利用体外产气法和CNCPS法对不同生育期虉草营养价值的评价研究

为了评定不同生育期虉草的营养价值,分析体外产气法和CNCPS法两种营养评价方法的相关性。采用体外产气法测定抽穗期、开花期、灌浆期、乳熟期4个生育期虉草的体外发酵产气量、发酵参数(a、b、c)、DM消化率、NDF和ADF降解率,用康奈尔净碳水化合物和蛋白质体系(cornell net carbohydrate and protein system,CNCPS)分析方法测定相应组分,评价其营养价值,并比较研究两种方法评价结果的相关性。结果表明,体外发酵至72 h结束,累计总产气量和产气参数a以开花期最高(122.68 和4.46 mL),显著高于各组(P<0.05),且从抽穗期、灌浆期、乳熟期有降低的趋势;抽穗期和开花期的DM消化率、NDF和ADF降解率分别显著高于灌浆期和乳熟期(P<0.05)。随着虉草生育期的延长,CP和SOLP呈递减趋势,即抽穗期和开花期显著高于灌浆期和乳熟期(P<0.05);NDF、ADF和ADL呈递增趋势,但NDFIP、ADFIP呈递减趋势。灌浆期和乳熟期的CHO含量显著高于抽穗期和开花期(P<0.05),而CA和CNSC含量则相反,且CC含量同样以灌浆期与乳熟期较高;蛋白质组分中,PA与CA及CC与PC组分呈现同样的趋势。虉草CNSC、CA、CB₁、CB₂、PA、PB₃组分含量与不同时间点产气量呈显著或极显著的相关性(P<0.05或P<0.01)。因此,开花期虉草营养价值最高,其次为抽穗期、灌浆期和乳熟期,产气法和CNCPS评价牧草具有相关性,用体外产气量估测牧草CNCPS组分是可行的。     

Rumen parameters of yaks (Bos grunniens) and indigenous cattle (Bos taurus) grazing on the Qinghai‐Tibetan Plateau

Yaks and indigenous Qaidam cattle and cattle‐yak crosses (C × Y) graze on the Qinghai‐Tibetan Plateau (QTP) throughout the year, but yaks are raised at higher elevations than cattle. Yaks do not receive supplementary feed whereas cattle require supplementary feed during harsh winter. We hypothesized that yaks would cope with the severe conditions of the QTP better than cattle and utilize the pasture more efficiently. We also hypothesized that differences between species would be pronounced in winter, when conditions are particularly harsh. To test these hypotheses, seasonal rumen fluid parameters of yaks, C × Y and cattle (n = 3 for each) were examined. Rumen fluid was collected in summer and winter from each genotype 2, 5 and 12 hr after a day of grazing. Concentrations of total volatile fatty acid (VFA), acetate, propionate, isobutyrate and isovalerate were greater in yaks than in cattle in summer (p < 0.05), while propionate concentration was lower in yaks than in cattle in winter (p < 0.05). Concentrations of ammonia and urea were greater (p < 0.001) in yaks than in cattle (p < 0.001) in summer, whereas, concentrations of free amino acids (AA) were greater in cattle than in yaks in summer and winter (p < 0.001). Concentrations of total VFA, acetate, propionate and butyrate decreased linearly (p < 0.05), whereas concentrations of isobutyrate and isovalerate increased linearly for yak and C × Y with sampling time (p < 0.05) in summer. In summer, concentrations of isobutyrate and isovalerate were greater in yaks than in cattle (p < 0.05). In conclusion, rumen fermentation characteristics of yaks showed that they coped better than cattle or C × Y in the harsh climate as we hypothesized. However, in contrast to our hypothesis, this emerged only in summer, when pasture was plentiful and not in winter, when pasture was scarce.     

添加剂对尖叶胡枝子青贮发酵品质及体外消化率的影响

以尖叶胡枝子为原料,添加乳酸菌制剂LaLsIL dry(0.000 5%,0.001 0%,0.002 0%和0.007 0%)和Micromanger H/M(0.001 135%,0.003 405%和0.004 450%),在实验室条件下制作小型塑料袋青贮,贮藏90 d,分析乳酸菌制剂对尖叶胡枝子青贮保存性能和消化率的影响。 结果表明,添加LaLsIL dry和Micromanger H/M可显著提高胡枝子青贮的乳酸含量(P<0.05),降低丁酸和氨态氮含量 (P<0.05)。通过Flieg青贮饲料评分方案得出尖叶胡枝子直接青贮饲料的发酵品质最差,等级为劣,而2种乳酸菌添加剂处理的青贮品质均明显好于对照组。LaLsIL dry和Micromanger H/M各处理均明显改善了尖叶胡枝子青贮饲料的发酵品质,同时也明显提高了尖叶胡枝子青贮饲料的干物质、粗蛋白质和中性洗涤纤维的体外消化率。     

