A metallo phosphatase activity present on the surface of Trypanosoma brucei procyclic forms

In this work, we describe how living cells of Trypanosoma brucei procyclic forms were able to hydrolyze extracellular p-nitrophenylphosphate (pNPP). These intact parasites, which had their viability determined by motility and the Trypan blue method, presented a low level of pNPP hydrolysis in the absence of any divalent metal (0.72+/-0.07 nmol pNP/mg min). Interestingly, in the presence of 5mM MgCl(2), ectophosphatase activity of 1.91+/-0.21 nmol pNP/mg min was observed. The ectophosphatase activity was also stimulated by MnCl(2), CoCl(2) and CuCl(2) but not by CaCl(2) and CdCl(2) and was inhibited by ZnCl(2). The addition of Mg(2+), Mn(2+), Co(2+) and Cu(2+) to extracellular medium increased the ectophosphatase activity in a dose-dependent manner. At 5mM pNPP, half-maximal stimulation of pNPP hydrolysis was obtained with 0.39+/-0.05 mM MgCl(2), 0.33+/-0.03 mM MnCl(2), 1.63+/-0.12 mM CoCl(2) and 2.04+/-0.33 mM CuCl(2). In the absence of any divalent metal (basal activity) the apparent K(m) for pNPP was 0.66+/-0.09 mM, while at saturating MgCl(2) concentrations the corresponding apparent K(m) for pNPP for Mg(2+)-stimulated phosphatase activity (difference between total minus basal phosphatase activity) was 0.27+/-0.03 mM. The Mg(2+)-stimulated pNPP hydrolysis was strongly inhibited by ZnCl(2) and vanadate, while the metal-independent basal phosphatase activity was less inhibited by these phosphotyrosyl phosphatase inhibitors.     

Hemolytic assay for goat (caprine) and swine (porcine) complement

Optimal conditions for assaying hemolytic complement of goat (caprine) and swine (porcine) sera were determined. Effects of the following were tested: pH, ionic strength, calcium and magnesium ion concentrations, time and temperature of incubation, and ethylenediamine tetracetate concentration. Guinea pig erythrocytes sensitized with goat or cattle antibodies were the most sensitive target cells for goat complement. Sheep and cattle erythrocytes sensitized with rabbit hemolysin were the best target cells for swine complement. Barbital buffer, pH 7.3, ionic strength of 90 nmM relative salt concentration, containing 0.5 mM CaCl2 and 1 mM MgCl2 was the best for swine complement assay. Goat complement lysed best in a barbital buffer, pH 8, ionic strength of 90 to 120 mM of relative salt concentration, in presence of 0.5 mM CaCl2 and 1 mM MgCl2. The optimal incubation temperature was 37 degrees C for both complements. The complement dependent lysis required 75 minutes to reach its maximum. Ethylenediamine tetracetate in 4 mM concentration completely inhibited lysis by both species complements. The CH50 for goat sera varied between 18 and 75 per ml, in swine sera between 75 and 210 per ml.     

瘤胃混合微生物对稻糠、玉米麸、麦麸饲料中植酸磷降解特性的比较研究

采用短期人工瘤胃发酵技术,研究瘤胃混合微生物对糠麸类饲料中植酸磷的降解作用。试验采集3头健康泌乳期荷斯坦奶牛瘤胃液,以不同来源稻糠、玉米麸、麦麸饲料为底物,测定厌氧发酵3、12、24、48和72 h饲料植酸磷的消失率。结果表明:1)糠麸类饲料植酸磷消失率均随发酵时间的延长而增加,并且在发酵至24 h时,植酸磷消失率趋于90%以上。2)不同来源的同种糠麸类饲料间植酸磷的潜在降解率和降解速率明显不同。3)3种糠麸类饲料比较发现,麦麸饲料的植酸磷潜在降解率可达到100%(P<0.000 1),而玉米麸(95.58%)与稻糠(96.82%)饲料间差异不显著(P>0.05),植酸磷降解速率呈现为稻糠>玉米麸>麦麸(P<0.000 1)。由此可见,瘤胃混合微生物能很好地降解富含植酸磷的糠麸类饲料,且降解作用在不同饲料间存在一定的差异。     

Characterization of an ecto-ATPase of Tritrichomonas foetus.

