尿液嘌呤法估测瘤胃微生物蛋白研究

瘤胃微生物蛋白质（MCP）是反刍家畜的重要蛋白质来源。近年来以尿液嘌呤衍生物估测反刍动物瘤胃微生物蛋白产量的方法正逐步取代传统的标记法，其优点是不用瘘管，无损害，操作简单；缺点是其值为相对值，尚需进一步标准化。     

尿嘌呤衍生物法估测瘤胃微生物蛋白质产量

尿嘌呤衍生物法具有非侵入性和操作方便等优点,是估测微生物蛋白质产量的一种有效方法.该方法能够准确估测瘤胃微生物蛋白质产量的变化,且其残差变异低于其他方法,估测结果的一致性较好.对其改良后的方法有点采样法和菊糖法.点采样法优点在于可以确定恰当的采样时间,保证尿嘌呤衍生物与肌酐的比例(PD/C)与尿嘌呤衍生物日排出量的高相...     

用尿中嘌呤衍生物估测瘤胃微生物蛋白产量的研究进展

反刍动物尿中嘌呤衍生物包括尿囊素、尿酸、黄嘌呤和次黄嘌呤,尿中嘌呤衍生物是一种很好的估测瘤胃微生物蛋白合成量的标记物。本研究就动物内源嘌呤排泄、尿中嘌呤衍生物排泄量与嘌呤吸收量关系模型建立、全收尿法和点尿法计算微生物蛋白供给以及最近对不同动物(包括绵羊、黄牛、山羊、水牛、牦牛、骆驼、驼羊)嘌呤衍生物排泄的研究等方面,阐述用尿中嘌呤代谢物估测瘤胃微生物蛋白产量的方法。     

微生物氮测定方法比较

定量测定瘤胃微生物细胞净合成量对研究反刍动物营养需要至关重要。目前所采用的估测微生物蛋白合成量的方法主要是借助外科造瘘手术，利用瘤胃微生物自身固有的物质或通过外源性同位素标记物标记来估测微生物蛋白合成量。近年来，以尿液嘌呤衍生物作为标记物估测瘤胃微生物蛋白产量的非损害方法也得到较快发展。笔者认为内外源标记结合的方法最合适。     

瘤胃微生物蛋白测定新进展—尿液嘌呤法

估测瘤胃微生物蛋白质(MCP)一直是反刍动物营养研究中的重要组成部分,其方法虽多但各有优缺点。近年来尿液嘌呤法以其无需瘘管、无损害、易操作的特点正逐步取代传统的标记法,许多研究者提出了不同的估算公式,适用于牛和羊的有所不同。本文就其原理、计算方法、局限性及发展前景作一综述。     

瘤胃微生物蛋白质合成量估测标记物的研究

确定瘤胃微生物细胞净合成量对研究反刍动物营养需要至关重要。目前所采用的估测微生物蛋白合成量的方法主要是借助外科造瘘手术，利用瘤胃微生物自身固有的物质或通过外源性同位素标记物标记来估测微生物蛋白合成量。近年来，以尿嘌呤衍生物作为标记物估测流入十二指肠的瘤胃微生物蛋白产量的非损害方法也得到较快发展。笔者认为内外源标记结合的方法最合适。     

尿液嘌呤衍生物法估测瘤胃微生物蛋白产量的研究进展

一种非伤害性方法-尿液嘌呤衍生物(PD)作为标记物估测瘤胃微生物蛋白产量近年来得到较快发展.本文就这种方法的原理依据,发展及应用进行了综述,分析了这种方法存在的一些问题,展望了将来的研究和发展方向.     

尿液嘌呤衍生物法估测瘤胃微生物蛋白产量的研究进展

一种非伤害性方法-尿液嘌呤衍生物（PD）作为标记物估测瘤胃微生物蛋白产量近年来得到较快发展。本文就这种方法的原理依据，发展及应用进行了综述，分析了这种方法存在的一些问题，展望了将来的研究和发展方向。     

用嘌呤衍生物估测牛关小肠消化吸收的微生物蛋白产量（综述）

综述了一种测定微生物蛋白（ＭＣＰ）的简单方法－－嘌呤衍生物法。其测值可用作估测是白的ＭＣＰ部分，对建立和完善反刍动物蛋白质新体系具有重要意义。     

用嘌呤衍生物估测牛羊小肠消化吸收的微生物蛋白产量(综述)

综述了一种测定微生物蛋白（ＭＣＰ）的简单方法——嘌呤衍生物法。其测值可用作估测代谢蛋白的ＭＣＰ部分，对建立和完善反刍动物蛋白质新体系具有重要意义。     

Urinary purine derivates excretion as an indicator of in vivo microbial N flow in cattle: A review

