Estimating purine derivatives and nitrogen compound excretion using total urine collection or spot urine samples in grazing heifers

The study aimed to evaluate the excretion of purine derivatives (PDs) and nitrogen compounds (NCs) and their ratios with creatinine in supplemented Zebu heifers kept on pastures by comparing total urine collection and spot sampling. Five Nelore heifers (400 ± 15 kg) were used in a 5 × 5 Latin square design. The treatments were the amount of concentrate (220 g of crude protein/kg dry matter) offered (0, 3, 6, 9 and 12 g/kg BW). In each period, the total urine collection was performed continuously for 3 days (subsampled at intervals of 4 h, 00:00–04:00 h, 04:00–08:00 h, 08:00–12:00 h, 12:00–16:00 h, 16:00–20:00 h and 20:00–24:00 h). The spot urine samplings were performed (in each period) for 24 h (0, 4, 8, 12, 16 and 20 h). Creatinine, total urinary nitrogen (UN), urea nitrogen (UreaN), allantoin and uric acid were analysed. Creatinine excretion was 23.01 ± 0.19 mg/kg BW and was not affected by collection day, treatment or their interactions (p > 0.05). Treatments affected (p < 0.05) PD excretions, however did not affect the ratio PD:creatinine (p > 0.05). Treatments and collection time affected (p < 0.05) NC excretion, whereas the UN:creatinine and UreaN:creatinine ratios were not affected (p > 0.05). Creatinine excretion and the PD:creatinine ratios in the urine samples estimated by the total or spot sampling were not different (p > 0.05). However, sampling method affected (p < 0.05) the UN:creatinine and UreaN:creatinine (p < 0.05) ratios. Creatinine can adequately estimate urinary excretion in grazing heifers, and a single spot urine sample at any time of the day can be used to estimate PD excretion in grazing heifers. But two spot urine samples are needed for proper NC excretion estimations in grazing heifers’ urine.     

Urinary excretion of advanced glycation end products in dogs and cats

The present study was conducted with privately owned dogs and cats to investigate whether a relationship exists between the dietary AGEs and the urinary excretion of AGEs, as indication of possible effective absorption of those compounds in the intestinal tract of pet carnivores. For this purpose, data were collected from both raw fed and dry processed food (DPF) fed to dogs and cats, through spot urine sampling and questionnaires. Raw pet food (RF, low in AGE diets) was fed as a primary food source to 29 dogs and DPF to 28 dogs. Cats were categorized into 3 groups, which were RF (n = 15), DPF (n = 14) and dry and wet processed pet food (DWF, n = 25). Urinary-free carboxymethyllysine (CML), carboxyethyllysine (CEL) and lysinoalanine (LAL) were analysed using ultrahigh-performance liquid chromatography (UHPLC)—mass spectrometry, and were standardized for variable urine concentration by expressing the AGE concentrations as a ratio to urine creatinine (Ucr) concentration (µg/µmol Ucr). Urinary excretion of CML, CEL and LAL in dogs fed with DPF was 2.03, 2.14 and 3 times higher compared to dogs fed with RF (p < .005). Similar to the dogs, a significant difference in CML:Ucr, CEL:Ucr and LAL:Ucr between the three diet groups was observed in cats (p-overall < 0.005, ANOVA), in which the RF fed group excreted less AGEs than the other groups. Linear regression coefficients and SE of CML:Ucr, CEL:Ucr and LAL:Ucr showed that body weight and neuter status were significantly correlated with CML and CEL excretion, but not to LAL excretion. Our results revealed a significant correlation between dietary AGEs and urinary excretion of free CML, CEL and LAL, and also showed that endogenous formation of these AGEs occurs in both dogs and cats under physiological conditions.     

