Allantoin net flux through the plasma allantoin compartment was determined in sheep given a roughage diet by means of a continuous infusion of [4,5-14C]allantoin for 17 h. Unlabelled allantoin was infused intravenously during the last 7 h of the tracer infusion to increase the allantoin flux by approximately 75 %. When unlabelled allantoin was infused, the specific radioactivity of allantoin in plasma and urine declined exponentially to approach a lower plateau some 2–3 h later. The estimate of net flux during the infusion of unlabelled allantoin, estimated from blood plasma and urine, was on average 79 and 90 % of expected values. Expected values of allantoin net flux during infusion of unlabelled allantoin were calculated as the sum of allantoin net flux pre-loading plus the known rate of infusion of the allantoin load. It is probable that endogenous allantoin synthesis was decreased by the infusion of allantoin: allosteric inhibition of uricase appears a plausible explanation for this observation, and for lower estimates of net flux. Appearance of labelled allantoin-C in ruminal or blood bicarbonate was negligible. Our results indicate that net flux of allantoin through blood plasma is a good predictor of the entry rate of allantoin into the primary compartment and should be a better predictor of rumen microbial outflow than urinary allantoin excretion. However, measurements of allantoin-specific radioactivity, during continuous infusion, should be taken after a period of 24 h, at which time the true plateau specific radioactivity value of allantoin in plasma would be attained.

A dynamic model was developed to assess insulin sensitivity and pancreatic response in lactating dairy cows. The model is based on a simultaneous analysis of insulin and glucose intravenous tolerance tests. It comprises five compartments corresponding to insulin in portal–hepatic plasma, and insulin or glucose in both systemic plasma and in interstitial fluid. Insulin secretion rate is a sigmoidal function of glucose in plasma. Insulin is cleared from hepatic plasma and from the interstitial fluid. The glucose entry rate is constant and glucose utilization rate is a sigmoidal function of insulin in the interstitial fluid. Six parameters were estimated: two for insulin secretion rate, two for insulin clearance, one for glucose entry rate and one for glucose utilization rate. After integration of the functions, the model yielded a relative estimate of the quantities of insulin secreted and cleared, as well as the glucose entering and utilized during each test. Using an experimental dataset composed of ten pairs of tolerance tests, the explained variations for plasma insulin and glucose concentrations were 96·0 and 98·3 % and standard errors of estimates were 0·032 nmol/l and 0·14 mmol/l respectively. Except in the early stages after injection, residual errors were low. A Jackknife analysis showed that the estimated parameters exhibited low statistical bias. This model simplifies the interpretation of both tests through a simulation based on six common parameters. Compared to a classical analysis of tolerance tests, it may improve the analysis of modifications in the key functions regulating glucose homeostasis in ruminants.

The aim of the present study was to evaluate a model of body composition for assessing total body protein (TBP) mass using dual-energy X-ray absorptiometry (DXA), with either measured or assumed total body water (TBW); it was intended to provide a less complex or demanding alternative technique to, for example, the four-component model (4-CM). The following measurements were obtained in healthy adults (n 46) aged 18–62 years, and children (n 30) aged 8–12 years: body weight (BWt), body volume (BV; under-water weighing), TBW (2H-dilution space or predicted using an assumed hydration fraction of fat-free mass (HFffm)), bone mineral content (BMC; DXA) and fat-free soft tissue (FFST; DXA). TBP was calculated using the 4-CM (TBP=3·050BWt-0·290TBW-2·734BMC-2·747BV) and the DXA model (TBP=FFST-0·2305BMC-TBW). DXA measurements were obtained using the Lunar DPX (Lunar Radiation Corporation, Madison, WI, USA) or Hologic QDR 1000/W (Hologic, Waltham, MA, USA). Precision of the DXA model for TBP with measured TBW (4·6–6·8 % mean TBP) was slightly worse than the 4-CM (4·0–5·4 %), whereas that modelled with assumed HFffm was more precise (2·4–5·2 %) because it obviated imprecision associated with measuring TBW. Agreement between the 4-CM and DXA model with measured TBW was also worse (e.g. bias, 15 % of the mean; 95 % limits of agreement up to ±39 % for adults measured on the Lunar DPX) than when a constant for HFffm was assumed (3·7 % and ±21 % respectively). Most of the variability in agreement between these various models was due to interpretation of biological factors, rather than to measurement imprecision. Therefore, the DXA model, which is less complex and demanding than the 4-CM, is of value for assessing TBP in groups of healthy subjects, but is of less value for individuals in whom there may be substantial differences from reference 4-CM estimates.

