Methodologic studies on the metabolically-oriented determination of lysine requirements in broiler chickens. 2. Effect of the lysine content of the diet on the lysine oxidation rate in a 15-hour feed withdrawal before 14C-lysine injection

This experiment was designed to narrow the estimated range for lysine requirement of broiler chickens determined by isotopic techniques. In addition the influence of a long-term feed withdrawal previous 14C-lysine-injection on the lysine catabolism was investigated. 120 male broiler chickens 7 to 21 days posthatching received a diet based on wheat and wheat gluten. Lysine content was varied from 8.3 to 16.0 g/kg DM (3.2 to 6.3 g/16 g N) at 8 levels by supplementing the basal diet with L-lysine-HCl. After the feeding period animals of each group were labelled with 14C-L-lysine by intravenous injection 5.5 and 15.5 hours after feed withdrawal, respectively. During the following 4 hours the excretion of 14CO2 and CO2 was measured. Highest body weight gain was observed in the group with 13.8 g lysine/kg DM. In case of 14CO2 excretion measurements starting 5.5 hours after feed withdrawal an increase of 14CO2 excretion was observed if the lysine content of the diet exceeded 11.6 g/kg DM. This estimated range for lysine requirement (11.6 to 12.7 g/kg DM with 26% CP in the DM) was lower compared with the lysine requirement estimated by the growth curve (12.7 to 13.8 g/kg DM). This discrepancy could be explained by the fact that the results of the metabolism oriented determination of lysine requirement represent the requirement at the actual age, while the feeding experiment reflects a mean lysine requirement of the previous period of 14 days. If the animals were labelled with 14C-lysine 15.5 hours after feed withdrawal no clear response in 14CO2 excretion and specific radioactivity of CO2 on the dietary lysine content was observed.     

Influence of dietary lysine concentration on the oxidation of an indicator amino acid by growing boars

The influence of dietary lysine concentration on the oxidation of 14C-phenylalanine by growing boars was determined. Forty-five crossbred boars (30 to 40 kg) were fed a ground corn diet fortified with crystalline L-lysine to provide .28, .50, .85, 1.00, 1.25 and 1.54% total lysine. All other essential amino acids were supplemented to provide 135% of NRC (1979) recommendations. Release of 14CO2 from L-[1-14C]-phenylalanine was measured for 1 h following a meal of the experimental diet, which contained 20 mu Ci 14C-phenylalanine. Increasing dietary lysine concentration from .28 to .85% decreased 14CO2 production. Regression analysis of the data using a two-phase linear regression crossover model indicated that phenylalanine oxidation was minimized at a dietary lysine concentration of .65%. It was concluded that a concentration of .65% lysine minimized the oxidation of amino acids and provided them as possible substrates for protein deposition. The oxidation of an indicator amino acid can, therefore, be used to determine the effect of dietary lysine concentration on the partition of amino acids between metabolic fuels and body protein.     

Effect of glutamic acid content in the diet on the catabolic rate of the isotope-labeled glutamic acid in rats. 4. Measurement of 14C-glutamic acid oxidation to 14CO2 with various energy sources in the diet

48 male rats (body weight 80-100 g) were fed with 2 diets different in the glutamic acid content (diet I 2.42 and diet II 6.24% glutamic acid in the dry matter). The mixture of the other synthetic L-amino acids was adapted to the egg protein pattern corresponding 10% crude protein in the diet. Each diet was fed either on 73% or 98 to 104% of the energy maintenance requirement. After 7 days of experimental feeding 14C-U-L-glutamic acid was given to each group by intragastric infusion (i.g.), intraperitoneal (i.p.) or subcutaneous injection (s.c.), respectively, followed by a measurement of the CO2-and 14CO2-excretion during two subsequent periods of 3 hours. The CO2-excretion was lower in animals with restricted energy intake especially during the first 3 hour-period, which was started 2 hours after feed intake. The relative 14CO2-excretion (percent of the dose) was neither significantly influenced by the level of energy intake nor by the amount of dietary glutamic acid. The highest degradation rates of 14C-glutamic acid to 14CO2 were measured after i.g. application (more than 50%), followed by the i.p. injection (nearly 50%) and the lowest values were observed after s.c. injection (about 40%). These differences were only evident during the first CO2-absorption period. Furthermore the s.c. injection caused a lower specific radioactivity of CO2 compared with the data after i.g. and i.p. application. The results suggest the high metabolic activity of the intestinal tissue for glutamic acid.     