豆科和蓼科牧草的组合发酵特性研究

应用体外发酵产气技术,评价了鲁梅克斯、三叶草和苜蓿分别以0∶100、25∶75、50∶50、75∶25和100∶0进行两两组合时的发酵特性.结果表明,鲁梅克斯与苜蓿组合时,不同组合比例间的发酵动力学参数差异不显著(P>0.05);苜蓿与三叶草组合时,理论最大产气量(A)及产气速率常数(b)以苜蓿占25%时最高,显著(P<0.05)高于苜蓿占75%的组合;鲁梅克斯与三叶草组合时理论最大产气量及产气速率常数分别以鲁梅克斯占25%和50%的组合最高,并显著高于鲁梅克斯占75%的组合.相同牧草组合时不同组合比例产气延滞时间之间的差异不显著(P>0.05).不同牧草组合时不同比例组合牧草各时间点的产气量表现出显著的正组合效应或正组合效应趋势,在发酵12和24 h时,产气量最易出现显著的正组合效应,不同牧草组合时产气量的组合效应量随发酵时间的延长而下降.理论最大产气量、产气速率常数及48 h产气量(GP)与OM、ADF、NDS含量及NDS/CP呈显著正相关关系,而与CP、NDF及HC的含量呈显著负相关关系.发酵延滞时间与OM(P<0.001)、ADF(P<0.001)含量及NDS/CP(P<0.01)呈显著负相关关系,而与CP(P<0.001)及HC(P<0.001)的含量呈显著正相关关系.结论认为,组合牧草体外发酵产气程度主要受非结构性碳水化合物与粗蛋白比例的影响.     

青贮剂对再生稻头季全株青贮品质和体外瘤胃发酵特性的影响

旨在探究青贮剂对再生稻头季全株青贮品质和体外瘤胃发酵特性的影响.以再生稻头季全株为青贮原料,在青贮中分别添加植物乳杆菌(60％)+纤维素酶(30％)+木聚糖酶(10％)、植物乳杆菌(70％)+粪肠球菌(20％)+纤维素酶(5％)+半纤维素酶(5％)、植物乳杆菌(30％)+粪肠球菌(60％)+纤维素酶(5％)+半纤维素酶...     

添加剂对扁蓿豆青贮饲料影响的研究

以结实期的扁蓿豆Medicago ruthenica为青贮原料,在实验室条件下采用单因素完全区组试验设计,调制扁蓿豆青贮饲料,研究商品菌制剂LaLsIL Dry(LD)和硫酸盐制剂Siloguard(SG)对扁蓿豆青贮饲料发酵品质和体外消失率的影响。结果表明:扁蓿豆直接青贮的丁酸含量较低,V-Score评分等级为良好;添加乳酸菌制剂未能改善扁蓿豆青贮饲料的发酵品质和提高其体外消失率;SG0.2%处理组的乳酸含量、粗蛋白含量和体外干物质消失率显著高于对照(P0.05),氨态氮占总氮比例显著低于对照(P0.05),说明SG0.2%处理可提高乳酸菌的发酵能力,降低了不良微生物对蛋白的降解,保存了大量可供动物体消化利用的营养成分;原料草的体外干物质和粗蛋白消失率显著高于青贮饲料(P0.05)。     

不同粗饲料组合对晋南牛瘤胃体外发酵特性的研究

旨在研究全株玉米青贮(WCS)、小麦秸秆(WS)和苜蓿青贮(AS)不同组合比例对晋南牛瘤胃体外发酵特性的影响,筛选不同饲料组合应用的最适比例.将全株玉米青贮、小麦秸秆、苜蓿青贮以不同比例组合(80:10:10,70:10:20,70:20:10,60:10:30,60:20:10,60:30:10,50:10:40,5...     

