Reducing dietary crude protein (CP) concentration while maintaining adequate amino acid (AA) supply by free AA inclusion can contribute to attenuate the negative environmental effects of animal farming. This study investigated upper limits of dietary free AA inclusions without undesirable effects including the dependence on asparagine (Asn) and glutamine (Gln) supply. Ten broilers were allocated to sixty-three metabolism units each and offered nine experimental diets from day (d) 7–21 (n 7). One diet (167 g CP/kg) contained 80 g soya protein isolate (SPI)/kg. In the other diets, 25, 50, 75 and 100 % of the digestible AA from SPI were substituted with free AA. Digestible Asn+aspartic acid (Asp) and Gln+glutamic acid (Glu) were substituted with Asp/Glu or 50/50 mixes of Asp/Asn and Glu/Gln, respectively. Total excreta were collected from d 11–14 and from d 18–21. Growth and nitrogen accretion were unaffected by 25 and 50 % substitution without and with free Asn/Gln, respectively, but decreased at higher substitution (P ≤ 0·024). Circulating concentrations of Asp, Glu and Gln were unaffected by treatment, while Asn decreased at substitution higher than 50 % when Asn/Gln were not provided (P ≤ 0·005). Blood gas analysis on d 21 indicated a compensated metabolic acidosis at substitution higher than 50 and 75 % without and with free Asn/Gln, respectively (P ≤ 0·017). Results suggest that adding Asn/Gln increased an upper limit for proportion of dietary free AA from 10 to 19 % of dietary CP and enabled higher free AA inclusion without affecting the acid–base balance.

Grape marc (GPM) is a viticulture by-product that is rich in secondary compounds, including condensed tannins (CT), and is used as a supplement in livestock feeding practices. The aim of this study was to determine whether feeding GPM to lactating dairy cows would alter the milk proteome through changes in nitrogen (N) partitioning. Ten lactating Holstein cows were fed a total mixed ration (TMR) top-dressed with either 1.5 kg dry matter (DM)/cow/day GPM (GPM group; n = 5) or 2.0 kg DM/cow/day of a 50:50 beet pulp: soy hulls mix (control group; n = 5). Characterization of N partitioning and calculation of N partitioning was completed through analysis of plasma urea-N, urine, feces, and milk urea-N. Milk samples were collected for general composition analysis, HPLC quantification of the high abundance milk proteins (including casein isoforms, α-lactalbumin, and β-lactoglobulin) and liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis of the low abundance protein enriched milk fraction. No differences in DMI, N parameters, or calculated N partitioning were observed across treatments. Dietary treatment did not affect milk yield, milk protein or fat content or yield, or the concentrations of high abundance milk proteins quantified by HPLC analysis. Of the 127 milk proteins that were identified by LC-MS/MS analysis, 16 were affected by treatment, including plasma proteins and proteins associated with the blood-milk barrier, suggesting changes in mammary passage. Immunomodulatory proteins, including butyrophilin subfamily 1 member 1A and serum amyloid A protein, were higher in milk from GPM-fed cows. Heightened abundance of bioactive proteins in milk caused by dietary-induced shifts in mammary passage could be a feasible method to enhance the healthfulness of milk for both the milk-fed calf and human consumer. Additionally, the proteome shifts observed in this trial could provide a starting point for the identification of biomarkers suitable for use as indicators of mammary function.

Soybean [Glycine max (L.) Merr. ‘Hark’] plants responded similarly to a sublethal application of metribuzin [2-amino-6-tert-butyl-3-(methylthio)-as-triazin-5(4H)-one] or to low light intensities. Both treatments increased soluble amino acid, ATP, nitrate reductase, and nitrate levels (up to 67, 140, 190, and 10,000%, respectively) and decreased soluble reducing sugars (33%), malate levels (86%), and the chlorophyll a/b ratio (24%). The similar response of light-limited and of chemically photosynthesis-inhibited plants is thought to be induced by a carbohydrate stress or another parameter related to a decreased rate of photosynthetic electron flow. However, during the induction process in shaded plants, light quality might play an important role in some cases. The results obtained with metribuzin and soybean plants are compared to and discussed together with (a) the results obtained by other authors working with s-triazine herbicides and (b) results obtained with methabenzthiazuron [N-(benzothiazol-2-yl)-N,N′-dimethylurea] in earlier experiments.