Microbial protein flow to the duodenum may be regarded as the most important and sensitive indicator to optimise rumen metabolism in high-yielding dairy cows. In this review, the methodology and the sources of variation to estimate the duodenal microbial N flow with urinary excretion of purine derivatives (PD) as a non-invasive method is discussed. The urinary PD excretion was linearly related with the amount of purine bases (PB) infused in the abomasum or duodenum, but the recovery of PB in urine differed between experiments. The main sources of variation in the relationship between microbial N flow and urinary PD excretion are dietary contribution of nucleic acids to duodenal flow, varying N:purine ratio in duodenal digesta, differences in intestinal digestibility of nucleic acids and infused PB, and endogenous contribution of PD to urinary excretion. The recycling of PD to the rumen is negligible, and does not explain the incomplete urinary recovery of PD. A large proportion of the total PD is excreted as allantoin in urine. In some experiments this proportion was constant, whereas in others it varied with diet or physiological state of the animal. The excretion of PD in milk is not a suitable indicator of microbial N flow, due to mammary purine catabolism to uric acid and due to the strong positive correlation between milk allantoin excretion and milk yield. Instead of total urine collection, the molar ratio between urinary PD and creatinine can be used to estimate microbial N flow. However, a substantial between-animal variation in this ratio was found, and effects of changes in energy balance of dairy cows on urinary creatinine excretion should be determined. The urinary excretion of total PD and of allantoin provided lower estimates of duodenal microbial N flow than with measurements in the omasum or duodenum, but they closely reflected the changes observed with these measurements.     

Effects of protein concentration and degradability on performance, ruminal fermentation, and nitrogen metabolism in rapidly growing heifers fed high-concentrate diets from 100 to 230 kg body weight

Twenty crossbred heifers (101 +/- 4.5 kg BW) were used to examine the effects of protein concentration and degradability on performance, ruminal fermentation, nutrient digestion, N balance, and urinary excretion of purine derivatives. Heifers were offered concentrate and barley straw for ad libitum consumption. Two protein concentrations (17 vs 14%, DM basis) and two protein sources differing in ruminal degradability (58 vs 42% of CP for soybean meal and treated soybean meal, respectively) were tested. The experiment was divided into four consecutive 28-d periods to evaluate the age (period) effect. Increasing protein concentration and degradability did not improve ADG or intake (P > .05). The increase in urinary N excretion (P < .001) in heifers fed 17% CP suggests that N was in excess of requirements. When the low-degradable protein source was supplemented and(or) CP concentration was low, ruminal NH3 N concentrations fell below 5 mg/100 mL. Urinary excretion of purine derivatives was not affected (P > .05) by protein concentration and degradability, suggesting that in high-concentrate diets NH3 N concentration was not limiting microbial growth. Total VFA concentration decreased (P < .001) and the acetate:propionate ratio increased (P < .01) with advancing period, suggesting an increase in ruminal absorption capacity and an increase in fiber fermentation. The decrease in ruminal NH3 N concentration in the last period suggests a greater use of NH3 N by microorganisms. This hypothesis is supported by the increase (P < .001) in urinary excretion of allantoin and estimated duodenal flows of purine bases and microbial protein with advancing period. Reducing CP concentration and increasing ruminal undegradable protein supply did not affect animal performance or estimated duodenal flow of microbial protein in rapidly growing heifers fed high-concentrate diets.     

Effect of N fertilisation rate, energy supplementation and supplementation strategy on efficiency of N utilisation in the sheep rumen.

The effect of nitrogen (N) fertilisation (200 vs. 400 kg N ha-1 year-1) of pasture cut in the beginning (end of May) and the end (end of August) of the grazing season and of simultaneous or separated feeding of maize and grass (400 kg N ha-1 year-1) on efficiency of N utilisation in the rumen has been studied using four rumen cannulated wethers. Doubling N fertiliser rate increased grass CP production by about 60%, but induced extensive excess of rumen degradable N, reflected in high urinary urea excretion (rpearson = 0.747). The latter was lower (74% at the maximum) when feeding less fertilised or older grass or when supplementing with maize silage. Although simultaneous feeding of maize silage with grass changed patterns of rumen ammonia concentrations, no change in urinary excretion of purine derivatives was observed. Hence, faecal or urinary N excretion was unaffected by the supplementation strategy. Microbial growth efficiency was estimated from urinary excretion of purine derivatives and fermented OM. The latter was calculated from total rumen CH4 production, based on rumen fermentation stoichiometry and taking into account proportional concentrations of individual volatile fatty acids. Higher levels of intake tended to improve rumen microbial growth efficiency slightly (rpearson = 0.406), which, however, could not compensate for the reduced effective rumen DM degradability (rpearson = -0.442). The latter was apparently associated with a partial shift of the fermentation from the rumen to the hindgut, as suggested from the negative correlation (rpearson = -0.745) between faecal RNA concentrations and rumen effective degradability.     

