16S rRNA定量分析技术对绵羊瘤胃固相纤维降解菌的研究

本试验选择3只安装永久性瘤胃瘘管、体重25￣30kg的羯羊,使用特异性16SrRNA目标探针进行斑点杂交研究绵羊瘤胃固相纤维降解菌的动态变化规律。结果表明:3种纤维分解菌的RNA占总菌数RNA的15.76%;在各时间点产琥珀酸拟杆菌的相对含量均高于黄化瘤胃球菌和白色瘤胃球菌相对含量。     

Detection and identification of rumen bacteria constituting a fibrolytic consortium dominated by Fibrobacter succinogenes

A fibrolytic consortium, dominated by the rumen cellulolytic bacterium Fibrobacter succinogenes, was artificially constructed on hay stems to detect and identify rumen bacteria that can potentially interact with F. succinogenes . Consortium-bacterial members were determined by DGGE and sequencing analysis targeted bacterial 16S rDNA. An artificial consortium was formed in a 2-step incubation of hay stems; the first step with group 1, 2 or 3 F. succinogenes strains, the second step with rumen fluid. After consortium formation, morphologically different bacteria were observed in association with F. succinogenes . DGGE exhibited more than 30 bands, the pattern of which depended on the F. succinogenes group. Sequencing suggested that Butyrivibrio fibrisolvens, Pseudobutyrivibrio ruminis , Clostridium sp., F. succinogenes group 2, Prevotella ruminicola and unclassified Bacteroides were prominent in the group 1 consortium and that Treponema bryantii , B. fibrisolvens , Acinetobacter sp, and Wolinella succinogenes were prominent in the group 2 consortium. However, in the group 3 consortium, F. succinogenes -like bacteria were microscopically undetectable, whereas cellulolytic Ruminococcus albus and F. succinogenes group 1 were prominent, suggesting that the group 3 cannot be a core member of this consortium. This study is the first attempt to identify bacterial members of a fibrolytic consortium dominated by a specific bacterium.     

Activity of fiber-degrading microorganisms in the pig large intestine

The large intestine is comparable to the rumen fermentation in many aspects; however, it is understood less well. Fiber in the form of cellulose and hemicellulose is one of the major substrates fermented in the large intestine. Various studies suggest that the pig can utilize fiber for growth, and up to 30% of its maintenance energy may be derived from volatile fatty acids produced in the large intestine. The total number of microorganisms in the pig large intestine do not change when a high fiber diet such as 50 or 80% alfalfa meal is fed. However, the fiber-degrading organisms increase and obviously replace others. The increase in fibrolytic bacteria normally coincides with an increase in enzyme activity (cellulase and xylanase), indicating that diet can be used to enhance fibrolytic activity. This is true for growing pigs and adult animals. The cellulolytic organisms in the pig, Bacteroides succinogenes and Ruminococcus flavefaciens, are similar to those in the rumen and are present at comparable numbers. This partly explains why adult pigs can maintain themselves by merely grazing on forage in pastures. Assuming other conditions are met, there is a significant potential for fiber degradation in the pig large intestine. Whether various genotypes such as the genetically selected obese and lean pigs have different abilities to degrade fiber is unknown. More work is required to understand the interaction of the fibrolytic organisms with the other organisms present in the large intestine, similar to that which has been done in the rumen, as well as the microbe-host interaction.     

羊草茎在奶牛瘤胃中降解特性及其对食糜纤维分解菌数量的影响

本试验旨在研究羊草茎降解特性和紧密吸附于茎的3种主要纤维分解菌的动态变化。选用羊草茎为试验材料,将其纵切6份后装入尼龙袋投入瘤胃中,分别在6,12,24,48和72 h取出,利用扫描电子显微镜观察超微结构变化;取粉碎后羊草茎进行尼龙袋试验,分别在0.5,2,6、12,24,48和72 h取出,测定不同时间点中性洗涤纤维(neutral detergent fiber,NDF)降解率和吸附在茎中3种主要纤维分解菌的数量变化。结果表明,薄壁组织和韧皮部可被瘤胃微生物降解,维管束会伴随薄壁组织的降解而发生脱落。羊草茎和食糜不同时间点纤维分解菌数量均为产琥珀酸丝状杆菌>白色瘤胃球菌>黄色瘤胃球菌,茎中产琥珀酸丝状杆菌和黄色瘤胃球菌数量在24 h达峰值,分别为10⁹和10⁵ copy/g羊草茎,白色瘤胃球菌在12 h达峰值,为10⁸ copy/g羊草茎。瘤胃食糜中3种纤维分解菌的数量在24 h内基本处在一个恒定的水平,而羊草茎NDF降解率在72 h内逐渐提高,羊草NDF降解率与瘤胃食糜中3种纤维分解菌数量不同步,这可能与纤维分解菌分泌的酶活力存在滞后有关。     

Low- and high-quality forage utilization by heifers and mature beef cows.