A total of 156 plant species from 35 botanical families collected from diversified grasslands in the French Massif Central were screened in vitro for their potential to combine high nutritive value for ruminants and a reduced impact on the environment. The vegetative part of plants were analyzed for their chemical composition and incubated in a batch system containing buffered rumen fluid at 39°C for 24 h. The gas production and composition were recorded, and the fermentation end-product concentrations in the incubation medium and the in vitro true organic matter digestibility (IVTOMD) were determined. The results were expressed relative to perennial ryegrass (PRG) values used as a reference. We observed that no relationship between methane (CH4) and volatile fatty acids (VFA) was evidenced for 12 plants, the fermentation of these plants producing significantly less CH4 for a similar level of VFA production. In all, 13 plants showed 50% less CH4 production per unit of organic matter truly digested (OMTD) than PRG. Among these plants, two reduced CH4 by more than 80% and four species had an IVTOMD higher than 80%. The underlying modes of action seem to be different among plants: some result in an accumulation of H2 in the fermentation gas, but others do not. In terms of nitrogen (N) use efficiency, the fermentation of 37 plants halved the ratio between ammonia (N–NH3) and plant N content compared with PRG, of which six showed a complete absence of N–NH3 in the medium. Among these plants, four maintained the IVTDMO at values not significantly different from PRG (P>0.05). Considering the multi-criteria selection, 16 plants showed simultaneously a reduction of more than 80% in N–NH3 production and 30% in CH4 emission per unit of OMTD compared with PRG, including three with an IVTOMD higher than 80%. Overall, the botanical families that reduced simultaneously CH4 and N–NH3 most efficiently were the Rosaceae, Onagraceae, Polygonaceae and Dipsacaceae. The Onagraceae also gave high values for IVTOMD.

Little nutritional information has been collected from domestic cats fed raw meat diets. The objective of the present study was to evaluate differences in N metabolism of domestic cats fed raw beef-based diet (66 % crude protein (CP) and 20 % fat), bison-based diet (49 % CP and 39 % fat), elk-based diet (79 % CP and 6 % fat) and horse-based diet (60 % CP and 26 % fat). A total of eight intact adult female cats were fed to maintain body weight in a cross-over design. Daily food intake, faecal and urinary outputs, and N metabolism were measured. Dietary N was highly digestible (96·8 (sem 0·7)) for all treatments. Urinary N accounted for a majority of total N excretion, and differences in total N excretion reflect differences in urinary N. Differences in N intake and N absorption were due to differences in CP levels among diets. N retention was similar to values reported in the literature for domestic cats fed purified and traditional extruded diets. Despite differences in protein concentrations and N intake, all raw meats tested maintained N metabolism.