Determination of lysine requirement in growing rats as based on the catabolism rate of 14-C- and 15-N-labeled lysine

Male Wistar rats (weighing some 80 g at the start of the experiment) were fed diets containing maize gluten as protein carrier and which was supplemented with amino acids (except lysine) in such way that their concentrations came up to the requirement norms. Lysine was gradually supplemented this resulting in 10 diets of different lysine content (1.6-10.6 g lysine/16 g N). On the 7th experimental day, 4 animals of each group were labelled with 14C-lysine and subjected to 2-hour measuring of 14CO2-excretion. On the following day, the animals were injected i.p. 15N-lysine, the urine being collected over 24 hours to determine 15N-frequency in urine. Both 14CO2-excretion and 15N-frequency in urine were found to remain constant at a lysine content of the diet up to 4.5 g/16 g N and rose steeply from 5.8 g lysine/16 N on. Under the experimental conditions chosen the lysine requirement is deduced to be 5 g/16 g N. This method of lysine requirement determination is highly sensitive and exact because it covers the catabolization of the amino acids under study and not so parameters that are known to be influenced by other factors such as growth, N-balance, total N-conversion or CO2-formation. The method can also be applied to metabolic situations not connected with productive performances.     

生长猪回肠氨基酸消化率的标准化测定方法

根据荷兰CVB(1996)和德国GfE(2002、2005)的最新规定,提出了生长猪回肠氨基酸消化率测定的一个标准化方法。本文从①试验动物、猪舍环境和饲喂条件,②消化糜的收集及处理,③回肠氨基酸消化率测定方法三个方面比较了这些规定之间的差异和相似性。     

Standardization of methods for the determination of ileal amino acid digestibilities in growing pigs

Based on current recommendations of the CVB (1996) in the Netherlands and the GfE (2002, 2005) in Germany, a standardization of methods for the determination of ileal amino acid digestibilities in growing pigs is proposed. Differences and similarities between these protocols in terms of (i) animals, housing and feeding conditions (ii), methods for digesta collection and handling, and (iii) approaches to determine ileal amino acid digestibilities are described.     

Estimate of the variability of the lysine requirement of growing pigs using the indicator amino acid oxidation technique

Although AA requirements for the mean in a population of growing pigs are well established, there are no direct estimates of their variability within the population. The indicator AA oxidation method allows repeated measurements in a short period of time so that the AA requirement can be determined for individual pigs. The objective was to determine the Lys requirement in individual pigs to derive a first estimate of the population mean requirement and its variability. Nine individually housed barrows (15 to 18 kg) were surgically implanted with venous catheters for isotope infusion. Pigs were offered, in random order, isonitrogenous and isoenergetic diets with one of seven Lys concentrations (4.8 to 15.5 g of Lys/kg diet, as-fed basis). The pigs were fed twice daily, except for study days when they received one-half of the daily allowance in eight equal hourly meals. After a validated minimum adaptation period, indicator (Phe) oxidation was determined for each dietary Lys level during a 4-h primed, constant infusion of L-[1-(14C)]Phe at a rate of 464 kBq/h. The Lys requirement was calculated using a two-phase linear regression crossover analysis within individual pigs. For each pig, Phe oxidation decreased linearly (P < 0.02) as the dietary Lys concentration increased until the requirement was reached; thereafter, Phe oxidation was not different. The true ileal digestible Lys requirement ranged from 7.5 to 10.6 g/kg of diet (as-fed basis) for the nine animals. The mean requirement for all pigs was 9.1 g/d (CV, 11.6%) or 93.9% (CV, 9.8%) of the predicted (NRC, 1998) requirement based on each pig's mean BW and energy intake. The measured and predicted requirements did not differ. The indicator AA oxidation method gave values for Lys requirement similar to conventional methods. The short (< 3 wk) experimental period allows, for the first time, the estimate of population variability, which provides for more accurate calculation of the effect of altering Lys intake on herd performance and production economics. This method is suitable to use with all dietary indispensable AA.     