Estimation of 24-h aldosterone secretion in the dog using the urine aldosterone:Creatinine ratio

ObjectivesOne potential method of evaluating renin–angiotensin–aldosterone system (RAAS) activation involves the quantification of urinary aldosterone excretion. While blood concentrations of aldosterone are easily obtained, results may be misleading because of minute-to-minute variation in aldosterone secretion and subsequent blood concentrations. Urinary aldosterone concentration measurement represents a more consistent “pooled” index of aldosterone secretion, but obtaining 24-h urine samples is time-consuming, difficult, and fraught with potential error. We postulated that the urinary aldosterone:creatinine ratio, measured from spot urine samples, would correlate well with 24-h urinary aldosterone excretion, and would provide a simple index of aldosterone excretion that would eliminate the need for 24-h urine collection.Animals, materials and methodsAfter validating an assay for aldosterone in canine urine, 24-h urinary aldosterone excretion was determined by radioimmunoassay from 8 normal, male beagle dogs under control conditions, after RAAS stimulation with amlodipine administration, and after RAAS attenuation with the addition of enalapril to amlodipine administration. Spot urine samples, each obtained at the same time of day, were used to determine the aldosterone:creatinine ratio during control conditions, RAAS stimulation, and RAAS attenuation.ResultsThe aldosterone:creatinine ratio from spot-checked urine samples correlated well with 24-h urinary aldosterone excretion (r = 0.77, P < 0.0001).ConclusionsA spot urinary aldosterone:creatinine ratio might be substituted for 24-h urinary aldosterone determination.     

High versus low protein diets to mink--postprandial plasma urea and creatinine response, osmotic load and pattern of nitrogen and electrolyte excretion.

Nitrogen balance, pattern of excretion of nitrogenous end-products, endogenous urinary N excretion, postprandial plasma urea and creatinine, osmotic load, urinary electrolyte excretion and water intake/output relationships were studied in 12 adult female mink fed a high protein diet (HP; n = 6) providing about 155 g protein/kg or a low protein diet (LP; n = 6) providing about 95 g protein/kg. Two balance periods of each 3 d were used and diets were fed raw or cooked. After the last balance period followed a 48 h fasting period. Postprandial plasma urea and creatinine were studied for 48 h following a test meal given after an overnight fast. Osmotic load was determined based on collection of non-acidified urine carried out during 48 h. Level of protein supply did not affect N balance, being close to zero, whereas slightly negative balances were achieved for fasting animals. Protein supply was clearly reflected in excretion of urinary urea and allantoin but not in creatinine and uric acid. Endogenous urinary N excretion was estimated by a second order regression equation giving an intercept of 280 mg/kg0.75. Post-prandial plasma urea concentrations were strongly influenced by protein supply, HP animals having substantially higher peaks than LP animals, but values returned to fasting values within 24 h after the test meal. Plasma creatinine followed a biphasic pattern with a peak about 2 h after feeding and a nadir approximately 6 h after feeding. Physical form of diet influenced postprandial urea, animals fed raw diets having a higher peak, but not creatinine. The HP diet provided almost the double osmotic load of the LP diet and a corresponding increase in urine volume. The resulting water balances were identical irrespective of diet, showing that water intake/output relationships are very accurately regulated.     

The rate and pattern of urea infusion into the rumen of wethers alters nitrogen balance and plasma ammonia

Changes in N balance, urinary excretion of purine derivative (PD), urea, creatinine and ammonia and plasma ammonia, glucose, urea, insulin and IGF‐1 were examined in four wethers (37 ± 2.6 kg BW). The animals were fitted with permanent ruminal catheters, fed lucerne hay (9.4 MJ/day; 23 g N/day; 7 g soluble N/day, 6 equal meals/day) and treated with contrasting rates of urea infusion into the rumen: first, a continuous infusion (CT), at 3.2 mg urea‐N/min for 10 days and then a discontinuous infusion (DT) at 156 mg urea‐N/min for 4 min; in 6 daily doses with the meals for 7 days. N balance was calculated from pooled samples of faeces and urine. Jugular blood samples were collected before and 1.5 h after the morning meal (M1) on days CT10, DT2, DT4 and DT6. N retention decreased during DT (p = 0.01) due to a significant increase of N excretion in urine (4 g/day; p = 0.009) and faeces (1 g/day; p = 0.02). Dry matter (p < 0.001) and N digestibility in vivo (p = 0.01) decreased significantly during DT. Urinary urea and PD excretion were not altered by treatment. Significant linear (p = 0.004) and quadratic (p = 0.001) effects were observed for plasma ammonia in M1 (from 170 CT10 to 235 μm DT2 and returned to 120 μm DT6). No changes were observed in plasma glucose, urea, insulin and IGF‐1. Results indicate that changes from CT to DT reduced N retention in sheep due to enhanced urinary N excretion, but it was not associated with changes in urinary urea or PD excretion; or plasma concentrations of insulin and IGF‐1. As the dry matter (DM) an N digestibility could account a 0.23 of the decrease in N retention; the largest fraction of the reduction in N retention remained unexplained by the results.     