The aims of the present study were to investigate (1), the potential anthelmintic properties and (2), the nutritional consequences of commercially available condensed tannins on parasitised sheep fed, ad libitum, either a high- or a low-protein food. For this purpose, forty-eight previously parasite-naïve sheep (n 12) were infected with 2000 Trichostrongylus colubriformis larvae/d for a 67-d experimental period. Two experimental foods were made: a low (L), formulated to be inadequate in meeting the requirements of growing sheep for metabolisable protein (MP), and based on wheat, citrus pulp, and oatfeed; a high (H), expected to be above the requirements of growing sheep for MP, based on similar ingredients but supplemented with protected soyabean meal. Two additional foods were made by adding 60 g Quebracho (a condensed tannins (CT) extract)/kg fresh matter to foods L and H (foods LQ and HQ respectively). This level of Quebracho supplementation has been previously shown to reduce the level of parasitism in restrictedly fed, parasitised sheep. The experiment was divided into two periods: period 1 (P1, day 1–38) and period 2 (P2, day 39–67), each one associated with different phases of an intestinal parasitic infection. Six sheep from each group were slaughtered at the end of P1, and the remaining sheep were slaughtered at the end of P2 (day 67). Although faecal egg counts (FEC; number of parasite eggs/g faeces) and total egg output were reduced in sheep offered the supplemented foods during P1 (P<0·05), worm burdens on day 38 were unaltered. Neither Quebracho supplementation nor food protein content during P2 affected FEC and worm burdens. Food intake and performance were higher in sheep offered food HQ compared with sheep offered food H (P<0·05); no differences were observed in sheep offered foods LQ and L throughout the experiment. The previously shown anthelmintic properties of CT were not observed following ad libitum intake of either low- or high-protein foods supplemented with Quebracho extract. Higher levels of CT supplementation may be required to reduce parasitism and consequently improve the performance of parasitised sheep, when fed ad libitum. Supplementation with CT conferred advantages on the performance of parasitised sheep on a high- but not on a low-protein food.

The aim of this present study was to identify the earliest point at which riboflavin deficiency affects post-weaning bowel development in rats. After weaning, eighty Wistar rats were weight-matched as pairs, one animal being fed a normal synthetic diet and the other being fed the same diet but deficient in riboflavin. Body weight, feeding and rates of growth were monitored and eight pairs of animals were taken for analysis at 45, 69, 93, 117 and 141 h. Riboflavin status was monitored by determining the erythrocyte glutathione reductase activation coefficient (EGRAC), and hepatic flavins were measured by a fluorescence assay. Changes to the number and dimensions of villi and crypts in the duodenum were determined, as well as crypt division (bifurcation) and the DNA synthesis index of the crypt epithelium by bromodeoxyuridine (BrdU) labelling. Riboflavin deficiency was established in the experimental rats, as demonstrated by a significant increase in EGRAC after 45 h (P<0·001) and decreased liver flavins after 96 h (P<0·001). After 96 h a significant increase in the size and cellularity of the crypts (P<0·001 in both cases) was seen in these riboflavin-deficient animals, with a decreased incidence of bifurcating crypts and of BrdU-labelled cells. No changes to villus number or size were observed. The present study has demonstrated that developmental changes to the duodenal crypt arise shortly after circulating riboflavin measurements show evidence of deficiency. These changes primarily affect cell proliferation and crypt bifurcation, and precede long-term changes such as the reduction of villus number.

