1. The heart-rate (HR) method for determining the energy expenditure of free-living subjects has been evaluated using a whole-body calorimete in which individuals lived continuously for 27 h while carrying out normal daily activities. Eight male volunteers each occupied the calorimeter on at least two occasions when HR and energy expenditure were measured continously.

2. After each session in the calorimeter a calibration was obtained using standard techniques by determining HR and heat production (HP) over periods of 10–15 min at several levels of activity. Energy expenditure in the calorimeter was then predicted, by each of five methods, from the mean HR in the calorimeter. Additionally, one session in the calorimeter was used to obtain a calibration and was used for predicting the subject's energy expenditure while in the calorimeter on other occasions.

3. Standard methods of prediction using one calibration point at rest and several points during activity were unreliable for predicting the energy expenditure of an individual. The 24 h HR was at the lower end of the calibration scale and there were considerable over-estimates or underestimates of energy expenditure, particularly during the night when the mean (±SD) difference between the actual and predicted HP was −66±38±6%. A linear regression fitted to points at the lower levels of activity improved the prediction of 24 h HP while a logistic plot reduced the error even further. The best estimate of energy expenditure was that obtained from a calibration over 24 h within the calorimeter; the mean (±SD) difference between the actual and predicted 24 h HP was +3+10.5% for light activity and −3±6.7% for moderate activity. Thus current procedures for calibrating subjects may lead to large errors which could be reduced by using a respiratory chamber.

1. Ascorbic acid was absorbed across the mucosa of the human mounth.

2. Omission of sodium ions from the medium decreased the absorption of ascorbic acid.

3. The presence of D-glucose, or 3-O-methyl-D-glucose, incrased the absorption of ascorbic acid but D-fructose had little effect and D-mannitol had no effect.

4. Calcium ions also increased ascorbic acid absorption probably by a secondary effect on Na+ fluxes.

5. Buccal mucosa was also premeable to dehydroascorbic acid and D-isoascorbic. acid

1. Rats were made oedematous by feeding them low-protein diets (protein: energy (P:E) 0.005) ad lib., and measurements were made of plasma and interstitial fluid colloid osmotic pressures (πp and πi respectively) and interstitial fluid hydrostatic pressure (Pi) before, and at the onset of, oedema formation. Taken together as (πp − πi + Pi) these forces oppose capillary pressure (Pc) and thus determine rates of transcapillary water filtration. Interstitial fluid was sampled, in non-oedematous and oedematous animals, from perforated capsules implanted subcutaneously for the measurement of Pi. Blood, plasma and interstitial fluid volumes were also determined.

2. In Expt I comparisons were made between animals fed on a control diet (P:E 0.210) and the lowprotein diet. In normal animals the ratio πp:πi was approximately 2, but in protein deficiency it was increased since reductions in the absolute value of πi matched those in πp. These changes were observed 2 weeks after the start of the experiment and became more exaggerated when oedema appeared (weeks 18–22).

3. Pi was normally negative with respect to atmospheric pressure but increased to values close to zero when oedema formation occurred.

4. Despite the reductions in πp that were seen in the protein-deficient animals the sum of the forces opposing filtration (πp − πi + Pi) did not change significantly during the experiment.

5. Plasma and interstitial fluid volumes expressed per kg body-weight (measured using 125I-albumin and 35SO42−) were unchanged as πp initially decreased in the protein-deficient animals but increased markedly with the onset of oedema.

6. In Expt 2 comparisons were made between animals fed the low-protein diet ad lib. and others fed on the control diet in restricted amounts so that weight loss was the same in the two groups of animals.

7. The wasting induced by restriction of the control diet did not produce reductions in πp or πi and values for Pi were normal. Changes in the animals fed on the low-protein diet were similar to those observed in Expt I. By using 51Cr-labelled erythrocytes it was shown that the expansion in plasma volume that occurred when oedema appeared in the protein-deficient animals was mainly due to a reduction in total erythrocyte volume. Blood volume did not increase significantly.

8. It was concluded that in the hypoproteinaemia induced in the experimental animals reductions in the value of πp, which might otherwise result in an imbalance of forces that would produce excessive rates of transcapillary water filtration, were compensated for by reductions in πi. Increases in Pi also compensated but were quantitatively less important.