Basal glucosuria in cats

Objective of this study was to demonstrate the ubiquitous presence of glucose in urine of euglycemic cats by a highly sensitive glucose assay. The local electronic database was searched for results of quantitative urine glucose measurements in cats. A total of 325 feline urine glucose measurements were identified, of which 303 (93%) had been submitted by one of the co‐authors working in a near‐by small animal practice. After the exclusion of patients with kidney disease (n = 60), hyperthyroidism (n = 15), diabetes mellitus (n = 11), multiple diseases (n = 9) or steroid treatment (n = 3), as well as serial measurements (n = 87) and outliers (n = 8), the final study population consisted of 132 cats. Urine creatinine concentration was unavailable in five patients. Whereas all but one cat had glucose concentrations above the detection limit of the assay (0.11 mmol/L, Gluco‐quant Enzyme Kit/Roche Diagnostics), no positive glucose dipstick test result (Combur 9‐Test, Roche Diagnostics) was observed. The median (range) of urinary glucose concentration and the glucose‐to‐creatinine ratio (UGCR) was 0.389 (<0.11–1.665) mmol/L and 0.0258 (0.007–0.517) respectively. The UGCR was not affected by age, gender, breed or leukocyturia, whereas cats with hematuria had slightly higher values. Data show that so‐called “basal glucosuria” is present in the majority of cats and by no means diagnostic for diabetes mellitus or renal glucosuria. This has to be considered when using bio‐analytical methods with a low limit of quantification.     

Determination of creatinine excretion and evaluation of spot urine sampling in Holstein cattle

Four experiments were carried out to determine urinary creatinine excretion in Holstein growing bulls, lactating cows, and replacement heifers. In addition, we evaluated the use of spot sampling technique to estimate purine derivatives (PD) excretion. In Experiment I, 15 lactating cows were used in a randomized block design to compare creatinine excretion obtained in different time-spans of urine collection (during 6, 9, 12, 15, 18, 21, and 24 h). In Experiment II, four bulls were allocated in a 4 × 4 Latin square to evaluate the effect of diet (levels of cottonseed hulls of 0, 10, 20, and 30% of the DM) on excretion of creatinine. In Experiment III, 15 lactating cows were used to evaluate the effect of milk production (ranging from 3.9 to 36.7 kg/d) on daily creatinine and PD excretions. In Experiment IV, 22 replacement heifers were utilized to evaluate the effect of body weight (BW, ranging from 107 to 545 kg) on daily creatinine and PD excretions. For all experiments, total urine collections were made over 24 h and daily creatinine and PD excretions were determined. Different time-spans for total urine collection had no effect (P = 0.70) on creatinine excretion compared to the 24-h collection period, indicating a constant excretion rate of creatinine. The roughage source did not influence (P = 0.64) creatinine excretion by bulls, averaging 0.248 ± 0.008 mmol/kg BW. Similarly, milk production did not affect (P = 0.82) creatinine excretion in cows, averaging 0.212 ± 0.004 mmol/kg BW. In contrast, the creatinine excretion (mmol/kg BW) decreased linearly (P < 0.001) as BW of heifers increased, suggesting that creatinine excretion might vary with the degree of maturity of growing animals. There were no differences (P > 0.14) between the 24-h total collection and spot sampling technique in estimating daily PD excretion. The spot sampling technique may be used to estimate the daily excretion of urinary PD in Holstein cattle under practical conditions.     

