The plasma-cholesterol-lowering effects of some dietary legumes are now well established from animal and human studies, but the mechanism is not completely understood. The present study investigated the effect of baked beans (Phaseolus vulgaris) on steroid metabolism of hypercholesterolaemic pigs. Three groups of four pigs were studied: baseline (BL), normal pigs (NP) and those previously prepared with an ileo-rectal anastomosis to nullify the function of the large intestine (IR). All three groups were given a semi-purified control diet, with about 40% energy as fat (polyunsaturated: saturated fatty acid (P:S) ratio 0.3), supplemented with 10 g cholesterol/kg, for 14 d. Then IR and NP pigs were fed for 28 d on a diet supplemented with 10 g cholesterol/kg and 300 g baked beans/kg (dry-matter basis), so that the 40% contribution to energy from fat was maintained (P:S ratio 0.3). Group BL was fed on the control diet throughout. The intact pigs (NP) fed on baked beans showed considerable differences compared with the other groups, as follows: (a) reduced plasma cholesterol (NS); (b) higher concentration of cholesterol in bile (NS); (c) higher concentration of bile acids, especially secondary bile acids, in bile (P < 0.05); (d) reduced elimination of bile acids in faeces, especially secondary bile acids (P < 0.05); (e) higher excretion of coprostanol and lower elimination of cholesterol in faeces (P < 0.05). From these findings it is proposed that a baked-bean-enriched diet potentiates bacterial fermentation and steroid degradation in the large intestine and enhances conservation of bile acids and cholesterol within the enterohepatic circulation. The high concentration of bile acids and cholesterol in bile may thus promote feedback inhibition of hepatic cholesterol synthesis, and hence, reduce plasma cholesterol.

The absorption of 59Fe from preparations of FeSO4 and the ferric hydroxypyranone complexes maltol and ethyl maltol was studied by whole-body counting in normal subjects and patients with Fe deficiency. Fe in the Fe3+ complexes was in general absorbed almost as well as Fe2+. It is concluded that the absorption of Fe3+ from hydroxypyranone complexes is much greater than that from simple Fe3+ salts; this may prove an efficient and less toxic form of Fe for the treatment of deficiency.

This study has investigated the influence of dietary fatty acid composition on mammary tumour incidence in N-ethyl-N-nitrosourea (ENU)-treated rats and has compared the susceptibility to dietary fatty acid modification of the membrane phospholipids phosphatidylinositol (PI) and phosphatidylethanolamine (PE) from normal and tumour tissue of rat mammary gland. The incidence of mammary tumours was significantly lower in fish oil- (29%), compared with olive oil- (75%; P < 0.04) but not maize oil-(63%; P < 0.1) fed animals. No differences in PI fatty acid composition were found in normal or tumour tissue between rats fed on maize oil, olive oil or fish oil in diets from weaning. When normal and tumour tissue PI fatty acids were compared, significantly higher amounts of stearic acid (18:0) were found in tumour than normal tissue in rats given olive oil (P < 0.05). A similar trend was found in animals fed on maize oil, although differences between normal and tumour tissue did not reach a level of statistical significance (P < 0.1). In mammary PE, maize oil-fed control animals had significantly higher levels of linoleic acid (18:2n−6) than either olive oil- or fish oil-fed animals (P < 0.05, both cases) and levels of arachidonic acid were also higher in maize oil- compared with fish oil-fed animals (P < 0.05). In tumour-bearing animals no differences in PE fatty acid composition were found between the three dietary groups. When normal and tumour tissue PE fatty acids were compared, significantly lower amounts of linoleic acid (18:2n-6; P < 0.01) and significantly greater amounts of arachidonic acid (20:4n−6; P < 0.05) were found in tumour than normal tissue of rats fed on maize oil. The present study shows that the fatty acid composition of PI from both normal and tumour tissue of the mammary gland is resistant to dietary fatty acid modification. The PE fraction is more susceptible to dietary modification and in this fraction there is evidence of increased conversion of linoleic acid to arachidonic acid in tumour compared with normal tissue. Lower tumour incidence rates in rats given fish oils may in part be due to alteration in prostanoid metabolism secondary to displacement of arachidonic acid by eicosapentaenoic acid, but PE rather than PI would appear to be the most likely locus for diet-induced alteration in prostanoid synthesis in this tissue. Effects of dietary fatty acids other than on the balance of n−6 and n−3 fatty acids, and on prostanoid metabolism, should also be considered. The significance of increased stearic acid content of PI in tumours of olive oil-fed animals and the possible influence of dietary fatty acids on the capacity for stearic acid accumulation requires further study.