Eight cows (7 to 9 yr old, 522 kg) and six heifers (10 mo old, 169 kg) were fed either alfalfa hay (18.7% CP) or mature brome hay (5.1% CP) to determine the effect of cattle age on apparent forage utilization. Cattle were fitted with ruminal and duodenal cannulas and were individually fed once daily (ad libitum intake, 1000). The split-plot design consisted of age (whole-plot) and two sampling periods feeding alfalfa or brome hay (subplot). Each period consisted of 28 d: d 1 to 13 for adaptation, d 13 to 20 for feed intake determination, and d 20 to 28 for sampling. Nylon bags containing NDF substrate from alfalfa or brome hay were incubated ruminally for 0, 3, 6, 12, 24, 48, 96, and 192 h to determine the rate and extent of fiber degradation. Ruminal liquid dilution rate and fermentation characteristics were conducted on d 27. Ruminal fill was determined by total evacuation at 0800 on d 28. Cows consumed more feed (BW.75; P<.01) and had greater ruminal OM fill (P = .04) but had similar fluid fill (P = .88) compared with heifers. Ruminal liquid dilution rate was greater in cows than in heifers (P<.01). The rate of in situ NDF degradation was 3 and .5% per hour greater in cows than in heifers when alfalfa and brome hay were fed, respectively (age x hay, P<.01). Ruminal NDF digestibility as a percentage of intake was greater in cows than in heifers (P<.01). Numbers of ruminal cellulolytic bacteria were not affected by treatment (P>.21). These data indicate that mature cows have a smaller ruminal fluid fill that turns over more rapidly, and this may be responsible for a faster rate of ruminal fiber degradation in cows than in young heifers.     

Quantitative comparisons of select cultured and uncultured microbial populations in the rumen of cattle fed different diets

ABSTRACT: BACKGROUND: The number and diversity of uncultured ruminal bacterial and archaeal species revealed by 16S rRNA gene (rrs) sequences greatly exceeds that of cultured bacteria and archaea. However, the significance of uncultured microbes remains undetermined. The objective of this study was to assess the numeric importance of select uncultured bacteria and cultured bacteria and the impact of diets and microenvironments within cow rumen in a comparative manner. RESULTS: Liquid and adherent fractions were obtained from the rumen of Jersey cattle fed hay alone and Holstein cattle fed hay plus grain. The populations of cultured and uncultured bacteria present in each fraction were quantified using specific real-time PCR assays. The population of total bacteria was similar between fractions or diets, while total archaea was numerically higher in the hay-fed Jersey cattle than in the hay-grain-fed Holstein cattle. The population of the genus Prevotella was about one log smaller than that of total bacteria. The populations of Fibrobacter succinogenes, Ruminococcus flavefaciens, the genus Butyrivibrio, and R. albus was at least one log smaller than that of genus Prevotella. Four of the six uncultured bacteria quantified were as abundant as F. succinogenes, R. flavefaciens and the genus Butyrivibrio. In addition, the populations of several uncultured bacteria were significantly higher in the adherent fractions than in the liquid fractions. These uncultured bacteria may be associated with fiber degradation. CONCLUSIONS: Some uncultured bacteria are as abundant as those of major cultured bacteria in the rumen. Uncultured bacteria may have important contribution to ruminal fermentation. Population dynamic studies of uncultured bacteria in a comparative manner can help reveal their ecological features and importance to rumen functions.     

