Influence of direct-fed fibrolytic enzymes on diet digestibility and ruminal activity in sheep fed a grass hay-based diet

Six rumen-fistulated Merino sheep were used in a crossover design experiment to evaluate the effects of an exogenous fibrolytic enzyme preparation (12 g/d; ENZ), delivered directly into the rumen, on diet digestibility, ruminal fermentation, and microbial protein synthesis. The enzyme contained endoglucanase and xylanase activities. Sheep were fed a mixed grass hay:concentrate (70:30; DM basis) diet at a daily rate of 46.1 g/kg of BW(0.75). Samples of grass hay were incubated in situ in the rumen of each sheep to measure DM and NDF degradation. The supplementation with ENZ did not affect diet digestibility (P = 0.30 to 0.66), urinary excretion of purine derivatives (P = 0.34), ruminal pH (P = 0.46), or concentrations of NH(3)-N (P = 0.69) and total VFA (P = 0.97). In contrast, molar proportion of propionate were greater (P = 0.001) and acetate:propionate ratio was lower (P < 0.001) in ENZ-supplemented sheep. In addition, ENZ supplementation tended to increase (P = 0.06) numbers of cellulolytic bacteria at 4 h after feeding. Both the ruminally insoluble potentially degradable fraction of grass hay DM and its fractional rate of degradation were increased (P = 0.002 and 0.05, respectively) by ENZ treatment. Supplementation with ENZ also increased (P = 0.01 to 0.02) effective and potential degradability of grass hay DM and NDF. Ruminal fluid endoglucanase and xylanase activities were greater (P < 0.001 and 0.03, respectively) in ENZ-supplemented sheep than in control animals. It was found that ENZ supplementation did not affect either exoglucanase (P = 0.12) or amylase (P = 0.83) activity. The results indicate that supplementing ENZ directly into the rumen increased the fibrolytic activity and stimulated the growth of cellulolytic bacteria without a prefeeding feed-enzyme interaction.     

添加外源纤维水解酶对羊草体外瘤胃发酵酶活性及特性的影响

试验通过分别添加纤维素酶与木聚糖酶0,10.0与50.0 mg水平,0与24 h两个预处理时间,每个处理3个重复,体外法评价纤维素酶与木聚糖酶复合处理羊草后与瘤胃液共培养对木聚糖酶与葡聚糖酶活性及发酵特性的影响。结果表明,添加纤维水解酶能提高0与8 h培养液中木聚糖酶和内切葡聚糖酶活性以及0 h培养液中外切葡聚糖酶活性,且具有添加剂量效应。当培养至24及48 h,添加外源酶制剂并不能提高培养液中相关酶活性。外源酶制剂显著增加培养24与48 h发酵液中乙酸含量,8,24与48 h 总挥发性脂肪酸产量 (P<0.05)以及48 h累积产气量(P<0.05),但对培养期内戊酸与异戊酸含量没有影响(P＞0.05)。结果提示添加外源酶制剂能提高早期培养液中木聚糖酶和葡聚糖酶活性、增加VFA产量和改善体外瘤胃发酵特性。     

Screening of exogenous enzymes for ruminant diets: relationship between biochemical characteristics and in vitro ruminal degradation

With the objective of developing a rational approach for the selection of feed enzymes for ruminants, 22 commercial enzyme products were examined in terms of protein concentration, enzymic activities on model substrates, and hydrolytic capacity, the latter determined from the release of reducing sugars from alfalfa hay and corn silage. An in vitro ruminal degradation assessment was carried out using the same substrates, untreated or treated with the 22 enzyme products at 1.5 microL/g forage DM. Stepwise regressions were then performed to establish relationships between these factors. Protein concentration and enzymic activities explained at least 84% (P < 0.01) of the variation in the release of reducing sugars from alfalfa and corn silage. Alfalfa DM degradation after incubation with ruminal fluid for 18 h was positively related to xylanase activity (R2 = 0.29, P < 0.01), but the same activity was negatively related to DM degradation of corn silage (R2 = 0.19, P < 0.05). Protease activity explained a further 10% of the alfalfa DM degradation (P < 0.10). Following sequential steps involving the determination of rate and extent of DM and fiber degradation, the best candidates for alfalfa and corn silage were selected. Enzyme products effective with alfalfa hay seemed to exert part of their effect during the pretreatment period, whereas enzymes effective with corn silage worked exclusively after ruminal fluid was added. This finding suggests that different modes of action of exogenous enzymes are attacking different substrates and may partly explain enzyme-feed specificity. In alfalfa, it seems that effective enzymes work by removing structural barriers that retard the microbial colonization of digestible fractions, increasing the rate of degradation. In corn silage, effective enzymes seem to interact with ruminal enzymes to degrade the forage more rapidly, which is consistent with previous findings of synergism between exogenous and ruminal enzymes.     