The aim of the present study was to evaluate the effect of diets rich in n-6 and n-3 fatty acids on adipose tissue metabolism. Starting at weaning, male Wistar rats were fed ad libitum, for 8 weeks with one of the following diets: C, rat chow; S, rat chow containing 15 % (w/w) soyabean oil; F, rat chow containing 15 % (w/w) fish oil; SF, rat chow containing 15 % (w/w) soyabean and fish oil (5:1, w/w). Casein was added to the fat diets to achieve the same 20 % (w/w) protein content as in the control chow. Food intake and body weight were measured weekly. The rats were killed by decapitation and the retroperitoneal (RET) and epididymal (EPI) white adipose tissues were removed and weighed. Tissue lipid and protein content, in vivo lipogenesis rate, uptake of diet-derived lipids, in vitro lipolytic rate, adipocyte area, lipoprotein lipase, ATP citrate lyase, and malic enzyme activities were evaluated. Carcass lipid and protein contents were also measured. Energy intake was reduced while carcass lipid content was increased in the three fat-fed groups. However, carcass protein and body weight gains were elevated only with diets F and SF. Lipolysis rate was diminished by diets F and SF, while the uptake of diet-derived lipids was elevated by the diet S in both RET and EPI tissues. These metabolic alterations may have contributed to the increase in in vivo lipogenesis rate in the presence of decreased ATP citrate lyase and malic enzyme activities induced by the three lipid diets. These results indicate that enrichment of the diet with polyunsaturated fatty acids causes changes in adipose tissue metabolism that favour fat deposition. Different metabolic pathways were preferentially affected by each type of fatty acid used.

Iron deficiency induces thymus atrophy in laboratory animals and very likely in humans by unknown mechanisms. The atrophy is associated with impaired cell-mediated immunity. In this study, we tested the hypothesis that thymus atrophy is a result of increased apoptosis and reduced thymocyte proliferation. Thymocytes were obtained from twenty-seven control, twenty-seven pairfed, twenty-seven iron-deficient (ID) mice; twelve and fourteen ID mice that received the control diet (0·9 mmol/kg versus 0·09 mmol/kg for the ID diet) for 1 d (repletion, R1) and 3 d (R3), respectively. Cell cycle analysis and apoptosis were studied by flow cytometry using propidium iodide staining and terminal deoxyuridine nick end labeling of DNA breaks assay respectively. When mice were killed, haemoglobin, haematocrit, and liver iron stores of ID, R1, and R3 mice were 25–40 % of those of control and pairfed mice (P<0·01). Absolute and relative thymus weights and thymocyte numbers were 19 to 68 % lower in ID, R1, and R3 than in control and pairfed groups (P<0·05). We found no significant difference among groups in the percentage of cells undergoing apoptosis. A higher percentage of thymocytes from ID and R1 mice than those of control, pairfed, and R3 mice were in the resting phase of the normal cell cycle (P<0·05). Conversely, a lower percentage of thymocytes from ID and R1 mice than those from control, pairfed, and R3 mice were in the DNA synthesis phase and late phase of DNA synthesis and onset of mitosis (G2-M) (P<0·05). Indicators of iron status positively correlated (r 0·3 to 0·56) with the percentage of thymocytes in the G2-M phase (P<0·05). Results suggest that reduced cell proliferation but not increased apoptosis is the cause of thymus atrophy associated with iron deficiency.

Bile acids derived from intestinal bacterial metabolism and transported to the breast in plasma may influence risk of breast cancer. The purpose of the present study was to test the hypothesis that fatty acid chain length and degree of unsaturation differ with regard to their influence on the postprandial release of cholecystokinin (CCK) and the subsequent increase in plasma bile acid concentrations that occur following a meal. A randomized crossover design was used to compare five high-fat test meals (50 g fat) with a low-fat test meal (15 g) on plasma bile acid and CCK concentrations in eighteen healthy premenopausal women. The high-fat meals were enriched in oleate or palmitate, or linoleate or medium-chain triacylglycerols (MCT) or a blend of oleate and long-chain n-3 fatty acids. The postprandial increase in plasma CCK concentration was lower on the MCT meal compared with all meals and was greater following the linoleate compared with the low-fat meal. Plasma bile acid concentrations increased 2–3-fold postprandially but the increase was lower following the MCT meal compared with the other meals and was greater on the linoleate meal compared with the low-fat meal. The postprandial increases in plasma chenodeoxycholic acid concentration showed a trend to rise with increasing unsaturation of the test meal. In conclusion, meals rich in linoleate are a potent stimulus for CCK release and lead to prolonged elevations of plasma bile acids and meals containing MCT inhibit CCK release and the subsequent increase in plasma bile acid concentrations.