瘤胃纤维降解的研究进展

瘤胃微生物群降解和发酵粗料细胞壁的结构性碳水化合物,为宿主动物提供挥发性脂肪酸和蛋白质。但是,瘤胃降解纤维的速度和程度受到微生物对底物的可及性、粗料的物理和化学特性及瘤胃消化动力学等因素的调控。本文对瘤胃纤维降解微生物降解纤维的机制和调控作一介绍,并阐明今后的研究方向。     

内蒙古白绒山羊瘤胃中内纤毛虫及三种纤维分解菌的定量研究

试验采用荧光定量PCR技术,在不同精粗比日粮条件下对内蒙古白绒山羊瘤胃内容物中内纤毛虫和3种主要纤维分解菌进行定量研究。结果表明:日粮中粗料比例增加,内纤毛虫(Ento-dinium)的数量降低,组间差异不显著(P>0.05);日粮中粗料比例增加,产琥珀酸丝状杆菌(Fibrobacter succinogenes)、黄色瘤胃球菌(Ruminococcus flavefaciens)和白色瘤胃球菌(Ruminococcus albus)的数量均增加,组间差异不显著(P>0.05)。     

In vitro bacterial growth and in vivo ruminal microbiota populations associated with bloat in steers grazing wheat forage

The role of ruminal bacteria in the frothy bloat complex common to cattle grazing winter wheat has not been previously determined. Two experiments, one in vitro and another in vivo, were designed to elucidate the effects of fresh wheat forage on bacterial growth, biofilm complexes, rumen fermentation end products, rumen bacterial diversity, and bloat potential. In Exp. 1, 6 strains of ruminal bacteria (Streptococcus bovis strain 26, Prevotella ruminicola strain 23, Eubacterium ruminantium B1C23, Ruminococcus albus SY3, Fibrobacter succinogenes ssp. S85, and Ruminococcus flavefaciens C94) were used in vitro to determine the effect of soluble plant protein from winter wheat forage on specific bacterial growth rate, biofilm complexes, VFA, and ruminal H2 and CH4 in mono or coculture with Methanobrevibacter smithii. The specific growth rate in plant protein medium containing soluble plant protein (3.27% nitrogen) was measured during a 24-h incubation at 39 degrees C in Hungate tubes under a CO2 gas phase. A monoculture of M. smithii was grown similarly, except under H2:CO2 (1:1), in a basal methanogen growth medium supplemented likewise with soluble plant protein. In Exp. 2, 6 ruminally cannulated steers grazing wheat forage were used to evaluate the influence of bloat on the production of biofilm complexes, ruminal microbial biodiversity patterns, and ruminal fluid protein fractions. In Exp. 1, cultures of R. albus (P < 0.01) and R. flavefaciens (P < 0.05) produced the most H2 among strains and resulted in greater (P < 0.01) CH4 production when cocultured with M. smithii than other coculture combinations. Cultures of S. bovis and E. ruminantium + M. smithii produced the most biofilm mass among strains. In Exp. 2, when diets changed from bermudagrass hay to wheat forage, biofilm production increased (P < 0.01). Biofilm production, concentrations of whole ruminal content (P < 0.01), and cheesecloth filtrate protein fractions (P < 0.05) in the ruminal fluid were greater on d 50 for bloated than for nonbloated steers when grazing wheat forage. The molecular analysis of the 16S rDNA showed that 2 different ruminal microbiota populations developed between bloated and nonbloated animals grazing wheat forage. Bloat in cattle grazing wheat pastures may be caused by increased production of biofilm, resulting from a diet-influenced switch in the rumen bacterial population.     

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.     

Effects of nitrogen fertilization and maturity of bromegrass on nitrogen and amino acid utilization by cows.

Four nonlactating Holstein cows (593 kg) with ruminal cannulas were fed bromegrass hay that was fertilized with 0 or 89 kg of N/ha and harvested in the late-boot or full-head stage of maturity. Total tract apparent digestibility of N was less for full-head than for late-boot bromegrass (48 vs 64%; P less than .01) and was greater for N-fertilized than for unfertilized bromegrass (60 vs 52%, P less than .01). True N digestibility was greater (P less than .01) by cows fed late-boot hay than by those fed full-head bromegrass (85 vs 81%). Maturity did not affect N retention. Nitrogen fertilization increased N retained, both grams per day (70 vs 20; P less than .01) and as a percentage of N intake (P less than .01). Nitrogen and amino acid disappearance kinetics were determined in situ. Rate constants for N disappearance within rapidly and slowly degraded pools were not different among treatments, but because late-boot had a larger proportion of N in the rapidly degraded pool, overall disappearance rate for N was faster (P less than .05) for late-boot than for full-head bromegrass (12.0 vs 7.1 %/h). Treatment effects observed for individual amino acids were similar to those found for N. Both composition and kinetic data were needed to explain differences in N digestion and balance. Distribution of N within the forage had profound effects on the N economy of the cows.     

