Measurement of energy expenditure with doubly-labelled water and of body composition and breast milk output with 2H or 18O requires accurate and precise techniques for measuring isotopic enrichments. The possibility of an inaccuracy in measurements of 2H and 18O isotopic enrichment arising from the matrix in biological fluids was investigated (1) by simulating a dilution experiment in both water and urine samples and (2) by reconstituting urine samples, ranging from 10 to 60 g/kg in solid concentration, from freeze-dried urinary solids mixed with either natural abundance or doubly-labelled water. Current techniques involved in measuring 2H and 18O isotopic enrichments were used (reduction of the samples to H2 gas with either Zn or U, and CO2/H2O equilibration or direct measurement of mass 20:18 ratios on water vapour for 18O analysis). All four methods accurately measured serial dilutions in both urine and water. Dilution space calculated from isotopic enrichments, compared with the water content of urine (determined by freeze-drying and accounting for exchangeable isotopes) was overestimated by about 2.4 % by the Zn technique whereas other methods were accurate. The urinary solids content of a water solution was related to that inaccuracy. The use of the Zn technique with biological samples is likely to create biases in 2H distribution space. Examination of recent literature supports this view. Caution should therefore be used when physiological conclusions have to be made from the relative size of 2H and 18O distribution spaces.

In a cohort (n 81) of healthy children aged 1.5–4.5 years, measurements of energy intake and energy expenditure were compared. Energy intake was calculated following a 4 d weighed record completed by the mother or guardian of the child. Total energy expenditure was measured using the doubly-labelled water technique. Mean energy intake and expenditure in the cohort were 4773 kJ/d and 4928 kJ/d respectively. The mean relative bias between the techniques was 154 kJ/d. In the older children (3.5–4.5 years) the mean relative bias was only 37 kJ/d. At the population level the measurements of energy intake and energy expenditure were extremely close, and the study has provided sufficient confidence in weighed intake methodology for it to be used in a major nationwide study of dietary intake and nutritional status of children aged 1.5–4.5 years.

The doubly-labelled water (2H218O) technique was used to assess the long-term rates of energy expenditure and, after accounting for any changes in body composition, the derived rates of energy intake in weight-stable ‘large-eating’ (n 6) and ‘small-eating’ (n 6) women. The self-reported energy intakes (approximately 11.2 v. 5.6 MJ/d) and energy expenditures (approximately 8.5 v. 12.4 MJ/d) for the ‘large-eating’ and ‘small-eating’ groups respectively, should not be sustainable without significant body-weight changes. 2H218O-assessed rates of energy expenditure for the ‘large-eaters’ (approximately 8.5 MJ/d) and ‘small-eaters’ (approximately 11.3 MJ/d) were in close agreement with the results obtained using 5 d, self-reported activity diaries but the derived rates of energy intake for the ‘large-’ (approximately 8.5 MJ/d) and ‘small-eaters’ (approximately 10.8 MJ/d) were markedly different from those obtained using self-reported, weighed food diaries. When two ‘small-eaters’ were supplied with their self-reported energy intakes (approximately 5 MJ/d) for up to 28 d both subjects lost about 0.75 kg body-weight/week. These results provide no support for the existence of ‘metabolically efficient’ women in the community.

There is a paucity of data on differences between methods for the assessment of body composition in elderly subjects. Studies on younger adults suggest that such differences are of some practical significance at the individual level. In the present study the following methods of estimating percentage body fatness (BF%) were compared in healthy elderly men and women (mean age 70 (SD 6) years: densitometry; skinfold thickness; total body water; bioelectrical impedance (BIA) using an age-specific predictive equation and the manufacturers' equation; body mass index (BMI). Though BF% estimates from the various methods tended to be highly correlated with those from densitometry and with each other, differences between methods at the individual level were marked. In particular, the age-specific equations based on BMI and BIA systematically overestimated BF% relative to the other methods. Biases between BF% estimates derived from densitometry, skinfolds, BIA (manufacturers' equation) and total body water were less marked, indicating little evidence of systematic differences between these methods in elderly subjects. Individual differences between methods were slightly greater than those reported in some studies of younger adults, but this may be of little practical significance, and may be considered inevitable in view of variability between and within subjects in the extent to which the underlying assumptions of these two-component methods are met in elderly subjects.

