In ruminants, Co is required for the synthesis of vitamin B12, which in turn is needed for the resynthesis of methionine by methylation of homocysteine and thus, cobalamin deficiency may induce hyperhomocysteinaemia which is brought into context with perturbations of the antioxidative–prooxidative balance. The present study was conducted to explore whether Co deficiency in cattle is also associated with homocysteine-induced disturbances of oxidative status. Co deficiency was induced in cattle by feeding two groups of animals on either a basal maize-silage-based diet that was moderately low in Co (83 μg Co/kg DM), or the same diet supplemented with Co to a total of 200 μg Co/kg DM, for 43 weeks. Co deficiency was apparent from a reduced vitamin B12 status in serum and liver and an accumulation of homocysteine in plasma which was in excess of 4·8 times higher in Co-deprived cattle than in controls. The much increased level of circulating homocysteine did not indicate severe disturbances in antioxidant–prooxidant balance as measured by individual markers of lipid peroxidation, protein oxidation, and the antioxidative defence system. There were no quantitative difference in plasma thiol groups, nor were there significant changes in concentrations of α-tocopherol, microsomal thiobarbituric acid-reactive substances and carbonyl groups in liver. However, there was a trend toward increased plasma carbonyl levels indicating a slight degradation of plasma proteins in the hyperhomocysteinaemic cattle. Analysis of the hepatic catalase (EC 1.11.1.6) activity revealed an 11 % reduction in Co-deficient cattle relative to the controls. These results indicate that long-term moderate Co deficiency may induce a severe accumulation of plasma homocysteine in cattle, but considerable abnormalities in oxidative status failed to appear.

The objectives of the present study were to examine the effects of (1) ingesting mandatory snacks v. no snacks and (2) the composition of isoenergetically-dense snacks high in protein, fat or carbohydrate, on food intake and energy intake (EI) in eight men with ad libitum access to a diet of fixed composition. Subjects were each studied four times in a 9 d protocol per treatment. On days 1–2, subjects were given a medium-fat maintenance diet estimated at 1·6 × resting metabolic rate (RMR). On days 3–9, subjects consumed three mandatory isoenergetic, isoenergetically dense (380 kJ/100 g) snacks at fixed time intervals (11.30, 15.30 and 19.30 hours). Total snack intake comprised 30 % of the subjects' estimated daily energy requirements. The treatments were high protein (HP), high carbohydrate (HC), high fat (HF) and no snack (NS). The order was randomized across subjects in a counterbalanced, Latin-square design. During the remainder of the day, subjects had ad libitum (meal size and frequency) access to a covertly manipulated medium-fat diet of fixed composition (fat: carbohydrate: protein, 40:47:13 by energy), energy density 550 kJ/100 g. All foods eaten were investigator-weighed before ingestion and left-overs were weighed after ingestion. Subjective hunger and satiety feelings were tracked hourly during waking hours using visual analogue scales. Ad libitum EI amounted to 13·9 MJ/d on the NS treatment compared with 11·7, 11·7 and 12·2 MJ/d on the HP, HC and HF diets respectively (F(3,21) 5·35; P = 0·007, sed 0·66). Total EI values were not significantly different at 14·6, 14·5, 15·0 and 14·2 MJ/d respectively. Snack composition did not differentially affect total daily food intake or EI. Average daily hunger was unaffected by the composition of the snacks. Only at 12.00 hours did subjects feel significantly more hungry during the NS condition, relative to the other dietary treatments (F(3,18) 4·42; P = 0·017). Body weight was unaffected by dietary treatment. In conclusion, snacking per se led to compensatory adjustments in feeding behaviour in lean men. Snack composition (with energy density controlled) did not affect the amount eaten of a diet of fixed composition. Results may differ in real life where subjects can alter both composition and amount of food they eat and energy density is not controlled.

