Transpupillary collagen photocrosslinking for targeted modulation of ocular biomechanics.

The central vision-threatening event in glaucoma is dysfunction and loss of retinal ganglion cells (RGCs), thought to be promoted by local tissue deformations. Here, we sought to reduce tissue deformation near the optic nerve head by selectively stiffening the peripapillary sclera, i.e. the scleral region immediately adjacent to the optic nerve head. Previous scleral stiffening studies to treat glaucoma or myopia have used either pan-scleral stiffening (not regionally selective) or regionally selective stiffening with limited access to the posterior globe. We present a method for selectively stiffening the peripapillary sclera using a transpupillary annular light beam to activate methylene blue administered by retrobulbar injection. Unlike prior approaches to photocrosslinking in the eye, this approach avoids the damaging effects of ultraviolet light by employing red light. This targeted photocrosslinking approach successfully stiffened the peripapillary sclera at 6 weeks post-treatment, as measured by whole globe inflation testing. Specifically, strain was reduced by 47% when comparing treated vs. untreated sclera within the same eye (n = 7, p=0.0064) and by 54% when comparing the peripapillary sclera of treated vs. untreated eyes (n = 7, p<0.0001). Post-treatment characterization of RGCs (optic nerve axon counts/density, and grading), retinal function (electroretinography), and retinal histology revealed that photocrosslinking was associated with some ocular toxicity. We conclude that a transpupillary photocrosslinking approach enables selective scleral stiffening targeted to the peripapillary region that may be useful in future treatments of glaucoma.

Prediction of sublayer depth in turbid media using spatially offset Raman spectroscopy.

We demonstrate experimentally the feasibility of monitoring the depth of optically thick layers within turbid media using spatially offset Raman spectroscopy (SORS) in combination with multivariate analysis. The method uses the deep penetration capability of SORS to characterize significantly thicker (by at least a factor of 2) layers than possible with conventional Raman spectroscopy. Typical relative accuracies were between 5 and 10%. The incorporation of depth information into a SORS experiment as an additional dimension allows pure spectra of each individual layer to be resolved using three-dimensional multivariate techniques (parallel factor analysis, PARAFAC) to accuracies comparable with the results of a two-dimensional analysis.

Non-invasive detection of cocaine dissolved in beverages using displaced Raman spectroscopy.

We demonstrate the potential of Raman spectroscopy to detect cocaine concealed inside transparent glass bottles containing alcoholic beverages. A clear Raman signature of cocaine with good signal-to-noise was obtained from a approximately 300 g solution of adulterated cocaine (purity 75%) in a 0.7 L authentic brown bottle of rum with 1 s acquisition time. The detection limit was estimated to be of the order of 9 g of pure cocaine per 0.7 L (approximately 0.04 moles L(-1)) with 1 s acquisition time. The technique holds great promise for the fast, non-invasive, detection of concealed illicit compounds inside beverages using portable Raman instruments, thus permitting drug trafficking to be combated more effectively.

Noninvasive detection of concealed liquid explosives using Raman spectroscopy.

We present a Raman spectroscopic method for the noninvasive detection of liquid explosives within bottles, and other packaging, of substantially higher sensitivity and wider applicability than that currently available via conventional Raman spectroscopy. The approach uses a modification of the spatially offset Raman spectroscopy (SORS) concept, which permits the interrogation of a wide range of containers, including transparent, colored, and diffusely scattering plastic and glass beverage, medicine, and cosmetic bottles, with no change in experimental geometry. The enhanced sensitivity is achieved by the technique's inherent ability to effectively suppress fluorescence and Raman contributions originating from the wall of the container. The application is demonstrated on the noninvasive detection of hydrogen peroxide solution, a critical component of a number of liquid explosives. In contrast to conventional Raman spectroscopy, the modified SORS concept enables the detection of concealed hydrogen peroxide solution in all the studied cases.

Effect of glucose supply on fasting nitrogen excretion and effect of level and type of volatile fatty acid infusion on response to protein infusion in cattle.

