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Evaluated the levels of rain tree (Samanea saman) pod meal (RTPM) (0, 10, 15, 20 and 25%) replacing maize in the dry matter of the diet on intake of DM, CP and ME, creatinine and total purine derivatives excretion in urine and microbial protein synthesis in lambs. Twenty-five uncastrated Bergamasca lambs were used, with an initial body weight of 24±5kg and an average age of 120 days. The experimental design was completely randomized, with five treatments and five replications. The trial lasted 84 days and the 24h and spot urine collections were performed in the last day of the experiment. The intake was not affected (P>0.05), the urine volume and daily creatinine excretion were influenced (P<0.05) by RTPM replacing maize, observing increasing linear effect and cubic variation (P<0.05), respectively. The cubic and quadratic components were significant (P<0.05) for the excretion of total purine derivatives, absorbed purines, and microbial synthesis. The cubic variation, with peaks at 5% and 18% replacement for urinary excretion of creatinine and purine derivatives, indicates that the levels used of RTPM affected the renal activity of lambs. Substitution of maize by RTPM up to 10% enhances efficiency of rumen microbial crude protein synthesis.

Avaliaram-se os níveis de farinha de vagem de Samanea saman (0, 10, 15, 20 e 25%) em substituição ao milho na matéria seca da dieta sobre o consumo de MS, PB e EM, a excreção urinária de creatinina e de derivados de purina totais e a síntese de proteína microbiana em cordeiros. Foram utilizados 25 cordeiros Bergamácia, não castrados, peso corporal inicial de 24±5kg e idade média de 120 dias. O delineamento experimental foi inteiramente ao acaso, com cinco tratamentos e cinco repetições. O experimento durou 84 dias e as coletas de urina spot e de 24h foram realizadas no último dia do experimento. O consumo não foi afetado (P>0,05), o volume de urina e a excreção diária de creatinina foram influenciados (P<0,05) por S. Saman substituindo o milho, observando-se efeito linear crescente e variação cúbica (P<0,05), respectivamente. Os componentes cúbicos e quadráticos foram significativos (P<0,05) para a excreção de derivados de purina totais, purinas absorvidas e síntese microbiana. A variação cúbica, com picos em 5% e 18% de substituição para a excreção urinária de creatinina e de derivados de purina, indica que os níveis utilizados de S. saman afetaram a atividade renal de cordeiros. Substituição do milho por farelo de vagem de S. saman até 10% aumenta a eficiência de síntese de proteína bruta microbiana.

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In this work, Amberlite XAD-1180 resin is used for on-line surfactant-mediated pre-concentration of sildenafil as a prior step for its fluorescent detection. In order to activate the column for sildenafil pre-concentration, the cationic surfactant (hexadecyltrimethylammoniunm bromide, HTAB) is adsorbed onto the resin. In these conditions, sildenafil is retained by HTAB-resin and then it is eluted with ethanol and analyzed by spectrofluorimetry. Drug-surfactant association produces a considerable fluorescence enhancement, increasing considerably the sensitivity of detection. Therefore, sildenafil can be pre-concentrated and quantitatively determined, with a detection limit of 0.2 ng mL(-1). The proposed method was successfully applied to the analysis of bulk drug, human urine, tablets, and local herbal medicine. Validation processes were performed by recovering studies and statistical analysis with satisfactory results.