Our objective was to determine whether dietary supplementation with phosphatidylcholine (PC) plus vitamin B12 could afford beneficial effects on biochemical and biophysical events in the brain of senescence-accelerated mouse (SAM) substrain SAMP8. We measured learning behaviour, hippocampal protein kinase C (PKC) activity, cerebral antioxidant status, phospholipid composition and fatty acid composition in 6-month-old SAMP8 and in age-matched controls (SAM substrain SAMR1). In comparison with SAMR1, SAMP8 showed a significant elevation in total grading score of senescence (P<0·05) and a significant decline in acquisition (P<0·05). SAMP8 had a lower hippocampal PKC activity and cerebral PKC-β mRNA abundance than SAMR1. SAMP8 had increased cerebral lipid peroxide levels and proportion of sphingomyelin, and a lower proportion of 20 : 4n-6 and 22 : 6n-3 in cerebral phosphtidylethanolamine than SAMR1. SAMP8 fed the PC combined with vitamin B12 diet had an increased PKC activity and a higher proportion of 22 : 6n-3 than SAMP8 fed the control diet. These results indicate the potential benefit of PC combined with vitamin B12 as a dietary supplement.

In study 1, four cows had a ruminal canula, a catheter in the right ruminal vein and an ultrasonic flow probe around the right ruminal artery; a catheter was placed in the auricular artery on experimental days. Blood samples were taken every 10 min from -20 to 60 min after ruminal infusion of 5·79 mmol pteroylmonoglutamic acid and cyanocobalamin. There was a net release of these vitamins across the rumen wall following the infusion (P=0·06). In studies 2 and 3, four cows had catheters in the portal, one hepatic and two mesenteric veins and one mesenteric artery. Plasma flow was determined using p-aminohippurate. In study 2, blood samples were taken before and every 30 min for 6 h after feeding 0 or 4 mg of pteroylmonoglutamic acid. Flow of folates through the portal-drained viscera (PDV) and the total splanchnic tissues (TSP) tended to increase with the ingestion of pteroylmonoglutamic acid (P=0·19). In study 3, blood samples were collected every 30 min for the first 3 h to calculate plasma flow and basal net fluxes of folates and vitamin B12. The cows were fed 2·6 g pteroylmonoglutamic acid and 500 mg cyanocobalamin; blood samples were taken every 2 h for 24 h. Vitamin supplements increased the net release of folates and vitamin B12 from PDV (P=0·04) and TSP (P=0·13). The present results demonstrate that, in dairy cows, at doses reported to improve animal performance, passage of pteroylmonoglutamic acid to the portal blood appears during the 6 h following its ingestion, whereas for cyanocobalamin, it is a slow process, not yet completed 24 h after its ingestion.

The accuracy of the multiple pass 24 h recall for assessment of habitual energy intake in pre-school children is unclear. The primary aim of this study was to assess its accuracy by comparison with measurement of total energy expenditure by doubly-labelled water in a representative sample of forty-one 3–4-year-olds. The recall method was well tolerated by subjects and was administered quickly and easily. However, it produced estimates of energy intake which significantly exceeded measures of total energy expenditure from doubly-labelled water, mean paired difference 660 kJ/d (P<0·01). Agreement between the two methods was poor at the individual level: limits of agreement 660+/-3018 kJ/d. Error and imprecision in both methods contribute to individual differences, but the present study suggests that the multiple pass 24 h recall does not provide accurate estimates of dietary energy intake in individual children.