Foetal life malnutrition has been studied intensively in a number of animal models. Results show that especially foetal life protein malnutrition can lead to metabolic changes later in life. This might be of particular importance for strict carnivores, for example, cat and mink (Neovison vison) because of their higher protein requirement than in other domestic mammals. This study aimed to investigate the effects of low protein provision during foetal life to male mink kits on their protein metabolism during the early post-weaning period of rapid growth and to investigate whether foetal life protein deficiency affects the response to adequate or deficient protein provision post weaning. Further, we intended to study whether the changes in the gene expression of key enzymes in foetal hepatic tissue caused by maternal protein deficiency were manifested post-weaning. A total of 32 male mink kits born to mothers fed either a low-protein diet (LP), that is, 14% of metabolizable energy (ME) from protein (foetal low – FL), n = 16, or an adequate-protein (AP) diet, that is, 29% of ME from protein (foetal adequate – FA), n = 16) in the last 16.3 ± 1.8 days of pregnancy were used. The FL offspring had lower birth weight and lower relative abundance of fructose-1,6-bisphosphatase (Fru-1,6-P2ase) and pyruvate kinase mRNA in foetal hepatic tissue than FA kits. The mothers were fed a diet containing adequate protein until weaning. At weaning (7 weeks of age), half of the kits from each foetal treatment group were fed an AP diet (32% of ME from protein; n = 8 FA and 8 FL) and the other half were fed a LP diet (18% of ME from protein; n = 8 FA and 8 FL) until 9.5 weeks of age, yielding four treatment groups (i.e. FA-AP, FA-LP, FL-AP and FL-LP). Low protein provision in foetal life lowered the protein oxidation post-weaning compared with the controls (P = 0.006), indicating metabolic flexibility and a better ability to conserve protein. This could not, however, be supported by changes in liver mass because of foetal life experience. A lower relative abundance of Fru-1,6-P2ase mRNA was observed (P < 0.05), being lower in 9.5-week-old FL than in FA kits. It can be concluded that foetal life protein restriction leads to changes in post-weaning protein metabolism through lower protein oxidation of male mink kits.

The objective of the present study was to delineate how interactions between feeding sunflower oil (SFO) to partially defaunate the rumen and altering dietary ruminally fermentable carbohydrate may alter urea-N kinetics and N metabolism in lambs. In a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments, four Suffolk ram lambs (61·5 (se 4·0) kg) were used. Treatments were 0 ( − SFO) v. 6 % (+SFO) SFO and dry-rolled barley (DRB) v. pelleted barley (PB). N balance was measured over 4 d, with concurrent measurement of urea-N kinetics using continuous intra-jugular infusions of [15N15N]urea. Feeding SFO decreased (P = 0·001) ruminal protozoa and NH3-N concentrations. Urinary N excretion was lower (P = 0·003), and retained N was higher (P = 0·002) in +SFO lambs compared with − SFO lambs. Endogenous production of urea-N (urea-N entry rate; UER) was similar across treatments. Urea-N transfer to the gastrointestinal tract (GIT) (GIT entry rate; GER), expressed as absolute amounts (16·4 v. 13·1 g/d) or as a proportion of the UER (0·693 v. 0·570), its anabolic use (9·0 v. 6·0 g/d) and microbial N supply (14·6 v. 10·9 g/d) were higher (P ≤ 0·001) in +SFO lambs compared with –SFO lambs. As a proportion of the UER, GER was higher, whereas urinary urea-N loss was lower, in lambs fed PB compared with those fed DRB (P = 0·01). In summary, feeding SFO increased urea-N recycling to the GIT and microbial non-NH3-N supply, thus providing new evidence that the improved efficiency of N utilization in partially defaunated ruminants could be partly mediated by an increase in urea-N recycling.

Malnutrition during foetal life can induce modifications in the phenotype of an individual. The present study aimed to observe effects of low foetal life protein provision on modifications of the phenotype and changes in the progeny of 1-year-old female mink (F1 generation) offspring of mothers fed a low-protein diet. Traits studied included reproductive performance, energy and protein metabolism, and key hepatic enzymes associated with glucose homeostasis and metabolic hormones. The F0 generation offspring were fed either a low-protein (14 % of metabolisable energy (ME) from protein – FLP1) or an adequate-protein (29 % of ME from protein – FAP1) diet for the last 17·9 (sd 3·6) d of gestation. The F1 dams were studied at birth and at 1 year of age, during their first reproductive cycle, after maintenance on an adequate diet from birth and thereafter. Metabolic traits during gestation and lactation were largely unaffected by foetal life protein provision, but birth weight in the F2 generation was higher (P = 0·003) among FLP2 kits than among FAP2 kits. Furthermore, the relative abundance of pyruvate kinase mRNA was significantly (P = 0·007) lower, and fructose-1,6-bisphosphatase mRNA tended (P = 0·08) to be lower in FLP2 foetuses than in FAP2 foetuses, showing some similar difference in the F2 generation and F1 generation foetuses, suggesting an effect on some hepatic enzymes affecting glucose homeostasis being transmitted from the F1 to the F2 generation. These findings indicate that even though energy and nitrogen metabolism displayed no effect of protein provision during early life, programming effects still appeared at the molecular level in the following generation.