Influence of supplementary fibrolytic enzymes on the fermentation of corn and grass silages by mixed ruminal microorganisms in vitro

This study was done to determine the effectiveness of supplementary enzymes at increasing the fiber digestion by ruminal microorganisms and to assess whether enzyme activity limits the rate of fiber digestion in ruminal digesta. In vitro comparisons of enzyme activities in two feed enzyme preparations (A and B) with enzyme activities extracted from ruminal fluid indicated that the addition of fibrolytic enzymes at the application rates recommended by the manufacturers would not be expected to increase significantly glycanase and polysaccharidase activities in ruminal fluid. Preparations A and B both increased (P < 0.001) the rate of gas production from freeze-dried corn and grass silages in in vitro incubations with ruminal fluid, but only at concentrations much higher than recommended application rates. Autoclaved controls had little or no effect. Ultrafiltration of enzyme B indicated that most stimulation was due to components >100 kDa, which is consistent with the cause of the stimulation being enzyme activity. Fibrolytic enzymes from other sources were also able to stimulate gas production: increased rates of gas production were observed in seven out of eight combinations of "cellulase" and corn or grass silage (P < 0.05). The comparison of glycanase and polysaccharidase activities with gas-stimulatory activity in the different enzyme preparations indicated that the highest correlation was between increased gas production and enzyme activity against microgranular cellulose (P < 0.05). In a wider range of fibrolytic enzyme preparations, those with endo-(beta-1,4)- or exo-(beta-1,4)-xylanase activity equal to that of preparation A did not produce similar increased rates of fermentation of corn silage when glucanase activity was low (P > 0.05). In contrast, preparations with glucanase activity similar to enzyme A gave at least as great (P < 0.05) an improvement in gas production than enzyme A, irrespective of xylanase activity. It was concluded that enzyme activity, probably a type of endo-(beta-1,4)-glucanase activity, limits the rate of fermentation of corn and grass silage in the rumen. Enzyme supplements of the type used in these experiments are unlikely to possess sufficient activity to overcome this limitation by direct application to ruminal digesta, implying that treatment of the ration prefeeding will be key to harnessing the potential of exogenous fibrolytic enzymes in ruminant nutrition.     

Effect of alkali pretreatment of wheat straw on the efficacy of exogenous fibrolytic enzymes

The effects of pretreating wheat straw with alkali on the efficacy of exogenous fibrolytic enzymes for improving straw digestibility were studied in vitro, in situ, and in vivo. In Exp. 1, untreated straw (US); alkali-treated (5% NaOH, wt/wt) straw (AS); and autoclaved, alkali-treated straw (AAS) were sprayed with 0 or 1.5 mg/g DM of enzyme mix (xylanase, beta-glucanase, carboxymethylcellulase, and amylase) and incubated for 30 h in buffered ruminal fluid (3 x 2 factorial arrangement). Enzymes increased (P < 0.001) gas production and the incorporation of 15N into microbial N at 4 h, more so with AS or AAS than with US (P < 0.001 for gas; P < 0.05 for 15N). In Exp. 2, US and AS were sprayed with enzymes at 0, 0.15, or 1.5 mg/g DM (2 x 3 factorial) and incubated ruminally in nylon bags for up to 80 h to determine the in situ DM disappearance (ISDMD). Interactive effects (P < 0.05) of pretreatment and enzymes were observed on all ruminal degradation parameters. Alkali increased the rate (P < 0.01) and extent (P < 0.001) of ISDMD irrespective of enzymes. Enzymes applied to US did not affect the extent of ISDMD, but they increased (P < 0.01) the extent of ISDMD when applied to AS. Substrates from Exp. 1 and 2 were incubated in acetate buffer for 24 h to measure the hydrolytic loss of DM and release of reducing sugars and phenolic compounds. Alkali pretreatment and enzymes each increased (P < 0.001) DM loss and the release of reducing sugars and, in combination, exerted synergistic effects (P < 0.001). Enzymes did not affect the release of phenolic compounds from the straw. In Exp. 3, total-tract digestibility of untreated and enzyme-treated (100 mL/kg DM) ammoniated straw was assessed using 32 beef cows in eight pens. Wrapped straw bales were injected with NH3 (3% [wt/wt], DM basis) 4 mo before the study; enzymes were applied immediately before feeding. Applying enzyme to ammoniated straw increased (P < 0.05) digestibilities of DM, OM, and total N but did not affect the intake of DM or digestibility of ADF. Pretreatment of straw with alkali enhanced the efficacy of exogenous enzymes, presumably by breaking esterified bonds and releasing phenolic compounds and/or by swelling the crystalline cellulose and enhancing enzyme penetration. Including enzymes that mimic alkali hydrolysis (e.g., esterases) in commercial feed additives could substantially improve the value of these products for ruminants.     