Two experiments were carried out on cattle nourished entirely by intragastric infusion, to determine the extent to which glucose or a glucose precursor determines the response to protein infusion in energy-undernourished animals. In order to determine the requirement for glucose in 1-year-old fasting cattle, glucose was infused at increments to supply 0, 1.5, 2.5, 3.5, 4.5, 5.5 and 6.5 g/kg metabolic body weight (W0.75) and the effects on plasma beta-hydroxybutyrate and N excretion were measured. At 5.5 g glucose/kg W0.75 plasma beta-hydroxybutyrate was reduced to a basal level of 1.65 mmol/l and fasting N excretion reduced from 529 to 280 mg N/kg W0.75. No further reduction was observed with the higher level of 6.5 g glucose/kg W0.75. In the second trial, three steers were used in a 3 x 3 Latin square design and infused with a volatile fatty acid mixture of 65, 27 and 8 mol acetic, propionic and butyric acids respectively/100 mol, either at an estimated maintenance energy level of 450 kJ/kg W0.75 and supplying a calculated glucose equivalent level of 13.0 g/kg W0.75 (M1A), or at 1.5 x maintenance supplying a glucose equivalent of 20 g/kg W0.75 (M1.5A). Another mixture infused at the maintenance energy level contained 49, 43 and 8 mol acetic, propionic and butyric acids respectively/100 mol but with a glucose equivalent of 20 g/kg W0.75 (M1P). Casein was infused at each of these energy treatments to supply 0, 200, 400, 800, 1600 and 2500 mg N/kg W0.75 daily, and N balance and blood metabolites were measured. N retention increased linearly (r 0.98) with casein infusion. The coefficients for N retention were 0.55, 0.57 and 0.64 for M1A, M1.5A and M1P respectively. The mean efficiency of N utilization was 0.58. The results suggest that provided the glucose need is met there is no relationship between energy supply and efficiency and level of protein retention. However, the results also indicate that glucose requirement in cattle may be higher than that previously observed in sheep.

The effects of acute and chronic protein depletion and accretion on plasma concentrations of insulin-like growth factor-1, fibronectin and total protein for ruminants nourished by intragastric infusion of nutrients.

The relationships of N input or protein status and the concentrations of serum insulin-like growth factor-1 (IGF-1), plasma fibronectin (FN) and total protein (TP) were examined in three experiments with steers and sheep nourished by intragastric infusion of nutrients. In Expt 1, three steers (340 kg live weight) were infused with three levels of volatile fatty acids (0, 300 and 600 kJ/kg metabolic weight (W0.75) per d) and six levels of casein (0, 200, 400, 650, 1500 and 2500 mg N/kg W0.75 per d). Each N treatment was imposed for 5 d. In Expts 2 and 3, five groups of sheep (about 35 kg live weight) were infused with casein at 500 mg N/kg W0.75 per d for 2 weeks followed by 1500, 500 or 50 mg N/kg W0.75 per d in Expt 2, and in Expt 3, with 100 mg N/kg W0.75 per d for 6 weeks or 10 mg N/kg W0.75 per d for 4 weeks. Non-protein energy was maintained constant at 500 kJ/kg W0.75 per d throughout. Daily N balance and total body N content at the end were measured, and protein status was defined as a percentage of cumulative N accretion or depletion in relation to the total body N content at maintenance. It was found that IGF-1 and FN responded rapidly and substantially to altered N input, and that when daily N input was maintained constantly at sub-maintenance, their continuous declines were related closely to progressive protein depletion in the sheep. Plasma TP concentration was independent of N input when N input was altered acutely in the steers, but declined significantly and gradually with severe, chronic body protein depletion in the sheep. Plasma content of TP in the sheep however reduced acutely with a reduction in N input. Plasma volume fell substantially over the first 2 weeks of protein depletion, compensating for the declines in TP content and maintaining TP concentration plateau. The possible implications of the changes in TP concentration and content (concentration x volume) to body protein loss in sheep are discussed.

Excretion of benzoic acid derivatives in urine of sheep given intraruminal infusions of 3-phenylpropionic and cyclohexanecarboxylic acids.