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Two experiments were conducted to assess the endogenous fraction of purine derivative (PD) excretion, urinary recovery, and intestinal digestibility of purines in Nellore heifers. For both experiments, 8 Nellore heifers fitted with ruminal and abomasal cannulas were allocated to two 4 × 4 Latin squares. The diets were based on corn silage and concentrate (60 and 40% DM basis, respectively); feces and urine samples were obtained by total collection, and abomasal DM flow was estimated using indigestible NDF as an internal marker. In Exp. I, 4 of the 8 heifers (BW 258 ± 20 kg) were also fitted with ileal cannula. The planned treatments were 4 different DMI: 1.2, 1.6, 2.0, and 2.4% of BW (DM basis). The endogenous losses and purine recovery as urinary PD were estimated using linear regression between daily urinary PD excretion (Y) and daily abomasal flow of purine bases (X), expressed in millimoles per kilogram of BW(0.75). In Exp. II, the same 8 Nellore heifers (BW of 296 ± 15 kg) were fed at 1.37% BW (DM basis). The treatments were the infusion of purines (RNA from torula yeast, type VI, Sigma) into the abomasum in increasing amounts (0, 33, 66, and 100 mmol/d). All statistical analyses were performed using the PROC MIXED procedure in SAS. In Exp. I, the DMI range was 1.16 to 1.84% of BW and did not affect (P > 0.05) the apparent RNA digestibility in the small intestine, which had a mean of 75.6%, and a true digestibility of 93.0%. The mean ratio of the N-RNA to the total-N in the ruminal bacteria was 0.137. The daily urinary PD excretion (Y, mmol/kg of BW(0.75)) was a function of RNA flow in the abomasum (X, mmol/kg of BW(0.75)): Y = 0.860X + 0.460, where 0.860 and 0.460 were the PD recovery of purines and the endogenous fraction (in mmol/kg of BW(0.75)), respectively. In Exp. II, the daily urinary PD excretion was a function of RNA flow in the abomasum: Y = 0.741X + 0.301, where 0.741 and 0.301 were the recovery of PD in urine of infused purines and the endogenous losses (in mmol/kg of BW(0.75)), respectively. In conclusion, our data suggest that in Nellore heifers the respective values of endogenous PD excretion (mmol/kg of BW(0.75)), urinary recovery of the purines absorbed in the abomasum, and true digestibility of RNA in the small intestine were 0.30, 0.80, and 0.93.

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Four experiments were performed to study the kinetics of purine metabolism and urinary excretion in Zebu crossbred cattle. Fasting excretion was established in Expt 1, using eighteen male Bos indicus x Bos taurus crossbred cattle (261 (SE 9.1) kg body weight), six of each of the following genotypes: 5/8 Bos indicus, 1/2 Bos indicus and 3/8 Bos indicus. No significant differences were observed among genotypes in fasting purine derivative excretion (277.3 (SE 35.43) micromol/metabolic body weight). In a second experiment we measured the xanthine oxidase activity, which was higher in liver than in duodenal mucosa (0.64 and 0.06 (SE 0.12) units/g wet tissue per min respectively; P>0.05) being in plasma 0.60 (SE 0.36) units/l per min. The kinetics of uric acid were measured by intravenous pulse dose of [1,3-15N]uric acid (Expt 3). The cumulative recovery of the isotope in urine was 82 (SE 6.69) %, and uric acid plasma removal, pool size and mean retention time were 0.284 (SE 0.051) per h, 5.45 (SE 0.823) mmol and 3.52 (SE 0.521) h, respectively. Allantoin was removed from plasma at an estimated fractional rate of 0.273 (SE 0.081) per h and mean retention was 3.66 (SE 1.08) h. In Expt 4, the relationship between urinary purine derivative excretion (Y; mmol/d) and digestible organic matter intake (X, kg/d) was defined by the equation: Y=7.69 (SE 4.2)+5.69 (SE 1.68) X; n 16, Se 1.31, r 0.67.