9. The significance of the results is discussed in terms of the pathogenesis of oedema in kwashiorkor and the concept of an oncotic threshold for oedema formation in hypoproteinaemia.

1. Long-term clinical and biochemical riboflavin status and dietary intake of riboflavin were monitored for 18 months in a group of twenty-three relatively-healthy elderly subjects living at home in the north of England.

2. Both dietary intake and biochemical status, as measured by the activation coefficient (stimulated: basal activity) of NAD(P)H2: glutathione oxidoreductase (EC 1.6.4.2), remained fairly constant for each individual and for most subjects the usual intake and biochemical status were characterized quite accurately by a single week's intake dietary record and a single measurement of the activation coefficient.

The expected relationship between biochemical status and dietary intake was present, but not strongly evident. There was a significant within-subject correlation between α values and the immediately preceding dietary intake; however the between-subject correlation between α values and dietary intake approached zero if the two subjects receiving long-term riboflavin supplements were omitted. Over the limited range of intakes observed in unsupplemented subjects, non-dietary factors evidently obscure the relationship.

4. Although eight subjects had average values above 1.2 for the activation coefficient, no excessively high values were observed, and no subject showed any clear-cut clinical deficiency symptoms.

1. Long-term clinical and biochemical vitamin C (ascorbic acid and dehydroascorbic acid) status and dietary intake of vitamin C were monitored for 18 months in twenty-three relatively-healthy elderly subjects living at home in the north of England.

2. Plasma vitamin C showed a strong positive correlation with buffy-coat vitamin C both cross-sectionally between subjects and longitudinally within subjects; plasma levels, therefore, were almost as good an index of long-term status as buffy-coat levels.

3. Vitamin C intake was strongly correlated with plasma and with buffy-coat levels both between subjects and within subjects. This contrasts with the poor correlation observed between riboflavin intake and biochemical riboflavin status in the same subjects.

4. Subjects with relatively high average intakes showed considerable seasonal variation, and several widely-spaced measurements would be needed to characterize their long-term status accurately. Those with low average intakes and blood levels showed little variation during the study.

5. The strongest within-subject correlation was obtained by relating the biochemical values to the 7 d dietary intake directly preceding the biochemical analysis. Blood and tissue levels therefore appear to be strongly related to the current dietary intake.

6. Although some subjects had blood vitamin C levels consistently within the region associated with biochemical deficiency, none showed clinical evidence of specific deficiency symptoms.

7. After the main study, fifteen of the subjects received supplementary vitamin C for 2 months. Plasma and buffy-coat levels rose sharphy, but fell to presupplementation levels within 1 month of withdrawal, emphasizing the transitory nature of increased tissue levels. No significant changes were detected in the following collagen-related urinary ratios: hydroxyproline: creatinine, proline: creatinine, proline: total amino-nitrogen and proline: hydroxyproline in hydrolysates either of whole urine or of various fractions. These variables thus appear to be insensitive to short-term changes in vitamin C status over the ranges encountered in this study.

1. Adipose tissue samples were obtained by needle biopsy from three subcutaneous sites (thigh, abdomen and upper arm) in twenty-two obese women. The fatty acid composition was determined using gas-liquid chromatography and the results presented relate to eleven component fatty acids.

2. The fatty acid composition of adipose tissue obtained from the arm and abdomen was remarkably similar, with the exception of the levels of lauric acid.

3. The analyses showed that the majority of the saturated fatty acids were present in smaller proportions whilist the majority of unsaturated fatty acids were present in larger proportions in the thigh than in the two other sites. Highly significant inter-site differences were demonstrated for six of the major fatty acids and also for both the total amounts of saturated and unsaturated fatty acids and their ratios.

4. No marked differences in the fatty acid composition of adipose tissue from obese subjects were revealed during this study when compared with previously reported results obtained from ‘normal-weight’ subjects.

1. Fermentation in the rumen and nitrogen dynamics in the body were studied in mature Merino sheep given a maintenance ration of a low-quality-roughage diet containing mainly chopped wheat straw.