In this work, we describe the ability of living Tritrichomonas foetus to hydrolyze extracellular ATP. The addition of MgCl(2) to the assay medium increased the ecto-ATPase activity in a dose-dependent manner. At 5mM ATP, half maximal stimulation of ATP hydrolysis was obtained with 0.46mM MgCl(2). The ecto-ATPase activity was also stimulated by MnCl(2) and CaCl(2), but not by SrCl(2). The Mg(2+)-dependent ATPase presents two apparent K(m) values for Mg-ATP(2-) (K(m1)=0.03 mM and K(m2)=2.01 mM). ATP was the best substrate for this enzyme, although other nucleotides such as ITP, CTP, UTP also produced high reaction rates. GTP produced a low reaction rate and ADP was not a substrate for this enzyme. The Mg(2+)-dependent ecto-ATPase activity was insensitive to inhibitors of other ATPase and phosphatase activities, such as oligomycin, sodium azide, bafilomycin A(1), ouabain, furosemide, vanadate, molybdate, sodium fluoride and levamizole. The acid phosphatase inhibitors (vanadate and molybdate) inhibited about 60-70% of the Mg(2+)-independent ecto-ATPase activity, suggesting that the ATP hydrolysis measured in the absence of any metal divalent could, at least in part, also be catalyzed by an ecto-phosphatase present in this cell. In order to confirm the observed Mg(2+)-dependent activity as an ecto-ATPase, we used an impermeant inhibitor, 4,4'-diisothiocyanostylbene-2',2'-disulfonic acid (DIDS) as well as suramin, an antagonist of P(2) purinoreceptors and inhibitor of some ecto-ATPases. These two reagents inhibited the Mg(2+)-dependent ATPase activity in a dose-dependent manner. This ecto-ATPase was stimulated by more than 90% by 50mM D-galactose. Since previous results showed that D-galactose exposed on the surface of host cells is involved with T. foetus adhesion, the Mg(2+)-dependent ecto-ATPase may be involved with cellular adhesion and possible pathogenicity.     

体外法研究异麦芽寡糖对生长绵羊瘤胃发酵功能的影响

采用批次培养的方法,研究了体外条件下日粮添加异麦芽寡糖对生长绵羊瘤胃发酵功能的影响。异麦芽寡糖的添加水平分别为0%(对照组)、0.20%、0.40%、0.60%、0.80%、1.00%。结果表明,日粮添加异麦芽寡糖可以提高培养液中挥发性脂肪酸(VFA)含量,降低培养液中的氨氮(NH3-N)含量和培养残渣中中性洗涤纤维(NDF)含量,但对培养液中菌体蛋白(BCP)含量的影响不确定。研究初步证明,日粮添加异麦芽寡糖可以在一定程度上提高生长绵羊的瘤胃发酵功能。     