Nine 5-month-old lambs were randomly allocated to two groups and were fed on either a Co-deficient whole-barley diet (n 5), or the same diet supplemented with Co (n 4). The lambs were fed on their respective diets for 28 weeks. Plasma vitamin B12 concentrations fell below the lower limit of normality after 6 weeks, and plasma methylmalonic acid (MMA) concentrations rose above the upper limit of normality after 10 weeks. However, plasma MMA concentrations fell to near normal levels towards the end of the experiment suggesting that diagnosis of more severe Co deficiency based on determination of plasma MMA concentrations may be of limited value. Analysis of tissue samples collected at slaughter revealed a marked reduction in the vitamin B12 concentration and the activity of methylmalonyl-CoA mutase (EC 5.4.99.2) in the tissues taken from the Co-deficient sheep, by comparison with the controls. Although tissue concentrations of MMA in the Co-deficient animals were not significantly different from those of the controls, we did detect increased concentrations of branched-chain fatty acids. This suggested that misincorporation of MMA, but not propionic acid, into fatty acids had occurred. The Co-deficient lambs did not develop any neurological signs, suggesting that accumulation of branched-chain fatty acids may not be involved in the development of neurological lesions.

Broiler chickens were fed on a control diet based on maize or a diet containing sugar-beet-pulp fibre (Beeffiber) at an inclusion level of 46 g/kg. Diets were provided ad lib. or at a restricted level either once daily or three times daily. On days 13 and 20, chickens fed on the ad lib. control and sugar-beet-pulp-containing diets generally weighed more and had poorer feed conversion ratios than chickens given the restricted control and sugar-beet-pulp-containing diets respectively. Furthermore, chickens given the restricted diets once daily had greater body weights and generally improved feed conversion efficiencies compared with chickens given the restricted diets three times daily. Generally, elevated plasma Lipid concentrations were observed amongst chickens given the restricted diets once daily compared with chickens fed ad lib. as well as the restricted diets three times daily, while chickens fed on restricted diets three times daily had plasma lipid concentrations intermediate between those fed ad lib. and once daily. In a plasma triacylglycerol response study on day 22, feeding of sugar-beet-pulp-containing diets generally reduced postprandial triacylglycerol concentrations and delayed triacylglycerol response relative to chickens given the control diets either ad lib. or restricted, which may indicate gastrointestinal adaptation to feeding of a fibre-rich diet. Postprandial triacylglycerol concentrations observed for chickens receiving restricted diets were increased compared with chickens given the respective ad lib. diets, indicating adaptation of chickens to reduced feed frequency. On day 25, feeding of sugar-beet-pulp-containing diets decreased digesta dry matter content and ileal organic matter digestibility. Chickens given sugar-beet-pulp-containing diets generally had, on day 25, increased caecal short-chain fatty acid (SCFA) concentrations in comparison with chickens given the ad lib. control diet, indicating increased fermentation of dietary components. It is also noteworthy that the greatest SCFA concentrations were observed amongst chickens given the control diet once daily, suggesting enhanced caecal fermentation capacity. This may have been a consequence of increased bacterial activity and caecal hypertrophy due to infrequent feeding of a low-fibre diet.

High intakes of Fe may impair Cu status, but the underlying mechanism is not known. Male rats, aged 7 weeks, were given purified diets adequate in Cu (8 mg Cu/kg) and containing either 7, 40 or 389 mg Fe/kg. After 6 weeks the concentrations of Fe in liver and spleen were positively related with dietary Fe level and those of Cu were negatively related with dietary Fe level. Increasing Fe intakes reduced apparent absorption and biliary excretion of Cu in a dose-dependent fashion. In individual rats, biliary Cu excretion showed a significant, positive correlation with Liver Cu concentration. It is concluded that increased Fe intakes depress Cu absorption which produces a decrease in plasma and organ Cu concentrations. As a result, biliary Cu excretion is lowered which contributes to achieving Cu balance at high Fe intakes. Because the concentrations of Cu in plasma and bile, and also plasma ceruloplasmin (EC 1.16.3.1) activities, showed much greater percentage reductions with increasing Fe intake than did the concentrations of Cu in organs, it is possible that increased Fe status interferes with the mobilization of Cu stores.