Effect of microbial inoculants on the fermentation characteristics,nutritive value,and in vitro digestibility of various forages

This study was aimed to examine the characteristics of three isolated lactic acid bacteria (LAB) strains (HG24, M1, and TG1), and to evaluate their effects on fermentation characteristics, nutritive value, and in vitro digestibility in a variety of substrates. The experiments were conducted with Italian ryegrass (Lolium multiflorum Lam.), tall fescue (Festuca arundinacea Schred.), and oat (Avena sativa L.) in laboratory silo (1 L) for ensiling 30 days. All the treated silages had significantly (p < 0.05) higher in vitro dry matter digestibility (IVDMD), and lower undesirable microorganism populations compared with controls. In Italian ryegrass and oat silages, all the inoculants improved the fermentation quality, evidenced by distinctly (p < 0.05) higher LA contents and ratios of LA to acetic acid, and lower pH, butyric acid and ammonia nitrogen contents. Strains HG24 and commercial inoculant G (Lactobacillus plantarum MTD‐1) performed better than strains M1 and TG1, indicated by obviously (p < 0.05) higher LA contents, LAB numbers, and IVDMD. In tall fescue silage, all the isolated LAB strains (HG24, M1 and TG1) markedly (p < 0.05) increased IVDMD compared with strain G. Strain HG24 is recommended as a good LAB inoculant for ensiling in various forages.     

Interactive effects of molasses by homofermentative and heterofermentative inoculants on fermentation quality,nitrogen fractionation,nutritive value and aerobic stability of wilted alfalfa (Medicago sativa L) silage

The effect of adding molasses (0, UM or 50 g/kg on DM basis, M) and two types of inoculant including homofermentative (HO) and a combination of homofermentative and propionate‐producing bacterial (HOPAB) inoculants on silage fermentation quality, nitrogen fractionation and aerobic stability of pre‐bloom, wilted alfalfa (AS) was determined in laboratory silos. The HOPAB inoculant was more effective than HO in reducing the alfalfa silage pH but increased propionate content in the absence of M (p < 0.05). Inoculation of HOPAB reduced (p < 0.01) acid detergent fibre (ADF) and increased (p < 0.01) lactate to acetate ratio compared with uninoculated AS. Acetate concentration was lower (p < 0.01) in HOPAB‐inoculated than other AS. This difference was more pronounced in M‐added AS (inoculants × M interaction, p = 0.01). Both inoculants reduced (p < 0.01) ammonia‐N content in AS added with M, whereas only HOPAB decreased (p < 0.01) ammonia‐N concentration in silage without M. Inoculants increased (p < 0.01) B₂ fraction in AS with M addition but had no effect on AS without M. Treating silages with HO‐UM increased (p < 0.05) C fraction (acid‐detergent insoluble‐N) but HOPAB decreased C fraction at two levels of M. Treating alfalfa crop with M and HOPAB improved aerobic stability by increasing the concentration of acetate and propionate of AS respectively. Adding M tended (p < 0.10) to increase short‐chain fatty acids (SCFA) and cumulative gas production (CGP). HOPAB alone increased DM disappearance at 24 h post‐incubation and effective degradability assuming outflow rate of 8%/h relative to untreated AS (p < 0.05). It was concluded that adding M had no pronounced effects on AS fermentation quality, but increased aerobic stability. HOPAB‐inoculated AS with no addition of M improved fermentation quality and increased DM degradability compared with HO.     

Effect of field pea (Pisum sativum L.) level on intake,digestion, ruminal fermentation and in vitro gas production in sheep fed maintenance diets

The objective of this study was the characterization of the nutritive value, ruminal fermentation and nutrient digestibility of field pea (Pisum sativum L.) hay (FPH) by‐product through in situ, in vitro and in vivo tests. A FPH diet was administered ad libitum to four sheep (live weight (LW) 38.0 ± 2.0 kg) at four inclusion levels (0, 25, 50 and 75%) over a conventional sorghum grain/oat straw ration in a 4 × 4 Latin square design. The in situ degradability of FPH was 67% dry matter (DM), 71% crude protein and 66% neutral detergent fiber (NDF). The inclusion of FPH did not affect (P ≥ 0.05) the DM (61 ± 0.4 g/LW^0.75/day) and organic matter (OM) intake (56 ± 0.7 g/LW^0.75/day). Total DM digestibility (68 ± 2.5%), OM (70 ± 3%), and NDF (60 ± 3%), were not different (P ≥ 0.05) between control and FPH inclusion diets. The diet had no effect on ruminal pH, ruminal ammonia concentration, total and individual volatile fatty acids. The chemical composition and sheep intake behavior indicate that FPH by‐product is medium quality forage when incorporated in maintenance rations.     