The quantitative relationship between the urinary excretion of benzoic acid (BA) and the uptake of 3-phenylpropionic (PPA) and cyclohexanecarboxylic (CHCA) acids was assessed. PPA and CHCA are produced in the rumen by microbial fermentation of lignocellulosic feeds and metabolized, after absorption, to BA which is excreted in the urine mainly as its glycine conjugate hippuric acid (HA). Four sheep nourished by intragastric infusions of all nutrients were given continuous ruminal infusions of PPA (8, 16 or 24 mmol/d) either alone or with CHCA (8 or 16 mmol/d) in a factorial experiment. The treatments were allocated to ten consecutive 6 d periods, with a control being repeated at periods 1, 5 and 10. PPA and CHCA ruminal absorption rates, estimated using the liquid-phase marker Cr-EDTA, were 0.78 (SD 0.29)/h and 0.88 (SD 0.28)/h respectively. For the control, HA excretion was only 0.22 (SD 0.33) mmol/d and free BA was absent. For the other treatments, both HA and free BA were present and HA accounted for 0.85 (SD 0.05) of total BA: The urinary excretion of total BA showed a significant linear correlation (r = 0.997, P < 0.001) with the amounts of PPA and CHCA infused. The urinary recovery of infused PPA and CHCA as total BA was 0.79 (SE 0.01). Faecal excretion of BA and its precursors was negligible. Results of this study show that urinary total BA is a potential estimator of the absorption of PPA + CHCA produced in the rumen.

Effects of long-term protein excess or deficiency on whole-body protein turnover in sheep nourished by intragastric infusion of nutrients.

The effect of long-term dietary protein excess and deficit on whole-body protein-N turnover (WBPNT) was examined in lambs nourished by intragastric infusions of nutrients. Ten sheep were given 500 mg N/kg metabolic weight (W0.75) per d from casein for 2 weeks and then either 50 (L), 500 (M) or 1500 (H) mg N/kgW0.75 per d for 6 weeks. Volatile fatty acids were infused at 500 kJ/kgW0.75 per d. Daily WBPNT was measured by continuous intravenous infusion of [1-13C]leucine 3 d before, and on days 2, 21 and 42 after the alteration in protein intake. Whole-body protein-N synthesis (WBPNS) was calculated as the difference between WBPNT and the protein-N losses as urinary NH3 and urea. Whole-body protein-N degradation (WBPND) was then estimated from WBPNS minus protein gain determined from N balance. Fractional rates of WBPNS and WBPND were calculated against fleece-free body N content. WBPNS rates at the L, M and H intakes were respectively 35.1, 41.5 and 63.7 g/d (P < 0.001) on average over the 6 weeks and WBPND rates were 39.5, 41.1 and 56.8 g/d (P < 0.001). The fractional rates of WBPNS were 5.01, 6.37 and 7.73% per d (P < 0.001) while those of WBPND were 5.64, 6.29 and 6.81% per d (P < 0.005) respectively. On days 2, 21 and 42, WBPNS rates at intake H were 54.0, 61.8 and 75.4 g/d (P = 0.03) respectively, and WBPND rates were 43.2, 56.4 and 70.9 g/d (P = 0.03); at intake L the amounts were 38.2, 34.2 and 32.8 g/d for WBPNS (P = 0.003) and for WBPND were 43.4, 38.0 and 36.9 g/d (P = 0.016) respectively. There were no significant (P > 0.05) differences in fractional rates of WBPNS and WBPND with time at either the L or H intake. We concluded that absolute protein turnover was affected both by dietary protein intake and body condition while the fractional rate of turnover was predominantly influenced by intake.

The use of intragastric nutrition to study saliva secretion and the relationship between rumen osmotic pressure and water transport.

Four sheep sustained by intragastric nutrition were used to study saliva secretion and the relationship between osmotic pressure in the rumen and net water transport across the rumen wall. Different concentrations of buffer were infused into the rumen to change the rumen osmotic pressure. Salivary secretion was estimated from entrance of P into the rumen. Net water transport across the rumen wall was calculated as the difference between water inflow and water outflow from the rumen. A negative linear relationship between the rumen osmotic pressure (X, mOsm/kg) and the water absorption across the rumen wall (Y, ml/h) was found: Y = (394 SE 8.3)-(1.22 SE 0.03) X, r2 0.83, (P < 0.001), and a positive linear relationship was found between the rumen osmotic pressure (X, mOsm/kg) and the outflow rate of rumen fluid (Y, ml/h): Y = (34.0 SE 8.0) + (0.97 SE 0.03) X, r2 0.56, (P < 0.001). The implication is that rumen osmotic pressure can be a key factor in the control of the net water transport across the rumen wall, the outflow of rumen fluid to omasum and the rumen liquid dilution rate. A method is suggested by which salivary secretion in sheep may be calculated from the water balance in the rumen.