Intraruminal supplementation with increasing levels of exogenous polysaccharide-degrading enzymes: effects on nutrient digestion in cattle fed a barley grain diet

The effects of supplying increasing ruminal doses of exogenous polysaccharide-degrading enzymes (EPDE) on rumen fermentation and nutrient digestion were studied using eight ruminally cannulated heifers, four of which were also duodenally cannulated, in a replicated Latin square. The heifers were fed a diet of 85.5% rolled barley grain and 14% barley silage (DM basis), and once daily they were given intraruminal doses of 0 (Control), 100, 200, or 400 g of a preparation containing polysaccharide-degrading enzymes. Enzyme treatment decreased ruminal pH (linear, P<.001) and increased ammonia N (quadratic, P<.001) concentration. The ruminally soluble fraction and effective degradability of feed DM in situ were increased (quadratic response, P<.001) by enzyme treatment. Ruminal administration of EPDE increased ruminal fluid carboxymethylcellulase and xylanase activities linearly (P<.001) and beta-glucanase activity quadratically (P<.01), decreased (quadratic response, P<.05) ruminal fluid viscosity, and did not affect (P>.05) ruminal fluid amylase activity. Elevated levels of fibrolytic activities in the rumen resulted in increased (quadratic, P<.001) carboxymethylcellulase, xylanase, and beta-glucanase (P<.01) activities in duodenal digesta. Duodenal amylase activity and reducing sugar concentration were also increased (quadratic responses, P<.001 and P<.05, respectively) by EPDE. Xylanase activity of fecal DM was increased linearly (P<.05) with increasing ruminal EPDE levels. Apparent digestibilities of DM, crude protein, and NDF were not affected by EPDE supplementation. Enzyme treatment did not affect (P>.05) urinary excretion of allantoin and uric acid, or concentrations of glucose and urea in blood.     

Resistance of feed enzymes to proteolytic inactivation by rumen microorganisms and gastrointestinal proteases.

Potential feed enzyme additives for ruminants were tested in vitro for their stability to ruminal microbial and gastrointestinal proteolysis. Four commercial preparations from Trichoderma longibrachiatum (A, B, C, and D) and one from an undisclosed source (E) were incubated up to 6 h with ruminal fluid taken from four lactating dairy cows before or 2 h after feeding. The stability of preparation B was also tested in the presence of pepsin at pH 3 and pancreatin at pH 7. Cellulase (EC 3.2.1.4), cellulose 1,4-beta-cellobiosidase (EC 3.2.1.91), beta-glucanase (EC 3.2.1.6), xylanase (EC 3.2.1.8), beta-glucosidase (EC 3.2.1.21), and beta-xylosidase (EC 3.2.1.37) activities were monitored throughout the incubations. Polysaccharidase activities of all enzyme preparations were remarkably stable in ruminal fluid taken after feeding. Ruminal fluid obtained before feeding inactivated the polysaccharidases in preparations B and D to a greater extent than ruminal fluid obtained after feeding. Cellulase and cellulose 1,4-beta-cellobiosidase activities were the least stable, declining (P < 0.05) by 35 and 60% for preparations B and D, respectively. Xylanase activity of preparation D decreased (P < 0.05) by up to 30% after 6 h of incubation, whereas beta-glucanase activity was not affected. The ability to degrade exogenous enzymes also differed among cows (P < 0.05). Pepsin and acid (pH 3.0) did not affect polysaccharidases in preparation B but decreased glycosidase activities by 10 to 15% (P < 0.05) after 1 h of incubation. Pancreatin, at the maximum concentration used, inactivated cellulase, cellulose 1,4-beta-cellobiosidase, and xylanase activities at a rate of 0.55, 1, and 0.45%/min, respectively. beta-Glucosidase and beta-xylosidase activities decreased by 1 and 0.75%/min, respectively. Partial proteolysis of cellulase, cellulose 1,4-beta-cellobiosidase, and xylanase by pancreatin produced a transient increase in activity. This twofold increase for cellulase and fourfold increase for cellulose 1,4-beta-cellobiosidase was directly proportional to pancreatin concentration. These results suggest that the enzyme feed additives tested were stable in the rumen of animals after feeding. Exogenous enzymes are likely to be more susceptible to the host gastrointestinal proteases in the abomasum and intestines than to ruminal proteases. However, exogenous polysaccharidases may survive for a considerable period of time in the small intestine and they probably maintain activity against target substrates in this environment.     