利用体外产气法研究不同瘤胃降解蛋白平衡对瘤胃代谢的影响

旨在探讨利用体外产气法研究不同瘤胃降解蛋白平衡日粮(RDPB,g·kg-1 DM)对瘤胃发酵、总产气量和甲烷产量的影响.试验选用美国Ankom产气系统,采用随机区组试验设计,配置6种不同瘤胃降解蛋白平衡日粮,分别为日粮1(-20 g· kg-1 DM),日粮2(-10 g· kg-1 DM),日粮3(0 g· kg-1 DM),日粮4(10g·kg-1DM),日粮5(20 g·kg-1 DM)和日粮6(30 g· kg-1 DM),每组2个重复,培养期为24 h.结果表明:(1)本研究中6种类型日粮发酵液的pH、乙酸/丙酸、产气量和CH4产生量表现为日粮3(P＜0.01)或日粮4(P＜0.01)值最低.(2)挥发性脂防酸乙酸、丙酸、丁酸以及氨氮的浓度随着瘤冒降解平衡值的增加而增加,表明改变日粮瘤胃降解蛋白平衡可以影响瘤胃发酵.而瘤胃降解蛋白平衡对总挥发性脂肪酸以及原虫数量无显著影响.(3)日粮瘤胃降解蛋白平衡值影响不同时间点甲烷的浓度和24 h甲烷产量.其中以日稂4(P＜0.05)甲烷总产量最低.由此可见,在体外培养条件下,改变日粮RDPB影响了瘤胃发酵液pH、氨态氮(NH3-N)及挥发性脂肪酸(VFA)浓度、总产气量、CH4产生量,综合考虑其对瘤胃发酵和甲烷产量等指标的影响,体外产气条件下适宜的RDPB水平为0～10g·kg-1 DM.     

利用体外法研究功能性寡糖对生长绵羊瘤胃发酵特性的影响

试验采用批次培养法研究了体外条件下日粮分别添加异麦芽寡糖、甘露寡糖及果寡糖对生长绵羊瘤胃发酵功能的影响。三种功能性寡糖的添加水平分别为0.0%(对照组)、0.2%、0.4%、0.6%、0.8%以及1.0%。结果表明:(1)日粮添加异麦芽寡糖可以提高培养液中的挥发性脂肪酸(VFA)含量,降低培养液中的氨态氮(NH3-N)含量和培养残渣中的中性洗涤纤维(NDF)含量,对培养液中菌体蛋白(BCP)含量的影响不确定。(2)日粮添加甘露寡糖可以提高培养液中的VFA和BCP含量,并以0.6%的添加量效果最佳;添加量在0.8%以上时,可以显著降低(P<0.05)培养液中的NH3-N含量,对培养残渣中的NDF含量没有显著影响(P>0.05)。(3)日粮添加果寡糖可以显著(P<0.05)提高培养液中的VFA含量;当添加量在0.6%以上时,可以显著(P<0.05)提高培养液中的BCP含量,显著(P<0.05)降低培养液中的NH3-N含量和培养残渣中的NDF含量;在本试验所设添加范围内,1.0%的果寡糖对瘤胃发酵影响的综合表现最优。日粮添加异麦芽寡糖、甘露寡糖和果寡糖有助于提高生长绵羊的瘤胃发酵功能,其中果寡糖对提高瘤胃整体发酵功能的作用效果最佳。     

硫酸钠对奶牛瘤胃发酵的影响

本文旨在研究不同添加水平的硫酸钠对泌乳中期奶牛瘤胃发酵的影响。试验选用4头体重、胎次、产奶量均相近的荷斯坦奶牛,采用4×4拉丁方设计,试验分4个处理组,添加不同水平的硫酸钠,即0%(对照组)、0.4%(试验1组)、0.8%(试验2组)与1.2%(试验3组),试验分为4期,每期18 d(预试期15 d,正试期3 d)。结果表明:(1)饲粮中添加硫酸钠对瘤胃液pH、NH3-N平均浓度无显著影响(P>0.05);(2)饲粮中添加硫酸钠后,瘤胃内原虫数均有所提高,其中试验2组和试验3组与对照组相比均差异显著(P<0.05);(3)添加硫酸钠的试验2组瘤胃液菌体蛋白浓度与对照组相比显著提高(P<0.05)。结果说明,日粮中添加适量的硫酸钠可以改变瘤胃发酵环境,提高瘤胃液菌体蛋白浓度。     

Palmitate metabolism by isolated sheep rumen epithelial cells.

