

The effect of policosanol, a mixture of high-molecular-weight aliphatic alcohols isolated from sugar-cane wax, on casein-induced hypercholesterolaemia in rabbits was studied. When policosanol was administered by the oral route once daily for 30 d (50mg/kg) the increases in plasma total cholesterol and LDL-cholesterol (LDC-C) were significantly reduced when compared with the control group. The incorporation of 3H2O into sterols in the liver was significantly depressed, suggesting inhibition of hepatic cholesterol biosynthesis. The oral administration of policosanol raised the rate of removal of 125l-labelled LDL from serum. Kinetic parameters calculated following injection of [125I]LDL showed than in casein-fed rabbits, the terminal half-life (t½) was significantly decreased after policosanol treatment. The hepatic LDL-binding activity was increased after policosanol administration which suggested that the enhanced clearance was due, at least in part, to increased receptor-mediated uptake of LDL by the liver. Considered together, these results suggest that policosanol can significantly reduce the increase of plasma LDL-C in rabbits fed on a wheat starch-casein diet by reducing cholesterol biosynthesis in the liver. Such an effect could account for the enhancement of LDL catabolism through the receptor-mediated pathway.

Inclusion of fermentable fibres in the diet can have an impact on the hindgut microbiome and provide numerous health benefits to the host. Potato fibre (PF), a co-product of potato starch isolation, has a favourable chemical composition of pectins, resistant and digestible starch, cellulose, and hemicelluloses. The objective of the present study was to evaluate the effect of increasing dietary PF concentrations on the faecal microbiome of healthy adult dogs. Fresh faecal samples were collected from ten female dogs with hound bloodlines (6·13 (sem 0·17) years; 22·0 (sem 2·1) kg) fed five test diets containing graded concentrations of PF (0, 1·5, 3, 4·5 or 6 % as-fed; Roquette Frères) in a replicated 5 × 5 Latin square design. Extraction of DNA was followed by amplification of the V4–V6 variable region of the 16S rRNA gene using barcoded primers. Sequences were classified into taxonomic levels using Basic Local Alignment Search Tool (BLASTn) against a curated GreenGenes database. Inclusion of PF increased (P< 0·05) the faecal proportions of Firmicutes, while those of Fusobacteria decreased (P< 0·05). Similar shifts were observed at the genus level and were confirmed by quantitative PCR (qPCR) analysis. With increasing concentrations of PF, faecal proportions of Faecalibacterium increased (P< 0·05). Post hoc Pearson's correlation analysis showed positive (P< 0·05) correlations with Bifidobacterium spp. and butyrate production and Lactobacillus spp. concentrations. Overall, increases in the proportion of Faecalibacterium (not Lactobacillus/Bifidobacterium, as confirmed by qPCR analysis) and faecal SCFA concentrations with increasing dietary PF concentrations suggest that PF is a possible prebiotic fibre.

The objective of the study was to investigate the ability of an NMR-based metabonomic approach, applied to biofluids, to explore and identify overall exogenous and endogenous biochemical effects of a short-time high intake of milk protein or meat protein given to prepubertal children, the aim being to compare relative differences and not an absolute quantification. A total of twenty-four 8-year-old boys were asked to take 53 g protein as milk (n 12) or meat daily (n 12). At baseline and after 7 d, urine and serum samples were collected and high-resolution 1H NMR spectra were acquired on these using a 800 MHz spectrometer. The milk diet reduced the urinary excretion of hippurate, while the meat diet increased the urinary excretion of creatine, histidine and urea. The NMR measurements on serum revealed minor changes in the lipid profile, which most probably should be ascribed to an increase in the content of SCFA in the blood after consumption of the milk diet. The meat diet had no effect on the metabolic profile of serum. The study for the first time demonstrates the capability of proton NMR-based metabonomics to identify the overall biochemical effects of consumption of different animal proteins. The urine metabolite profile is more susceptible to perturbations as a result of short diet interventions than the serum metabolite profile. The milk diet-induced reduction in urinary excretion of hippurate suggests alterations in gut microflora, which may be useful information for further studies elucidating the effects of bioactive components in milk.