Studies on the 14C-15N-acetamide turnover in sheep. 1. Studies on the 14C turnover]

4 male sheep (average weight: 53.5 kgs) were fed a semisynthetic diet containing acetamide as sole source of nitrogen. At the beginning of the trial twin-labelled 15N-14C-acetamide was administered by way of a ruminal fistula. The curve pattern of 14C activity in the TCE-soluble fraction of the ruminal fluid showed a synchronous behaviour in all animals beginning at 3 hours after the beginning of the trial. A half-life of 2 1/2 hours for the rate of absorption of 14C acetamide and deaminated 14C acetate was established from the decline in 14C activity observed in the TCE-soluble fraction of the ruminal fluid. The peak level of 14C labelling in ruminal proteins was reached after 6 hrs. The specific 14CO2 activity in respiratory air reached its maximum level after 4 hrs, and was then found to decline continuously. 56% of the administered amount of 14C was excreted over a period of up to 50 hrs after beginning of the trial. The very fact that the peak level of 14C activity was observed to appear in the TCE-soluble fraction of the blood plasma as early as after 1 hr seems to indicate that acetamide is also absorbed through the ruminal wall. The half-life of decline in the 14C activity of this fraction was 5.7 hrs. Analysis by thin layer chromatography showed that 75% of this amount of activity is present in 14C acetamide. The rate of 14C incorporation into blood plasma proteins reached a plateau region after 21 hrs, which was also maintained on the 2nd day of the experiment. 6.5% of the administered amount of 14C activity was excreted in the urine until the 7th day of experiment. 76.6% of the amount of urinary 14C activity excreted within a period of 48 hrs were voided as 14C acetamide. 3.8% of the administered amount of 14C activity was excreted with the faeces within the first 6 days of experiment.     

Using the indicator amino acid oxidation technique to study threonine requirements in horses receiving a predominantly forage diet

Threonine has been reported to be the second limiting amino acid in typical equine diets, but its actual requirement has not been determined in horses. To evaluate amino acid metabolism and requirements, the indicator amino acid oxidation (IAAO) method has been successfully used in other species. The objective of this research was to estimate threonine requirements in mature horses fed timothy hay and concentrate in 4:1 ratio using the IAAO method. Six Thoroughbred mares (579.9 ± 46.7 kg) received each of 6 levels of threonine intake, 41, 51, 61, 70, 80 and 89 mg/kg BW/day, in a randomly determined order. Each study period was 7‐day long, and on day 6, blood samples were collected before and 90 min after feeding to measure amino acid concentrations using HPLC. On day 7, horses underwent IAAO procedures, which included a 2‐hr primed, constant intravenous infusion of [¹³C]sodium bicarbonate to measure total CO₂ production and a 4‐hr primed, constant oral administration of [1‐¹³C]phenylalanine to estimate phenylalanine oxidation to CO₂. Blood and breath samples were collected to measure blood [¹³C]phenylalanine, using GC‐MS analysis and breath ¹³CO₂ enrichment, using an infrared isotope analyser. Increasing threonine intake levels did not affect plasma phenylalanine oxidation by the ANOVA test (p > 0.05) but resulted in a linear decrease in phenylalanine oxidation (p = 0.04) without a breakpoint by the orthogonal linear contrast. This study is the first attempt to evaluate threonine requirements in horses by the IAAO method; however, threonine requirements are still unknown in mature horses at this time.     