Fermentable energy in insoluble dietary fibre (DF) sources was evaluated by in vivo and in vitro methods using rats. Test diets contained 50 and 100 g maize husk or organic-acid-treated maize husk/kg diet. Soluble fractions were removed from both the DF sources by washing. The acid treatment increased digestibility by a microbial hemicellulase from 12.7% to 32.6%. The fermentabiliry of DF was evaluated by measurement of the production rate of short-chain fatty acids (SCFA) in a short-term in vitro incubation of the caecal contents of rats fed on test diets for 22 d. The production rates of the major SCFA, acetic, propionic and butyric acids, were increased by feeding both DF sources, and these production rates in the acid-treated DF group were significantly higher than those in the untreated DF group. The production rate of a minor SCFA, isovaleric acid, was decreased by feeding both diets. The production rate of total SCFA in rats given the acid-treated maize husk was 32.6% higher than that in rats given the untreated maize husk. The fermentable energy in DF was estimated in vivo by subtracting the faecal excretion of DF energy from ingested DF energy. The fermentable energy in DF was increased by the acid treatment (32·5% in maize husk and 63.4% in acid-treated maize husk), which agreed with the SCFA production rate predicted in the caecum. These results indicate that a short-term incubation of caecal contents is a useful method for evaluation of the fermentability of DF sources, and that acid treatment can increase the fermentability of an insoluble DF source.

To investigate diurnal changes in large-bowel metabolism rats were fed once daily on a cooked maize-based diet without or with 200 g wLeat bran/kg diet and containing Cr2O3 as an indigestible marker. After 17 d four rats on each diet were killed at 4, 10, 16 and 22 h postfeeding. Emptying of dry matter (DM) from the stomach occurred by an apparently zero-order process at about 0·5 g/h in both diet groups. Feeding wheat bran had little effect on caecal pH or total short-chain fatty acid (SCFA) concentration but was associated with a marked increase in molar proportion of butyrate and a fall in propionate. There were substantial changes in caecal total SCFA concentration and in the molar proportions of individual SCFA throughout the day but no evidence of an interaction between diet and time interval after feeding. Caecal transit time (TT) was significantly reduced by feeding wheat bran whilst colonic TT was unaffected. Although when averaged across both diets there were no significant time effects on caecal or colonic TT, there was a significant diet × time interaction for caecal TT. The extent of coprophagy was measured. The proportion of stomach DM derived from ingested faeces increased with time interval after feeding but it was always a minor contribution so that in these circumstances coprophagy is unlikely to result in significant bias in estimates of digesta flow-rates or TT.

The kinetics of the response in the blond of piglets to physiological oral intakes of galactose and glucose, and intravenous administration of galactose are described. Following the intravenous administration of galactose to 2- and 10-d-old piglets (n 7), the half-life was 7.98 (SD 0.75) and 7.99 (SD 1.89) min respectively, and efficient elimination rate was 9.09 (SD 2.15) and 8.75 (SD 0.79) % per min respectively. The turnover of galactose in the piglets was 100-3 μg/min per kg body weight. These observations demonstrate that galactose was rapidly removed from the blood of the piglets. While the dosing and sampling procedures stimulated hyperglycaemia, they had no effect on the concentration of galactose in the peripheral plasma. The galactose area under the curve (adjusted to the plasma volume of the animal) following a dose of either galactose or galactose plus glucose was 1.75 (SD 0.15) and 1.95 (SD 0.14) arbitrary units respectively in 2-d-old piglets and 1.96 (SD 0.26) and 1.98 (SD 0.10) arbitrary units respectively in 10-d-old piglets. Since the presence of glucose did not lower the adjusted area under the curve for galactose in the peripheral blood, the effect of glucose on the metabolism of galactose in piglets was more like that reported for rats than that for man, guinea-pigs or mice. It is suggested that the galactose moiety of lactose may make an important contribution to the replenishment of liver glycogen in the neonatal piglet.