Intensive Grazing Management of Smooth Bromegrass With or Without Alfalfa or Birdsfoot Trefoil: Heifer Performance and Sward Characteristics

Over 3 yr, 324 (n=108 per yr) Holstein heifers (226±26 kg; ±SD) ranging in age from 5 to 7 mo were assigned to a 3×2 factorial arrangement of treatments to determine the effects of forage and grazing management system on animal performance, forage availability, and forage quality. The forage treatments were: 1) alfalfa and smooth bromegrass (ALF-BR), 2) birdsfoot trefoil and smooth bromegrass (BFT-BR), or 3) smooth bromegrass plus nitrogen (BR). The grazing management systems were: 1) a four-paddock system in which cattle were rotated based on time (4-p) or 2) a 12-paddock system in which cattle were rotated based on forage availability (12-p). Stocking rate was held constant at 3.75 animals per hectare. The ALF-BR and BR pastures yielded more available forage than BFT-BR pastures (9.86 and 10.04 vs 9.14 metric tons/ha). The ALF-BR and BFT-BR pastures, when compared to BR pastures, supported greater animal average daily gain (0.93 and 0.97 vs 0.83 kg/d) and increased animal gain per hectare (497.8 and 516.4 vs 443.3 kg/d). Legume-grass pastures also had lower neutral detergent fiber content than BR pastures. There were no differences in either animal gain or gain per hectare due to grazing system. Three years after seeding, BFT-BR and ALF-BR had a similar percentage of legume present within their respective pastures. Birdsfoot trefoil appears to be an acceptable substitute for alfalfa in properly managed grass-legume mixed pastures.     

Synergistic fibrolysis in the rumen by cellulolytic Ruminococcus flavefaciens and non-cellulolytic Selenomonas ruminantium: Evidence in defined cultures

To investigate the rumen bacterial interaction between cellulolytic Ruminococcus flavefaciens and non‐cellulolytic Selenomonas ruminantium, fiber digestibility and fermentation products were determined in defined cultures consisting of these two species. Avicel, orchardgrass hay, rice straw and alfalfa hay were used as substrates for 72 h incubation to monitor digestibility, volatile fatty acids, succinate, lactate and bacterial number. In monoculture, R. flavefaciens digested the fiber sources at 21–32%, while S. ruminantium strains did not. When R. flavefaciens was cocultured with one of three different strains (GA192, S137 and S150) of S. ruminantium, fiber digestion exceeded the value recorded by R. flavefaciens alone. In particular, cocultures with S. ruminantium S137 showed significantly higher digestibility for all the fiber sources than R. flavefaciens alone (P < 0.05). Propionate production and growth of S. ruminantium was notable in all cocultures but not in monocultures. Succinate was accumulated in monoculture of R. flavefaciens, while the accumulation was not observed in cocultures. These results indicate that R. flavefaciens provides fiber hydrolysis products to S. ruminantium as growth substrates. In addition, S. ruminantium could activate R. flavefaciens by rapidly consuming the products. Such cross‐feeding between cellulolytic and non‐cellulolytic bacteria could enhance fiber digestion, although the extent of the enhancement may depend on strain combinations.     

Evaluation of hay-type and grazing-tolerant alfalfa cultivars in season-long or complementary rotational stocking systems for beef cows