Obesity is the nutritional disorder which has shown the greatest increase in prevalence, even in those countries in which deficiency diseases represent a severe public health problem. The goal of the present study was to analyse the anthropometric profile of a community living in the outskirts of Maceió, capital of Alagoas (northeastern Brazil), and to investigate the hypothesis of a coexistence of undernutrition and obesity in a very low-income population. The survey was conducted on 315 families (1247 individuals). Among the children (aged ≤10 years), the prevalence of wasting, stunting and wasting plus stunting was 3·8, 8·3 and 8·7 % respectively. Wasting (10·2 %) was the most prevalent form of undernutrition among adolescents; nonetheless, a higher frequency of stunting (11 %) and overweight–obesity (5·5 %) was seen specifically in girls, in agreement with trends found in other studies. Adults exhibited a high prevalence of overweight–obesity (25 %), but stunting was also present (22 %). Of the stunted individuals, 30 % were overweight–obese and 16·3 % were underweight. There were eighty-six families with at least one parent who was underweight (27 %) and 104 families with at least one parent who was overweight (33 %). Underweight and overweight–obesity were both present in ninety-six households (30 %). These results may indicate that better living conditions in urban areas in a population ‘adapted’ to chronic famine might increase the susceptibility to obesity. Considering the harm caused by the cumulative effect of these two conditions (undernutrition in childhood and obesity in adult life) there is a clear need for new studies to uncover the determinant factors so that preventive measures can be implemented.

The aim of this study was to evaluate the effect of a synbiotic containing Lactobacillus acidophilus ATCC 4962, fructooligosaccharide, inulin and mannitol on plasma lipid profiles and erythrocyte membrane properties in hypercholesterolaemic pigs on high- and low-fat diets. Twenty-four white male Landrace pigs were randomly allocated to four treatment groups for 8 weeks (n 6). Treatment factors were the supplementation of synbiotic (with and without) and dietary fat (5 and 15 %). The supplementation of synbiotic reduced plasma total cholesterol (P = 0·001), TAG (P = 0·002) and LDL-cholesterol (P = 0·045) for both dietary fats. A higher concentration of esterified-cholesterol in HDL of pigs supplemented with synbiotic than the control regardless of dietary fat (P = 0·036) indicated that cholesterol was reduced in the form of cholesteryl esters. Reduced concentration of cholesteryl esters (P < 0·001) and increased concentration of TAG (P = 0·042) in LDL of pigs on synbiotic suggested that LDL-cholesterol was reduced via the hydrolysis of smaller and denser LDL particles. The erythrocytes of pigs without any synbiotic showed more prevalence of spur cells than those given the synbiotic, as supported by the higher cholesterol: phospholipid ratio in erythrocytes (P = 0·001). Also, membrane fluidity and rigidity were improved as supported by the decreased fluorescence anisotropies in the Hb-free erythrocyte membrane of pigs given synbiotic (P < 0·001). The administration of the synbiotic reduced plasma TAG, total cholesterol and LDL-cholesterol in hypercholesterolaemic pigs, possibly in the form of cholesteryl esters, via the interrelated pathways of lipid transporters (VLDL, LDL and HDL). The synbiotic also reduced deformation of erythrocytes via improved membrane fluidity and permeability.