Osmotic pressure, water kinetics and volatile fatty acid absorption in the rumen of sheep sustained by intragastric infusions.

The effects of changing rumen osmotic pressure (OP) upon water kinetics and volatile fatty acid (VFA) absorption in the rumen of sheep were studied in two 4 x 4 Latin square experiments, each using four lambs with a rumen cannula and an abomasal catheter. In both experiments the lambs were sustained by the intragastric infusion of all nutrients (VFA, Ca, P, Mg and a buffer solution into the rumen, and casein, vitamins and trace elements into the abomasum). On experimental days, which were at least 1 week apart, drinking water and the casein infusion were withdrawn, and the ruminal OP was changed and held constant for 9.5 h, by incorporating NaCl at different concentrations in the buffer solution being infused. In Expt 1 the target OP values were 300, 340, 380 and 420 mosmol/kg, and in Expt 2 were 261 (no saline addition), 350, 420 and 490 mosmol/kg. Using soluble non-absorbable markers (PEG in continuous infusion and Cr-EDTA injected in pulse doses) rumen volume, liquid outflow rates, apparent water absorption through the rumen wall and VFA absorption rates were estimated at six sampling times corresponding to the 1.5 h intervals during the last 7.5 h following the change in rumen OP. Liquid outflow rate (F; ml/h) showed a significant and positive linear relationship with the rumen OP (mosmol/kg), resulting in the equation F = 1.24 OP (SE 0.096)-36.5 (SE 36.6) (r2 0.96). Similarly, water absorption rate (W; ml/h) was significantly affected by rumen OP, and this relationship was given by W = 395 (SE 39.9)-1.16 OP (SE 0.105) (r2 0.95), which means that for an OP of 341 mosmol/kg the net movement of water across the rumen wall would be zero, and either a net efflux or a net influx of water would be observed with lower or higher OP respectively. In Expt 2 there was a significant linear effect of OP on rumen volume (P < 0.01), with higher OP being associated with increases in rumen liquid contents of about 10-20%. As rumen OP was increased there was also a decline in the absorption rate of VFA (from 232 mmol VFA/h for OP 350 to 191 mmol/h for OP 490 mosmol/kg), resulting in the accumulation of VFA (especially acetate) in the rumen and a consequent fall in rumen pH. Rumen OP seems to be important in defining water movement across the rumen wall and, hence, partitioning between absorption and outflow.

Effect of level of input of different proportions of volatile fatty acids on energy utilization in growing ruminants.

Four steers were maintained wholly by intragastric infusion of volatile fatty acids (VFA) and protein, together with a mineral-vitamin supplement. The infusion was given at three levels of energy, namely 450, 675 and 900 kJ/kg live weight0.75, calculated to supply energy at 1.0, 1.5 or 2.0 times that required for maintenance. The VFA provided 0.837 and the protein 0.163 of the energy infused. The molar proportions of individual VFA were varied so that the infusate contained 0.36-0.91 of acetic acid, 0.56-0.01 of propionic acid and a constant 0.08 of butyric acid. Heat production was measured in respiration chambers. Urine was analysed for N, urea, beta-hydroxybutyrate and VFA. Blood plasma was analysed for beta-hydroxybutyrate, free fatty acids, insulin and glucose. As the proportion of acetic acid was increased, and propionic acid reduced, there was no change in blood or urine metabolites or in heat production until acetic acid exceeded a proportion of about 0.75. At higher proportions beta-hydroxybutyrate increased in plasma and urine, blood glucose and insulin tended to fall and urinary N excretion rose. At a proportion of acetic acid of > 0.80, acetate appeared in the urine and at > 0.86 heat production declined. The effect of level of infusion on the molar proportion at which plasma and urine metabolites changed was less clear. There was a tendency for the increase in beta-hydroxybutyrate to occur at a slightly lower proportion of acetic acid at the highest level of infusion. It is concluded that differences in heat production that are observed between diets are probably not caused by differences in rumen VFA proportions.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of different volatile fatty acids mixtures on energy metabolism in cattle.