Data on N utilisation by sheep-fed diets containing foliages were analysed to develop prediction equations for N excretion in faeces (FN) and urine (UN), and to determine endogenous N excretion. Overall, 218 dietary treatments from forty-four publications were compiled in the database. This database was split into three subsets: without foliage in the diets (FL-0); foliage levels (FL) in between 0 and 310 g/kg (FL-L); FL in between 310 and 800 g/kg diets (FL-H) to study the effects of foliages on metabolic faecal N (MFN) and endogenous urinary N (EUN). Nitrogen intake (NI) as single independent factor was the best predictor of FN (R2 0·75), UN (R2 0·81) and total N excretion (R2 0·86). Addition of dietary N concentration and FL for FN (R2 0·82), dietary N concentration and foliage NI for UN (R2 0·85), and FL and foliage NI for total N excretion (R2 0·92) as supporting predictors to this relationship slightly increased R2 values. The monomolecular and exponential models slightly improved the prediction of N excretion with NI as a predictor compared with the linear model. The excretion of MFN was greater for FL-H compared with FL-0, but was similar between FL-0 and FL-L, and FL-L and FL-H. However, EUN decreased in FL-H compared with FL-0 and FL-L, but was similar between FL-0 and FL-L. In conclusion, using NI as the primary predictor produced an accurate prediction of N excretion. Inclusion of foliages in the diets may shift N excretion from urine to faeces and increase the excretion of MFN and EUN.

Abstract A study was designed to determine if pig performance and lysine conversion ratio would be improved by phase feeding a series of diets with either the same, or with reduced overall mean crude protein (lysine) content, compared with providing a single diet throughout the growing-finishing period. Four hundred and forty-eight group-penned pigs (no. = 32 groups) were assigned at random to one of four treatments: single diet (SD: 11·1 g lysine per kg), high lysine (HL: 12·2, 11·6, 11·1, 10·5 and 10·0 g lysine per kg), medium lysine (ML: 11·1, 10·5, 10·0, 9·5 and 8·9 g lysine per kg) and low lysine series of diets (LL: 10·0, 9·5, 8·9, 8·4 and 7·8 g lysine per kg). The mean lysine contents of the treatments were 11·1, 11·1, 10·0 and 8·9 g/kg for SD, HL, ML and LL respectively. There were five 2-week phases from 38·3 kg to slaughter at 97·3 kg live weight. LL pigs had lower carcass average daily gain (709 v. 742 g/day; P < 0·05) and poorer carcass FCR (P < 0·05: 2·95 v. 2·84 kg/kg) than SD pigs. Daily lysine intake was lower for pigs on HL (P < 0·05: 22·5 g/day), ML (P < 0·001: 20·9 g/day) and LL (P < 0·001: 18·2 g/day) compared with SD (23·4 g/day). Live weight lysine conversion ratio was better for HL (P < 0·05: 26·9 g/kg), ML (P < 0·001: 24·5 g/kg) and LL (P < 0·001: 22·6 g/kg) pigs compared with SD pigs (27·6 g/kg), but carcass lysine conversion ratio was only better for pigs on the ML (P < 0·001: 27·9 g/kg) and LL (P < 0·001: 25·7 g/kg) treatments compared with SD (31·6 g/kg). Nitrogen intake was lower for LL pigs than SD pigs (P < 0·001: 3·45 v. 4·03 kg). Estimated nitrogen excretion was lower for ML (P < 0·01: 2·16 kg) and LL (P < 0·001: 1·85 kg) pigs than SD pigs (2·47 kg). Nitrogen deposition rate was lower for pigs on the LL compared with the SD treatment (P < 0·05: 22·3 v. 23·2 g/day). Although phase feeding diets with the same mean lysine content (11·1 g/kg) as a single diet over the growing-finishing period resulted in similar pig performance, reduced overall daily lysine intake and improved lysine conversion ratio, there was no effect on carcass characteristics or carcass lysine conversion ratio. Reducing the overall mean lysine content to either 10·0 or 8·9 g/kg improved lysine conversion ratio and reduced nitrogen excretion. However, reducing the overall mean lysine content of the diet to 8·9 g/kg had a negative effect on pig growth performance.