The interaction between essential dietary components and changes in tissue nutrient reserves, egg quality and egg composition, were studied from 60 d before and during the spawning of Sparus aurata broodstock. Fish were given isonitrogenous (550 g/kg dry weight) and isolipidic (100 g/kg dry weight) diets, based on protein and lipid extracts of squid meal. Diets differed in the levels of n−6 (10–30 mg/g dry weight) and n−3 (0–10 mg/g dry weight) essential fatty acids. The effects of these diets on biochemical and fatty acid composition of body tissues, and the subsequent effects on egg composition and egg viability were measured. Dietary essential fatty acids were mostly incorporated into the liver, ovaries, digestive tract and associated adipose tissues. The lipid composition of these tissues reached an equilibrium with dietary lipid composition within 15 d of feeding on any given diet. Muscle and gill cartilage tissues did not show any significant changes in their biochemical and fatty acid composition, even after 60 d feeding. Egg viability decreased significantly within 10 d of feeding the broodstock with a diet deficient in n−3 highly unsaturated fatty acids (n−3 HUFA). The levels of n−3 HUFA in both polar and neutral fractions of egg lipid were directly correlated with their levels in the broodstock diet. When the total amount of egg n−3 HUFA dropped below 17 mg/g dry weight, egg viability and larvae hatching rate decreased by 53% and 47 % respectively. These results suggest that the biochemical composition of organs involved in S. aurata reproduction are highly sensitive to the nutritional value of the diet, which affects egg and larval quality rapidly.

The effect of dietary protein content (25, 60, 95, 130, 165 and 200 g crude protein (N × 6.25)/kg diet) on the apparent and true ileal digestibilities of N and amino acids in meat-and-bone meal given to the growing rat was investigated. Semi-synthetic diets in which meat-and-bone meal was the sole protein source were given to 180 g body-weight rats for 14 d. On the fourteenth day the rats were fed and then killed 4 h after the start of feeding and digesta were sampled from the terminal 200 mm ileum. Endogenous amino acid excretion was determined for eighteen rats given an enzymically hydrolysed casein (EHC)-based diet and with subsequent treatment of the digesta using ultrafiltration. The EHC-fed rats were killed 3 h after the start of feeding and digesta were collected from the terminal 200 mm ileum. True ileal digestibility values determined with reference to Cr as a marker were higher than the corresponding apparent estimates. Apparent digestibility values of N and amino acids increased significantly (P < 0.001) with increasing dietary protein level; however, dietary protein content had no significant (P > 0.05) effect on the true ileal digestibilities of N and amino acids. The mean apparent ileal digestibility of N in meat-and-bone meal ranged from 65.6 to 75.3%. The corresponding range for the true ileal digestibility of N was 76.9 to 78.2%. True ileal digestibility, unlike apparent digestibility, appears to be independent of dietary protein level and may allow feed ingredients to be compared accurately even if they are ingested in different quantities.

The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein, or hydrolysed soya-bean protein at a level of 9.1 g/kg BW per d. The diets, which were isoenergetic with the same carbohydrate: fat ratio, were given as a continuous intragastric infusion for at least 4 d. During the last 19 h 15N-glycine (a primed continuous infusion) was given intragastrically and 15N was recovered from urinary ammonia and urea during isotope steady state for measurement of protein synthesis and protein degradation. Compared with starvation the protein-free diet decreased N excretion by 75%, probably by increasing the rate of reutilization of amino acids from endogenous proteins for protein synthesis. The protein diets produced a positive N balance which was independent of the protein source. Intact and hydrolysed casein increased protein synthesis 2.6- and 2.0-fold respectively, compared with the protein- free diet. Protein degradation increased 1.4- and 1.2-fold respectively. Hydrolysed soya-bean protein did not increase protein synthesis but decreased protein degradation by 35% compared with the protein-free diet. Compared with the hydrolysed soya-bean protein, intact casein resulted in 2.2- and 2.8-fold higher rates of protein synthesis and degradation respectively. These results are not easily explained by known sources of misinterpretation associated with the 15N-glycine method. Hydrolysed casein and hydrolysed soya-bean protein produced similar concentrations of insulin-like growth factor-1, insulin, glucagon, and corticosterone. The difference in amino acid composition between the dietary proteins was reflected in plasma amino acid composition and this is suggested to be responsible for the different effect on protein turnover. Preliminary results from this study have previously been published in abstract form (Nielsen et al. 1991).