Resistant starch and inulin are complex carbohydrates that are fermented by the microflora and known to increase colonic absorption of minerals in animals. The fermentation of these substrates in the large bowel to short-chain fatty acids is the main reason for this increase in mineral absorption. The purpose of the present study was to examine the potential synergistic effect of a combination of these two fermentable carbohydrates. For this purpose, thirty-two adult male Wistar rats weighing 200 g were used in the present study. The rats were distributed into four groups, and fed for 21 d a fibre-free basal purified diet or diet containing 100 g inulin, or 150 g resistant starch (raw potato starch)/kg diet or a blend of 50 g inulin and 75 g resistant starch/kg diet. After an adaptation period of 14 d, the rats were then transferred to metabolic cages and dietary intake, faeces and urine were monitored for 5 d. The animals were then anaesthetized and caecal Ca and Mg absorption were measured. Finally, the rats were killed and blood, caecum and tissues were sampled. Ca and Mg levels were assessed in diets, faeces, urine, caecum and plasma by atomic absorption spectrometry. Our results confirmed that inulin and resistant starch ingestion led to considerable caecal fermentation in the three experimental groups compared with the control group diet. Moreover, both carbohydrates significantly increased the intestinal absorption and balance of Ca and Mg, without altering the plasma level of these two minerals. Interestingly, the combination of the studied carbohydrates increased significantly (P<0·05) the caecal soluble Ca and Mg concentrations, the apparent intestinal absorption and balance of Ca, and non-significantly the plasma Mg level. In conclusion, a combination of different carbohydrates showed synergistic effects on intestinal Ca absorption and balance in rats. Further studies with other types of carbohydrate combinations should be carried out to extend these findings.

The small intestines of hatching chicks undergo rapid developmental changes in the immediate post-hatch period when the birds are making the transition from endogenous nutrient supply from yolk to dependence on exogenous feed. This transition usually only begins 48 h or more after hatching, owing to logistical considerations of production. The effects of fasting for 48 h at different times during this critical period on small intestinal development and enterocyte dynamics were examined by morphometric determinations and use of staining for proliferative-cell nuclear antigen and 5-bromo-2-deoxyuridine. The effects of fasting were specific to both time of fasting and the intestinal segment examined. Decreased development was found in the duodenum and jejunum, but was less apparent in the ileum. Fasting between 0 and 48 h decreased crypt size in the duodenum and jejunum, the number of crypts per villus, crypt proliferation, villus area and the rate of enterocyte migration. Fasting at later times resulted in smaller effects, although the jejunum appeared to be the most sensitive of the intestinal segments. Growth was correlated with the number of cells in the crypts, the number of cells along the villus and the segment surface area. The common practice whereby feed is first available to chicks more than 48 h post-hatch may depress subsequent development.

Red kidney beans (Phaseolus vulgaris) processed to differ in distribution and content of indigestible carbohydrates were used to study hindgut fermentability and production of short-chain fatty acids (SCFA). Bean flours with low or high content of resistant starch (RS), mainly raw and physically-inaccessible starch, were obtained by milling the beans before or after boiling. Flours containing retrograded starch and with a high or low content of oligosaccharides were prepared by autoclaving followed by freeze-drying with or without the boiling water. Six diets were prepared from these flours yielding a total concentration of indigestible carbohydrates of 90 or 120 g/kg (dry weight basis). The total fermentability of the indigestible carbohydrates was high with all diets (80–87 %). Raw and physically-inaccessible starch was more readily fermented than retrograded starch (97–99 % v. 86–95 %; P<0·05). Non-starch glucans were fermented to a lesser extent than RS, but the fermentability was higher (P<0·05) in the case of autoclaved (50–54 %) than boiled beans (37–41 %). The distribution between acetic, propionic and butyric acid in the caecum was similar for all diets, with a comparatively high percentage of butyric acid (approximately 18). However, with diets containing the high amounts of RS, the butyric acid concentration was significantly higher in the distal colon than in the proximal colon (P=0·009 and P=0·047 for the high- and low-level diets respectively), whereas it remained constant, or decreased along the colon in the case of the other diets. Furthermore, the two diets richest in RS also promoted the highest percentages of butyric acid in the distal colon (24 and 17 v. 12 and 12–16 for the high- and low-level diets respectively).