Ruminal palmitate metabolism was examined using an isolated cell system. Palmitate oxidation to 14CO2 by rumen epithelial cells isolated from the rumens of mature sheep was linear during the course of a 2-h incubation (11.1 nmoles.million cells-1.2 h-1) and 3.6 times the rate of palmitate oxidation by cells isolated from neonatal rumen (3.1 nmoles.million cells-1.min-1). Subsequent experiments were conducted with mature rumen epithelial cells. Neither acetate (50 mM), propionate (10 mM), dibutyryl cAMP (.2 mM), nor insulin (10 mU/mL) altered palmitate oxidation to CO2. However, butyrate (10 mM) addition reduced (P less than .05), and ammonia (15 mM) tended to reduce (P less than .10), palmitate oxidation (51.6 and 82.0% of control, respectively), whereas addition of glucose (2.5 mM) increased (P less than .05) palmitate oxidation (151% of control). Of the compounds tested, only propionate, butyrate, and ammonia reduced palmitate oxidation to total acid-soluble metabolites. Propionate (10 mM) addition completely abolished palmitate oxidation to acid-soluble metabolites. Succinate addition (5 to 50 mM) increased palmitate oxidation to CO2 but exhibited no consistent effect on palmitate oxidation to either acid-soluble metabolites or beta-hydroxybutyrate. Propionate completely abolished palmitate oxidation to beta-hydroxybutyrate, suggesting that propionate-induced inhibition of palmitate oxidation is not mediated via succinate. The data indicate 1) that rumen epithelium is capable of oxidizing palmitate, 2) that ruminal palmitate oxidation may be subject to regulation by developmental factors, and 3) that palmitate metabolism seems to be influenced more by ruminally derived metabolites than by factors derived exclusively from the general circulation.     

非纤维性碳水化合物水平对活体外瘤胃发酵和产生共轭亚油酸的影响

在底物粗蛋白质和粗脂肪水平相同的条件下,研究非纤维性碳水化合物(NFC)水平(20%、40%、60%和80%)对活体外瘤胃发酵24h后的发酵参数和发酵液中共轭亚油酸(CLA)浓度的影响。结果表明:底物不同的NFC水平影响活体外瘤胃发酵和CLA的产生。随底物NFC水平的提高,瘤胃微生物的活体外发酵程度提高;同时发酵液中CLA的浓度随底物NFC水平的提高呈线性增加(L,P<0.0001)的变化趋势。     

Biosynthesis of arginine from citrulline and related compounds by mixed ruminal bacteria, protozoa and their mixture in vitro

Biosynthesis of arginine (Arg) from citrulline (Cit), ornithine (Orn), proline (Pro), and 5-aminovaleric acid (5AV) by mixed rumen bacteria (B), protozoa (P), and their mixture (BP) was quantitatively investigated in an in vitro system from the standpoint of protein nutrition in ruminants. Rumen microorganisms, collected from ruminally fistulated goats, were anaerobically incubated with or without 1 mmol/L each of substrates at 39°C for 12 h. Arginine and other related compounds, produced in both supernatants and acid-hydrolyzates of microorganisms in B, P, and BP suspensions, were analyzed by high-performance liquid chromatography. Arginine production from Cit in BP, when expressed with a unit of 'μmol/g microbial nitrogen', was approximately 70% and 94% higher than that in B and P, respectively, in a 12-h incubation period. In the case of Orn, the values were approximately 30% and 75%. Both rumen bacteria and protozoa could produce Cit and Orn from Pro, so it is assumed that they can produce Arg from Pro. Rumen protozoa were unable to degrade 5AV and it was the final product in the metabolism of Cit, Orn and Pro in P suspension. A trace amount of Orn and Pro produced from 5AV in B and BP suspensions indicated that the reversible reaction of 5AV formation was performed only by rumen bacteria. This is the first quantitative report on Arg biosynthesis from its precursors by rumen microorganisms.     