Knowledge of levels of fluoride ingestion and excretion is important in planning optimum fluoride therapy for young children. In previous literature, it has been assumed that only about one-third of ingested fluoride is excreted in young children. The aims of the present study were (a) to measure total fluoride intake, urinary fluoride excretion and fluoride balance, and (b) to investigate the effect of air temperature on fluoride intake and urinary fluoride excretion, in young children. Children (4 years old) living in a city, a small town and rural areas of Fars province, Iran, where drinking water contained 0·30–0·39 mg F/l, were invited to participate. Selection of subjects was by random sampling of kindergartens or health centres. The children were surveyed twice, once in summer and once in winter. Diet was obtained by 3 d diaries with interview. Samples of most foods and drinks were analysed for fluoride content. Ingestion of fluoride from toothpaste was estimated for each child. Each child's urine was collected over 24 h and analysed for fluoride content. Seventy-eight of the 116 volunteers completed all aspects of the study, which was conducted in 1995–6. For all children, the mean fluoride ingestion from diet was 0·390 (sd 0·122) mg/d or 0·028 (sd 0·008) mg/kg body weight per d. Fluoride ingestion from diet was higher in summer and higher in rural areas. The mean ingestion of fluoride from all sources was 0·426 (sd 0·126) mg/d and the mean fluoride urinary excretion was 0·339 (sd 0·100) mg/d. The difference between ingestion and urinary excretion was +0·087 (sd 0·143) mg, equivalent to 80 % excretion. Faecal excretion was not estimated. The results indicate fluoride retention at 4 years to be much lower than previously assumed.

Vegetarians have a lower incidence of many chronic diseases than omnivores. However, vegetarian diets could potentially result in lower intakes of some minerals, particularly Zn. In a cross-sectional study, dietary Zn intake was measured using 12 d weighed records in ninety-nine vegetarians (ten vegans) aged 18–50 years and forty-nine age- and sex-matched omnivores. In men, the mean daily Zn intake and Zn density values were similar in omnivores, ovolactovegetarians and vegans, but in women they were significantly lower in vegetarians (mean intake 6·8 mg v. 8·4 mg in omnivores) and few achieved the recommended intake. Significantly more vegetarian than omnivorous women had a daily Zn intake < 6 mg (44 % v. 13 %). Mean serum Zn concentrations were similar in female omnivores and vegetarians, despite the differences in intake. However, omnivorous men had a lower mean serum Zn concentration (0·85 μg/ml v. 0·95 μg/ml) and more subjects had levels below the reference range of 0·72–1·44 μg/ml than ovolactovegetarians (P < 0·01). Overall more women than men had low Zn concentrations; and these women generally had intakes below 6 mg/d. There was a significant correlation between serum Zn concentration and dietary Zn density in vegetarians, especially females (P < 0·001), but not in omnivores. Ovolactovegetarians did not have a significantly greater risk of low Zn status than omnivores.

The energy cost of locomotion of four Iberian pigs was measured in two experiments conducted when the animals averaged 41·3 (se 0·1) kg (first experiment) and 84·1 (se 0·1) kg (second experiment). The heat production of the pigs was determined when standing or walking at a speed of 0·555 m/s on a treadmill enclosed in a confinement-type respiration chamber, on different slopes (-10·5, 0, and +10·5 % in the first experiment, and -5·25, 0 and +10·5 % in the second experiment). The energy costs of locomotion, estimated from the coefficients of linear regressions of heat production per kg body weight (BW) on distance travelled, were in the first experiment 2·99, 3·31 and 5·88 J/kg BW per m for -10·5, 0, and +10·5 % inclines respectively, and 2·56, 2·84 and 7·13 J/kg BW per m for -5·25, 0 and +10·5 % inclines respectively, in the second experiment. The net energy cost of locomotion on the level appeared to be independent of live weight, attaining a value of 2·98 J/kg BW per m. Also, it was found that within experiments the net energy cost of walking on negative slopes was similar to that for locomotion on the level, indicating that no energy was recovered on vertical descent. Mean values were 3·11 and 2·72 kJ/kg BW per m for the light and heavy pigs respectively. The energy cost of raising 1 kg BW one vertical metre was found to be 27·1 J/kg BW per m in the first experiment and 40·0 J/kg BW per m in the second experiment. Correspondingly, the calculated efficiency for upslope locomotion appeared to decline with increasing BW, resulting in average values of 36·2 and 24·5 %.

