The passage of nitrogenous compounds through the wall of perfused sheep rumen (author's transl)]

Experiments were performed by the method of the extracorporeal perfusion of sheep rumen lasting 150 minutes. After 60 minutes of perfusion, 20 g of enzymatic casein hydrolyzate were applied to the rumen. Ammonia, urea, and total nitrogen were determined in the samples of perfusate, and ammonia and pH were determined in the rumen content. Considerable amounts of ammonia accumulated in the perfusate in the course of the experiment. At the end of perfusion, the concentration of NH3-N reached an average value of up to 12 mg per 100 ml. The level of urea in the perfusate decreased only in the first phase of perfusion (before the application of casein hydrolyzate to the rumen) and remained unchanged in the subsequent phase. It is assumed that part of urea-N from the perfusate after urea hydrolysis, taking place already in the rumen wall, returned to the perfusate in the form of ammonia. In the first phase of perfusion the passage of nitrogen from the blood into the rumen took place, in the second phase its absorption from the rumen into the blood was observed. The urea-N ratio from the passage of total nitrogen into the rumen ranged between 4.63% and 13.84%, but the concentration of total nitrogen in the perfusate decreased by 7-15% and that of urea nitrogen by 37-42%. It follows from the results that a major part of endogenous nitrogen passing from perfusate to the rumen was represented by proteins and/or their peptidic splits.     

Absorption of amino acids from the perfused ovine rumen]

The experiments with extracoroporeal perfusion of sheep rumen were performed [Leng et al., 1977]. Bovine plasma, diluted in a 1:1ratio with an isotonic solution of sodium chloride, was used for four perfusions, and autologous blood was used for two perfusions in the course of 150 minutes. After 60 minutes perfusion 20 g enzymatic casein hydrolyzate were applied to the rumen. The levels of free amino acids in the perfusate were recorded after 60 minutes' perfusion [the first phase of perfusion] and at the end of the experiment [the second phase]. The levels of lysine, aspartic acid and glutamic acid increased after perfusions with bovine plasma during the first phase, the levels of glutamic acid, phenylalanine, and in one case of alanine, increased after perfusions with autologus blood. Simultaneously the level of valine decreased after perfusions with bovine plasma, and after perfusions with blood the levels of arginine and valine, and/or lysine, dropped. During the second phase of perfusion, the levels of all the observed amino acids except methionine [bovine plasma], and/or orginine and methionine [blood] rose in the perfusate. The experiments showed that the level of amino acids in the rumen content presented a decisive factor affecting amino acid absorption from the rumen into the blood. Transformation of the amino acids during their passage through the remen wall may be assumed, and glutamic acid is one of the chief products of this process.     

Amino acid production in the isolated and perfused rumen wall of sheep

The metabolism of amino acids and urea in an isolated perfused wall of sheep rumen was studied by the method of organ perfusion, using 3 kinds of perfusate. In experiments with semisynthetic perfusate containing urea (SPurea) the levels of lysine, valine, glutamic acid and alanine increased at a simultaneous decrease of levels of leucine, arginine, aspartic acid, threonine and methionine. In perfusions with the semisynthetic medium without urea (SP) the levels of lysine, valine, phenylalanine, isoleucine, glutamic acid, glycine and alanine increased, the level of threonine dropped. In perfusions with whole autologous blood (KP) the levels of valine, glutamic acid and alanine increased, the level of arginine decreased. The level of urea decreased sharply at using SPurea and KP, however, the equal cumulation of ammonia (200-400 mumol x 1(-1)) was found in all three types of perfusate after 2-hr perfusion. Moreover, in experiments with SP a low level of urea was found already after 5-min. of perfusion. The results of the experiments suggest that the rumen wall is capable of producing amino acids also without the contents of rumen, and that independently on the presence or absence of urea as a nitrogen source.     

In situ disappearance of neutral detergent insoluble nitrogen from alfalfa and eastern gamagrass at three maturities.