Effects of low-crude protein diets supplemented with rumen-protected lysine and methionine on fattening performance and nitrogen excretion of Holstein steers

The objective of this experiment was to investigate the effects of low-crude protein (CP) diets supplemented with rumen-protected lysine and methionine on growth performance, nitrogen excretion, and carcass traits in Holstein steers. Steers consumed the following diets: (1) 17.2% CP on a dry-matter basis during the early period (from 7 to 10 months of age) and 14.5% CP during the late period (from 10 to 18 months of age; CON, n = 4, initial body weight [BW] 238 kg), and (2) 14.4% CP during the early period and 11.4% CP during the late period (AA, n = 4, initial BW 243 kg). The AA diet contains rumen-protected lysine and methionine. Except for CP intake, feed intake and body weight gain were not affected by dietary CP content. Total nitrogen excretion per metabolic BW tended to be lower (p < .10) in the early period and significantly lower (p < .05) in the late period with decreasing the feed CP content. Plasma urea nitrogen concentrations were lower in AA than CON. Carcass traits and total free amino acid contents of the longissimus thoracis muscle were not affected by dietary CP content. Adding rumen-protected lysine and methionine to a low-CP diet would reduce nitrogen excretion in fattening Holstein steers without affecting productivity.     

Estimation of Quantitative Enzymuria in Dogs With Gentamicin-Induced Nephrotoxicosis Using Urine Enzyme/Creatinine Ratios From Spot Urine Samples

The correlation between 24-hour urinary excretion of N -acetyl-β- d -glucosaminidase (NAG) and γ-glutamyl transferase (GGT) with urine NAG and GGT/creatinine ratios was assessed in dogs with gentamicin-induced nephrotoxicosis. Eighteen 6-month-oid male Beagles with normal renal function were randomly divided into 3 groups of 6. Each group was fed a different concentration of protein (high protein, 27.3%; medium protein, 13.7%; and low protein, 9.4%) for 21 days. After dietary conditioning, gentamicin was administered at a dose of 10 mg/kg IM tid for 8 days and each group was continued on its respective diet. Endogenous creatinine clearance and 24-hour urinary excretion of NAG and GGT were determined after dietary conditioning (day 0) and on days 2, 4, 6, and 8 of gentamicin administration. In addition, urine NAG and GGT/creatinine ratios (IU/L ± mg/dL) were determined from catheterized spot urine samples obtained between 7 and 10 am on the same days. The correlation between 24-hour urinary enzyme excretion and urine enzyme/creatinine ratio in the spot urine samples was evaluated by simple linear regression analysis. Spot sample urine enzyme/creatinine ratios were significantly correlated with 24-hour urinary enzyme excretion through day 4 for dogs on low dietary protein, through day 6 for those on medium protein, and through day 8 for those on high dietary protein. Mean ± SD baseline values for urine NAG/creatinine ratio and 24-hour urinary NAG excretion were 0.06 ± 0.04 and 0.19 ± 0.14 IU/kg/24 hr, respectively. Baseline values for urine GGT/creatinine ratio and 24-hour urinary GGT excretion were 0.39 ± 0.18 and 1.42 ± 0.82 IU/kg/24 hr, respectively.     

Effect of controlled alterations in maternal dietary retinol on foetal and neonatal retinol status and pregnancy outcome in pigs

The objective of this experiment was to examine the effects of feeding a vitamin A deficient diet (VAF) to pigs at different times on day 100 of foetal and days 0, 1 and 2 of neonatal development. Three treatments included a control (n = 12), VAF for 100 days before mating and during the first month of pregnancy (n = 13; VAF–control), and VAF during the oestrous cycle before mating and throughout pregnancy (n = 13; control–VAF). On day 100 of pregnancy, maternal liver and plasma retinol concentrations were reduced in both groups of gilts fed a vitamin A free diet compared to controls (P < 0.001 and P < 0.01, respectively). Day 100 foetal liver retinol concentrations were not affected by dietary treatment, whereas foetal plasma concentrations were higher in foetuses carried by gilts fed the VAF–control diet (P < 0.05). Piglets born to mothers fed the control–VAF, but not the VAF–control diet had consistently lower hepatic and plasma liver retinol concentrations (P < 0.05). Moderate reductions in maternal vitamin A at either stage of pregnancy did not affect pregnancy rate, litter size, progesterone secretion or the allometric relationships between foetal or neonatal organ and total body size. Reduced vitamin A during conception and early pregnancy, but not during later pregnancy, was associated with increased within-litter uniformity in birth weight (P < 0.05) and a tendency for fewer low birth weight piglets, but this needs to be confirmed in a greater number of sows. The mechanism underlying this effect is not known, but appears to not involve an alteration in progesterone production.     