In three experiments [amide-15N]glutamine was infused intravenously in male volunteers. After 4–8 h of infusion acidosis was achieved by an oral dose of CaCl2 (1 mmol/kg). In one subject acidosis was maintained for 5 d. The acid load produced an approximately 3-fold increase in urinary NH3 excretion, with a small (approximately 20%) and transient increase in the isotope abundance of urinary NH3 Estimates of glutamine production rate (flux) were obtained in two experiments. There was no evidence that it was increased in acidosis. The extra NH3 production by the kidney represented only a very small part, about 3%, of the total glutamine production rate.

Effects of dietary manipulations on the biliary bile acid glycine: taurine (G: T) ratio and the activity of hepatic bile acid-Co A: amino acidN-acyltransferase (EC 2.3.1) in the post-mitochondrial fraction of liver homogenates were examined in the rat. The G:T ratio in rats fed on the diet containing 100 g pectin/kg (2.18) was markedly higher than that in the animals fed on the diet containing 100 g cellulose/kg (0.09). The diets containing either 10 g cholesterol/kg or 5 g sodium cholate/kg, especially the latter, also increased the G:T ratio (0.77 and 2.33 respectively) compared with a control diet free of these steroids (0.34). When the saturating concentrations of taurine (20 mM) and glycine (100 mM) were the substrates, dietary pectin relative to cellulose significantly increased the activity of both taurine- and glycine-dependent bile acid-CoA: amino acidN-acyltransferase, but neither dietary bile acid nor cholesterol influenced it. In spite of the marked difference in the G:T ratio among the rats given various types of experimental diet, the bile acid-CoA: amino acid N-acyltransferase reaction produced taurine- but little glycine-conjugated bile acid when both taurine and glycine coexisted at physiological concentration ranges in the assay media. Dietary manipulations modified the hepatic taurine concentrations and the changes were inversely correlated with those in the G: T ratio. However, hepatic concentration of taurine (1.67–4.82 μmol/g) in rats given various types of experimental diet was comparable with or even higher than the reported Michaelis constant (Km) value ofN-acyltransferase for this compound (0.8–2.5 mM). In contrast, glycine concentrations (1.81–2.58 μmol/g) were much lower than the Km value for this amino acid (35–40 mM) under various dietary conditions. Thus, neither the substrate specificity of the bile acid conjugation enzyme nor the alteration in the hepatic concentration of taurine or glycine accounted for the change in the G: T ratio in the present study.

The changes in lipoprotein metabolism which follow the ingestion of a large fat load have been well described. The hypothesis was tested that similar changes in lipoprotein metabolism would occur after a relatively normal meal. Plasma and lipoprotein triacylglycerol, cholesterol and apolipoprotein concentrations were determined in twenty subjects (ten female) given a mixed meal containing approximately one-third of the daily intake of major nutrients in the typical Western diet. Fasting plasma triacylglycerol concentrations (range 0.38–2.70 mm/l) and the postprandial rise in plasma triacylglycerol varied considerably between subjects and were significantly associated (P < 0.01). The rise in plasma triacylglycerol corresponded to marked increases in the triacylglycerol concentration of the triacylglycerol-rich lipoproteins (TRL; chylomicrons and very-low-density lipoproteins). TRL cholesterol also increased after the meal. An increase in high-density-lipoprotein (HDL)-triacylglycerol following the meal was accompanied by a decrease in HDL-cholesterol concentration, presumably due to the action of the cholesteryl-ester transfer protein. The increases in HDL-triacylglycerol and in TRL- cholesterol were correlated with the postprandial rise in triacylglycerol in the TRL (P < 0.01). We conclude that potentially adverse changes occur in both triacylglycerol-rich and high-density lipoproteins following a typical mixed meal, as they do after large fat loads. The changes are exaggerated in those subjects with greater fasting plasma triacylglycerol concentrations.

The relationships between thiamin intake, erythrocyte transketolase (EC 2.2.1.1) activity coefficient (ETK-AC) and total erythrocyte thiamin were investigated in a group of adolescents (13 to 14 years old; nineteen boys, thirty-five girls). Thiamin intakes were calculated from 7 d weighed records, using food composition tables, and compared with those obtained by direct analysis of duplicate diets. Average 7 d calculated thiamin intakes were significantly lower than analysed intakes for both sexes. On an individual basis, calculated intakes ranged from 30 to 143 % of corresponding analysed values. Analysed and calculated intakes were significantly correlated when expressed as mg/d; however, when expressed in terms of energy intake, the correlation was significant for males only. Thiamin intake appeared largely adequate when compared with current UK dietary recommendations (Department of Health, 1991), but the limitations of such comparisons are considered. The major food groups contributing to thiamin intake were examined and showed breakfast cereals to contribute more than 25% of dietary thiamin. A proportion of the subjects had ETK-AC values in ranges usually associated with marginal or severe thiamin deficiency. There was, however, no statistically significant relationship between erythrocyte thiamin and basal or stimulated transketolase activity, or between thiamin intake and either of the methods used to assess status. The need to re-evaluate indices of thiamin status is discussed.