In vitro fermentation characteristics of different carbohydrate sources in two dog breeds (German shepherd and Neapolitan mastiff)

Few studies have been published on the normal intestinal biota of canines unlike the wealth of information regarding livestock animal species. The in vitro gas production technique (IVGPT) including measurements of accumulating gas during fermentation and end‐product determinations allows obtaining a complete picture of microbial activity kinetics. The aim of this study was to study the in vitro fermentation characteristics of different carbohydrate sources using inocula from two dog breeds (German Shepherd and Neapolitan mastiff). Faeces sampled from rectum of two GS and NM adult dogs, fed the same dry food, were used as inocula. The samples, diluted and filtered, were incubated at 39 °C under anaerobic condition with nine substrates different for carbohydrate composition (rice, corn, potato, spelt, pure cellulose, beet pulp, wheat bran, inulin and fructo‐oligosaccharide). Gas production was recorded 17 times using a manual pressure transducer. After 48 h, the fermentation was stopped and fermenting liquor was analysed for pH and volatile fatty acids (VFA). Organic matter digestibility (OMD) was calculated as difference after burning the residuals. OMD, gas production and end‐products were significantly correlated with chemical composition of substrates, in particular carbohydrate fractions (total dietary fibre and starch), confirming the effectiveness of the IVGPT in evaluating dog feeds. Concerning the comparison between breeds significant differences (p < 0.01) were found for OMD, gas production, fermentation kinetic parameters and end‐products, suggesting a different pathway of fermentation and consequently, a different anaerobic population.     

晾晒、添加物及不同温度对象草青贮发酵品质和微生物的影响

华南地区气候潮湿多雨,象草收获时水分含量高,单独青贮发酵品质较差。通过晾晒（水分含量由83.19%降至75.11%）、添加10%玉米粉或2%蔗糖,探究象草在不同温度（20、30和40 ℃）下青贮的发酵特性和微生物组成。研究结果表明：温度对象草各处理的青贮饲料的发酵品质有显著影响。晾晒后原材料的水溶性碳水化合物含量显著降低,青贮后发酵品质变差,尤其在30 ℃,pH达到5.85,乳酸含量只有1.56% DM,乙酸含量为2.10% DM,NH₃-N含量高达25.09% TN。对照和添加物处理的青贮饲料均随青贮温度升高,乙酸含量增加。晾晒后在20和30 ℃时青贮,产气量显著高于40 ℃或未晾晒处理。此外,象草青贮前的细菌多样性较丰富,青贮后主要是肠杆菌属和乳球菌属,尤其是20 ℃青贮的相对丰度较高,且晾晒处理在20和30 ℃青贮肠球菌属的相对丰度高于40 ℃。因此,在华南地区,象草原料中添加玉米粉或蔗糖可以降低环境温度对象草青贮发酵品质的不良影响,阴湿条件下晾晒未发挥积极效果。     

混贮模式对高丹草青贮发酵品质及体外产气动力学特性的影响

为了解决高丹草因水分高造成青贮发酵品质不佳的问题,通过添加不同种类干草(玉米秸秆、小麦秸秆和苜蓿干草)及干草添加量(12.5,25.0,37.5和50.0kg/t)对混贮高丹草营养价值、青贮发酵品质及体外产气动力学特性进行了研究。结果表明,单独青贮高丹草的丁酸含量较高,弗氏评分等级仅为"可",添加干草混贮可显著提高青贮高丹草的发酵品质,从添加干草的种类来看,添加小麦秸秆组青贮发酵品质最高,添加苜蓿干草组营养价值最高,苜蓿干草组的体外72h干物质消失率(IVDMD)、产气速率(c)和达到最大产气量1/2时的产气速率(AGPR)均为最高,3种干草在72h累积产气量、理论最大产气量以及产气延滞时期方面差异不显著(P0.05);从干草的添加量来看,添加25.0kg/t干草的青贮发酵品质最优,达到产气量1/2所需要时间也最长,添加50.0kg/t干草的营养价值和IVDMD最高,添加37.5kg/t干草的产气速率和AGPR最大,添加不同重量干草对混贮高丹草的72h累积产气量、理论最大产气量以及产气延滞时期无显著影响(P0.05)。综合考虑青贮发酵品质和饲料营养价值,得出最佳的混贮模式为在高丹草中添加37.5kg/t小麦秸秆,添加50.0kg/t苜蓿干草混贮高丹草的体外干物质消失率最高,添加37.5kg/t苜蓿干草组产气速率最快,添加50.0kg/t小麦秸秆组的72h累积产气量和理论最大产气量最高。     