An experiment was conducted to investigate the effects of branched-chain amino acid (BCAA) interactions on their utilization by growing pigs and the effects of excessive amounts of BCAA (leucine, isoleucine, valine) on the utilization of methionine. A semipurified diet containing 100 g crude protein/kg with a balanced amino acid pattern was prepared using casein supplemented with free amino acids. Three further diets were made by reducing the concentration of methionine + cyst(e)ine, valine or isoleucine by 20 %. Each of these four diets was then supplemented with leucine (50 % excess) or a mixture of BCAA (50 % excess of each but excluding the limiting amino acid). All diets were isoenergetic and were made isonitrogenous by replacement of glutamic and aspartic acids. The twelve diets were given to twenty-four growing pigs (30–40 kg) in three periods according to a randomized block design. Each period lasted 8 d and N retention was measured during the last 5 d of each period. Reducing dietary methionine, valine or isoleucine reduced the utilization of N (N retained/N digested) by approximately 20 % (P < 0·05). Adding leucine to the isoleucine-limiting diet decreased the utilization of N by 9 % (P < 0·05). This was reversed by simultaneous addition of valine. Excess leucine in a valine-deficient diet did not significantly reduce N utilization. In methionine-limiting diets an excess of either leucine alone or of all three BCAA increased the utilization of N by 8 % (P < 0·05).

The present experiment was designed to elucidate the mechanism of the methionine-sparing effect of excess branched-chain amino acids (BCAA) reported in the previous paper (Langer & Fuller, 2000). Twelve growing gilts (30–35 kg) were prepared with arterial catheters. After recovery, they received for 7 d a semipurified diet with a balanced amino acid pattern. On the 7th day blood samples were taken before (16 h postabsorptive) and after the morning meal (4 h postprandial). The animals were then divided into three groups and received for a further 7 d a methionine-limiting diet (80 % of requirement) (1) without any amino acid excess; (2) with excess leucine (50 % over requirement); or (3) with excesses of all three BCAA (leucine, isoleucine, valine, each 50 % over the requirement). On the 7th day blood samples were taken as in the first period, after which the animals were killed and liver and muscle samples taken. Plasma amino acid and branched-chain keto acid (BCKA) concentrations in the blood and branched-chain keto-acid dehydrogenase (BCKDH; EC 1.2.4.4) activity in liver and muscle homogenates were determined. Compared with those on the balanced diet, pigs fed on methionine-limiting diets had significantly lower (P < 0·05) plasma methionine concentrations in the postprandial but not in the postabsorptive state. There was no effect of either leucine or a mixture of all three BCAA fed in excess on plasma methionine concentrations. Excess dietary leucine reduced (P < 0·05) the plasma concentrations of isoleucine and valine in both the postprandial and postabsorptive states. Plasma concentrations of the BCKA reflected the changes in the corresponding amino acids. Basal BCKDH activity in the liver and total BCKDH activity in the biceps femoris muscle were significantly (P < 0·05) increased by excesses of leucine or all BCAA.

Rumen microbial degradation is an important route for detoxification of secondary plant compounds encountered in the diets of free-grazing ruminants. Exposure to diets containing particular secondary plant compounds can lead to increased rates of secondary compound degradation in the rumen. An experiment was conducted to determine whether rumen adaptation to oxalic acid would influence the diet selection of goats offered choices between plant species differing in their oxalic acid content. Twelve adult female goats were divided into two groups of six animals each. One group received a daily oral dose, in gelatin capsules, of 0·6 mmol oxalic acid/kg live weight per d throughout the experiment while the other group received placebos consisting of empty gelatin capsules. After an adaptation period of 8 d, the animals were allowed to graze a mixture of spinach (rich in oxalic acid) and cabbage (low in oxalic acid) for 7 h/d on two consecutive days per week during four consecutive 1-week periods. Intervening days were spent on grass pasture. Diet composition and intake were measured using cuticular wax n−alkanes as internal markers. Results showed that adapted goats included a higher proportion of spinach in their diet (P < 0·05) although absolute intakes of spinach were the same for the two groups. Goats in the oxalic-acid-adapted group consumed less cabbage than control animals (P < 0·05) suggesting that adaptation to oxalic acid at the rumen level may have interfered with detoxification of cabbage-derived secondary plant compounds. Voluntary intake increased progressively through the four experimental periods (P < 0·001) with a tendency for higher intakes among control than among adapted animals (P < 0·1). The experiment demonstrates how differences in the rate of degradation of secondary plant compounds may influence diet selection in ruminants.