2. Intake of metabolizable energy was 3.49 MJ/d and of total N 6.2 g/d.

3. From measurements of volatile fatty acid (VFA) production rates and stoichiometric principles, it was calculated that 75% of the digestible organic matter intake was fermented in the rumen, making an estimated 44 g/68d microbial dry matter available to the animal.

4. The total flux of ammonia through the rumen NH3 pool, estimated by 15NH3 dilution methods, was 8.2 g N/d of which 3.5 g N/d was irreversibly lost; thus 4.7 g N/d was recycled, partly within the rumen (approximately 3.8 g N/d) and partly via endogenous secretions (approximately 0.9 g N/d). The extensive recycling of NH3-N within the rumen indicated that turnover of microbial N was considerable, and the total production of micro-organisms was at least twice the net outflow.

5. The proportion of the N in rumen bacteria derived from rumen ammonia was 62% and thus 38% was derived from other nitrogenous compounds such as peptides and amino acids.

6. The rates of transfer of blood urea into the rumen, estimated from the appearance of 14CO2 or 15NH3 in the rumen after intravenous single injections of [14C]-and [15N]urea, did not differ significantly and the mean transfer was 2.3 urea-N/d.

7. Estimates of the rate of irreversible loss of urea-C (i.e. urea synthesis in the body) were obtained by analysis of samples of either blood or urine obtained after a single, intravenous injection of [14C]urea. The two methods gave results that did not differ significantly. The estimated rate of urea synthesis in the body was 5.3 g N/d. Urea excretion rate was relatively low, i.e. 1.2 g N/d, and thus transfer of urea to the digestive tract was approximately 4.1 g N/d. Approximately 53% of the latter was transferred to the rumen, and 47% to the rest of the digestive tract. These results are discussed in relation to similar studies with sheep given other diets.

8. Various aspects of isotope-tracer methods and the errors that could occur in this type of study are discussed.

1. The effects of defaunation of the rumen of lambs given low-protein-high-energy diets were studied using animals fed ad lib. a basal diet of sugar and oaten chaff which was supplemented with fish meal at various levels. These diets supported relatively large populations of protozoa in the rumen.

2. A nonly phenol ethoxylate (15 g, Teric GN9; ICI Australia Ltd) given intraruminally, was shown to be an effective means of defaunating the rumen and the lambs were maintained protozoa-free by isolation.

3. In lambs given low levels of fish meal in the diet, the absence of rumen protozoa increased growth rate and improved the food conversion efficiency (g dry matter intake/g growth). These effects were not apparent at high levels of protein supplementation.

4. These results suggested that the absence of rumen protozoa resulted in an increased availability of nutrients supplying both energy and amino acids since the efficiency of food utilization and the rate of wool growth (which is sensitive to amino acid supply) were increased in lambs on the low level of protein supplementation without apparently increasing food intake.

1. The minced carcases of twenty-seven lambs, ranging from 18 to 69 kg in live weight, and twenty-five calves (30–90 kg) were analysed for copper, iron, manganese and zinc. The lambs were weaned whereas the calves were reared exclusively on milk.

2. Mean concentrations of Fe, Mn and Zn for groups of lamb carcases fell within the ranges 52.6–75.1, 0.7–1.2 and 20.8–25.6 mg/kg fresh carcase weight respectively. The concentrations of Fe and Mn decreased while that of Zn increased slightly with age at slaughter. The concentrations of Fe, Mn and Zn in calves were close to thosein lambs.

3. For both species, the concentration of Cu in the carcase varied erratically: variation in hepatic Cu storage was implicated. In an additional study of ten full-term foetuses from Cu-depleted or Cu-supplemented ewes, a dietary Cu supplement (10 mg/kg dry matter (DM)) increased foetal Cu status 10-fold, due largely to an increase in foetal liver Cu.

4. The mean retentions of trace elements in the lamb carcases (%intake) were approximately: Cu 2.0, Fe 1.3, Mn 0.08, Zn 4.0. The corresponding values for the milk-fed calves were all probably much higher (Cu 23, Fe 43.7, Mn 4.9, Zn 34.0) but Cu intake was not accurately measured.