In situ digestion kinetics of neutral detergent insoluble nitrogen (NDIN) from alfalfa (Medicago sativa L.) harvested at one-tenth bloom and eastern gamagrass (Tripsacum dactyloides L.) harvested at the boot (GGB), anthesis (GGA), and physiological maturity (GGM) stages of growth were determined with nonlinear regression techniques. Whole-plant tissue and associated leaf and stem fractions were incubated in the ventral rumen simultaneously. On a wholeplant basis, potential extents of degradation were particularly high (> or =904 g/kg NDIN) for GGB and GGA, relative to those of GGM and alfalfa (772 and 658 g/kg NDIN, respectively). For all plant parts, degradation rates of NDIN were faster (P<.05) for alfalfa than for all gamagrass forages. Degradation rate of NDIN did not differ (P>.05) across maturities for any gamagrass tissue type. These results indicate 1) that phenological development and lignification do not limit the rate of NDIN degradation in gamagrass forages but do markedly limit the potential extent of NDIN availability and 2) that most of the NDIN in these forages is potentially available in the rumen and can contribute to the ruminal N supply. Our secondary objective was to compare estimates of N escaping ruminal degradation that were determined on the basis of NDIN degradation kinetics (NDIN method) with those determined traditionally, on the basis of total residual N. The NDIN method mathematically eliminates all neutral detergent soluble N from consideration as part of the pool of dietary N potentially escaping the rumen intact. Estimates of rumen escape nitrogen determined on the basis of degradation rates of NDIN were consistently less than corresponding estimates that were determined on the basis of total residual N. When ruminal escape N that was determined with the NDIN method was regressed on corresponding estimates with the total residual N method, the slopes of the regression lines were .53 and .66 for assumed passage rates of .02 and .06 h(-1), respectively. For the forages evaluated in this study, these results indicate that neutral detergent soluble N may make important contributions to the pool of N escaping ruminal degradation.     

Clinical and pathologic changes in acute bovine aflatoxicosis: rumen motility and tissue and fluid concentrations of aflatoxins B1 and M1

Effects of aflatoxin on bovine rumen motility were determined by radiotelemetric techniques. Aflatoxin altered amplitude and/or frequency of rumen contractions in steers given dosages of 0.2, 0.4, 0.6, or 0.8 mg of aflatoxin/kg of body weight. Effects of aflatoxin on rumen motility were dose dependent. An increase in elimination time of aflatoxin from rumen contents was observed in steers given the aflatoxin dosages of 0.4 to 0.8 mg/kg. The increase in elimination time of this toxin facilitates diagnostic capabilities for detecting bovine aflatoxicosis by obtaining rumen contents for analysis for aflatoxin. Aflatoxin M1 was detected in rumen contents from steers at 2 hours after aflatoxin was administered. Thus, intraruminal metabolism of aflatoxin B1 to M1 may occur.     

Elution of bacteria adhering to the epithelium in the rumen in sheep and their detection using electron microscopy

A method of preparing the pure concentrate of epithelium-adherent bacteria in sheep rumen was worked out and it was tested in 24 samples of the rumen wall, obtained from 12 slaughtered sheep. The purity of the bacterial eluate was checked by transmission electron microscopy and negative staining, followed by electron-microscopic evaluation. Besides bacterial cells no residues of feed, epithelium and other undesirable impurities were found out in the eluate. The percent yield of this method was determined by scanning electron microscopy and it made 93.3%. An isolated pure concentrate of the mixture of bacterial cultures, previously adhered to the rumen epithelium, was prepared which can be used for morphological and biochemical studies of this interesting group of rumen bacteria.     

用绵羊小肠液冻干粉测定饲料过瘤胃淀粉小肠消化率方法的研究

本试验旨在以玉米为样本筛选用绵羊小肠液冻干粉测定精饲料瘤胃非降解残渣淀粉小肠消化率的最佳培养条件。分别研究了小肠液冻干粉用量(0.2、0.3、0.4、0.5和0.6g)对16h玉米瘤胃非降解残渣干物质和淀粉小肠消化率的影响;离体培养时间(4、8、12、16、20和24h)对16h玉米瘤胃非降解残渣干物质和淀粉小肠消化率的影响。结果表明,在本试验条件下,绵羊小肠液冻干粉测定玉米瘤胃非降解残渣淀粉小肠消化率的最佳用量为0.45g小肠液冻干粉/0.56g玉米瘤胃非降解残渣,最佳培养时间为12h。利用绵羊小肠液冻干粉测定常用饲料过瘤胃残渣淀粉小肠消化率的方法是可行的。     