The optimum pattern and limiting order of some essential amino acids for growing-furring blue foxes were assessed from nitrogen (N) retention responses. Total tract digestibility and N balance trials were carried out on 24 weaned blue fox males in an 8 ✕ 5 cyclic change-over experiment. Eight experimental diets were prepared by removing proportionately about 0·4 of each of the amino acids studied – methionine + cystine, lysine, threonine, tryptophan and histidine – successively from the amino acid control diet. The main source of protein in the amino acid control diet was casein and an amino acid mixture was added to bring the calculated crude protein (CP) content up to the level of 170 g/kg dry matter (DM). Low-protein (CP 95·7 g/kg DM) and high-protein (CP 166·6 g/kg DM) diets, the protein proportion of which was casein protein, served as negative and positive control diets, respectively. The reduction in N retention when one amino acid in turn was deleted from the amino acid control diet was calculated, and a regression analysis was made between N retention and relative amino acid intake. Data on the animals’ intake of each limiting amino acid and those on the amino acid control diet were used. The optimum amino acid pattern, expressed relative to lysine = 100, proved to be: methionine + cystine 77, threonine 64, histidine 55 and tryptophan 22. The first-limiting amino acids were methionine + cystine. Blue fox responses (N retention, weight gain) to deletion of methionine + cystine from the diet were very severe and exceeded those to deletion of any other amino acid. Moreover, removing methionine + cystine from the diet significantly impaired the apparent digestibility of organic matter, reducing it to a level even lower than that of the low-protein diet. After methionine + cystine, the next-limiting amino acid in casein-based diets was threonine, followed by histidine and tryptophan. The results show the importance of verifying the sufficiency of dietary methionine + cystine in the practical feeding of blue foxes.

Sixteen early to mid lactation Finnish Ayrshire dairy cows were used in a cyclic change-over experiment with four 21-day experimental periods and a 4 5 2 factorial arrangement of treatments to evaluate the effects of heat-treated rapeseed expeller and solvent-extracted soya-bean meal protein supplements on animal performance. Dietary treatments consisted of grass silage offered ad libitum supplemented with a fixed amount of a cereal based concentrate (10 kg/day on a fresh weight basis) containing 120, 150, 180 or 210 g crude protein (CP) per kg dry matter (DM). Concentrate CP content was manipulated by replacement of basal ingredients (g/kg) with either rapeseed expeller (R; 120, 240 and 360) or soya-bean meal (S; 80, 160 and 240). Increases in concentrate CP stimulated linear increases (P < 0·05) in silage intake (mean 22·5 and 23·8 g DM per g/kg increase in dietary CP content, for R and S, respectively) and milk production. Concentrate inclusion of rapeseed expeller elicited higher (P < 0·01) milk yield and milk protein output responses (mean 108 and 3·71 g/day per g/kg DM increase in dietary CP content) than soya-bean meal (corresponding values 62 and 2·57). Improvements in the apparent utilization of dietary nitrogen for milk protein synthesis (mean 0·282 and 0·274, for R and S, respectively) were associated with higher (P < 0·05) plasma concentrations of histidine, branched-chain, essential and total amino acids (35, 482, 902 and 2240 and 26, 410, 800 and 2119 µmol/l, respectively) and lower (P < 0·01) concentrations of urea (corresponding values 4·11 and 4·52 mmol/l). Heat-treated rapeseed expeller proved to be a more effective protein supplement than solvent-extracted soya-bean meal for cows offered grass silage-based diets.