毛茛科植物提取物添加量对瘤胃微生物体外动态发酵的影响

试验采用活体外产气量法研究毛茛科植物提取物不同添加水平对瘤胃微生物体外发酵的影响。试验采用单因子完全随机化试验设计,以小麦面粉为底物进行体外发酵。瘤胃液供体动物为3头装有永久性瘤胃瘘管的本地黄牛,日粮精料水平为30%(日饲喂2次)。植物提取物添加水平分别为0、100、200、300和400mg/L。结果表明:随着毛茛科植物提取物添加水平的提高,小麦面粉24h的DM消化率呈线性下降(P=0.0002),活体外产气速度呈二次曲线规律降低(P=0.0001),而产气延滞期线性增加(P=0.0001),理论最大产气量呈二次曲线规律增加(P=0.0001)。随着植物提取物添加水平的提高,发酵液pH值呈二次曲线增加(P<0.024)。各培养时间点乳酸含量均较低(P<1mmol/L),且各处理之间差异不显著(P>0.10)。提高植物提取物添加水平导致tVFA产量呈二次曲线趋势下降(P<0.01)。通过本试验可得出:毛茛科植物提取物可以通过降低瘤胃发酵的产气速度、提高发酵液pH值以及乙酸和丁酸摩尔比例来调控瘤胃微生物体外发酵,其中植物提取物的适宜添加水平为200～300mg/L。     

微生物接种剂和酶制剂对玉米秸组成成分和瘤胃有效降解率的影响

通过玉米秸发酵试验及半体内法 ,评价了复合产乳酸菌 (植物乳杆菌 保加利亚乳杆菌 粪尿链球菌 嗜酸乳杆菌 )、酶制剂 (纤维素酶 淀粉酶 糖化酶 )、液体发酵剂 (益生菌 )单独或联合应用对玉米秸组成成分及瘤胃有效降解率的影响。结果表明 ,从整个发酵期来看 ,高剂量固体发酵剂组 (每吨秸杆添加复合产乳酸菌 2 0 g 纤维素酶 10 0 g 淀粉酶 30 0 g 糖化酶 30 0 g)、高剂量液体发酵剂组 (每吨秸杆添加益生菌 5 0 0 m L )、中性洗涤纤维 (NDF)含量略低于对照组 (P <0 .0 5 ) ;高剂量的固体发酵剂和液体发酵剂明显提高了玉米秸快速降解的干物质 (DM) (P <0 .0 5 )、NDF成分 (P <0 .0 1) ,显著降低了慢降解的 DM、NDF成分 (P <0 .0 1) ;从有效降解率来看 ,高剂量液体发酵剂组的DM有效降解率明显高于对照组及纤维素酶组 (P <0 .0 5 ) ,NDF有效降解率则比其他各组均有明显提高 (P <0 .0 5 )。     

Effect of exogenous xylanase on rumen in vitro gas production and degradability of wheat straw

The objective of this study was to determine effects of xylanase on in vitro gas production (GP) and in sacco degradability of wheat straw. Rumen fluid was obtained from three Mongolian native goats fitted with permanent rumen cannulas. The trial consisted of five doses (0, 0.5, 1.0, 1.5, 2.0 μL/g of substrate) of a commercial xylanase (Dyadic^® xylanase PLUS, Dyadic International, Inc., Jupiter, FL, USA). For the in sacco degradability, different levels of xylanase enzyme were added directly onto 2 g of wheat straw in nylon bags and incubated in the rumen for 3, 6, 12, 24 and 48 h to estimate degradability of wheat straw. Total GP increased (P < 0.001) at all times of incubation at intermediate levels of xylanase. Methane production had a similar pattern at 3 and 12 h of incubation; increased linearly at 24 h of incubation, and was unaffected at 6 and 48 h of incubation. Rumen NH₃‐N concentration increased linearly at 3 h and the highest values were observed with intermediate enzyme levels. All ruminal volatile fatty acids increased linearly with intermediate levels of the fibrolytic enzyme. The in sacco rate of dry matter degradation decreased linearly (P = 0.020) with increasing enzymes. Intermediate levels of xylanase improved rumen kinetic fermentation and degradability. The outcome of this research indicated that the application of xylanase enzyme could improve in vitro GP fermentation of wheat straw.     