白酒糟对黔北麻羊瘤胃发酵及血浆生化指标的影响

本试验旨在研究白酒糟(产自贵州美酒河流域)对黔北麻羊瘤胃发酵及血浆生化指标的影响。以6只年龄、体重基本一致的黔北麻羊为试验动物,采用3×3拉丁方试验设计,将其分成3组,每组2个重复,每个重复1只羊。对照组饲喂基础饲粮,试验1组和试验2组以白酒糟分别替代基础饲粮中20%、40%的精料。试验期45 d,分3期,每期15 d,包括10 d预试期和5 d正试期。正试期采集血浆和瘤胃液样品,测定瘤胃液pH、缓冲力、氨态氮和挥发性脂肪酸浓度,瘤胃液微晶纤维素酶、羧甲基纤维素酶、纤维二糖酶、木聚糖酶活性以及血浆葡萄糖、尿素氮、总蛋白、白蛋白、总胆固醇、肌酐含量。结果显示:1)3组黔北麻羊瘤胃液pH、缓冲力差异不显著(P0.05);氨态氮浓度均处于正常范围,试验1组氨态氮浓度显著高于试验2组(P0.05)。2)试验1组瘤胃液木聚糖酶活性显著高于对照组(P0.05),各组间瘤胃液微晶纤维素酶、羧甲基纤维素酶、纤维二糖酶活性差异不显著(P0.05)。3)试验1组瘤胃液乙酸浓度最高,显著高于试验2组(P0.05);瘤胃液总挥发性脂肪酸浓度也最高,显著高于对照组和试验2组(P0.05);3组瘤胃液丙酸和丁酸浓度、乙酸/丙酸差异均不显著(P0.05)。4)各组血浆生化指标无显著差异(P0.05)。由此可见,白酒糟替代40%以内精料不会对黔北麻羊瘤胃发酵与血浆生化指标产生不良影响,相比较而言,以20%水平较优。     

Effects of protozoa on bacterial nitrogen recycling in the rumen

The effects of protozoa on ruminal NH3-N kinetics and bacterial N recycling were measured in five sheep (57.6+/-7.1 kg BW, x +/- SD) with ruminal and duodenal cannulas in naturally faunated, defaunated, and refaunated periods. The sheep were fed a diet of 239 g of alfalfa haylage and 814 g of barley concentrate per day (DM basis) divided into 12 equal portions and allocated at 2-h intervals. A pulse dose of 300 mg of 15N as [15N]NH4Cl was administered into the rumen (on d 1 and 15) and 300 mg of 15N as [15N]urea was administered intravenously to the blood (d 8). Enrichment of 15N was measured in ruminal NH3-N, bacterial N, and plasma urea N over a period of 35 h. Total collection of urine was made for 5 d and analyzed for purine derivatives to calculate the flow of microbial N. Ruminal parameters and nutrient digestibilities were also measured. Sheep were defaunated using a rumen washing procedure 50 d prior to measurements in the defaunated period. Sheep were refaunated with ruminal contents from a faunated sheep receiving the same diet. Measurements began 26 d following refaunation, at which time protozoal numbers had returned to those in the originally faunated sheep. Data reported in parentheses are for faunated, defaunated, and refaunated sheep, respectively. Total culturable and cellulolytic bacterial numbers were unaffected by defaunation, but there was an increase in flow of microbial N from the rumen (10.8, 17.3, and 11.1 g N/d; P < .05) in the defaunated period. Flux, irreversible loss, and intraruminal recycling of NH3-N and recycling of NH3-N from plasma urea N were not affected by defaunation. Defaunation had no effect on reducing the absolute amount (13.8, 10.0, and 11.3 g N/d; P > .20) of bacterial N recycling and the percentage of N flux through the bacterial N pool. Total-tract digestion was reduced in defaunated compared with faunated sheep by 8, 17, 15, and 32% for OM, N, NDF, and ADF, respectively. In conclusion, defaunation improved ruminal N metabolism through the enhancement of bacterial protein synthesis, and improvement in the flow of microbial protein to the host animal.     