不同处理对王草青贮品质及其体外消化率的影响

 试验以热研4号王草为青贮原料,研究不同处理对王草青贮饲料发酵品质和体外消化率的影响,共设置鲜草直接青贮(C)、半干青贮(W)、添加粗饲料降解剂(3%,6%)(BA)、甲酸(FA)、蔗糖(S)等6个处理。结果表明：王草直接青贮品质不高;晾晒和添加粗饲料降解剂、甲酸、蔗糖等处理能显著降低pH值和提高乳酸含量,两个水平的粗饲料降解剂无显著差异;添加蔗糖处理能极显著降低pH值、提高乳酸含量和体外消化率。综合考虑,晾晒和添加剂处理能明显改善青贮品质和体外消化率,其中半干王草添加2%蔗糖效果最好。     

Effects of grass hay proportion in a corn silage‐based diet on rumen digesta kinetics and digestibility in dairy cows

In this study, we aimed to evaluate the effects of six levels of orchardgrass hay (GH) proportion (0%, 10%, 20%, 30%, 40% or 50% of dry matter) in finely chopped corn silage (CS)‐based diets on digesta kinetics of CS and GH in the rumen. Six non‐lactating, rumen‐cannulated Holstein cows were used in a 6 × 6 Latin square design. Ruminal digesta kinetics was measured by ruminal dosing of feed particle markers (dysprosium for CS, erbium for GH) followed by fecal sampling. The increase of GH proportion had a quadratic effect (P < 0.01) on total tract digestibility of neutral detergent fiber (NDF) and acid detergent fiber. The proportion of GH did not affect the particle size distribution of rumen digesta, total weight of dry matter or NDF in the rumen. The rates of large particle size reduction in the rumen for CS tended to increase linearly with increasing GH proportion (P = 0.077). A quadratic effect (P < 0.05) was found with increasing the GH proportion for the ruminal passage rate of small GH particles, but not for CS particles. The results suggested that associative effects between CS and GH could be generated on rumen digesta kinetics when cows were fed a CS‐based diet with an increased proportion of GH.     

Digestibility of common reed (Pharagmites communis Trin.) silage as ruminant feed and effects of inclusion levels in the diet of breeding cows on feed intake,ruminal fermentation and blood metabolites

To examine the digestibility of common reed (Pharagmites communis Trin.) silage and effects of inclusion levels in the diet of breeding cows on nutrient intakes, ruminal fermentation and nutrient status, two digestion trials using four sheep and cows, respectively, at the maintenance stage were conducted. In the trial using sheep, common reed silage contained 20% crude protein (CP) and 50% total digestible nutrients (TDN) on a dry matter basis. In the trial using cows, treatments were the control (92.4% Sudangrass hay and 7.6% soybean meal) and CS25, CS50 and CS80 (replacing 25%, 50%, and 80% of the control with silage, respectively). The intake and sufficiency rate of CP increased by increasing the silage level in the diet (P < 0.05), and the CP requirements of cows at maintenance was satisfied in CS80. In contrast, the sufficiency rate of TDN decreased in CS80, although it was 99.8%. Ruminal pH and NH₃‐N concentration changed among the treatments (P < 0.05); however, the blood metabolites were not affected by silage. The results indicated that including common reed silage of up to 80% in the diet of breeding cows was possible and did not have adverse effects on the blood metabolites.     