Effect of increasing taurine and methionine supplementation on urinary taurine excretion in a model insectivore,the giant anteater (Myrmecophaga tridactyla)

The giant anteater (Mymercophaga tridactyla) is a highly specialized insectivore for which nutrient requirements are not clearly established, making diet formulation challenging for this species. Multiple clinical reports suggest anteaters have an obligate dietary taurine (TAU) requirement. Sulphur amino acid (SAA) metabolism in adult anteaters was evaluated using noninvasive methods to measure TAU synthesis potential from dietary methionine (MET) and a basal diet containing on a dry matter (DM) basis 1.7 mg TAU/kg DM and 6.9 g MET/kg DM. Urinary equilibrium times for TAU excretion were determined by feeding the basal diet with or without 1.5 g/kg DM supplemental TAU (crossover design; n = 4). Effects of supplemental dietary TAU (1.7, 2.0, 2.4, 2.7, 3.0, 3.3 g/kg DM) or MET (6.9, 9.0, 11.2 g/kg DM) on urinary TAU were evaluated (randomized block trials; n = 5 or 4 respectively). All urinary values (TAU, MET, unbound inorganic sulphate) were normalized to creatinine (CRT). Results indicate urinary TAU equilibrium in anteaters requires at least 2 weeks of feeding. Urinary ratio of TAU to CRT (TAU:CRT) increased as dietary TAU content increased from 1.7 to 3.0 g/kg DM, consistent with renal homoeostatic modulation of TAU excretion. Our data indicate that TAU needs were met by TAU in the basal diet or by de novo synthesis. Supplemental MET resulted in ~five‐ to eightfold increases in urinary TAU:CRT excretion, further supporting existence of mechanisms for TAU synthesis from dietary SAA in anteaters. Adult anteaters appear able to synthesize TAU when diets contain adequate SAA, but dietary TAU may be critical if protein intakes are low or of poor quality. This study may provide guidance on choice of domestic canids vs. felids as suitable physiologic models for improved nutrition in giant anteaters, and also outlines a noninvasive method for assessing TAU status/metabolism that may be useful across species.     

Comparison between the effects of feeding copper sulphate-treated and untreated rapeseed cake containing high glucosinolates on rumen fermentation,nutrient digestion and nitrogen metabolism in steers

Two trials were carried out to study the effects of copper sulphate (CuSO₄) on detoxifying glucosinolates (GLS) in rapeseed cake (RSC) and compare the effects of feeding CuSO₄-treated and untreated RSC on nutrient digestion and nitrogen (N) metabolism in steers. In Trial 1, different concentrations of CuSO₄ solution (1.6 vs. 3.2 g CuSO₄·5H₂O L⁻¹), soaking temperatures (25 vs. 60°C) and drying methods (air drying at 60°C vs. freeze drying) were allocated in a 2 × 2 × 2 factorial arrangement in vitro. In Trial 2, six steers and dietary inclusions of untreated RSC (control), CuSO₄-treated RSC and CuSO₄-added RSC were assigned in a replicated 3 × 3 Latin square design. CuSO₄ treatment in vitro decreased the contents of GLS and thiocyanate (TC) in RSC (p < 0.001). The total amount of GLS and TC decreased by 62.7–68.5% for all treatments. The animal trial showed that CuSO₄-treated RSC inclusion decreased ruminal concentration of valerate (p < 0.01), whereas it did not affect ruminal pH, ammonia N and total volatile fatty acids. Compared with the control, feeding CuSO₄-treated or CuSO₄-added RSC had no effect on plasma concentrations of triiodothyronine and thyroxine, N excretion and N retention. CuSO₄-treated RSC tended to increase neutral detergent fibre digestibility (p = 0.072) and urinary excretion of urea (p = 0.056). Urinary excretion of purine derivatives (p = 0.076) and rumen microbial N supply (p = 0.084) tended to decrease when feeding CuSO₄-treated RSC versus control. TC was found to be the only metabolite of GLS in rumen fluid, plasma and urine. It was feasible to detoxify GLS in RSC using low CuSO₄ at room temperature. However, feeding CuSO₄-treated or CuSO₄-added RSC had minor effects on rumen fermentation, nutrient digestion and N metabolism in steers. CuSO₄ treatment on RSC for feeding steers seems to be unnecessary.     