Effect of various levels of imbalance between energy and nitrogen release in the rumen on microbial protein synthesis and nitrogen metabolism in growing double-muscled Belgian Blue bulls fed a corn silage-based diet

Seven double-muscled Belgian Blue bulls (initial BW: 341 +/- 21 kg) with cannulas in the rumen and proximal duodenum were used in an incomplete replicated Latin square. The study examined the effect of imbalance between energy and N in the rumen on microbial protein synthesis and N metabolism by giving the same diet according to 3 different feeding patterns. The feed ingredients of the diet were separated into 2 groups supplying the same amount of fermentable OM (FOM) but characterized by different levels of ruminally degradable N (RDN). The first group primarily provided energy for the ruminal microbes (12.5 g of RDN/kg of FOM), whereas the second provided greater N (33.3 g of RDN/kg of FOM). These 2 groups were fed to the bulls in different combinations with the aim of creating 3 levels of imbalance (0, 20, and 40 g/ kg of DM) between energy and N supplies in the rumen. Imbalance was measured by the variation of the degradable protein balance (OEB value in the Dutch system) of the diet between the 2 meals each a day. Diurnal variations in ruminal NH3-N concentrations and plasma urea concentrations were greatly influenced by the feeding patterns of the diet. Introduction of imbalance affected neither microbial N flow at the duodenum (P = 0.97) nor efficiency of growth (P = 0.54). The feeding patterns of the diet had no negative impact on NDF degradation in the rumen (P = 0.33). Nitrogen retention was not affected by imbalance (P = 0.74) and reached 49.7, 52.0, and 51.3 g/d, respectively for 0, 20, and 40 g of OEB/kg of DM imbalance. It seems that introduction of an imbalance between energy and N supplies for the ruminal microbes by altering the feeding pattern of the same diet does not negatively influence the microbial activity in the rumen nor N retention of the animal. Nitrogen recycling in the rumen plays a major role in regulating the amount of ruminally available N and allows a continuous synchronization of N and energy-yielding substrates for the microorganisms in the rumen. Therefore, imbalance between dietary energy and N created over a 24-h interval was not detrimental to rumen microbial growth for the animal as long as the level of imbalance did not exceed 40 g of OEB/kg of DM. Thus, these feeding patterns of the diet can be used under practical feeding conditions with minimal impact on the performance of ruminant animals for meat production.     

茶皂素对奶牛瘤胃发酵及瘤胃微生物区系的影响

本试验旨在研究茶皂素对奶牛瘤胃发酵及瘤胃微生物区系的影响。试验选用12头健康状况良好,体重相近的荷斯坦奶牛,随机分为4组,均饲喂基础饲粮,各组分别灌服0(对照)、20、30、40 g/d茶皂素,茶皂素与水混匀灌服,进行预试期14 d,正试期35 d的饲养试验。正试期期间每隔7 d,在晨饲前1 h用口腔采样器采集瘤胃液测定瘤胃发酵指标,用实时定量PCR(qRT-PCR)法测定瘤胃微生物含量。结果表明:1)与对照组相比,茶皂素显著降低了瘤胃液p H(30、40 g/d组)、氨态氮的浓度(20、30、40 g/d组)(P0.05),但均未超过正常范围值;茶皂素显著提高了微生物蛋白(30、40 g/d组)、丙酸(20、30、40 g/d组)和丁酸浓度(20、30、40 g/d组)(P0.05),30 g/d组的微生物蛋白浓度提高了20.20%;但茶皂素对总挥发性脂肪酸和乙酸浓度的影响不显著(P0.05)。2)与对照组相比,茶皂素各组的瘤胃液原虫、溶纤维丁酸弧菌含量均显著降低(P0.05),甲烷菌、白色瘤胃球菌、产琥珀酸丝状杆菌、黄色瘤胃球菌、真菌含量没有显著变化(P0.05)。综上所述,补饲茶皂素改善了奶牛瘤胃发酵模式,并显著影响了奶牛瘤胃微生物区系,30 g/d的剂量对奶牛较为适宜。     