不同蛋白和中性洗涤纤维水平对瘤胃发酵、消化和微生物蛋白合成的影响

以3头装有瘤胃瘘管的干奶牛作为瘤胃液供体,用体外培养法研究不同日粮蛋白和中性洗涤纤维(NDF)水平对瘤胃发酵、消化和微生物蛋白合成的影响。采用2×3双因子完全随机设计,粗蛋白水平14%、16%和18%,NDF水平为32%和36%。体外发酵后2、4、6、12、24 h取瘤胃液测定pH、NH3-N浓度和微生物蛋白含量,发酵结束后测定饲料中干物质(DM)、NDF和酸性洗涤纤维(ADF)消化率。结果表明:随着蛋白水平升高,瘤胃发酵pH有增加趋势(P=0.079),显著提高NDF和ADF消化率和细菌蛋白合成量(P<0.05),但不影响DM消化率和原虫蛋白的合成量(P>0.05);日粮中NDF水平显著影响瘤胃pH、NH3-N浓度和ADF消化率(P<0.05),但不影响DM和NDF消化率以及微生物蛋白的合成量(P>0.05)。本试验条件下,日粮中16%CP和32%NDF组可在一定程度上促进瘤胃发酵,提高纤维消化率以及微生物蛋白的合成量。     

Effects of exogenous cellulase supplementation on microbial growth and ruminal fermentation of a high-forage diet in Rusitec fermenters

Two incubation runs were carried out with a Rusitec system to investigate the effects of 2 exogenous pure cellulases on ruminal microbial growth and fermentation of a 70:30 grass hay:concentrate (DM basis) substrate. The substrate was sprayed with buffer (control; pH = 6.5), a cellulase from Trichoderma longibrachiatum (TRI), a cellulase from Aspergillus niger (ASP), or a 1:1 mixture of both cellulases (MIX) 24 h before being placed in the fermenters. Enzymes were applied at a rate of 30 endoglucanase units/g of substrate DM. Treating the substrate with enzymes reduced substrate NDF and ADF content (P < 0.001 to P = 0.002) and increased DM, NDF, and ADF disappearance after 6 and 24 h of incubation (P < 0.001 to P = 0.004) but not after 48 h of incubation. Daily VFA production was increased (P = 0.004) by 15, 9, and 15% for TRI, ASP, and MIX, respectively, with half of the increase being due to production of acetate. All enzyme treatments augmented (P = 0.009) methane production, but none of them altered the methane:VFA ratio (P = 0.70). There were no differences (P = 0.80) among treatments in the daily flow of solid-associated microorganisms, as measured using 15N as a microbial marker. Although the TRI and MIX treatments increased (P < 0.05) the daily flow of liquid-associated microorganisms and the proportion of microbial N in the solid residue after 48 h of incubation, no effects were observed (P = 0.92 and P = 0.95, respectively) for the ASP treatment. The results show that the TRI and MIX treatments enhanced in vitro fermentation by increasing substrate fiber degradation, VFA production, and ruminal microbial growth. The lack of differences between TRI and MIX in most of the measured variables indicates that treating the substrate with a mixture of both cellulases did not further improve the effects of the TRI treatment.     

不同来源木聚糖酶及其组合对木聚糖水解产物组成、细菌增殖与大肠杆菌黏附性的影响

本试验旨在研究不同来源木聚糖酶及其组合对木聚糖的水解效果,并研究不同木聚糖水解产物对细菌增殖及大肠杆菌对肠道上皮细胞黏附性的影响。试验用木聚糖酶A和木聚糖酶B分别来源于毕赤酵母和米曲霉。采用木聚糖酶A、木聚糖酶B、组合酶1(木聚糖酶A∶木聚糖酶B=3∶7)、组合酶2(木聚糖酶A∶木聚糖酶B=1∶1)、组合酶3(木聚糖酶A∶木聚糖酶B=7∶3)分别水解木聚糖,然后测定木聚糖水解产物对大肠杆菌、枯草芽孢杆菌和乳酸杆菌增殖和大肠杆菌对肠道上皮细胞黏附性的影响。结果表明:1)2种木聚糖酶有一定的组合效应,木聚糖酶A、木聚糖酶B、组合酶1、组合酶2、组合酶3组木二糖和木三糖的总含量分别为95.70%、86.79%、93.11%、94.55%和87.55%,其中木聚糖酶A组木二糖含量最高,组合酶2组木三糖含量最高。2)培养20 h时,5种木聚糖水解产物对大肠杆菌的增殖(以菌液吸光度值表示)没有产生显著影响(P0.05);培养20和30 h时,5种木聚糖水解产物显著促进枯草芽孢杆菌的增殖(P0.05);培养13和17 h时,5种木聚糖水解产物显著促进乳酸杆菌的增殖(P0.05)。3)5种木聚糖水解产物均可以显著降低大肠杆菌对肠道上皮细胞的黏附率(P0.05)。由此可见,通过不同来源木聚糖酶及其组合水解木聚糖,可以产生以木二糖和木三糖为主要组分的水解产物,从而起到促进枯草芽孢杆菌和乳酸杆菌增殖、减少大肠杆菌对肠道上皮细胞黏附的作用。     