Pipecolic acid in rumen fluid and plasma in ruminant animals

A survey was conducted to investigate the physiological levels of pipecolic acid (Pip) in rumen fluid and plasma of ruminants such as goats and cattle in the presence or absence of rumen protozoa. The concentration of Pip was determined using HPLC. Basal Pip levels in the rumen fluid and plasma of normal faunated animals were 21 ± 8 and 2.3 ± 1.3 µM, respectively, and levels increased 1–2 h after feeding. The Pip levels in the rumen fluid and plasma of faunated goats and cattle were significantly higher than those of defaunated goats and unfaunated cattle. A small amount of Pip was also found in the rumen fluids of the defaunated and unfaunated animals; this appeared to be derived from feeds such as hay cube and corn silage. The results obtained in the present study suggest that a significant amount of rumen‐produced Pip is likely to be absorbed into the plasma of the host animals and that rumen protozoa significantly enhance the concentration of Pip in the rumen fluid and plasma of ruminant animals.     

Studies on rumen fermentation with special reference to the volatile fatty acid production rate in ewes during reproduction. 2. Production rate of volatile fatty acids and rumen volume

Methods and results of the measuring of the rumen volume and the passage rate as well as the dependence of the production of volatile fatty acids on the parameters mentioned are described. The correlative relations between rumen volume on the one hand and feed passage on the other hand to the production quotas are investigated. Passage rates and rumen volumes were determined with the help of the isotope dilution method. In the course of reproduction rumen volumes of 7.81, 6.41, 8.91, 7.11 in experiment 1 and 5.41, 5.81, 9.51 and 4.71 in experiment 2 were measured in the sequence early stage of gestation, late stage of gestation, lactation and the period between pregnancies. The average total production of volatile fatty acids within 12 hours amounted to 475 mmol, 265 mmol, 313 mmol and 351 mmol in experiment 1 and 155 mmol, 175 mmol, 271 mmol and 144 mmol in experiment 2 in the same sequence. The average passage duration of the feed through the digestive tract of the ewes was 32.4 h in experiment 1 and 33.6 h in experiment 2. The relatively high feed intake of the ewes resulted in a short passage duration and could be a cause of the low production quotas of the volatile fatty acids.     

Ruminal 15N-straw degradation in sacco and the in vivo digestibility of 15N from straw in the sheep and pony

The nylon-bag method was applied for determination of the rumen degradation of dry matter and nitrogen of 15N-labelled wheat straw. For the experiment 4 wethers fitted with a rumen cannula were used. The bags containing 15N straw were introduced into the rumen and withdrawn 3, 6 or 12 h after incubation. In a second experiment the apparent 15N-digestibility of the same straw was determined in wether and pony. The dry matter disappearance varied between 6 and 23%. For 15N-labelled straw the disappearance of 15N was higher than that of total N. 12 h after incubation 71% of 15N and only 25% of total N were disappeared. It was calculated that after incubation rumen microbial-N in the nylon bag increased from 31% (3 h) to 45% (6 h) and 61% (12 h) resp. The apparent 15N digestibility amounted 53 +/- 2% for wethers and 51 +/- 2% for ponies.     

棉粕与豆粕在反刍动物瘤胃内降解动力学的评定分析

本试验旨在评定棉粕与豆粕的干物质（DM）、有机物（OM）、粗蛋白质（CP）在绵羊瘤胃内的降解动力学参数变化及瘤胃降解前后有效赖氨酸的含量变化。选用3只安装有永久性瘤胃瘘管的体重为（28.0±2.97）kg的育肥哈萨克公羊,代谢笼内单笼饲喂,每天饲喂棉籽壳200 g、精料500 g、小麦秸500 g,余料不超过10%。采用尼龙袋技术评定棉粕与豆粕DM、OM及CP的降解动力学参数,计算有效降解率（ED）及瘤胃停留时间（RRT）,染料结合法（DBL）测定有效赖氨酸的含量。结果表明,在瘤胃固相食糜的平均外流速度为1.84%/h条件下,豆粕DM、OM及CP的瘤胃降解率及有效降解率均高于棉粕,而瘤胃停留时间均短于棉粕。豆粕经瘤胃内培养16 h后有效赖氨酸含量略有升高,而棉粕无明显变化。该试验结果为进一步评定棉粕及豆粕养分的小肠消化率提供参考。     

Continuous automated recording of ruminal pH in sheep nourished entirely by intragastric infusions

A method is described whereby the rumen pH in sheep entirely nourished by infusion of volatile fatty acids was determined continuously. The electrodes used were standard laboratory combination electrodes connected to a digital pH meter, a multi-channel chart recorder and an isolation module. The correlation between pH readings from the bench samples and from electrodes immersed in the rumen for periods of up to 28 days was 0.980 +/- 0.015 for 250 observations. There were fairly regular patterns of change in ruminal pH. The pH rose and fell by approximately 0.1 to 0.2 units over a period of about one hour. These changes were thought to be associated with periods of pseudorumination.     