饲粮不同NFC/NDF对肉用绵羊瘤胃pH、氨态氮和挥发性脂肪酸的影响

本试验旨在研究饲粮不同非纤维性碳水化合物/中性洗涤纤维(NFC/NDF)对肉用绵羊瘤胃pH、氨态氮和挥发性脂肪酸的影响.选用(47.21±1.01)kg安装有瘤胃瘘管的杜泊羊(♂)×小尾寒羊(♀)杂交1代肉用公羊12只,采用12×4不完全拉丁方设计,试验分4期进行,每期16 d,分别饲喂NFC/NDF为0.25、0.34、0.36、0.52、0.60、0.80、0.87、1.13、1.30、1.58、2.17和2.49的12种饲粮.结果表明:随着NFC/NDF的增加,试验羊瘤胃pH极显著线性降低(P＜0.01),氨态氮浓度极显著线性增加(P＜0.01),瘤胃总挥发性脂肪酸及丁酸比例呈显著三次曲线变化(P＜0.05),总挥发性脂肪酸中的丙酸、戊酸和异戊酸比例极显著线性增加(P＜0.01),乙酸比例和乙酸/丙酸极显著线性降低(P＜0.01).由此可见,饲粮NFC/NDF对瘤胃pH、氨态氮和挥发性脂肪酸具有显著影响.     

Production of aromatic amino acids and related compounds from p-hydroxyphenylacetic acid by rumen microorganisms in vitro

Suspensions of mixed rumen bacteria (B), protozoa (P), and mixed rumen microorganisms (BP) prepared from rumen contents of fistulated goats were anaerobically incubated with 1 mM p‐hydroxyphenylacetic acid (HPA) at 39°C for 24 h. Tyrosine (Tyr), phenylalanine (Phe), tryptophan (Trp) and other related compounds in both supernatants and hydrolyzates of microbial cells in all incubations were analyzed by HPLC. Large amounts of Tyr (32.1, 42.7 and 36.1% of disappeared HPA in B, P and BP, respectively) were produced from HPA during a 12 h incubation period. The formation of Tyr in P (178.6 µmol/g MN) was 1.5 and 2 times higher than in B and BP, respectively. Phe (7–11% of the disappeared HPA) and Trp (3–6% of the disappeared HPA) were also synthesized from HPA in B, P, and BP. Phe synthesis in P (46.3 µmol/g MN) was 1.7 times higher than in B but, in contrast, Trp synthesis in B, was 1.6 times higher than in P. The metabolites p‐hydroxyphenylpyruvic acid (in the range of 5–14% of disappeared HPA), phenylacetic acid (1–11%), p‐hydroxybenzoic acid (3–7%) and benzoic acid (1–6%) were produced from HPA in B, P and BP. Phenylpropionic acid (6% of the disappeared HPA) was produced only in B and BP.     

Effect of heat treatment and subsequent urea supplementation of sunflower meal on the in vitro ruminal degradability of crude protein content and its postruminal digestibility

A three-phase laboratory procedure was used for predicting the degradability of the protein of extracted sunflower meal before (SFM) and after heat treatment (HSFM). The rumen fluid degraded 69.1% and 67.1% of the SFM and HSFM protein, respectively. The digestibility values of rumen undegraded protein (UDP) were 57.3% (SFM) and 57.0% (HSFM) with pepsin and 17.6% (SFM) and 15.9% (HSFM) with pancreatin. Urea supplementation practically did not alter the rumen degradability of HSFM protein, while the pepsin digestibility of UDP decreased to 47.2%. Four fractions (NH3, dissolved amino acids, oligopeptides and proteins) of rumen degradable crude protein (RDP) were also determined in vitro: 81 to 92% of the degraded crude protein was found in the fractions tested. Heat treatment reduced free NH3 content but did not alter the other three fractions. Urea supplementation decreased the quantity of NH3, peptides and proteins as well.     