没食子酸和三甲胺N-氧化物对体外瘤胃发酵和三甲胺代谢的影响

试验旨在研究没食子酸(GA)和三甲胺N-氧化物(TMAO)对体外瘤胃发酵和三甲胺(TMA)代谢的影响.通过瘤胃体外模拟试验分析瘤胃底物消失率、总产气量、发酵参数和TMA代谢情况,设置对照组、15 mg GA/g DM组、5 mg TMAO/g DM组、5 mg TMAO+15 mg GA/g DM组,每组4个重复,培养...     

不同乳酸菌添加量和发酵时间对全株玉米青贮营养价值及发酵品质的影响

本试验以登海605玉米品种为材料,于蜡熟期2/3乳线时进行刈割,乳酸菌制剂的添加量分别为0,10,20和30 mg·kg^-1,4个处理,每个处理4个重复,在室温条件下发酵45和90 d,取样测定营养成分、发酵品质和瘤胃降解率等指标,旨在研究不同乳酸菌添加量和发酵时间对全株玉米青贮营养价值的影响。结果表明:干物质(DM)受乳酸菌添加量和发酵时间的影响较小,差异不显著(P>0.05);随乳酸菌添加量的增加,发酵90 d的中性洗涤纤维(NDF)含量显著线性降低(P=0.018),而相对饲喂价值(RFV)显著线性增加(P=0.006)。发酵90 d的酸性洗涤纤维(ADF)含量和pH值较45 d显著降低(P<0.01),而乳酸和乙酸含量显著增加(P<0.05)。随乳酸菌添加量的增加,乳酸含量显著线性增加(P<0.05)。24 h的DM和NDF消化率受乳酸菌添加量和发酵时间的影响较小,差异不显著(P>0.05)。发酵90 d时,48 h DM降解率随乳酸菌添加量的增加显著线性增加(P=0.034),48 h NDF降解率较45 d显著增加(P=0.022)。发酵90 d时,20 mg·kg^-1组的RFV、总可消化养分(TDN)和有机酸含量最高,而pH值最低。综上所述,乳酸菌添加量和发酵时间对全株玉米青贮的营养成分含量、发酵品质和DM瘤胃降解率均有显著影响,全株玉米青贮在发酵90 d且乳酸菌添加量为20 mg·kg^-1的营养价值和发酵品质最优。     

Dietary copper effects on lipid metabolism, performance, and ruminal fermentation in finishing steers

Sixty Angus steers (391.1+/-6.1 kg) were used to determine the effects of dietary Cu concentration on lipid metabolism and ruminal fermentation. Steers were stratified by weight and randomly assigned to treatments. Treatments consisted of 0 (control), 10, or 20 mg of supplemental Cu (as CuSO4)/kg diet DM. Steers were housed in pens equipped with individual electronic Calan gate feeders. On d 86 and 92, ruminal fluid was collected from two steers/treatment for IVDMD determination. Equal numbers of steers per treatment were slaughtered after receiving the finishing diets for 96 or 112 d. Gain, feed intake, feed efficiency, IVDMD, and ruminal VFA molar proportions were not affected by Cu supplementation. Copper supplementation increased (P < .05) liver Cu concentrations, and steers supplemented with 20 mg Cu/kg DM had higher (P < .05) liver Cu concentrations than steers supplemented with 10 mg Cu/kg DM. Serum total cholesterol concentrations were reduced by d 56 and at subsequent sampling dates in steers receiving supplemental Cu. Longissimus muscle cholesterol concentrations were lower (P < .10) in steers supplemented with Cu. Backfat depth was less (P < .05) in steers receiving supplemental Cu, but marbling scores were similar across treatments. Unsaturated fatty acid composition of longissimus muscle was increased (P < .05) and saturated fatty acid composition tended (P < .12) to be reduced in Cu-supplemented steers. Polyunsaturated fatty acid concentrations were higher (P < .05) in steers receiving Cu. These results indicate that addition of 10 or 20 mg Cu/kg to a high-concentrate diet containing 4.9 mg Cu/kg DM alters lipid and cholesterol metabolism in steers but does not affect ruminal fermentation.     