Summary 221

I. Introduction 221

1. The role of sucrose in plants 221

2. C4photosynthesis 222

II. Pathway of sucrose synthesis in C4plants 223

III. Control of sucrose synthesis in C4plants 225

1. The diurnal cycle in C3plants 225

2. The diurnal cycle in C4plants 227

3. Abiotic stress and sucrose synthesis 230

4. Integration of sucrose synthesis with other metabolic pathways 232

IV. Future prospects 233

Acknowledgements 234

References 234

Sucrose has a central role in the metabolism of all higher plants. Here we discuss the location of sucrose synthesis in the leaves of C4 plants, the control of flux through the pathway and its response to environmental conditions, and finally some of the prospects for the genetic manipulation of sucrose metabolism in C4 plants. Much of our knowledge about sucrose synthesis in plants comes from C3 species; it is evident, from those studies of C4 species that are available, that there are many similarities between sucrose synthesis in C3 and C4 plants. However, some of the basic regulatory mechanisms common to both C3 and C4 plants seem to have been adapted for the specialized photosynthetic metabolism of C4 plants. There are also some other important differences: for example, some C4 species possess photosynthetic cells with little or no capacity for sucrose synthesis. For these reasons it is not always appropriate to extrapolate directly from C3 to C4 plants. Even where data are available from one C4 species, usually maize, it should not be assumed that all C4 plants are necessarily the same in every respect. Although C4 plants constitute a small fraction of the plant kingdom as a whole, they include a disproportionate number of agriculturally important species. C4 plants also have some advantages over C3 plants for studies of the regulation of cell-specific gene expression and the control of metabolism. For these reasons we suggest that they are worthy of more attention by researchers.

Egg-yolk protein hydrolysate (YPp) is an alternative protein source in formulas for infants with intolerance to cow's milk or soyabean protein, or for patients with intestinal disorders. However, the nutritional value of YPp has never been investigated. YPp was prepared by enzymic hydrolysis of delipidated yolk protein, which led to an average peptide length of 2.6 residues. Three experiments were performed. In Expt 1, we compared the intestinal absorption rate of YPp and soyabean protein hydrolysate (SPp) in rats. YPp and SPp solutions were injected into the duodenum of anaesthetized rats and blood samples were taken from the portal vein at 7, 15, 30, 60, and 120 min. A higher amino acid concentration in the serum of the YPp group demonstrated that YPp was absorbed faster than SPp. In Expt 2, the effects of dietary YPp and SPp on body-weight gain, protein efficiency ratio (PER) and feed efficiency ratio (FER) were determined. At the end of the experiment, body weight had increased in both groups, while PER and FER were significantly higher in rats fed on YPp. In Expt 3, to investigate the effects of dietary YPp and SPp on N metabolism, we determined the biological value and net protein utilization. Yolk protein was the reference protein. Biological value and net protein utilization values were very similar between animals fed on yolk protein and YPp diets, and significantly higher than in rats fed on the SPp diet. The present findings demonstrate that there is no adverse effect of hydrolysis of yolk protein on N utilization, and that the nutritive value of YPp is similar to that of yolk protein and superior to that of SPp.