Evaluation of alfalfa-corn cob associative action. III. The effect of mechanically separated alfalfa fractions on intake, digestibility and ruminal characteristics of ammonia-treated corn cob diets fed to sheep

The influence of mechanically separated alfalfa fractions on intake, digestibility and rate of ruminal passage was investigated using 48 lambs (32 kg) in a digestion trial. Whole plant pre-bloom alfalfa (25% crude protein) or fractions (presscake, dehydrated presscake, protein coagulum, dehydrated protein coagulum, whole juice or deproteinized juice) were added to a 3% ammonia (NH3)-treated corn cob negative control diet at levels equal to 20% wholeplant alfalfa dry matter (DM); eight treatments, six lambs/treatment. Ad libitum intake was greater (P less than .05) for alfalfa fiber (presscake) or juice supplemented diets compared with 20% direct cut alfalfa. Dry matter intake, digestibility of DM and cell walls, and rate of passage were highly correlated when diets were fed ad libitum. At equal DM intakes, dehydrated vs wet presscake increased (P less than .05) DM and cell wall digestibility. Heating of the protein may have reduced degradation rate and consequently a slower release of nutrients for microorganisms in the rumen. Whole vs deproteinized juice increased digestibility of cob DM (P = .11) and cell walls (P = .13), suggesting a response to level of degradable alpha-amino N. Whole and deproteinized juice increased cell wall digestibility compared with the negative control by 23.0 and 18.5 percentage units, respectively, suggesting that degradable alpha amino-N and cell solubles or other nutrients interacted to maximize microbial fiber digestion. Total and branch-chain volatile fatty acids measured at 6, 12 and 18 h post-feeding were highly correlated with nutrient digestibility. Ruminal NH3-N measured at 18 h was negatively correlated with dry matter (r = -.74) and cell wall (r = -.72) digestibility, showing that alfalfa supplies nutrients required by ruminal microorganisms for NH3 assimilation and fiber digestion. The mode of alfalfa associative action in high fiber diets is in supplying ruminal microorganisms with degradable protein and (or) other nutrients, rather than altering ruminal retention time.     

不同精粗比下添加维生素B12对体外瘤胃发酵和微生物酶活力的影响

本试验通过体外培养法,研究在不同精粗比饲粮中添加维生素B12对体外瘤胃发酵和微生物酶活力的影响。试验采用3×3双因子试验设计,即3个底物精粗比(玉米∶羊草=35∶65、50∶50和65∶35)和3个维生素B12添加量(0、40和90 ng/mL)。体外试验用瘤胃液取自3只安装有永久性瘤胃瘘管的湖羊。体外培养24 h后测定体外瘤胃发酵参数和微生物酶活力。结果显示:1)随着底物精粗比和维生素B12添加量的提高,体外培养24 h的产气量、潜在产气量和有机物消化率极显著地增加(P<0.01),且维生素B12添加量与上述指标存在线性剂量效应(P<0.01)。2)当底物精粗比为50∶50和65∶35时,添加维生素B12显著提高了发酵液中氨态氮、微生物蛋白、总挥发性脂肪酸、乙酸和丙酸浓度(P<0.05),但对丁酸浓度和乙酸/丙酸无显著影响(P>0.05)。3)当底物精粗比为35∶65和50∶50时,添加40 ng/mL维生素B12使发酵液中羧甲基纤维素酶、木聚糖酶活力显著提高(P<0.05);当精粗比为65∶35时,添加90 ng/mL维生素B12使发酵液中羧甲基纤维素酶、木聚糖酶活力显著提高(P<0.05)。结果提示,底物精粗比和维生素B12添加量影响体外瘤胃发酵。添加维生素B12可增加瘤胃微生物酶的活力,从而提高有机物消化率以及微生物蛋白和总挥发性脂肪酸的产量。当底物精粗比(玉米∶羊草)较高(50∶50和65∶35)时,维生素B12的添加效果更明显,并且具有剂量依赖效应。