5. After allowing for tissue storage of Fe and Mn, values of 55, 0.85 and 24 mg/kg carcase gain were taken to represent the approximate net growth requirements of lambs for Fe, Mn and Zn respectively: the corresponding value for Cu was probably < 1.0 mg/kg. Values for calves were similar to those for lambs.

6. It was concluded that the total net requirements of ruminants for Fe and Zn shouldbe considered in terms of daily intakes of the metans rather than dietary concentrationsbecause of the relatively large and constant contribution of the growth component to thetotal requirement.

1. Two experiments were conducted to investigate the effect of intestinal coccidiosis (Eimeria acervulina) on blood plasma and tissue ascorbic acid (AA) concentrations in chicks, and a third experiment was carried out to examine the role of dietary AA in this infection.

2. Experimental infection with intestinal coccidiosis (Eimeria acervulina) brought about a depletion in the concentration of AA in blood plasma, duodenum, jejunum, ileum, liver and the adrenal glands. Generally, these changes were obtained after the prepatent period of 4–5 d of the infection.

3. Dietary AA at 1000 mg/kg prevented the depletion in plasma and tissue AA levels but did not significantly affect adrenal weight.

1. Alkaline phosphatase (EC 3.1.3.1) from many rat tissues was separated into two or three bands by electrophoresis on polyacrylamide gel. Ten of the bands had different electrophoretic mobilities, but some were present in more than one tissue.

2. Bands from the corresponding tissues of magnesium-deficient and control rats were qualitatively similar, but there were quantitative differences in the distribution of enzyme activity between them.

3. Mg deficiency had differential effects on the absolute activity of individual bands, but two main types of response were observed. There was an increase in the activity of the first bands from liver and kidney, the second band from femur and both bands from spleen, whereas the first band from femur, the first and second bands from intestinal mucosa and the second bands of serum, liver and kidney all decreased in activity during the deficiency.

4. The change in total alkaline phosphatase activity of a tissue during Mg deficiency depended on the ratio between the enzyme components within it.

1. Balance studies were carried out with rats and golden hamsters (Mesocricetus auratus) given diets high and low in calcium (10.8 and 5.1–5.2 g Ca/kg respectively) and high and low in phytate (3.4 and 1.6 g phytate-phosphorus/kg respectively) in a 2 × 2 factorial experiment.

2. Values for Ca absorption and phytate availability (relative to the amount ingested) and for urinary excretion of Ca and P were significantly higher in the hamster than in the rat for all four diets.

3. Phytate availability was significantly greater on the low-Ca than on the high-Ca diets with both species.

4. Ca absorption was greater on the low-phytate than on the high-phytate diets but the differences were significant only for the hamster.

5. Absorption of both Ca and P was poorly controlled in the hamster compared to the rat.

1. The Bowman-Birk soya-bean trypsin inhibitor (BBTI) begins to cause pancreatic enlargement and increased enzymic activity in the pancreas of chicks after a minimum of 7 d of feeding.

2. The active inhibitory site of BBTI against trypsin is the factor involved in the pancreatic enlargement and increase of pancreatic nzesyme activity in chicks.

1. An experiment was carried out on growing male rats to study the effects of a phosphorus deficiency on voluntary energy intake, estimated by the separate-feeding technique, as well as the consequences on growth and metabolic utilization of energy, protein and minerals. After a preliminary period of P deprivation, three groups of twelve animals were fed separately and simultaneously two dietary rations. A protein ration provided 1.2 g protein/d to which was added one of three levels of P in the form of monosodium phosphate: a normal level of 35.4 mg/d (treatment A), half the normal level (19.1 mg/d, treatment B) or one-quarter the normal level (9.6 mg/d, treatment C). Another protein-free ration was fed ad lib.