Alfalfa (Medicago sativa L.) persistence and forage and cow-calf production were evaluated on pastures containing smooth bromegrass with or without grazing-tolerant or hay-type alfalfa cultivars rotationally stocked in either a season-long or complementary system. In 1997, six 2.02-ha pastures were seeded with smooth bromegrass, a mixture of a grazing-tolerant alfalfa (Amerigraze variety) and smooth brome-grass, or a mixture of a hay-type alfalfa (Affinity variety) and smooth bromegrass to be used in season-long stocking systems. Four 2.02-ha pastures were seeded with smooth bromegrass on 1.21 ha of each pasture, and mixtures of either the grazing-tolerant or hay-type alfalfa cultivars and smooth bromegrass on the 0.81 ha of each pasture to be used in complementary stocking systems. All 10 pastures were divided into 10 paddocks and rotationally strip-stocked at 1.98 cow-calf units/ha with crossbred cows and calves for 120 and 141 d starting May 18, 1998 (yr 1), and May 6, 1999 (yr 2), respectively. Each year, first harvest forage was harvested as hay from 40% of all 10 pastures, this being the portions of the pasture seeded with the alfalfa-smooth brome-grass mixtures for pastures with the complementary stocking systems. In yr 1 and 2, the remaining 60% of each pasture was grazed for the first 44 and 54 d, and 100% of each pasture was grazed on d 45 to 120 and d 55 to 141, respectively. Proportions of alfalfa in the live dry matter of pastures seeded with the grazing-tolerant and hay-type alfalfa cultivars decreased by 70 and 55% in paddocks stocked season-long and by 60 and 42% in paddocks used for complementary stocking (alfalfa cultivar, P < 0.05; stocking system, P < 0.05) in yr 1, but decreased by 72% across cultivars and stocking systems in yr 2. Total (P < 0.08) forage masses in September of yr 1 and in August of yr 2 were greater in pastures in which alfalfa paddocks were stocked season-long than in those with complementary alfalfa stocking. Grazing of alfalfa in grass mixtures increased calf and total cow/calf weight gains in comparison with grazing of smooth bromegrass, but alfalfa persistence, measured as a proportion of the live dry matter, was not affected by alfalfa cultivar.     

Preliminary observations of interaction between bacteriophages and Streptococcus bovis bacteria on ruminal epithelium primoculture.

Five Streptococcus bovis strains (47/3, 59/2, 4/1, 46/2 and 44/9) isolated from calf ruminal fluid samples were examined for the adherence to cultured ruminal epithelium cells. Four strains (47/3, 59/2, 4/1 and 46/2) were able to attach to the cultured epithelial cells. However, S. bovis 47/3 strain attached to the target cells in significantly greater numbers than the other strains. Strain 44/9 did not adhere to cells of ruminal epithelium. The adherent bacteria were observed on the surface of differentiated (mainly keratinized) cells of ruminal epithelium primoculture only. The different effect of F4, F5 and F6 bacteriophages was ascertained on S. bovis bacteria adhering to rumen epithelial primoculture. A significant decrease in the number of adherent bacteria was shown after cultivation of strains 47/3 and 4/1 with F6 bacteriophage and of 47/3 strain with F4 phage. The F5 bacteriophage had no significant effect on these bacteria.     

Evaluation of alfalfa-corn cob associative action. II. Comparative tests of alfalfa hay as a source of ruminal degradable protein

The effect of ruminal degradable protein source in roughage diets on nutrient digestibility and animal growth was evaluated in two trials using lambs. In trial 1, two qualities of alfalfa and smooth brome hays replaced 0, 15, 30 or 100% of an ammonia (NH3)-treated corn cob negative control diet in a digestion trial using 26 mixed breed wethers (31.8 kg). Fifteen or 30% inclusion of hay increased (P less than .01) dry matter (DM) intake, regardless of hay type or quality. Cell wall intake was highest for 100% high quality brome hay. Positive associative action on digestibility of DM and cell walls occurred with 30% of each hay tested when diets were fed ad libitum. Measured at equal intakes, DM and cell wall digestibilities were affected by forage type X level and forage quality X level interactions (P less than .01), which showed that the magnitude of associative action was greater for 30% of high vs low quality and alfalfa vs brome hay, respectively. Magnitude of associative response on cell wall digestibility was more highly correlated to degradable N (r = .88) than cell solubles (r = .64) content of hay. In trial 2, 72 young growing lambs were allotted to three sources of supplemental ruminal degradable N (NH3, casein, corn steep liquor) superimposed upon two levels of alfalfa hay (0 or 30% of diet DM). Diets containing 0% alfalfa were supplemented with ruminal escape protein equivalent to that supplied by 30% alfalfa hay.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of microbial fermentation on functional specific gravity of small forage particles.