Use of fibrolytic enzymes additives to enhance in vitro ruminal fermentation of corn silage

Two experiments were conducted to evaluate the effect of four enzyme additives on ruminal fermentation of corn silage using a 48 h batch culture in vitro assay with buffer and ruminal fluid. Experiment 1 (Exp. 1) and Experiment 2 (Exp. 2) were conducted as completely randomized designs each with two runs and four replicates. The enzyme additives (E1, E2, E3, and E4) were commercial products that provided a range in endoglucanase, exoglucanase, and xylanase activities. For both xylanase (birch wood and oat spelt substrate) and endoglucanase (carboxymethylcellulose substrate), the enzyme products (per ml) were ranked E4>E1>E2>E3. In Exp. 1, the four enzymes were added at 0, 2, 4, and 8 μl/g of corn silage dry matter (DM), whereas in Exp. 2 enzymes were added at 0, 0.5, 1, 2, and 4 μl/g DM. Gas production (GP) was measured at 3, 6, 12, 18, 24, and 48 h after incubation. Disappearance of DM (DMD), neutral detergent fiber (NDFD), and acid detergent fiber (ADFD), and volatile fatty acid concentrations (VFA; total and individual molar proportions) were determined after 24 and 48 h. In Exp. 1, E1 and E2 had higher NDFD and ADFD at 24 and 48 h of incubation (P<0.001) compared with E3 and E4. Increasing dose rate increased NDFD and ADFD for all enzymes (except ADFD for E4 at 48 h), with the optimum dose rate dependant on the enzyme additive (dose×enzyme; P<0.01). There were some treatment effects on DMD and total GP at 24 and 48 h, but these responses were not consistent with responses in NDFD and ADFD. Experiment 2 was conducted to confirm the effects and optimum dose rate of each enzyme additive. In Exp. 2, DMD was not affected by enzyme after 24 and 48 h incubation. There were no enzyme×dose interactions for DMD, NDFD, or ADFD after 24 or 48 h of incubation (except for ADFD at 48 h). After 24 h, DMD, NDFD, and ADFD increased linearly with increasing dose (P<0.05); after 48 h DMD increased linearly, whereas NDFD increased quadratically with increasing enzyme dose (P<0.05). The ADFD increased linearly after 48 h for E3 and E4, but after 48 h ADFD increased quadratically for E1 and E2. Total GP was consistently lowest for E4 at both incubation times (P<0.05). There were no enzyme×dose interactions (P>0.05) for any of the fermentation variables at either 24 or 48 h of incubation in Exp. 2. There were differences amongst the additives for total VFA at 24 and 48 h (P≤0.05); increasing enzyme dose decreased total VFA after 24 h but increased total VFA at 48 h, such that all doses were higher than the control (P<0.001). Overall, the enzyme additives increased NDFD and ADFD of corn silage in vitro; however, E1 and E2 were more effective than E3 or E4. Responses to increasing dose of enzyme were generally linear or curvilinear, and the optimum dose rate differed amongst the products evaluated. Evaluation of the enzymes at 24 and 48 h generally led to the same ranking of the additives, and the degradation of NDF and ADF was more useful in differentiating the enzymes compared with DM and total GP.     

Effects of enzyme supplementation of a total mixed ration on microbial fermentation in continuous culture, maintained at high and low pH

A dual-flow continuous culture system was used to investigate the effects of pH and addition of an enzyme mixture to a total mixed ration (TMR) on fermentation, nutrient digestion, and microbial protein synthesis. A 4 x 4 Latin square design with a factorial arrangement of treatments was used, with four 9-d periods consisting of 6 d for adaptation and 3 d for measurements. Treatments were as follows: 1) high pH with control TMR, 2) high pH with TMR treated with enzyme, 3) low pH with control TMR, and 4) low pH with TMR treated with enzyme. Ranges of pH were 6.0 to 6.6 and 5.4 to 6.0 for high and low, respectively. Fermenters were fed twice daily a TMR consisting of 30% alfalfa hay, 30% corn silage, and 40% rolled corn (DM basis). The silage was milled fresh and the TMR was fed to the fermenters in fresh form (64% DM). The enzyme mixture was a commercial product of almost exclusive protease activity; it was applied daily to the fresh TMR and stored at 4 degrees C for at least 12 h before feeding. Degradability of OM, NDF, ADF, and cellulose was decreased (P < 0.05) by low pH. Hemicellulose and protein degradation were not affected by pH. Enzyme addition increased (P < 0.01) NDF degradability (by 43% and 25% at high and low pH, respectively), largely as a result of an increase in hemicellulose degradation (by 79% and 51% at high and low pH, respectively). This improvement was supported by an increase (P < 0.05) in the xylanase and cellulase activities in the liquid phase of the fermenter contents. Total VFA were decreased (P < 0.05) by low pH, but were not affected by enzyme addition. Total bacterial numbers were increased (P < 0.03) at low pH and tended (P < 0.13) to increase with enzyme addition. Cellulolytic bacteria in effluent fluid were decreased (P < 0.02) at low pH but were unaffected by enzyme addition. Despite a large increase (P < 0.001) in protease activity, protein degradation was only numerically increased by enzyme addition. Microbial protein synthesis was higher (P < 0.10) at high pH but was not affected by enzyme addition. Methane production, expressed as a proportion of total gases, was decreased (P < 0.001) at low pH but was not affected by enzyme addition. It is concluded that it is possible to adapt the continuous culture system to use fresh feeds instead of dried feeds. Overall, the results indicate that the enzyme product used in this study has a potential to increase fiber degradability without increasing methane production.     