2. It was only when the level of P represented one quarter the normal level that a significant decrease in growth rate was noticed, accompanied by a definite decrease in daily energy consumption. The food conversion ratio (g dry matter intake/g body-weight gain) increased whereas the protein efficiency ratio (g body-weight gain/g protein intake) was lower. The nitrogen and energy retentions changed in the same way: both dectreased with treatment C compared to treatment A and B. Independently of the protein supply, the voluntary energy intake was closely related to the intensity of protein retention, which depended on the dietary level of P. The daily retention of P only slightly decreased at the lowest ingestion level (9.6 mg/d in treatment C), compared to that of the higher levels. On the other hand, the amount of calcium retained regularly decreased with the decreasing supply of P. The result of this was a progressive reduction in retained Ca:P as the level of P decreased.

3. From these results it appeared that the primary effect of a P deficiency in the growing rat was a decrease in bone mineralization. At a more advanced stage, the tissue P levels were affected and the resulting metabolic alterations reduced protein deposition and consequently the voluntary energy intake. The level of energy consumption, in separate-feeding, is more generally dependent on the level of protein deposition allowed by the limiting factor for growth, either protein, minerals or vitamins.

1. Three sheep fitted with duodenal re-entrant cannulas and three with large rumen fistulas were given red clover, perennial ryegrass and a 2:1 (w/w) mixture of grass and clover in two Latin square arrangements. Measurements were made of voluntary intake, digestibility, flow of nutrients into the duodenum and the flow of Cr-EDTA marker through the reticulo-rumen.

2. Organic matter (OM) digestibility was similar for the three feeds but the voluntary intake decreased in the order mixture > red clover > perennial ryegrass. There was an increase in the rate of marker flow from the rumen and a decrease in retention time of the same order. Rumen volume did not change significantly.

3. There was a reduction in the mean particle size of rumen contents in the order perennial ryegrass > clover > mixture. The in vitro digestibility of particles decreased with size; the reduction being more rapid for clover than perennial ryegrass.

4. The proportion of ingested digestible OM appearing at the duodenum increased from 18.4% to 26.7% to 30.0% for perennial ryegrass, clover and the mixture respectively.

5. A higher proportion of digestible cellulose and hemicellulose disappeared over the stomach for the perennial ryegrass feed compared to the clover and the mixture but over 96% of water soluble carbohydrates and starch disappeared over the stomach for all three feeds.

6. The apparent digestibility of nitrogen was similar for all three feeds but the proportion of undigested feed N appearing at the duodenum was calculated to be greater by a factor of 1.71 and 2.52 for clover and mixture feeds compared to grass.

7. It was concluded that the higher nutritive value of red clover compared to perennial ryegrass was due to an increased rate of flow of nutrients througth the reticulo-rumen and an increase in the proportion of digestible OM digested post ruminally.

1. Sixteen first-calf Friesian heifers were used in a continuous treatment design experiment. For 2 weeks after calving they were given a 750 g concentrate, 250 g hay/kg ration with 169 g crude protein (nitrogen × 6.25; CP)/kg dry matter (DM). They were then divided into two groups of eight and given a high-protein (223 g CP/kg DM) or low-protein (107 g CP/kg DM) ration at the rate of 10.8 kg concentrates+3.6 kg hay for 8 weeks.

2. Milk yield and composition, live weight and blood composition were monitored throughout. A digestibility trial was carried out with six animals on each treatment.

3. The low protein ration reduced DM, organic matter, energy and fibre digestibility significantly (P < 0.001) so that intakes of digestible energy were not equal and the low-protein group lost more weight than the high-protein group.

4. Milk yield and the fat content of milk were lower in heifers given the low-protein ration (P < 0.01). The lactose content of the milk was not affected and protein content only slightly reduced (P < 0.1) by low-protein feeding. When the heifers were all changed onto an adequate protein (190 g CP/kg DM) ration in mid-lactation, those which had previously been under-fed protein appeared to recover in milk yield to the point they might have been expected to reach if given an adequate-protein ration throughout.

5. Concentrations of urea (P < 0.001) and albumin (P < 0.05) were reduced by underfeeding protein, but albumin concentration was affected less by diet than by stage of lactation. Blood concentrations of total protein, glucose, sodium, potassium, calcium, inorganic phosphate, iron, copper, haemoglobin and packed cell volume were unaffected by treatment. Blood magnesium concentration was slightly lower (P < 0.1) with low-protein feeding.