宏基因组学揭示瘤胃微生物多样性及功能

反刍动物瘤胃内栖息着庞大和复杂的微生物群体,这些微生物与宿主的消化吸收、营养代谢和免疫功能息息相关,宿主及其微生物共同组成了一个"超级生物体"。由于绝大部分瘤胃微生物不可培养,因此以厌氧培养为基础的传统研究方法存在明显的弊端。宏基因组学通过高通量的测序方法,能够全面展示微生物多样性,准确发现新的功能基因。此外,宏基因组学揭示了宿主基因和微生物组之间的互作关系。随着组学技术的不断发展,宏基因组学在瘤胃微生物组研究方面具有广阔的应用前景。     

Modification by 6-hydroxydopamine (6-OHDA) of the inhibition of extrinsic rumen contractions in sheep produced by beta-endorphin

In sheep, beta-endorphin (1 and 2 micrograms/kg) administered into the third cerebral ventricle caused a significant inhibition of the frequency of rumen contractions. The amplitude of the first rumen contractions, following immediately after the end of infusion, and the average amplitude of primary rumen contractions, were inhibited. Beta-endorphin caused general psychomotor excitability. These results suggest that an inhibitory mu and delta opioid system is involved in the control of forestomach motility and general behaviour in sheep. All effects of beta-endorphin were completely prevented by i.c.v. 6-hydroxydopamine (6-OHDA, 18.2 micrograms/kg) pre-treatment. These results suggest that beta-endorphin-induced inhibition of rumen motility is due to central noradrenergic system activation. The exact location of this noradrenergic system remains to be determined.     

The estimation of utilizable amino acids (uAA) of feeds for ruminants using an in vitro incubation technique

The in vitro incubation technique of Zhao and Lebzien (2000; Arch. Anim. Nutr. 53, 293-302) was used for the estimation of utilizable amino acids (uAA) (sum of amino acids from undegraded feed protein and microbial protein, when N is not limiting) of feeds for ruminants. The rumen fluid from a cow fed only with hay (Expt. 1) and that from a sheep fed with a mixed ration (Expt. 2) was compared with respect to estimation of uAA. In Expt. 1, 30 feeds and feed mixtures were tested and in Expt. 2, 33 feeds and feed mixtures were tested. A close linear relationship was found between the utilizable crude protein (uCP=undegraded feed protein + microbial protein) (X, g/kg) calculated from in vivo experiments and the uAA (Y, g/kg) estimated from in vitro incubations both in Expt. 1: y= 0.95 x-1.39, r2=0.85, p<0.001,n=30; and in Expt. 2: Y=0.85X-6.67,r2= 0.85,p<0.001,n=33. Statistical analysis indicates that there was a significant regressive relationship between uAA determined with the rumen fluid of a sheep (X, g/kg) and uAA determined with the rumen fluid of a cow (Y, g/kg): Y=1.06X+12.4,r2= 0.80,p<0.001,n=27. The results indicate that the in vitro incubation technique of Zhao and Lebzien (2000) can be used for the estimation of uAA of feeds for ruminants. As a rumen-fistulated cow is more expensive than a rumen-fistulated sheep, it is suggested to use a sheep fed a mixed ration as the donor of rumen fluid for the estimation of uAA of feeds with in vitro incubation. Further experiments should be performed to standardize the method and to test the most valid length of the incubation period.     