This article describes the principles and limitations of methods used to investigate reactive oxygen species (ROS) protective properties of dietary constituents and is aimed at providing a better understanding of the requirements for science based health claims of antioxidant (AO) effects of foods. A number of currently used biochemical measurements aimed of determining the total antioxidant capacity and oxidised lipids and proteins are carried out under unphysiologcial conditions and are prone to artefact formation. Probably the most reliable approaches are measurements of isoprostanes as a parameter of lipid peroxidation and determination of oxidative DNA damage. Also the design of the experimental models has a strong impact on the reliability of AO studies: the common strategy is the identification of AO by in vitro screening with cell lines. This approach is based on the assumption that protection towards ROS is due to scavenging, but recent findings indicate that activation of transcription factors which regulate genes involved in antioxidant defence plays a key role in the mode of action of AO. These processes are not adequately represented in cell lines. Another shortcoming of in vitro experiments is that AO are metabolised in vivo and that most cell lines are lacking enzymes which catalyse these reactions. Compounds with large molecular configurations (chlorophylls, anthocyans and polyphenolics) are potent AO in vitro, but weak or no effects were observed in animal/human studies with realistic doses as they are poorly absorbed. The development of -omics approaches will improve the scientific basis for health claims. The evaluation of results from microarray and proteomics studies shows that it is not possible to establish a general signature of alterations of transcription and protein patterns by AO. However, it was shown that alterations of gene expression and protein levels caused by experimentally induced oxidative stress and ROS related diseases can be normalised by dietary AO.

1. Three cows were given 3, 5 or 7 kg hay once daily in a Latin-square design, and samples of digesta from four sites in the rumen and one site in the reticulum were taken at six times within the 24 h period after the feed. Dry-matter content and distribution of particle size and of particle density were measured for each sample. The changes in these measurements with time were studied. The incidence of rumination was also recorded.

2. Dry-matter contents of samples ranged from 15 to 3%. Values for samples from the dorsal sacs of the rumen were considerably higher than those for samples from the ventral sites; they also changed more with time after feeding and with level of feeding.

3. Particles were fractionated by sieving into six size groups with mean dimensions (mm) of 9·3 × 0·8, 4·4 × 0·6, 2·6 × 0·3, 1·6 × 0·25, 0·5 × 0·1 and smaller than 0·5 × 0·1. Coarse particles occurred at highest concentrations in the dorsal sacs of the rumen and responded to effects of time and level of feeding; smaller particles showed less response.

4. Particle densities ranged from 800 g/l to 1500 g/l. The proportions of low-density particles were higher in samples from the dorsal sacs of the rumen than in samples from ventral sites; the latter samples had higher proportions of high-density particles than of low-density particles. The changes which occurred are discussed.

5. The density of coarse particles tended to be low and that of fine particles tended to be high.

6. Rumination started at the time of maximum concentration of particles of low density and minimum concentration of particles of high density. Conversely, rumination ended when the concentration of the low-density particles was a minimum and that of the dense particles a maximum.

7. The results are discussed in relation to the possible movement of particles within the reticulo-rumen and the kinetics of particle breakdown.

The aims of the present study were to examine the serum amino acid profiles in obese and non-obese women and investigate the relationships between the serum amino acids and inflammation and oxidative stress in a human case–control study. Serum amino acids, inflammatory biomarkers (C-reactive protein and IL-6) and oxidative biomarkers (superoxide dismutase, malondialdehyde and glutathione peroxidase) were measured and compared in 235 obese women and 217 non-obese controls. The relationships between serum amino acids and inflammatory and oxidative biomarkers were examined using multiple linear regression. Among the amino acids determined, serum histidine, arginine, threonine, glycine, lysine and serine were found to be significantly lower in obese women as compared to non-obese controls (P < 0·001). The difference was the greatest for histidine (P < 0·001). In obese women, both histidine and arginine were negatively associated with inflammation and oxidative stress. In non-obese controls, histidine was negatively associated with oxidative stress. The findings in this study indicate that the metabolism of amino acids is abnormal in obese women in whom histidine and arginine have close relationships with inflammation and oxidative stress.