The authoritative talk by Professor Fowler (Fowler et al., 1998), emphasized the huge increase in the rate of NOx (NO and NO2) emissions into the atmosphere due to fossil fuel combustion, from 1 Tg N y−1 to over 20 Tg N y−1 during the 100 yr between 1880 and 1980. He went on to predict that this rate of emission from anthropogenic sources would increase to 46 Tg N y−1 by the year 2025. In addition, NO can also be released from the soil following microbial action, a process that is very dependent upon soil temperature, nitrogen availability and water content. Later in the meeting, Professor Raven (Raven & Yin, 1998) pointed out that terrestrial plants, though not necessarily each individual species, have over the past 450 million yr coped with large changes in nitrogenous compounds in the environment. Nevertheless, this is no basis for complacency about the current situation because the rates of change caused by man's activities are probably unprecedented. Furthermore, the fact that terrestrial plant life in some form can continue, despite massive changes in environmental chemistry, does not necessarily indicate that the systems on which we ourselves are dependent will be conserved.

The green alga Chlamydomonas reinhardtii can use the ureides allantoin and allantoate as sole nitrogen sources. Once the uptake systems for allantoin and allantoate were induced, the uptake and growth rates were identical for the two ureides. However, the enzymatic activities involved in the degradation of the two ureides (allantoinase and allantoicase) were regulated differently. Allantoinase seems to be constitutive, since it was detected in all the nitrogen sources studied, while allantoicase behaved as an inducible enzyme, since it was present only in cells cultured in ureides or any metabolic precursor of these compounds. Neither allantoinase nor allantoicase activities were repressed by ammonium in the presence of ureides. Allantoicase activity was not induced under nitrogen starvation conditions, while it was induced in cells that had been cultured with allantoin or allantoate in the dark. Allantoin uptake showed a pattern similar to that of allantoate under all nutritional and environmental conditions tested. Inhibition of allantoin and allantoate uptake by N-ethylmaleimide suggests that thiol (SH–) groups are involved in both uptake systems. The use of both allantoin and allantoate was similarly inhibited by the metabolic poisons tested (cyanide, azide, 2,4-dinitrophenol and 3′-(3,4-dichlorophenyl)-1′,1′-dimethyl urea) but only at very high concentrations. The possibility that uptake of allantoin and allantoate might take place through two independent systems is discussed.

The dynamics of N metabolism in mature horses were investigated when they were fed on a low-N diet or the same diet supplemented with sufficient urea or soyabean meal to meet their theoretical N requirements. There were no differences in DM, organic matter or neutral-detergent-fibre digestibilities for the three diets. N digestibilities and digestible-N intakes were similar for the urea- and soyabean-supplemented diets and very low for the low-N diet. For all three diets plasma urea was degraded in the digestive tract to NH3 which was utilized by the bacterial population as a N source. NH3 was absorbed from the large intestine into the blood and converted to urea. NH3 was also incorporated into plasma proteins. The horses fed on the low-N diet degraded a greater proportion of endogenous urea in the digestive tract than did horses fed on the urea- or soyabean-supplemented diets. However, the horses fed on the urea diet had the highest degradation rate of urea. The quantity of urea degraded in the digestive tract of horses fed on the urea-supplemented and the low-N diets could not compensate for a lack of dietary crude protein in these diets. The horses were in a negative N balance when fed on the low-N and urea-supplemented diets and a positive N balance when fed on the soyabean-supplemented diet. Dietary urea supplementation did not benefit the horses.