The present study has examined the possibility that the positional distribution of fatty acids on dietary triacylglycerol (TAG) influences the postprandial response to a liquid meal in adult subjects. Postprandial TAG, non-esterified fatty acids (NEFA), ketones, glucose, insulin and gastric inhibitory polypeptide (GIP) responses were monitored in sixteen normal adult male subjects over 6 h following consumption of test meals containing dietary TAG in which palmitic acid was predominantly on the sn-1 (Control) or sn-2 positions (Betapol). Plasma total TAG, chylomicron-rich TAG and chylomicron-poor TAG concentrations were identical in response to the two test meals. The peak increase (mean (sd)) in chylomicron TAG was 0 85 (0 46) mmol/l after the Control meal and 0 85 (0 42) mmol/l after the Betapol meal. Plasma glucose, insulin, GIP, NEFA and ketone concentrations were also very similar following the two meals. It is concluded that dietary TAG containing saturated fatty acids on the sn-2 position appear in plasma at a similar level and over a similar timescale to TAG in which saturated fatty acids are predominantly located on sn-1 or sn-3 positions. The results reported in the present study demonstrate that the positional distribution of fatty acids on dietary TAG is not an important determinant of postprandial lipaemia in adult male subjects, but do not exclude the possibility that different responses may occur when these dietary TAG are given long term.

Bone density and Ca intake were assessed in fifty-six healthy premenopausal women, aged 21–47 years. Bone density was measured at the spine (lumbar vertebrae 1–4, anterio-posterior view), the non-dominant femur (neck, Ward's triangle and trochanter) and radius (33 % distal and ultradistal) using dual energy X-ray absorptiometry (Lunar DPX-L). The mean values (SD) for bone density (g/cm2) were 1·18 (0·10) at the lumbar spine,0·81(0.10) at the trochanter and for Ca intake 783 (329) mg. Bone density values were close to published normal values for young women and the Ca intakes were close to the recommended levels for the UK. However, there was a wide range especially in dietary Ca intake, and 27% of the group were at or below 500 mg/d. Correlations between Ca intake and bone density were highly significant at all three femoral sites (neck rO·41; Ward's triangle r 0·40, P < 0·01; trochanter r 0·47, P < 0.001), significant at the spine (r 0·27, P < 0.05) but not found at the radius. These correlations were independent of body mass. The low levels of Ca intake found in a substantial proportion of this selected group of young women and their association with low femoral bone density are cause for concern.

Infants born to atopic parents have been found to be at high risk of allergy development. The present study investigated the effect of a maternal milk-free diet during late pregnancy and lactation on the immune response and allergy incidence in at-risk and control infants. Atopic mothers were randomly allocated into an intervention group (n 12) or an unrestricted-diet group (n 14) and compared with non-atopic mothers following an unrestricted diet (n 12). The intervention involved a maternal milk-free diet during late pregnancy and lactation. Infants were followed up for 18 months postnatally. A significant fall in maternal serum β-Iactoglobulin (β-Lg)–immunoglobulin G (IgG) antibody levels (P < 0.05) was observed after a 7-week milk-exclusion diet. In maternal and cord serum samples the levels of β-Lg–IgG and α-casein–IgG antibodies were significantly correlated (r 0.89, P < 0.0001 and r 0.71, P < 0.001 respectively). Higher levels of β-Lg–IgG (P < 0.05) were observed in the cord serum samples compared with paired maternal serum samples. Single-blind allergy assessment by a paediatrician at 12 and 18 months showed that the infants born in the non-atopic group had a significantly lower allergy incidence compared with the infants born in the atopic group following an unrestricted diet (P < 0.008 and P < 0.02 respectively). The allergy incidence in the infants born in the atopic diet group was significantly lower compared with that of the atopic group following an unrestricted diet (P < 0.04). It was observed that the atopic nature of the parents significantly affected the allergy incidence in their children. A trend towards a beneficial effect of a maternal milk-free diet during late pregnancy and lactation was also observed in infants born to atopic parents.

Relationships between micronutrient intake and status, and micronutrient status and performance in tests of intelligence were investigated in a group of adolescents (13–14 years old). Dietary intakes were assessed using a 7 d weighed dietary record method, coupled with the collection of duplicate diets. Vitamin and trace mineral intakes calculated using food composition tables were compared with those obtained by direct analysis of duplicate diets. Micronutrient status was judged via a range of biochemical indices measured in blood samples taken after a 12–15 h fast. Blood samples were taken both before and after a 16-week period of vitamin and trace mineral supplementation. Individual tests of verbal and nonverbal intelligence were also performed pre- and post-supplementation. The results of this study indicate that the use of food table data may lead to substantial over- or underestimation of the intake of several micronutrients. In general, the total calculated or analysed amount of a specific micronutrient consumed did not adequately predict status, as judged by a range of biochemical indices. There were significant changes in status measurements over the 16-week study period, irrespective of supplementation, and these changes were markedly influenced by the initial status of the subject. There was no effect of supplementation on performance in tests of intelligence. However, there was a significant association between plasma ascorbic acid and initial non-verbal intelligence quotient (IQ) in the boys, and between whole blood glutathione peroxidase (EC 1.11.1.9) activity and non-verbal and verbal IQ in both sexes. These findings are discussed in relation to other recent studies of the influence of micronutrient supplementation on the psychological performance of children.