酶制剂对水稻秸秆青贮发酵品质及体外消化特性的影响

为探讨纤维素酶、木聚糖酶及两种酶组合添加对水稻秸秆青贮过程中结构性、水溶性碳水化合物组分含量及体外消化特性和发酵品质的影响,试验设4个处理组:1)0.3%蒸馏水(对照组,CO);2)0.3%纤维素酶(CE);3)0.3%木聚糖酶(XE);4)0.15%纤维素酶+0.15%木聚糖酶(组合酶组,CX),分别于青贮3、7、14、30 d后取样分析。结果表明,与CO相比,CE、XE和CX组显著提高了乳酸、葡萄糖、果糖和蔗糖含量,显著降低了pH值、氨态氮、总挥发性脂肪酸、酸性洗涤纤维、中性洗涤纤维、纤维素和半纤维素含量,减少了干物质损失(P<0.05)。酶制剂显著提高了水稻秸秆青贮饲料24、48和72 h时的累积产气量和干物质体外消化率(P<0.05)。青贮末期CX组有最高的乳酸含量(34.13 g·kg^-1DM)、体外产气量(68.27 mL)、干物质体外消化率(61.31%)和最低的pH值(4.36)。与CE和XE相比,CX组水溶性碳水化合物(葡萄糖、果糖和蔗糖)含量更高。综上所述,添加酶制剂可促进结构性碳水化合物的降解,提高水溶性碳水化合物的含量,改善水稻...     

大豆油不同添加水平对瘤胃体外发酵和产气的影响

本试验通过体外培养,研究在精粗比为30:70(偏粗型)日粮类型条件下,不同大豆油添加水平对产气量、甲烷产量、pH值和纤维降解率的影响.结果表明:随着大豆油添加水平的提高,产气量和甲烷产量逐渐降低,且各添加组显著低于对照组;pH值随着培养时间的延长,各添加水平在不同培养时间点与对照组相比均上升,其中添加量为4%和6%与对照组相比差异显著(P<0.05),添加量为8%时,与对照组相比差异极显著(P<0.01);培养24 h后.纤维降解率的抑制潜力8%水平最大,与对照组相比差异极显著(P<0.01),其次是6%的添加水平.与对照组相比差异显著(P<0.05).而2%和4%添加水平对纤维降解率的抑制程度与对照组相比差异不显著(P>0.05).通过产气量、pH值和纤维降解率综合考察大豆油对瘤胃体外发酵和产气的影响,本试验得出大豆油的最适添加量为4%.     

体外产气法评价青海高原燕麦青干草与天然牧草组合效应

本研究采用体外产气法评价了青海高原反刍家畜补饲用燕麦青干草分别与藏嵩草、金露梅+珠芽蓼、线叶嵩草3种天然牧草均按0∶100,25∶75,50∶50,75∶25,100∶0的比例两两组合后的发酵产气特性。结果表明,48 h产气量、理论最大产气量与NDS(P＜0.01)呈正相关关系,而与NDF(P＜0.01)、HC(P＜0.01)呈负相关关系;产气速率常数与HC(P＜0.01)呈负相关关系;产气延滞时间与ADF(P＜0.001)、NDF(P＜0.05)呈正相关关系,而与CP(P＜0.01)、NDS(P＜0.05)呈负相关关系。不同牧草组合在发酵12～24 h的正组合效应较高,且随着发酵时间的延长,各组合均呈组合效应量逐渐减弱的变化趋势;且以燕麦青干草作为补饲草进行该地区天然草地反刍家畜的冷季营养补饲时,燕麦青干草分别与藏嵩草、金露梅+珠芽蓼、线叶嵩草草地型牧草均以50∶50比例组合较为合适。因此,做好牧草间的科学搭配组合是提高青海高原天然草地冷季牧草利用率的有效途径之一。     

Effects of supplementary mother liquor,by‐product of monosodium glutamate,on in vitro ruminal fermentation characteristics

Mother liquor (ML) is monosodium glutamate by‐product and contains much crude protein (CP). The objective of this study was to investigate the effect of supplementation of two types of MLs having different chemical compositions on in vitro ruminal fermentation characteristics. ML1 had 74% organic matter (OM) and 70% CP and ML2 had 85% OM and 83% CP on a dry matter (DM) basis. The experiments were conducted using 0.5 gDM rolled barley or ryegrass straw as substrates. Urea and freeze‐dried MLs were added at three levels of nitrogen to be isonitrogenous. The ML1 treatment increased gas production compared with the ML2 treatment in barley substrate condition (p < 0.05). The supplementary ML2 increased gas production compared with the supplementary urea, and the digestibilities of DM and neutral detergent fiber expressed exclusive of residual ash (NDFom) were higher for the supplementary ML2 than for the supplementary ML1 in straw substrate condition (p < 0.05). In both the conditions, ammonia nitrogen concentrations of the ML1 and ML2 treatments were lower than that of the urea treatment (p < 0.05). The results indicated that the supplementary ML1 and ML2 activated in vitro ruminal fermentation particularly in concentrate and roughage substrate conditions respectively.