Response to lysine in a wheat gluten diet in adult minipigs after short-and long-term dietary adaptation as assessed with an indicator amino acid oxidation and balance technique

An experiment was conducted to examine the response to wheat gluten (WG)-based diets at two lysine levels in adult minipigs (23 kg BW) using the indicator AA oxidation (IAAO) approach and N balance. Twenty minipigs (n = five per group), fitted with reentrant ileoileal cannulas allowing collection of ileal effluents, were fed restrictively two WG-based diets (WG and WG + Lys; 2.7 and 6.6 g of lysine/kg, respectively) for adaptation periods of 10 and 100 d. On d 7 and 9, for pigs fed the diets for 10 d, and on d 97 and 99, for pigs fed the diets for 100 d, primed i.v. fasted/fed tracer protocols with [(13)C]bicarbonate, and [(13)C]leucine were performed. With the WG diet, [(13)C]bicarbonate recoveries (%) were lower irrespective of the adaptation period, and higher during the fed period (fasted: WG + Lys = 82.5, and WG = 69.1; fed: WG + Lys = 90.6, and WG = 85.9; P < 0.05). Leucine oxidation rate was higher with the lower lysine intake (WG = 194.6 vs. 109.5 mg/[kg BW x d]; P < 0.05). Wheat gluten feeding resulted in a negative leucine balance independent of the adaptation period (WG = -29.1, and WG + Lys = 48.2 mg/[kg BW x d]; P < 0.05). In contrast with the IAAO method, N balance did not differ between the two lysine intakes, possibly because of an underestimation of N losses. The finding of a lower (13)C bicarbonate recovery with the lower dietary lysine intake suggests that caution should be taken in using a single recovery factor for all AA oxidation studies.     

测定哺乳母猪总含硫氨基酸相对于赖氨酸的需要量

本试验通过163头母猪,测定母猪在哺乳期中相对于赖氨酸的总含硫氨基酸(total sulfur amino acids,TSAA)需要量。所有试验日粮均以玉米-大豆粕型日粮为基础,通过配制使日粮的回肠赖氨酸真消化率达0.88%(总赖氨酸含量为0.97%)。试验日粮含0.37%L-赖氨酸盐酸,通过添加其它晶体氨基酸以使TSAA成为日粮第一限制性氨基酸。经过配制,日粮回肠可消化含硫氨基酸分别为0.44%、0.48%、0.53%、0.57%和0.62%,并使日粮赖氨酸含量分别达到50%、55%、60%、65%和70%。试验母猪分成6个产仔组,随后根据胎次将其随机分入各个日粮试验组,喂给总含硫氨基酸/赖氨酸比率不同但逐步提高的试验日粮。结果在整个哺乳期中,各组试验母猪在日平均采食量(ADFI)、失重和背膘损失及血浆尿素氮含量上并无显著差异(P>0.14)。同时,提高母猪日粮的总含硫氨基酸/赖氨酸比率,仔猪断奶体重或断奶前死亡率均未受到影响(P>0.25)。总之,逐步提高母猪日粮的总含硫氨基酸/赖氨酸比率,不会影响其仔猪的窝生长性能。这些结果表明,哺乳期母猪对回肠真可消化总含硫氨基酸的需要量不超过赖氨酸50%。体组织的动用可以作为总含硫氨基酸一种可能的来源,从而可降低哺乳母猪对日粮总含硫氨基酸的需要量;高氨基酸摄入量,同时伴随着体组织的动用,由此产生的作用还需作进一步的研究。     

Studies on the 14C-15N-acetamide turnover in sheep. 2. Studies on 15N turnover and comparative studies on the 14C turnover]

Three fistula sheep with average weights of 52.2 kgs were given 37.9 g of 15N and 14C labelled acetamide (= 1.09 mg 15N' and 0,95 mCi14C) which were administered directly through the fistula. The half-life period of 15N retention in the ruminal fluid (TCE soluble portion) was found to be 4 hrs. 18 hrs after 15N administration increasing amounts of 15N were carried back to the rumen by way of the rumino-hepatic circulation. The 15N concentration in the blood (TCE soluble portion) rapidly increased up to a peak value and, from 3 hrs after isotope administration, the 15N concentration was found to decline continuously, with a slight discontinuation at about the 10th hr of experiment. The rate of 15N incorporation into the protein fraction (TEC soluble portion) of the blood was delayed by 4 hrs, relative to the rate of 15N incorporation into ruminal proteins. An average of 43.1% of the administered amount of 15N was excreted in the urine within 7 days. Up to the 4th day of experiment the half-life period of urinary 15N excretion was 19 hrs. An average of 15% of the administered total amount of 15N was excreted in the faeces. In this process, the peak values in both TCE fractions were observed to occur on the 2nd day of experiment. The proportion of isotope in the TCE soluble fraction was found to increase continuously compared with the total amount of the isotope excreted in the faeces. Isotope concentrations between 0.03 and 0.13 atom% of surplus 15N were found in organ and muscle tissues of a sheep that had been slaughtered 7 days after administration of the isotope. The results obtained are discussed on the basis of comparisons made with the analogous behaviour of 14C activity.     