The effect of eating chocolate containing sugar alcohols as sweetening agents on colonic fermentation has been investigated by monitoring breath H2 levels. Levels were compared with those occurring after the consumption of normal, sugar-containing chocolate. Ten healthy volunteers aged 19 to 21 years ingested equal amounts of either sorbitol, isomalt or sucrose incorporated into standard chocolate bars. Breath H2 levels after consumption of chocolate containing either sorbitol or isomalt were significantly higher than those after consumption of chocolate containing sucrose (P < 0.001). After consumption of chocolate containing sorbitol, double the mean estimated volume of breath H2 was produced over 6 h compared with that produced after eating chocolate containing isomalt. Taken together with results relating to the incidence of intolerance symptoms, these findings demonstrate that sorbitol is associated with greater colonic fermentation compared with isomalt.

A stable isotope technique has been developed which uses 57Fe and 58Fe as labels and which enables the simultaneous measurement of Fe absorption from two test meals in infants. The method was evaluated by measuring Fe absorption from a commercial whey-adjusted infant formula in nine healthy infants aged 13–25 weeks. Each infant was fed 210 ml formula, labelled with either 57Fe or 58Fe, on four consecutive mornings, in random order. The total Fe content in each feed was 2.5 mg Fe; either as 2.5 mg 57Fe, or 0 6 mg 58Fe plus 1.9 mg Fe with normal isotopic composition. Isotopic enrichment of Fe in erythrocytes was measured by thermal ionization mass spectrometry 14 d after the last administration, and Fe absorption was calculated based on isotope ratio shifts, total circulating Fe and intake of each isotope. Geometric mean absorption for the 57Fe and 58Fe labels was 6.72 and 658% respectively, and the absorption of the two isotopes was not significantly different (Student's paired t test). By this technique, paired comparisons of Fe absorption can be obtained and systematic studies of the influence of dietary factors on Fe absorption during infancy can he conducted.

The general acceptance of NaFe3+EDTA for food Fe fortification has been partly restricted by concern over the influence of EDTA on the metabolism of other nutritionally important trace elements and minerals. We have investigated the influence of NaFe3+EDTA, and of increasing dietary levels of Na2EDTA, on Zn, Cu and Ca metabolism in rats fed on Zn-sufficient and Zn-deficient soya-bean-isolate- based diets. With the Zn-deficient diets, changing the dietary Fe compound from FeSO4 to NaFe3+- EBTA significantly (P < 0·05) increased mean apparent Zn absorption from 50·2 to 67·4%. urinary Zn excretion from 2·0 to 4·0% of intake, and Zn retention from 48·2 to 63·4%. Increasing the dietary EDTA level to 1000 mg/kg further increased Zn absorption to 78·1%, urinary Zn excretion to 15·6% of intake and Zn retention to 62·5%. Increased Zn retention was accompanied by a significant increase in weight gain indicating that the extra Zn was available for normal metabolic processes. With rats fed on the Zn-sufficient diet, NaFe3+EDTA and Na2EDTA similarly increased the absorption, urinary excretion and retention of Zn but to a lesser extent. NaFe3+EDTA, however, had no influence on the absorption, urinary excretion and retention of Cu and Ca, and additional Na2EDTA caused only minor increases in Cu absorption and retention and in the urinary excretion of Ca. We conclude that using NaFe3+EDTA as a food fortificant would have no detrimental effect on the metabolism of Zn, Cu and Ca and, in some situations, could improve Zn absorption and retention from low-bioavailability diets.