We measured daily dietary Cr intake in southern Spain by sampling duplicate diets for seven consecutive days in different population groups. Cr was determined by electrothermal atomization–atomic absorption spectrometry. The samples were mineralized in a digestion block with HNO3, HClO4 and V2O5. A total of 161 duplicate diets from twenty-three subjects were analysed, and mean levels of Cr intake ranged from 9·39 to 205·16 μg/d. Mean Cr intake (100 μg/d) was similar to levels found for most other countries, and was within the range recommended by the National Research Council for a safe and adequate daily intake (50–200 μg/d). Chromium intake correlated significantly with energy, protein and carbohydrate intake, and with the daily intake of Zn, Fe, Mg, K, Na, Ca and nicotinic acid in the diets analysed.

Incubations were carried out with a batch culture system to study the effects of different N sources on the fermentation by ruminal micro-organisms from Merino sheep of two fibre substrates derived from feedstuffs that differed in their fermentation rate. The substrates were neutral-detergent fibre (NDF) from barley straw and sugarbeet pulp. N sources were ammonia (NH4Cl) and peptides (Trypticase). Three treatments were made by replacing ammonia-N with peptide-N at levels of 0 (AMMO), 33 (PEPLOW) and 66 % (PEPHIGH) of total N. There were no differences (P>0·05) between treatments in NDF degradation for both the barley straw and the sugarbeet pulp. Peptides increased (P<0·05) total volatile fatty acids daily production for both substrates, with greater values (P<0·001) for PEPHIGH than for PEPLOW for the sugarbeet pulp. The presence of peptides also increased (P<0·05) microbial N synthesis compared with AMMO, with PEPHIGH supporting more growth (P<0·001) than PEPLOW when the sugarbeet pulp NDF was fermented. The presence of peptides increased (P<0·01) the amount of solids-associated micro-organisms (SAM)-N for both the barley straw and the sugarbeet pulp fibres, values in the PEPHIGH treatment being higher (P<0·001) than those in PEPLOW. The proportion of SAM-N in the total microbial N was not affected (P>0·05) by the presence of peptides compared with the AMMO treatment, but values were greater for the PEPHIGH compared with the PEPLOW N source, reaching statistical significance (P<0·05) only for the sugarbeet pulp. For liquid-associated micro-organisms, the AMMO treatment resulted in the greatest (P<0·05) proportion of N derived from ammonia for both substrates, with a further decrease (P<0·01) for the PEPHIGH treatment compared with the PEPLOW for the sugarbeet pulp, indicating preferential uptake of peptides when they were available. Microbial growth efficiency (g microbial N/kg NDF degraded) was not affected (P>0·05) by N source. These results indicate that N forms other than ammonia are needed for maximal growth of fibre-digesting ruminal micro-organisms.

Many Indonesian infants have an inadequate nutritional status, which may be due in part to inadequate maternal nutrition during pregnancy. This study was designed to investigate whether infant nutritional status could be improved by maternal vitamin A and Fe supplementation during gestation. Mothers of these infants from five villages had been randomly assigned on an individual basis, supervised and double-blind, to receive supplementation once weekly from approximately 18 weeks of pregnancy until delivery. Supplementation comprised 120 mg Fe and 500 μg folic acid with or without 4800 retinol equivalent vitamin A. Mothers of infants from four other villages who participated in the national Fe and folic acid supplementation programme were also recruited; intake of tablets was not supervised. Anthropometric and biochemical parameters of infants and their mothers were assessed approximately 4 months after delivery. Infants of mothers supplemented with vitamin A plus Fe had higher serum retinol concentrations than infants of mothers supplemented with Fe alone. However, the proportion of infants with serum retinol concentrations <0·70 μmol/l was >70 % in all groups. Maternal and infant serum retinol concentrations were correlated. Fe status, weight and length of infants were similar in all groups. Fe status of girls was better than that of boys, but boys were heavier and longer. We conclude that supplementation with vitamin A in conjunction with Fe supplementation of women during pregnancy benefits vitamin A status of their infants. However, considering the large proportion of infants with marginal serum retinol concentrations, it may still be necessary to increase their vitamin A intake.