Effect of glutamic acid content of the diet on the catabolic rate of isotope-labeled glutamic acid in rats. 2. Time course of 14CO2 excretion following subcutaneous administration of 14C-glutamic acid

40 rats with a body weight of 100 g received 7 semisynthetic diets with different contents of glutamic acid and one diet contained whole egg. A L-amino acid mixture corresponding to the pattern of egg protein was the protein source of the semisynthetic diets. Glutamic acid was supplemented successively from 0 to 58 mol-% of the total amino acid content. On the 8th day of experimental feeding the animals were labelled by subcutaneous injection of 14C-U-L-glutamic acid. Subsequently the CO2- and the 14CO2-excretion were measured for 24 hours. In this period 64 to 68% of the injected radioactivity were recovered as 14CO2. The curve pattern of 14CO2-excretion indicates two different processes of 14CO2-formation. One characterizing the direct degradation of glutamic acid to CO2 with a high rate constant and a second one with a lower rate constant characterizing the 14CO2-formation via metabolites of glutamic acid. 77% of the total 14CO2-excretion in 24 hours resulted from the direct oxidation of glutamic acid and 23% from the oxidation of intermediates. When 14CO2-formation was measured 10 to 24 hours after injection of 14C-glutamic acid a positive correlation to the content of glutamic acid in the diet was observed. The intestinal tissue contribute considerably to the catabolization of glutamic acid, however, there seems to exist an upper limit for this capacity.     

如何根据氨基酸配比平衡日粮营养

1氨基酸配比与猪日粮营养的关系 动物对饲料中蛋白质的吸收程度取决于饲料中必需氨基酸赖氨酸、蛋氨酸、苏氨酸和色氨酸的配比．因为这些氨基酸在猪体内不能合成。如果动物日粮中缺乏上述任何一种营养,都会影响新陈代谢．并降低动物的生产性能．     

家禽氨基酸消化率的测定方法

家禽日粮中氨基酸消化率的测定方法包括:体外法(化学法、酶法、微生物发酵法)、体内法(生长或消化率试验)。体外法似乎是提供某些蛋白源的热破坏信息的唯一有效方法,而体外酶法(IDEATM)和体内法都可以用来测定蛋白质的消化率。生长试验是唯一能够直接确定氨基酸生物利用率的手段,但由于其费时而且昂贵,所以实际的应用也极为有限。目前,测定氨基酸消化率普遍采用体内消化率试验法。尽管如此,在进行试验和计算(表观或真消化率)时还是有差异的,这些差异也使得氨基酸消化率的测定值有所变化。体内氨基酸消化率测定值会因是使用排泄物还是回肠末端消化物而不同,这是因为末段肠道(盲肠)微生物的发酵作用对排泄物中氨基酸消化率的影响,还会因为是采用表观还是真消化率(修正内源损失)计算而有所差异。如果考虑了内源性损失并计算了真消化率,那么如何计算分析内源性损失就成了主要问题,因为在怎样获取精确的内源性损失值上仍然存在争论。     

Effects of excess arginine with and without supplemental lysine on performance, plasma amino acid concentrations and nitrogen balance of young swine