In the present study the effect of pyridoxine deficiency on the utrastructure and morphology of bone and its metabolism was examined in the rapidly growing chick. Pyridoxine-deficient animals had tibias of reduced dry weight and cortical thickness. Histomorphometry demonstrated a disproportionately high eroded surface, lower amount of osteoid tissue and reduced mineralized trabecular width. Anterior–posterior radiographs of the tibiotarsometatarsal joint showed reduced secondary ossification centres and coarse trabeculation. Decalcified metaphyseal cartilage showed irregular trabeculas and a markedly reduced amount of Fast-green counterstain matrix suggesting that there is less collagen present and in turn less availability for matrix to be laid down for later calcification. Plasma activity of the bone alkaline phosphatase isoenzyme (EC) 3. 1.3.1) was decreased. Plasma Ca and PO4 levels did not vary. The present bone study referring to a pseudo-lathyritic state in which collagen maturation is not completely achieved supports the hypothesis that pyridoxine is an essential nutrient for the connective tissue matrix.

The postprandial glycaemic response following a meal is reduced with the addition of soluble dietary fibre. The reductions in the glycaemia are thought to be due largely to increased viscosity of the gastrointestinal (GI) contents retarding digestion and absorption. The aims of the present study were to determine the effect that the GI tract has on the viscosity of meals containing different soluble fibres and to determine whether the glycaemic response of a meal (containing the soluble fibre) was predicted by the viscosity of the digesta in the small intestine. High carbohydrate diets containing 70 g soluble fibre (guar gum, xanthan gum or methylcellulose)/kg or 70 g insoluble fibre (wheat bran)/kg were diluted in water to a final fibre concentration of 18 g/kg. Following dilution the wheat bran diet had no measurable viscosity, while the viscosities of the soluble fibre diets were elevated. When the diets were fed to male Sprague–Dawley rats for 2 weeks the viscosities of the stomach and small intestinal digesta were not predicted by the viscosity of the diets measured before ingestion The action of the GI tract on the viscosity of the soluble fibres was investigated in vitro by dilution of the diets with acidic and neutralizing solutions, mimicking gastric and duodenal secretions. Dilution of diets with either acidic and neutralizing solutions or saline control significantly lowered the viscosity of all diets, while alterations in the pH of the diets had little impact on the resultant viscosity. When fasted rats were orally administered with the differing diets (0.25 g carbohydrate/kg body weight), the postprandial glucose response was reduced following the soluble-fibre-containing meals when compared with the wheat bran-supplemented meal, although the reduction in glycaemia only reached statistical significance with xanthan supplementation. These results indicate that there are large changes in the viscosity of a meal containing soluble fibre following ingestion, and that dilution of the diet by GI secretions is important in determining the resultant viscosity in the small intestine. Furthermore, the large differences in viscosity of the GI contents following consumption of the diets containing the soluble fibres were not predictive of the postprandial glycaemic response.

The present study aimed to determine whether lysine and/or methionine are absorbed in nutritionally significant amounts from the proximal colon of milk-formula-fed piglets (15–32 d old; 2.0–7.4 kg liveweight). Piglets, surgically prepared with simple catheters which allowed infusion into the proximal colon, were randomly allocated to one of two milk-formula diets which were either 40 % deficient in lysine (L– diet) or 60% deficient in methionine and 40% deficient in cysteine (S – diet), yet balanced for all other amino acids. The piglets were individually bottle-fed the milk-formula diets seven times daily at 2 h intervals between 08.00 and 20.00 hours. Physiological saline (9 g NaCl/l) or an isotonic solution containing the deficient amino acid was infused via the catheter at each feeding. The experimental procedure followed a cross-over design. Total daily excretions of urinary urea and total N were determined. There were no significant differences (P > 0.05) in urinary N metabolite excretion for piglets infused with amino acids compared with those infused with saline. Lysine and methionine do not appear to be absorbed in nutritionally significant amounts from the proximal colon of the milk-fed piglet.

Thirty-two calves were studied to determine the effects of level of nutrition (400 and 1000 g (air-dry matter) milk substitute per d) and age at weaning (5, 9 and 13 weeks) on humoral immune responses measured in serum and bronchoalveolar washings (BAW). All calves were immunized with Keyhole limpet haemocyanin (KLH) at 3 weeks of age, and with horse erythrocytes (HRBC) 1 d after weaning. Feeding the higher level of nutrition compared with the lower level decreased anti-HRBC titres and serum IgG2 and IgA responses to KLH (P < 0·05). Weaning at 5 compared with 9 weeks of age decreased serum anti-HRBC responses (P < 0·05), but weaning age had no effect on anti-KLH responses (P > 0·5). Feeding the higher level of nutrition increased total protein (P < 005) and IgG2 concentrations (P < 0·01) in BAW. The results showed that variation in husbandry conditions that is within conventional limits affects humoral immune responses in young, artificially-reared calves.