Urinary excretion of purine derivatives and plasma allantoin level in sheep and goats during fasting

The relationship between blood plasma level and urinary excretion of allantoin (AN) was examined in sheep and goats during fasting to investigate the possible use of purine derivatives (PD) in urine and/or plasma for estimating the microbial protein production in the rumen, and the further digestion in the lower guts of ruminants. Urinary AN excretion decreased markedly during fasting (0.13 mmol/kgW^0.75 per day), although urinary levels of other PD, hypoxanthine + xanthine and uric acid did not differ irrespective of the feeding condition, that is, feeding, fasting and refeeding in both species. The AN concentration in blood plasma also decreased drastically in the starvation period, and was suddenly increased on refeeding in sheep and goats, and these phenomena were very similar to those of urinary AN excretion. Therefore, there was a high positive correlation between plasma AN level and urinary AN excretion, and the coefficient of correlation was statistically significant (P < 0.01). These results clearly indicate that changes in urinary AN reflect change in plasma AN, which is induced by the catabolism of purine base in the body.     

饲粮不同采食水平下肉羊氮沉积和尿中嘌呤衍生物排出规律的研究

本试验旨在研究饲粮不同采食水平下肉羊氮沉积和尿中嘌呤衍生物(PD)的排出规律.试验选用12只平均体重为(41.3±2.8)kg的杜寒杂交绵羊公羔,随机平均分为3组,按照自由采食、自由采食量的70％和自由采食量的40％3个干物质采食水平饲喂,试验期为12d,其中预试期7d,正试期5d.结果表明:随着饲粮采食水平的降低,营养物质消化率均显著上升(P＜0.05),粪氮和尿氮排出量均显著降低(P＜0.05),沉积氮与摄入氮之间存在线性相关(R2=0.92).尿中PD排出量和微生物氮(MN)产量均随采食水平的降低而显著降低(P＜0.05),嘌呤氮指数(PNI)与氮沉积率和氮吸收率均表现出相同变化趋势.由此可见,PD排出量与可消化有机物采食量(DOMI)以及MN产量之间均存在线性相关,相关方程分别为y=21.41X-1.81(R2=0.94)和Y=0.91X+2.57(R2=0.94);PNI能够将氮平衡和PD排出量有机结合起来,用于评价饲粮氮的利用效率.     

Metabolism of microbial nitrogen in ruminants with special reference to nucleic acids

Characteristically the metabolism of microbial nitrogen (N) compounds in ruminants involves the degradation of dietary N and synthesis of microbial protein (MP), compounds including a small amount of peptides and free amino acids, which may account for 75–85% of total N and the remainder are nucleic acids (NA: DNA and RNA). Generally rumen microbes contain 10–25% NA‐N of the total N while 70–80% is in the form of RNA. This paper describes the degradation and synthesis of NA in the rumen and their fate in the lower digestive tracts. Their physiological and nutritional significance in different types of ruminant animals is also discussed. The research works on NA metabolism in ruminants has been mainly on metabolism of purines after rumen microbial digestion and absorption in the lower gut. Subsequently, the fate of absorbed purines has been intensively investigated to assess the extent of MP synthesis in the rumen. The method for predicting ruminal synthesized MP and subsequently digested MP has been proposed using urinary purine derivative (PD) excretion in sheep and cattle fed on ordinary feed. The latter approach has now been adopted for calculation of protein supply in some feeding standards, although there are still difficulties in predicting representative samples of rumen microbes, and also uncertainties in variations of non‐renal and endogenous purine losses.     

Effect of corn supplementation on purine derivatives and rumen fermentation in sheep fed PKC and urea-treated rice straw

This study investigated the effect of different levels of corn supplementation as energy source into palm kernel cake–urea-treated rice straw basal diet on urinary excretion of purine derivatives, nitrogen utilization, rumen fermentation, and rumen microorganism populations. Twenty-seven Dorper lambs were randomly assigned to three treatment groups and kept in individual pens for a 120-day period. The animals were subjected to the dietary treatments as follows: T1: 75.3% PKC?+?0% corn, T2: 70.3% PKC?+?5% corn, and T3: 65.3% PKC?+?10% corn. Hypoxanthine and uric acid excretion level were recorded similarly in lambs supplemented with corn. The microbial N yield and butyrate level was higher in corn-supplemented group, but fecal N excretion, T3 has the lowest level than other groups. Lambs fed T3 had a greater rumen protozoa population while the number of R. flavefaciens was recorded highest in T2. No significant differences were observed for total bacteria, F. succinogenes, R. albus, and methanogen population among all treatment. Based on these results, T3 could be fed to lambs without deleterious effect on the VFA and N balance.