Two experiments were designed to determine the effect of gas production during in vitro digestion on functional specific gravity (FSG) of forage particles. In Exp. 1, FSG of ground alfalfa hay decreased from 1.123 to 1.049 between 3 and 9 h of incubation and increased thereafter to reach a plateau at 1.309 after 30 h of incubation. Gas production peaked at 6 h, but gas associated with particles increased until 9 h of incubation. Gas associated with solid residue was correlated to gas production (r = -.67) but also was influenced by gas holding capacity and rate of escape from the particles. In Exp. 2, measurements were performed on ground alfalfa hay, alfalfa silage, and bromegrass hay containing 42.6, 35, and 66.4% NDF, respectively. Gas production seemed to be related to the amount of readily available substrate. Although at 9 h of incubation more gas was produced by alfalfa silage (.235 mL.min-1.g of DM-1) than by bromegrass hay and alfalfa hay (.087 and .187 mL.min-1.g of DM-1, respectively), gas associated with particles was greater for alfalfa hay (.416 mL/g of DM) than for bromegrass hay and alfalfa silage (.256 and .281 mL/g of DM, respectively). The increase in FSG was more rapid for alfalfa silage than for the hays. After 27 h of digestion, gas associated with particles (milliliters per gram of DM) and FSG were .164, 1.226; .147, 1.235; and .001, 1.467 for bromegrass hay, alfalfa hay, and alfalfa silage, respectively. Gas produced during fermentation delayed the increase in specific gravity of forage particles.(ABSTRACT TRUNCATED AT 250 WORDS)     

尼龙袋法评价香蕉叶单宁对瘤胃降解特性的影响

旨在探究香蕉叶单宁对奶牛瘤胃降解特性的影响和评价香蕉叶的饲喂价值。样品分别为苜蓿干草组(100%的苜蓿干草)、苜蓿干草和香蕉叶混合组(50%苜蓿干草+50%香蕉叶)、香蕉叶组(100%香蕉叶)、苜蓿干草组(100%的苜蓿干草)+6%聚乙二醇(6% PEG)、苜蓿干草和香蕉叶混合组(50%苜蓿干草+50%香蕉叶)+6%聚乙二醇(6% PEG)、香蕉叶组(100%香蕉叶)+6%聚乙二醇(6% PEG)。测定样品中的营养成分含量,使用3头安装了瘤胃瘘管的荷斯坦奶牛进行半体内试验,用尼龙袋法测定上述样品在瘤胃内0、2、4、8、12、24、48、72 h的干物质(DM)、粗蛋白质(CP)、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)的降解率,得到各营养物质的动态降解参数。结果表明,相较于苜蓿干草,香蕉叶干物质和粗蛋白的快速降解部分、慢速降解部分、慢速降解部分的降解速率、有效降解率是显著低于苜蓿干草的(P<0.001),香蕉叶和苜蓿的NDF的有效降解率没有显著差异。添加了PEG后,香蕉叶和香蕉叶苜蓿混合组的快速降解部分和有效降解率显著高于未添加PEG的两组(P<0.001)。结果表明香蕉叶中的单宁会抑制香蕉叶的瘤胃降解率,因此在使用香蕉叶调制反刍动物的饲料时需要控制单宁的含量。     

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.     

Effects of forage source on ruminal microbial nitrogen metabolism and carbohydrate digestion in continuous culture

Eight single-flow, continuous culture fermentors were used in Exp. 1 to study the effects of forage source on ruminal bacterial N metabolism and carbohydrate digestion. Forages included alfalfa, cicer milkvetch, birdsfoot trefoil and sainfoin with respective CP concentrations of 26.0, 28.7, 26.3 and 20.0%. Each forage provided 100% of the substrate for microbial metabolism and supplied 2.6 g N/d. Ammonia-N, protein degradation and efficiency of ruminal bacterial protein synthesis were lowest (P less than .05) for sainfoin. Protein degradation and efficiency of bacterial protein synthesis were higher (P less than .05) for birdsfoot trefoil than for alfalfa. Effluent flow of amino acids was highest (P less than .05) for sainfoin. Total nonstructural carbohydrate digestion tended to be highest for sainfoin and birdsfoot trefoil, whereas structural carbohydrate digestion was highest (P less than .05) for alfalfa and cicer milkvetch. In Exp. 2, mixed diets were supplied to dual-flow, continuous culture fermentors with alfalfa, cicer milkvetch, birdsfoot trefoil and sainfoin contributing 85% of the total dietary CP. Each diet contained approximately 12.9% CP. Ammonia-N concentration in the effluent and CP degradation tended to be lowest with the sainfoin diet and highest with the birdsfoot trefoil diet. Effluent flow of amino acids tended to be highest with the cicer milkvetch diet and lowest with the alfalfa and birdsfoot trefoil diet. Total structural and nonstructural carbohydrate digestion was not different (P greater than .05) among forages. Results from these experiments indicate that bacterial degradation of protein was lower for sainfoin than for alfalfa. Birdsfoot trefoil and cicer milkvetch appear to be comparable to alfalfa with regard to metabolism of N and carbohydrates by ruminal bacteria.