Four ruminally and duodenally cannulated multiparous Finnish Ayrshire cows were fed on diets consisting of grass silage (0·6 kg/kg DM) and one of four concentrates: barley, barley + urea, barley + rapeseed meal and barley + rapeseed cake. The objective of the present study was to compare omasal canal and duodenal digesta flows. Values for digesta flow into the omasal canal and duodenum were determined using a triple-marker method based on Co-EDTA, Yb-acetate and indigestible neutral-detergent fibre (NDF) markers. Microbial non-NH3 N (NAN) flow was assessed by purine flow. Microbial samples to determine the bacterial purine: N ratio were harvested from the rumen, omasum and duodenum. Organic matter flow was significantly lower into the omasum than the duodenum, indicating an endogenous organic matter secretion into the abomasum. In contrast, NDF and acid-detergent fibre flows were significantly higher into the omasum indicating digestion of fibre in the omasum. Microbial NAN flows were significantly different (P < 0·001) when estimates were based on bacterial samples harvested from different sites. Differences in total NAN, microbial NAN and dietary NAN flows entering the omasal canal and duodenum were non-significant. The results indicated that the omasal sampling technique provides a promising alternative to the duodenal sampling technique to investigate forestomach digestion in dairy cows and offers an alternative means to study rumen N metabolism.

Intrauterine growth retardation associated with maternal undernutrition is proposed to play a significant role in the aetiology of hypertension and CHD. Animal experiments suggest that the kidney, which is extremely vulnerable to the adverse effects of growth-retarding factors, may play an important role in the prenatal programming of hypertension. Maintenance of renal haemodynamic functions following structural impairment in fetal life is proposed to require adaptations which raise systemic blood pressure and promote a more rapid progression to renal failure. Rats were fed on diets containing 180 g casein/kg (control) or 90 g casein/kg (low protein) during pregnancy. The offspring were studied in terms of blood pressure, creatinine clearance, blood urea N, plasma and urinary albumin, renal morphometry and metabolic activity at 4, 12 and 20 weeks of age. Blood pressure was elevated at all ages in the low-protein-exposed offspring, relative to control rats. Rats (4 weeks old) exposed to the low-protein diet had smaller kidneys which were shorter and wider than those of control animals. Creatinine clearance was significantly reduced in 4-week-old rats exposed to the low-protein diet. Renal morphometry and creatinine clearance at older ages were not influenced by prenatal diet. Blood urea N, urinary output and urinary albumin excretion were, however, significantly greater in low-protein-exposed rats than in control rats at 20 weeks of age. These findings are suggestive of a progressive deterioration of renal function in hypertensive rats exposed to mild maternal protein restriction during fetal life. This is consistent with the hypothesis that adaptations to maintain renal haemodynamic functions following impairment of fetal nephrogenesis result in an accelerated progression towards glomerulosclerosis and increased intrarenal pressures mediated by rising vascular resistance.

The present study measured changes in antioxidant enzyme activity in, and the phospholipid fatty acid composition of the ageing rat brain and tested whether dietary supplementation with thyme oil or thymol could provide beneficial effects. There were significant declines in superoxide dismutase (EC 1.15.1.1) and glutathione peroxidase (EC 1.11.1.9) activities and the total antioxidant status in the untreated rats with age, while thyme-oil- and thymol-fed rats maintained significantly higher antioxidant enzyme activities and total antioxidant status. The proportions of 18: 2n−6, 20: 1n−9, 22: 4n−6 and 22: 5n−3 in the brain phospholipids resulting from all three dietary treatments were significantly higher in 28-month-old rats than in 7-month-old rats. Only 20: 1n−9 levels in 28-month-old thyme-oil- and thymol-treated rats were significantly higher than in the age-matched control. The proportion of 22: 6n−3 in brain phospholipids, which declined with age in control rats, was also significantly higher in rats given either supplement. This latter finding is particularly important as optimum levels of 22: 6n−3 are required for normal brain function. These results highlight the potential benefit of thyme oil as a dietary antioxidant.