Pharmacodynamic assessment of diuretic efficacy and braking in a furosemide continuous infusion model

Introduction

Diuretic failure is a potential life-ending event but is unpredictable and poorly understood. The objectives of this study were to evaluate pharmacodynamic markers of furosemide-induced diuresis and to investigate mechanisms of diuretic braking in dogs receiving constant rate infusion (CRI) of furosemide.

Dairy cow defecation and urination frequency and spatial distribution in relation to time-limited grazing

The objective of this paper was to investigate the effect of limited grazing time on urination and defecation frequency, spatial distribution of excrement in the paddock, and the resulting nitrogen balance at animal and field level. During a 6-week period in early summer, 60 Holstein Frisian dairy cows (31.0 ± 5.4 kg ECM) were randomly allocated to three different treatments, with grazing at clover-grass pasture during daytime for 4, 6.5 or 9 h daily. Indoor feeding, with a mixture of roughage and concentrates (13% crude protein), was restricted for treatment 4 and 6.5 h to the amount the 9-h treatment could eat. Cows allowed grazing at pasture for 4 h moved more rapidly during pasture, moved longer distance per active hour and used a higher proportion of the time eating, both at pasture and indoor, than the cows allowed longer time at pasture. Limiting the grazing time had no influence on the urination (mean = 0.26) and defecation (mean = 0.37) frequency per cow per hour during pasture. Even though the proportion of time active (eating, drinking, standing or walking), and the actual time active during pasture was different for the treatments, the frequency of urination and defecation per active hour was also unaffected by the treatments. Urine and faeces were distributed in the pasture, without specific hot-spots. The estimated daily N-balance at animal level showed increased N excretion with time at pasture. Assuming that excretion follows the active periods during the day and 7000 kg DM foliage is available on yearly basis, this would result in total excretion at field level of 58, 86 and 108 kg N per ha respectively for treatment 4, 6.5 and 9 h. The results of this experiment show that it is possible to reduce the nitrogen excretion in a grazing system by restricting the grazing time of dairy cows together with restricted indoor feeding while maintaining high foliage intake.     

Alternative methodologies to estimate ingestion of caecotrophes in growing rabbits

This study was carried out in order to estimate caecotrophe intake in growing rabbits by three existing procedures: caecotrophes collection after collar fitting, urinary purine derivatives (PD) excretion and microbial ¹⁵N-lysine incorporation. In a first experiment sixteen New Zealand White male rabbits were divided in three groups receiving the same diet, but supplemented with ¹⁵NH₄Cl in the first group (T₁: 6 rabbits). The second group (T₂: 6 rabbits) was also fed the labelled diet but only during the last ten days of the fattening period when animals were fitted a neck collar to prevent caecotrophy. The third group (T₃: 4 animals) received the basal diet and was used as control. In two additional trials the daily contribution to urinary excretion of endogenous purine compounds (469 ± 50.8 μmol/W^0.75) and creatinine excretion (807 ± 127.6 μmol/W^0.75) were determined. The highest estimation of microbial nitrogen recycling was obtained by the urinary PD method (0.79 ± 0.096 g/d), whereas caecotrophes collection and ¹⁵N-lysine incorporation methods showed similar values (0.49 ± 0.049 and 0.45 ± 0.015 g/d, respectively). Our results seem to indicate an overestimation of microbial nitrogen recycling in growing rabbits by PD methodology, while neck collar fitting procedure gave similar results, although more variable than microbial ¹⁵N-lysine incorporation.     