Influence of cobalt concentration on vitamin B12 production and fermentation of mixed ruminal microorganisms grown in continuous culture flow-through fermentors

An experiment was conducted to determine the effects of dietary concentrations of Co on vitamin B12 production and fermentation of mixed ruminal microbes grown in continuous culture fermentors. Four fermentors were fed 14 g of DM/d. The DM consisted of a corn and cottonseed hull-based diet with Co supplemented as CoCO3. Dietary treatments were 1) control (containing 0.05 mg of Co/kg of DM), 2) 0.05 mg of supplemental Co/kg of DM, 3) 0.10 mg of supplemental Co/kg of DM, and 4) 1.0 mg of supplemental Co/kg of DM. After a 3-d adjustment period, fermentors were sampled over a 3-d sampling period. This process was repeated 2 additional times for a total of 3 runs. Ruminal fluid vitamin B12 concentrations were affected by Co supplementation (P < 0.01), and there was a treatment x day interaction (P < 0.01). By sampling d 3, cultures fed the basal diet supplemented with 0.10 mg of Co/kg had greater (P < 0.05) vitamin B12 concentrations than those supplemented with 0.05 mg of Co/kg of DM, and increasing supplemental Co from 0.10 to 1.0 mg/kg of DM increased (P < 0.01) ruminal fluid vitamin B12 concentration. Ruminal fluid succinate also was affected (P < 0.10) by a treatment x day interaction. Cobalt supplementation to the control diet greatly decreased (P < 0.05) succinate in ruminal cultures on sampling d 3 but not on d 1 or 2. Molar proportions of acetate, propionate, and isobutyrate, and acetate:propionate were not affected by the addition of supplemental Co to the basal diet. However, molar proportions of butyrate, valerate, and isovalerate increased (P < 0.05) in response to supplemental Co. The majority of long-chain fatty acids observed in this study were not affected by Co supplementation. However, percentages of C18:0 fatty acids in ruminal cultures tended (P < 0.10) to be greater for Co-supplemented diets relative to the control. Methane, ammonia, and pH were not greatly affected by Co supplementation. The results indicate that a total (diet plus supplemental) Co concentration of 0.10 to 0.15 mg/kg of dietary DM resulted in adequate vitamin B12 production to meet the requirements of ruminal microorganisms fed a high-concentrate diet in continuous-flow fermentors.     

In vitro effects of the ionophore lysocellin on ruminal fermentation and microbial populations.

Batch and continuous culture techniques were used to evaluate the effect of the ionophore lysocellin on ruminal fermentation and microbial populations. In batch culture, .5 and 1 ppm (of the fluid) lysocellin markedly decreased (P less than .01) the acetate:propionate ratio without affecting fiber digestion, ammonia concentration, or culture pH. Greater concentrations of lysocellin had negative effects (P less than .05) on fiber digestion and increased (P less than .05) culture pH. In continuous culture, a low level of lysocellin (33 ppm of the diet DM or about .7 ppm of the fluid) decreased pH (P less than .05) and methane (P less than .05) production but had no effect on fiber digestion. Lysocellin tended to increase (P less than .05) OM digestion in corn-based diets but decreased OM digestion in barley-based diets (starch source x lysocellin interaction, P less than .05). In addition, the molar proportion of propionate was increased more in barley- than in corn-based diets. Total anaerobes and amylolytic and lactate-utilizing microorganisms were not affected by the ionophore. In continuous culture, cellulolytic and lactate-producing organisms were insensitive to lysocellin, but, in lysocellin-treated media, cellulolytic organisms were resistant, whereas lactic acid producers were sensitive to lysocellin at 4 ppm. In summary, the ionophore lysocellin alters ruminal fermentation by decreasing ruminal methane production and increasing the molar proportion of propionate; however, effects varied depending on whether corn or barley served as the primary starch source.