Four Friesian steers (mean BW = 282 kg) were given mixtures of VFA and casein by intragastric infusion to give a total energy input of 675 kJ/kg BW.75. Casein supplied 16.3% of the energy and 777 mg N/kg BW.75. The molar proportion of butyric acid was held constant at 8 mol/100 mol, and the acetic and propionic acids varied inversely. Acetic acid was varied in 12 increments from 11 to 91 mol/100 mol and propionic acid proportion varied inversely. Heat production, blood (urea, insulin, beta-hydroxybutyrate, free fatty acids) and urine metabolites (urea, N, VFA) were measured. There were no differences (P greater than .05) in heat production until the acetic acid proportions exceeded approximately 90 mol/100 mol, at which point there was a decrease in heat production (P less than .05) accompanied by a considerable excretion of acetic acid in the urine. Above 80 mol/100 mol acetic acid, beta-hydroxybutyrate was greatly elevated, accompanied by a small decrease in blood glucose and blood insulin together with an increase in blood free fatty acid concentration. There was also an elevation of N excretion in the urine. When the proportion of propionic acid exceeded 76 mol/100 mol there were some metabolic disturbances resulting in blood hemolysis, an increase in N excretion in the urine, and nervous disposition of the animals. It is concluded that differences in heat production between roughage and concentrate diets are not likely to be a result of differences in the energetic response to different proportions of VFA. Differences in activity during standing, feeding, and ruminating may, therefore, be more important.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects on plasma insulin of intermittent infusions of propionic acid, glucose or casein into the alimentary tract of non-lactating cows maintained on a liquid diet.

1. An experiment was conducted using three non-lactating cows completely maintained by infusions of volatile fatty acids into the rumen, and casein into the abomasum. Plasma insulin responses to propionic acid, glucose or casein were recorded. Further information was obtained using protein-free infusions. 2. When part of the propionic acid was infused into the rumen in a twice-daily 3 h dose and the remainder infused continuously with acetic and butyric acids and casein, there were large increases in the concentrations of propionic acid and insulin in the jugular blood. When glucose, corresponding in energy to that supplied by the intermittent propionic acid infusions was similarly infused, the plasma levels of glucose and insulin were increased. Glucose appeared to stimulate a greater increase in insulin than did propionic acid. Casein infused into the abomasum in intermittent doses produced a rise in plasma insulin, but smaller than that observed with propionic acid or with glucose. 3. The protein-free infusion was characterized by a lower concentration of insulin in the blood plasma, a reduction in plasma urea and free amino nitrogen and unchanged plasma glucose.

Undernutrition in sheep. Nitrogen repletion by N-depleted sheep.