抑制瘤胃甲烷排放的酵母菌的筛选及培养条件优化

减少瘤胃甲烷排放措施的研究已成为当前的研究热点之一。本研究以酒、面包和畜禽专用酿酒酵母产品为原材料,从中分离到三株酵母菌,将其制成冻干粉,通过体外产气试验,筛选出了一株抑制瘤胃甲烷排放效果最为显著的酵母菌株。然后以该酵母菌株作为研究对象,进行分子生物学鉴定;采用分光光度计比浊法测定该酵母菌株的生长曲线,确定菌体生长规律;并以此为依据,设计单因素试验考察该酵母菌株最适宜的生长条件。结果表明,分离得到的酵母菌株均为酿酒酵母saccharomyces cerevisiae;抑制瘤胃体外甲烷排放效果最显著的1株酵母菌分离自面包中;该酿酒酵母菌株的对数生长期为8～24h,其最佳培养条件为:生长温度30℃,pH 5.5,培养基中葡萄糖的浓度4.0%。     

Research note: a method for studying local differences in ruminal fermentation in dairy cattle.

A method was developed for studying local differences in ruminal fermentation. The developed sampler consisted of an acrylic glass container (460 cm3) with an aperture for digesta sampling, which could be opened and closed by the scaled "T" rod. The scale was a reference for defined rumen layers: top, middle, 5 to 10 cm and 25 to 35 cm beneath the top of particles mat, respectively, and bottom 5 to 10 cm above the rumen floor. The repeatability of the method was proved in two rumen cannulated cows. Particle/fluid ratio, pH and sample amount were measured 2 to 2 1/2 h after morning feeding in four replicates each day (over 5 days), rumen layer and animal. No significant differences between replicates were observed. The coefficients of variation (CV) of the particle/fluid ratio varied between 8.7% and 13.6%. Top layer had higher CV than middle and bottom layer. CV of pH ranged between 0.59% and 1.27%. The developed method of sampling showed satisfactory repeatability for investigation of digesta properties and fermentation in different rumen layers.     

奶牛瘤胃液中浸麻类芽孢杆菌的分离鉴定

采用平板划线分离法从健康奶牛瘤胃液中分离得到1株细菌Y-1,体外模拟瘤胃环境增殖Y-1,进行形态学观察、生化反应和16S rDNA基因同源性比较,并对其进行系统发育树的构建等系统鉴定。结果表明,该细菌为浸麻类芽孢杆菌(Paenibacillus macerans),本研究还对瘤胃液中存在该菌的原因做了分析。     

慢性瘤胃酸中毒状态下奶山羊瘤胃细菌内几种相关细菌数量变化的研究

研究慢性瘤胃酸中毒（subacute rumen acidosis,SARA）状态下,相关瘤胃微生物数量的变化。将6只体重相近,体况良好,并安装有永久性瘤胃瘘管的泌乳期关中奶山羊作为试验对象,以逐渐增加精料的方式诱导动物发生SARA。试验分4期进行,4期日粮的精粗比分别为5∶5、6∶4、7∶3和8∶2。分别采用16S rRNA 探针杂交法和传统培养法对SARA状态下瘤胃内微生物数量变化进行测定。随着日粮精料比例的增加,牛链球菌、乳酸杆菌、反刍兽新月单胞菌、埃氏巨型球菌和淀粉分解菌的数量出现不同程度的增加,而3种主要纤维分解菌的数量有所下降,其中淀粉分解菌的数量极显著增加（P<0.01）。在SARA状态下瘤胃细菌总数、乳酸产生菌和乳酸利用菌数量均有所增长,同时3种主要纤维分解菌的数量明显下降。     

不同种类牧草对山羊瘤胃挥发性脂肪酸变化规律的研究

研究山羊瘤胃内放入装有不同种类牧草的尼龙袋后,瘤胃挥发性脂肪酸浓度的动态变化规律。选用带有瘤胃瘘管的山羊为实验动物,结合尼龙袋法测定3类牧草在山羊瘤胃中的挥发性脂肪酸浓度。结果表明,添加装有不同牧草的尼龙袋后,山羊瘤胃乙酸、丙酸、丁酸、总挥发性脂肪酸（TVFA）浓度及乙酸／丙酸值均呈先上升后下降的规律,并在4h达到峰值,其中乙酸浓度约为70．49％、丙酸浓度约为15．38％,瘤胃的发酵类型总体上偏向于乙酸-丙酸型．3类牧草均适用于山羊的科学饲养。由此可见,添加装有不同种类牧草的尼龙袋均会对山羊瘤胃挥发性脂肪酸产生影响,其原因主要与原料的蛋白质含量有关。