The effect of vitamin C on the common cold has been the subject of several studies. These studies do not support a considerable decrease in the incidence of the common cold with supplemental vitamin C. However, vitamin C has consistently decreased the duration of cold episodes and the severity of symptoms. The benefits that have been observed in different studies show a large variation and, therefore, the clinical significance may not be clearly inferred from them. The biochemical explanation for the benefits may be based on the antioxidant property of vitamin C. In an infection, phagocytic leucocytes become activated and they produce oxidizing compounds which are released from the cell. By reacting with these oxidants, vitamin C may decrease the inflammatory effects caused by them. Scurvy, which is caused by a deficiency in vitamin C, is mostly attributed to the decreased synthesis of collagen. However, vitamin C also participates in several other reactions, such as the destruction of oxidizing substances. The common cold studies indicate that the amounts of vitamin C which safely protect from scurvy may still be too low to provide an efficient rate for other reactions, possibly antioxidant in nature, in infected people.

1. Mn and Cu intake and retention in twenty full-term infants and six preterm infants were studied on the basis of 72 h balances. The age of the infants was 2–16 weeks and the gestational age of the preterm infants (triplets) 34 and 36 weeks. Three nutrition schemes were pursued: breast-fed, formula-fed with unsupplemented adapted formula and formula-fed with trace element supplementation.

2. The mean Mn concentration of all breast-milk samples (n 2339) was 6·2 μg/1. The two formulas had similar Mn concentrations (77 and 99 μg/1) but had different Fe, Cu (121 and 619 μg/1), Zn and I contents. The mean Cu concentration in mother's milk was 833 μg/1.

3. The following mean daily Mn intakes and retentions (μg/kg) respectively were measured: breast-fed fullterm 1·06 (sd 0·43) and 0·43 (sd 0·65), formula-fed full-term 14·2 (sd 3·1) and 2·8 (sd 4·8), formula-fed preterm 15·0 (sd 2·2) and 0·06 (sd 5·87). The results for Cu were 114·5 (sd 22·3) and 88·0 (sd 46·5) μg/kg in breast-fed, 19–8 (sd 4·2) and 4·6 (-11·5–9·6) in the unsupplemented formula-fed and 106·4 (sd 18·9) and 55·5 (sd 20·3) in the supplemented formula full-term infant group. No significant influence of the trace element contents of the formulas on the relative retention of Mn or Cu was found.

4. Young preterm infants, and to some degree young full-term infants, often had negative Mn balances caused by a high faccal excretion. The formulas with a Mn concentration below 100 μg/l gave a sufficient supply of Mn. Preterm infants fed on the unsupplemented formula had a marginal Cu supply and their first balances were negative (-3·8 (sd 1·8) μg/kg).

5. In accordance with the estimated safe and adequate daily dietary intakes (recommended dietary allowances), formula-fed infants receive much more Mn than breast-fed infants and their absolute retention is higher.

6. Cu from breast-milk had a significantly better biological availability than that from cow's milk formula. If retentions similar to those in breast-fed infants are intended, we conclude, therefore, that cow's milk formula should be fortified with Cu up to a level of at least 600 μg/l.

The aims of the present study were to review the validity of dietary methods used to measure the usual long chain (LC) omega-3 polyunsaturated fatty acid (n-3 PUFA) intake of a population and to assess the usefulness of different biomarkers of n-3 PUFA in healthy humans. Two systematic literature searches were conducted until May 2011 to update previous systematic reviews. The first literature search aimed to find studies validating the methodology used for measuring the dietary intake of n-3 PUFA. The second search aimed to find human intervention studies in which n-3 PUFA status changed after 2 weeks of n-3 PUFA supplementation. Sixteen studies were identified for inclusion in the first review. Correlation coefficients between fatty acids in subcutaneous fat or blood lipids and dietary intake of n-3 PUFA from different questionnaires were similar. Subcutaneous fat has been reported as the best reference method for some authors, and these studies showed moderate correlation coefficients with no dietary intake method being superior to any other. As for the evaluation of biomarkers of docosahexaenoic acid (DHA, 22 : 6 n-3) and eicosapentaenoic acid (EPA, 20 : 5n-3) status in response to supplementation, the new search reaffirmed and reinforced the evidence supporting that plasma phospholipid DHA, erythrocyte DHA, and platelet DHA were all effective and robust biomarkers of DHA status. Our findings only confirmed earlier studies and did not provide evidence for reaching new conclusions.