Wether lambs of 29-44 kg live-weight, totally nourished by the infusion of volatile fatty acids (VFA) into the rumen and casein into the abomasum, were given five treatments in consecutive periods. The treatments were (daily amounts per kg live weight (W)0.75): (a) high-protein for 7 d (2500 mg nitrogen, 650 kJ VFA); (b) low-protein for 7-15 d (525 mg N, 650 kJ VFA); (c) N-free for 7 d (no N, 450 kJ VFA); (d) very-low-protein for 24-28 d (300 mg N, 400 kJ VFA); (e) high-protein for 40 d (2500 mg N, 650 kJ VFA). Nine lambs were subjected to treatments (a), (b) and (c) (Expt 1) and four of the lambs additionally received treatments (d) and (e) (Expt 2). In Expt 1 all nine lambs had a positive N retention on treatment (a) but abrupt change to treatment (b) resulted in substantial negative N balances initially, and a period of approximately 5 d adaptation was required before N equilibrium was re-established. Animals again exhibited negative N balances when the N-free infusion (treatment c) was introduced and during that period there was no evidence of adaptation. Basal urinary N excretion was estimated to be 356 (SE 12) mg N/kg W 0.75. In Expt 2 all four lambs were depleted of N when receiving the very-low-protein treatment (d). The progressively decreasing N losses recorded during days 1 to 12 of the treatment period were slightly greater than those recorded during days 13 to 28 but the difference between the means was not significant (P greater than 0.05). There was no evidence of an adaptation in N retention between days 13 and 28 of the treatment. As assessed during days 13 to 28 of the treatment the efficiency of utilization of infused casein N was 1.0; this compared with a value of 0.66 recorded during treatment (b) in Expt 1. Live weight loss during the period of N depletion was 101 (SE 27) g/d. When lambs were given treatment (e) during the last period of Expt 2, N repletion was rapid and complete within a few days. Ten days after the introduction of the treatment the rate of N retention was estimated to be 1019 (SE 38) mg/kg W 0.75 per d and this value declined at a rate of 9.5 (SE 1.9) mg N/kg W 0.75 per d for the following 30 d.(ABSTRACT TRUNCATED AT 400 WORDS)

Flow of nitrogen from the rumen and abomasum in cattle and sheep given protein-free nutrients by intragastric infusion.

1. Three experiments were conducted to determine the flow of nitrogen through the rumen and abomasum when cows, steers and lambs were totally nourished on volatile fatty acids infused into the rumen. 2. In two dairy cows (650-700 kg) and two large steers (370-405 kg) the daily flow of non-ammonia-N (NAN) from the rumen was 50.7 and 58 mg/kg live weight (W)0.75 respectively. 3. The flows of NAN through the rumen and abomasum in four young steers (240-315 kg) were 85.0 (SE 21.0) and 195 (SE 7.0) mg/kg W 0.75 respectively. 4. In the third experiment the effects of altering rumen pH and osmotic pressure on flow of NAN through the rumen and abomasum were investigated in lambs. While rumen pH and osmotic pressure influenced rumen volume and outflow they had no significant effect on NAN flow. The mean values for NAN outflow from the rumen and abomasum were 76 and 181 mg N/kg W 0.75 respectively. 5. Abomasal NAN flow increased with increasing abomasal pH. When osmotic pressure was greater than about 330 mosmol/l in the rumen there was a net inflow of water, while below this value there was net loss of water. 6. For all experiments the flow of N both from the rumen and abomasum was highly variable; this has to be considered if a constant value is used for endogenous N in estimating dietary N in the abomasum. 7. With N-free infusion the rumen NH3 concentration varied from 50 to 120 mg NH3-N/l. 8. The amino acid composition of rumen and abomasal N was also determined. Relative to tissue N it contained a higher proportion of cysteine.

The effect of intragastric infusion of glucose, lipids or acetate on fasting nitrogen excretion and blood metabolites in sheep.

Two experiments are reported in which the effect of the intragastric infusion of non-protein energy on fasting nitrogen losses was studied. Expt 1 was a preliminary trial with two 35 kg lambs given 0, 144, 288 or 432 kJ/kg live weight (W)0.75 per d as lipid or glucose infused into the abomasum for periods of 3 d. Expt 2 was of a 4 X 4 Latin square design with four sheep of about 30 kg live weight. The four treatments were control (fasted with water infusion), or the infusion of 144 kJ/kg W0.75 per d as glucose or lipid into the abomasum or as acetic acid into the reticulo-rumen. Compared with the fasted control, glucose infusion reduced (P less than 0.05) N excretion to about 0.6 of that of the control, increased (P less than 0.05) plasma glucose, decreased (P less than 0.05) plasma urea and beta-hydroxybutyrate, and was without effect on plasma amino-N or creatinine excretion. Lipid and acetate infusions were without statistically significant effect on N or creatinine excretion or any of the blood indices measured, with the exception of plasma glucose which was reduced (P less than 0.05) with acetate infusion.

The effect of changes in the amount of energy infused as volatile fatty acids on the nitrogen retention and creatinine excretion of lambs wholly nourished by intragastric infusion.