Ruminal ammonia load affects leucine utilization by growing steers

Six ruminally cannulated Holstein steers (initial BW = 189 +/- 11 kg) housed in metabolism crates were used in a 6 x 6 Latin square to study effects of ruminal ammonia load on Leu utilization. All steers received a diet based on soybean hulls (2.7 kg of DM/d), ruminal infusions of 200 g of acetate/d, 200 g of propionate/d, and 50 g of butyrate/d, as well as an abomasal infusion of 300 g of glucose/d to provide energy without increasing microbial protein supply and an abomasal infusion of a mixture (238 g/d) of all essential AA except Leu. Treatments were arranged as a 3 x 2 factorial and included Leu (0, 4, or 8 g/d) infused abomasally and urea (0 or 80 g/d) infused ruminally. Abomasal Leu infusion linearly decreased (P < 0.05) both urinary and fecal N excretions and linearly increased (P < 0.05) retained N, but the decreases in urinary N excretion in response to Leu tended (P = 0.07) to be greater, and the increases in retained N in response to Leu were numerically greater in the presence of the urea infusion. Although urea infusions increased (P < 0.05) plasma urea concentrations, urinary N excretions, and urinary urea excretions, retained N also was increased (P < 0.05). The efficiency of deposition of supplemental Leu ranged from 24 to 43% when steers received 0 or 80 g of urea/d, respectively. Under our experimental conditions, increasing ammonia load improved whole-body protein deposition in growing steers when Leu supply was limiting.     

Nitrogen efficiency of dairy cows as affected by diet and milk yield

The aim of this study, which was part of the EU-financed project Life Ammonia, was to evaluate the effects of dietary components and milk production on nitrogen efficiency of dairy cows. The study included examining the effects of decreased crude protein (CP) concentration in a grass-clover silage based diet and results of mixing whole-crop barley silage (WCBS) with grass-clover silage in the diet, on feed intake, milk production and nitrogen efficiency. Rations were formulated and milk production data were registered individually each month for 42 cows of the Swedish Red Cattle breed during four indoor periods from 1999 to 2003. The range in nitrogen efficiency by the cows, 11 to 398 days in milk, was 18 to 40%, when fed a diet containing 135 to 184 g CP/kg DM, 44 to 56% of NDF as rumen degradable fibre (RDF) and milking 13 to 57 kg of ECM daily. The average CP concentration of the diet, containing mainly grass-clover silage and concentrate, was decreased from 168 g/kg DM (170 g in early lactation) in the control treatment period to 160 g/kg DM (163 g in early lactation) during the following treatment period. The CP concentration was 170 g/kg DM (171 g in early lactation) during the third treatment period, when the grass-clover silage was fed in a mixture with WCBS. Using the whole data set (n = 284 for primiparous, n = 440 for multiparous cows based on measurements each month) resulted in models, in which total DM intake, ECM yield, dietary CP concentration and RDF were the most important factors affecting nitrogen utilisation of primiparous and multiparous cows. Increases in both average DM intake and milk yield by multiparous cows and no changes in average intake and milk yield by primiparous cows fed the low CP diet or the normal CP diet containing WCBS, compared to cows fed the normal CP diet, resulted in similar nitrogen efficiencies among the treatments. Hence, dietary CP concentrations of 160 to 170 g/kg DM can be used for cows in early lactation in commercial herds to improve nitrogen utilisation without causing a simultaneous decrease in milk yield.     