Changes in splanchnic energy and N metabolism were studied in sheep, prepared with vascular catheters across the portal-drained viscera (PDV) and the Liver, and maintained on supramaintenance intakes of either grass or grass + barley pellets. The animals were challenged, on both diets, with 4 d intra- mesenteric vein infusions of NH4CI (25 µmol/min) plus NH4HCO3 (at either 0 or 125 µmol/min). On the final day of each treatment the natural abundance NH4Cl was replaced with 15NH4Cl over a 10 h infusion while over the same period [l-13C]leucine was infused via a jugular vein. Measurements were made of blood flow plus mass transfers of NH3, urea, free amino acids and O2, across the PDV and liver. Enrichments of [14N15N]urea and [15N15N]urea plus [15N]glutamine, aspartate and glutamate were also monitored. Whole-body urea flux was determined by infusion of [14C]urea. At the end of the study the animals were infused for 3 h with 15NH4CI, killed and liver samples assayed for intracellular free amino acid enrichments and concentrations. Blood flows across the splanchnic region were unaffected by either diet or level of ammonium salt infusion. At the lower ammonium salt infusion there was a trend for greater absorption of NH3 across the PDV (P<0·10) with grass + barley than with the grass diet, while removal of urea was unaltered. At the higher ammonium salt infusions there was a significantly greater appearance of NH, across the PDV and this exceeded the extra infused. Urea-N removal, however, was also elevated and by more than that required to account for the additional NH3. The PDV contributed 19–28% to whole-body O2 consumption and the liver 23–32%. Hepatic extraction of absorbed NH3 was complete on all treatments and systemic pH remained constant. The fractions of urea-N apparently derived from NH3, were similar on the grass (0·59–0·64) and grass + barley (0·64–0·67) diets. Hepatic production of urea agreed well with urea flux measurements. Between the two levels of ammonium salt infusion and within diets the additional NH3 removed across the PDV was accounted for by the increased urea-N production. The [14N15N]: [15N15N] ratio of the urea produced was 97:3, while the enrichment of hepatic intracellular free aspartate was lower than that of [14N15N]urea. Glutamine enrichments were 0·23–0·37 those of [14N15N]urea, indicating a minor role for those hepatocytes (probably perivenous) which contain glutamine synthetase (EC 6.3.1.2). Leucine kinetics, either for the whole body or splanchnic tissues, were not different between diets or level of ammonium salt infusion, except for oxidation which was less on the grassfbarley ration. Amino acid concentrations were lower on the grass + barley diet but net PDV absorptions were similar. The pattern of essential amino acids absorbed into the PDV showed good agreement with the published composition of mixed rumen microbial protein. Fractional disappearances of absorbed free essential amino acids across the liver varied from 0·4 (branched chains) to near unity (histidine, phenylalanine)

N metabolism was studied in young rats fed on lipid-rich, isonitrogenous, purified liquid diets, a convenient and easy technique for inducing voluntary overfeeding of energy and lipids under controlled nutritional conditions. Overfed rats showed a marked N retention at the expense of a reduced production of urea. The capacities of isolated hepatocytes to synthesize urea and glucose from added precursors were greatly diminished. The activities of the urea cycle enzymes and several enzymes involved in the availability of NH3, for this pathway were concomitantly reduced in overfed animals. Therefore, our results showed an improved N metabolism in overfed rats promoted by the overfeeding of lipids that could be due to an enhanced biosynthetic utilization and a reduced catabolism of amino acids. In addition, the versatile and accurate technique for inducing overfeeding in young rats used in the present study could have many advantages for nutritional studies.

Heat production (HP) and the intake and retention of energy and nitrogen were measured in growing broiler fowl kept at 32° and given diets with metabolizable energy contents from 8 to 15 MJ/kg and crude protein (N × 6·25; CP) contents of 130 and 210 g/kg. The temperature of 32° was chosen for comparison with earlier measurements at 20° to minimize heat produced for the maintenance of body temperature. The effects of diet composition were observed when the same birds were taken from 20 to 32°. The tendency for energy intake to increase with dietary energy concentration was less at 32 than at 20°. The lower heat increments measured for the high-fat diets did not, therefore, confer an increased ability to sustain higher energy intake at 32°. HP was about 17% lower at 32 than at 20°; the change in HP between 20 and 32° was not significantly influenced by diet composition. The absence of significant effects of diet composition on HP, combined with the significant trend in energy intake, produced significant differences (related both to dietary energy and dietary protein concentrations) in total energy retention and in the partition of retained energy between protein and fat. As at 20°, variation in energy retention and in the composition of retained energy were the main responses to variation in dietary CP concentration and energy intake; a significantly higher energy cost of unit protein accretion on the low-CP diets was insufficient to produce an elevation in total HP because the higher unit energy cost was balanced by a lower absolute rate of protein accretion.