Three experiments were conducted to determine the effects of excess arginine on performance, plasma amino acid levels and N balance of young pigs (initial weights 6.9, 7.0 and 10.3 kg, respectively). In a 28-d growth trial, various amounts of arginine (0 to 1.6%) were added to a conventional starter diet. Addition of arginine decreased (P less than .01) average daily feed intake (ADFI) and gain (ADG), but had no effect on feed efficiency (G/F). Plasma urea, arginine and ornithine concentrations were elevated (P less than .001) by the increasing dietary arginine levels at d 14 and 28. Plasma histidine levels were reduced (P less than .01) at d 28. Plasma lysine levels exhibited a cubic response (P less than .05) at d 14, but were not affected by excess arginine at d 28. In a second growth trial ADFI and ADG were decreased (P less than .05), but G/F was not affected by the addition of 1.6% dietary arginine. Lysine supplementation (0, .15 or .30%) increased performance in the absence of excess arginine, but the main effect of lysine was not significant for any performance criteria. As in the first experiment, plasma concentrations of urea, arginine and ornithine were increased (P less than .001) by the addition of arginine. Plasma histidine was not affected by either arginine or lysine. Plasma lysine levels were reduced (P less than .001) by dietary arginine and increased (P less than .001) by lysine. In a N balance experiment, addition of 1.6% dietary arginine increased N digestibility, but decreased apparent biological value. Nitrogen balance was not affected by added arginine. Lysine addition did not improve any of these three indices of N utilization. The inability of lysine supplementation to alleviate any of the adverse effects of excess arginine in young swine indicates that the reduced performance is caused by a generalized amino acid imbalance, and not by a specific interference with lysine utilization in the manner of a classical arginine-lysine antagonism.     

Studies on the protein and amino acid metabolism of laying hens using 15N labeled casein. 4. Fecal excretion of nitrogen and amino acids, digestibility of crude protein and absorption of basic amino acids]

Generally, the faeces of laying hens fed 15N casein rations were found to contain equal proportions of TCE-precipitable and TCE-soluble nitrogen. Considerable variations were observed to occur between the 64 samples investigated (27%-75%) and no explanation was found to account for this fact. The content of basic amino acids in faecal proteins was found to differ considerably from that of the proteins in the intestinal contents. A high lysine content was found after the feeding of wheat. The present trial substantiated this result, provided the casein contained a certain proportion of non-available lysine. The apparent and true digestibility of dietary N was 88% or 91%, that of 15N (2nd and 6th day of experiment) and 92%. During the feeding of labelled casein a higher level of N labelling was found in the TCE-soluble portion of the faeces, whereas on the 8th to 12th day a higher level of labelling was observed in the TCE-precipitable portion of the faeces. The peak of 15N excretion occurred on the 3rd day of experiment. When 15N administration terminated the atom% 15N in the faeces and in urine was found to decrease rapidly approximating the initial level of labelling asymptotically.     

Studies on the incorporation of 14 C-labelled amino acids into egg proteins

The incorporation of ¹⁴C‐labelled amino acids into egg white and yolk proteins has been studied. When the labelled amino acids were given intravenously as a hydrolysate of [U‐¹⁴C]‐protein from Chlorella, 10 per cent and 7 per cent of the ¹⁴G were recovered in the whites and yolks respectively of the first nine eggs laid. Differences in the specific activities of the conalbumin, the ovalbumins, ovomucoid, “postalbumins “ and lysozyme, isolated from the first active egg, were related to differences in amino acid composition. The specific activity of each amino acid prepared from the proteins was similar between different proteins, although within each protein specific activities of different amino acids varied widely. The proportionate rate of decrease of specific activity in the plasma of amino acids essential for egg production (except glycine) was constant or almost so, but it rapidly decreased for the non‐essential ones (except serine and proline). The specific activities of the amino acids, excepting aspartic and glutamic acids, in each protein were proportional to their mean specific activities in the plasma throughout the 24‐h period in which the egg white proteins were synthesised. It is concluded that these different proteins are synthesised from a shared amino acid pool, derived from plasma, at a rate which probably remains constant throughout the egg‐laying cycle and which is proportional to their content in the albumen.     

Studies on the true amino acid digestibility in pigs of various wheat varieties from the Soviet Union]

Studies were carried out with pigs for determining, with classical methods, the true digestibility of amino acids contained in 5 intensively grown wheat varieties imported from the Soviet Union and in one wheat variety grown in the GDR. Of all the varieties tested the variety Mironovskaya 808 exhibited the best characteristics. An extremely low true lysine digestibility was established for the variety Avrora (68%). If this variety is to be more extensively grown and used for feeding purposes further studies will have to be made to see whether this low lysine digestibility is really characteristic of this variety.