Twelve groups of eight rats and two control groups of sixteen rats were given semisynthetic diets with 40% energy as fat for a period of 76 d. All diets contained a minimum of 3% energy as linoleic acid and comparable basal levels of D-α- and D-γ-tocopherol. The diets varied in fat composition and in the content of DL-α-tocopheryl acetate. The diets high in polyunsaturated fatty acids (PUFA) were either rich in fish oil (FO; groups 1–4; 10% energy as fish oil PUFA), linseed oil (LN; groups 1–4; 10% energy as α-linolenic acid) or sunflower seed oil (SF; groups 1–4; 10 + 3% energy as linoleic acid). The control groups were given a diet high in monounsaturated fatty acids (MUFA; CO 1; 10 + 13% energy as oleic acid) or a diet with an ‘average’ linoleic acid content (CO 2; 8.5% energy as linoleic acid). Of each high PUFA diet three groups were supplemented with graded levels of DL-α-tocopheryl acetate. Steatitis, a sensitive histopathological indicator of vitamin E deficiency in animals fed on diets rich in fatty acids with three or more double bonds, was observed only in the adipose tissue of the FO groups, even in the group with the highest DL-α-tocopheryl acetate supplementation. Liver and serum α- tocopherol levels were found to be positively correlated and liver and serum γ-tocopherol levels negatively correlated with dietary DL-α-tocopheryl acetate. The groups on the FO diets had significantly reduced liver and serum tocopherol levels in comparison with the groups on the other high-PUFA diets. With the supplementation scheme used for the FO groups the liver α-tocopherol levels of both control groups were reached but the serum control levels were not.

Untreated wheat straw (UWS) or ammoniated wheat straw without (AWS) or with (AWSP) a supplement of potato protein of a low rumen degradability was fed to three steers according to a 3 × 3 Latin square design. All rations were supplemented with sugar-beet pulp and minerals. Voluntary organic matter intake (OMI, g/kg0.75 per d) was 67.8, 76.0 and 80.1 for whole rations (51.1, 59.7 and 59.2 for straw) for UWS, AWS and AWSP respectively, which was significantly higher for AWS and AWSP than for UWS. Organic matter digestibility (OMD, g/kg) was 561, 596 and 625 for the respective rations UWS, AWS and AWSP, also significantly higher for AWS and AWSP than for UWS. The increased voluntary intake and digestion of ammoniated wheat-straw-based rations were associated with a significantly higher potentially degradable fraction (D) of neutral detergent fibre (NDF) in offered straw (556 and 661 g/kg for untreated and ammoniated wheat straw respectively) and in the rumen pool (469, 555 and 554 g/kg for UWS, AWS and AWSP respectively). Isolated small rumen particles (retained on sieves with a pore size < 1.25 and > 0.041 mm) had a significantly lower D of NDF (average 588 g/kg) than isolated large rumen particles (average 663 g/kg). Fractional rates of degradation of NDF did not differ significantly either between untreated and ammonia-treated wheat straw offered (2.9 and 2.6%/h respectively) or between rumen pools (1.8, 1.7 and 2.1 %/h for UWS, AWS and AWSP respectively). Rations based on ammoniated wheat straw had a significantly higher rumen NH3-N concentration than UWS. Although the rumen pool size of total contents differed significantly between treatments, those of dry and organic matter and of cell wall constituents were not significantly different. The proportion of rumen dry matter passing through a sieve with a pore size of 1.25 mm averaged 0.684 over rations (not significantly different between rations). Daily rumination (96 min) and eating (52 min) times/kg NDF ingested did not differ between rations. The rate of comminution of large particles estimated from the disappearance of indigestible NDF in large rumen particles from the rumen of animals without access to feed was 4.1, 6.3 and 7.1 %/h for UWS, AWS and AWSP respectively. These values were not significantly different. The fractional rate of passage estimated from the faecal excretion of Cr-NDF was 5.4, 6.1 and 6.3%/h for UWS, AWS and AWSP respectively (significantly higher for AWS and AWSP than for UWS) but the turnover rate of indigestible NDF did not differ between treatments.