Influence of sow dietary fatty acid composition on the behaviour of the piglets

Polyunsaturated fatty acids (PUFA) are essential for the development of the nervous system in animals, and increased concentration of n−3 PUFA in maternal diet improves the cognitive development of mammalian foetuses. In this study the effect of maternal diet fatty acid composition in pigs on the development of the central nervous system, monitored as behaviour of piglets, was investigated using three behavioural tests: recognition of the mother's faeces, back test, and hidden door test.Twenty-seven multiparous Yorkshire sows were split into four groups and fed diets with different content of fat and PUFA throughout pregnancy and lactation. LF (n = 6) was fed a standard diet, HFS (n = 4) a high fat and low PUFA diet, HFO (n = 7) a high n−6 PUFA diet, and HFL (n = 10) a high n−3 PUFA diet.Three behavioural tests were performed on 5–7 randomly chosen piglets per litter (n = 167). Recognition of the mother's faeces was tested in a maze two days after birth. Back test was performed twice (2–4 d and 4 w) and a hidden door test was performed at 4 w. In addition, the brain content of docosahexaenoic acid (22:6 n−3, DHA) of the newborn piglets was determined in treatment groups. Data from the tests were analysed with linear mixed models for each of the tests.Piglets from HFL treatment had significantly higher content of DHA (P < 0.001) and the ratio of n−6/n−3 PUFA was significantly lower in brain tissue (P < 0.001), compared to piglets from the other treatments. In parity 3, means for recognition for mother's faeces were for diets LF, HFS, HFO and HFL; 22.2, 37.0, 26.4 and 18.0%, respectively (P < 0.05), but no other significant effect of diet was found. Piglets in HFS treatment had the shortest latency to make escape attempts and HFO piglets the longest latency in the back test (P = 0.030). No significant effect of sow diet was found on piglet performance in the hidden door test, but intermediate piglets weighing 1410–1619 g had a lower probability of success in hidden door test than piglets weighing < 1410 g (P = 0.028), and ≥ 1875 g (P = 0.027), respectively.It was found that sow diet influenced the DHA content in the piglet brains, but there was no clear effect of sow diet on piglet behaviour. In order to draw any conclusions about possible enhancements of the behavioural development of the piglet more studies need to be performed.     

Proof-of-concept study: Evaluation of plasma and urinary electrolytes as markers of response to L-asparaginase therapy in dogs with high-grade lymphoma

Response to chemotherapy is one of the most important prognostic factors in dogs with lymphoma. The objective of this feasibility study was to evaluate if clinical responses to a specific cytotoxic agent (L-asparaginase) could be anticipated by measuring analyte concentrations in plasma and urine concentrations of lymphoma-bearing dogs. We hypothesized that potassium and phosphate concentrations in plasma and urine would be higher in dogs that completely responded to therapy. Plasma and urine samples of dogs with lymphoma were obtained before 12 and 24 hours after intramuscular L-asparaginase injections. Peripheral lymph node volumes were evaluated according to the Veterinary Cooperative Oncology Group standardized criteria. Plasma and urine electrolyte, calcium, phosphate, creatinine, urea, total protein, and albumin concentrations were measured, and the fractional excretions of each electrolyte were calculated. Statistical analyses compared complete vs partial responders using a linear regression model. Contrast analyses were also performed to differentiate the mean of each group, with adjustments made with the Benjamini-Hochberg procedure. Fourteen dogs were included, eight with complete responses, and six with partial responses. Plasma phosphate concentrations were significantly higher at 12 hours (P = .0003) and 24 hours (P = .009) after complete responses to therapy. This study demonstrates the potential use of plasma and urine analyte monitoring after chemotherapy induction. Plasma phosphate measurements represent a potential indicator of early responses to L-asparaginase therapy. Larger population studies are warranted to confirm these preliminary results.     

Estimation of urine flow rate in mares

To determine the rate of urine flow and thus urinary excretion in the horse from untimed urine samples alone, the flow rate, creatinine concentration, osmolarity, and refractive index of 228 quantitatively collected urine samples were determined in 53 experiments on 12 healthy Thoroughbred mares. Forty samples were collected after water-induced diuresis; 11 samples were collected after furosemide-induced diuresis. Flow rates, which ranged from 1.2 to 84.5 ml/min, could be predicted from the urinary creatinine concentration. Correlation of urinary flow with urinary creatinine concentration accounted for 94% of the variability in the urinary flow rates. Phenylbutazone was administered before collection of 168 urine samples. Urine flow rates that were predicted from urinary creatinine concentration were used to estimate phenylbutazone excretion. Urine flow could be estimated without quantitative urine collection.