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Three multiparous, ruminally and duodenally cannulated Holstein-Friesian milking cows (558 +/- 14 kg BW) with a mean milk yield of 19.9 +/- 1.4 kg/d in their 4th mo of lactation were fed a mixed diet of forage and concentrate at 100, 85, and 75% of ad libitum intake in a 3 x 3 Latin square design. Duodenal digesta flow was estimated using the dual-phase technique in which Cr-EDTA and Yb-acetate were used as liquid and solid markers, respectively. Microbial N (MN) was estimated using the duodenal flow of purine bases (PB); bacterial isolates from the rumen liquid and solid phases were used as references. Additionally, duodenal flow of PB and MN were estimated indirectly using the excretion of purine derivatives (PD) in urine and milk. Duodenal flow of PB and derived MN tended to decrease with feed restriction (from 258 to 154 mmol/d and 123.5 to 74.4 g/d, respectively). Estimates of PB and MN based on urinary PD showed the same trend, and decreases in PB (from 314 to 266 mmol/d, using LAB) were statistically significant. Using LAB, efficiencies of microbial protein synthesis in the ad libitum treatment were 12.9 and 17.0 g of MN/g of organic matter apparently digested in the rumen when estimated using duodenal PB and urinary excretion of PD, respectively. Urinary excretion of PD closely reflected changes in duodenal flow of PB as a result of feed restriction.

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Twenty-four multiparous dairy cows (eight with ruminal cannulae) were blocked by days in milk and assigned to six balanced 4 x 4 Latin squares with 21-d periods. The four diets, formulated from alfalfa silage plus a concentrate mix based on ground high moisture ear corn, contained (dry matter basis): 1) 20% concentrate, 80% alfalfa silage (24% nonfiber carbohydrate; NFC), 2) 35% concentrate, 65% alfalfa silage (30% NFC), 3) 50% concentrate, 50% alfalfa silage (37% NFC), or 4) 65% concentrate, 35% alfalfa silage (43% NFC). Soybean meal and urea were added to make diets isonitrogenous with equal nonprotein nitrogen (NPN) (43% of total N). Total urine was collected with indwelling Folley catheters for 24 h during each period. There was no effect of diet on urinary creatinine excretion (average 29 mg/kg of BW/d). There were quadratic effects of diet on total urinary ecretion of allantoin, uric acid, and purine derivatives (allantoin plus uric acid), and on ruminal synthesis of microbial N estimated from purine derivatives; maxima occurred at about 35% dietary NFC. Urinary excretion also was estimated with spot urine samples from creatinine concentration and the mean daily creatinine excretion. Daily excretion of allantoin, uric acid, and purine derivatives estimated from spot urine sampling followed the same pattern as that observed with total collection; differences between measured and estimated urine volume were significant only for 35% dietary concentrate. Spot urine sampling appeared to yield satisfactory estimates of purine derivative excretion. Maximal urea N excretion was estimated to occur at about 31% dietary NFC. Milk allantoin secretion increased linearly with concentrate and accounted for 4 to 6% of the total purine derivative excretion. Microbial yield was maximal at 35% dietary NFC, suggesting that this was the optimal level for utilization of dietary NPN from alfalfa silage and other sources.

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La presente revisión tuvo como objetivo recopilar y presentar información actualizada sobre la técnica de los derivados de purina en rumiantes domésticos. La técnica de los derivados de purina representa una alternativa simple y no invasiva para el estudio del aporte de N microbial a nivel intestino delgado en animales rumiantes. Existe información y modelos cuantitativos de respuesta para bovinos y ovinos que permiten el uso de esta técnica, y numerosos métodos analíticos publicados para el análisis de los derivados de purina. El método de los derivados de purina tiene la ventaja de no requerir animales quirúrgicamente preparados, pero tiene la desventaja de requerir colecta total de orina. El uso de submuestras de orina colectada a lo largo del día(s) junto con el índice `derivados de purina:creatinina' puede ser una alternativa a la colección total de orina, pero se necesita más información sobre este punto. La medición de los derivados purínicos en leche no parece ser una alternativa viable a la colección total de orina. Deben de validarse algunos de los parámetros en uso para el cálculo del flujo de nitrógeno microbial al intestino como son; la recuperación de purinas microbiales absorbidas como derivados purínicos en orina y la digestibilidad de la purina en intestino delgado en animales bajo condiciones de alimentación normal. No obstante, comparada con otras téncicas convencionales, la medición de los derivados de purina en orina representa una herramienta simple para profundizar en el estudio y entendimiento de la dinámica ruminal

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