The nitrogen balance and creatinine excretion of wether lambs of 30-48 kg, wholly nourished by the intragastric infusion of nutrients, were measured in two experiments. Four lambs were used in each experiment. In Expt 1 a constant amount of casein was infused into the abomasum (640 mg N/kg body-weight (W)0.75 per d) and the amount of volatile fatty acids (VFA) infused into the rumen ranged from 0 to 670 kJ/kg W0.75 per d as six increments. Expt 2 was of similar design but two levels of casein were infused (530 and 1060 mg N/kg W0.75 per d) and, with each level of casein, VFA infused ranged from 0 to 700 kJ/kg W0.75 per d as seven increments. Daily creatinine excretion was not constant, but varied between 2-d means with standard deviations of between 7.1 and 16.5% (average 13.1%) of the over-all means. There was an apparent correlation between creatinine excretion and the amount of energy infused in six out of eight lambs. There was no effect of the amount of casein infused. In both experiments N balance was negative only when the amount of energy infused was substantially below published values for energy maintenance. In Expt 1, N equilibrium was calculated to be achieved at a gross (VFA plus protein) energy infusion level of 162 (SE 29) kJ/kg W0.75 per d. In Expt 2 it was observed that, at a given level of energy infusion, N retention was greater when the amount of energy had been increased from the previous level, than when it had been decreased. It is concluded that the animal appears to allocate available amino acids to protein synthesis, rather than to oxidation, even when in negative energy balance. It is also concluded that enhanced N retention observed when the amount of energy infused had been increased represented compensation after a period of energy restriction.

Basal urinary nitrogen excretion and growth response to supplemental protein by lambs close to energy equilibrium.

Two experiments are reported. In Expt 1, five male lambs of 26-33 kg were used to measure basal nitrogen excretion when the lambs were entirely sustained by an intraruminal infusion of 450 kJ/kg body-weight 0.75 per d of volatile fatty acid (VFA) and were receiving no protein. In Expt 2, which was a conventional growth trial, the response to fish meal (66 or 132 g dry matter/d) of lambs given a control diet of sodium-hydroxide-treated barley straw was measured. In Expt 1 the mean basal N excretion of the lambs was 429 (SE 21) mg N/kg body-weight 0.75 per d. This exceeds current UK standards for the amino acid N of microbial origin which would be made available to the normally-fed host animal at a maintenance level of metabolizable energy intake. In Expt 2 there was a clear growth response to the fish meal, which was greater (P less than 0.05, single-tailed test) than that to be predicted from the energy content of the fish meal. There was no effect of fish meal on the voluntary intake of the basal diet, but there was a suggestion that the digestibility of the basal diet was improved. It is concluded from Expt 1 that the basal requirement for amino acid N by lambs is three- to fourfold that currently recommended in the UK. This higher basal N requirement should have resulted in a marked response to supplemental protein in Expt 2. The fact that the growth response in Expt 2 was less than anticipated may have been due to a combination of a slightly lower basal N excretion than that found in Expt 1, a higher yield of amino acids of microbial origin than current UK standards predict, and possibly to a change in the body composition of the lambs.

Investigation of nitrogen balance in dairy cows and steers nourished by intragastric infusion. Effects of submaintenance energy input with or without protein.

Two dairy cows were maintained by intragastric infusion of volatile fatty acids and casein. Except when fasting, the casein-nitrogen was held constant, while total gross energy supply was varied from zero during fasting to 650 kJ/kg body-weight (W)0 . 75. One cow was estimated to attain zero N balance at an energy intake of 255 kJ/kg W0 . 75 and the other at 307 kJ/kg W0 . 75, which was calculated to be substantially below the estimated energy required for zero energy balance. When the cows were later given an N-free infusion for a period preceding the trial, N balance occurred at 98 kJ/kg W0 . 75 for one cow and 115 kJ/kg W0 . 75 for the other. Four steers were similarly nourished by intragastric infusion and the energy nutrient increased from 0 at fasting to 450 kJ/kg W0 . 75. The protein was held constant at 1 g N/kg W0 . 75 except at fasting. The energy level at which N balance occurred was 154 (SE 38) kJ/kg W0 . 75 or approximately equal to the energy content of the protein. The practical implications of these findings are discussed.