Diet and procedures used to detach particle-associated microbes from ruminal digesta influence chemical composition of microbes and estimation of microbial growth in Rusitec fermenters

Four different detachment methods were evaluated for their ability to remove particle-associated microorganisms (PAM) from ruminal digesta in semicontinuous fermenters fed two diets differing in their forage:concentrate ratio (80:20 [C20] and 20:80 [C80]). In the methylcellulose method, ruminal digesta was incubated at 38 degrees C for 15 min with saline solution containing 0.1% methylcellulose before being stored at 4 degrees C for 24 h. In the other procedures, samples were incubated with 0.1% methylcellulose before storage for 24 h at 4 degrees C in different solutions (pH = 2): 1) saline solution with 0.1% Tween 80; 2) saline solution with 0.1% Tween 80 and 1% tertiary butanol; and 3) saline solution with 0.1% Tween 80, 1% methanol, and 1% tertiary butanol. Common to all treatments was subsequent homogenization, followed by filtration and resuspension of the residue five times in the treatment solutions. Microbial removal was estimated indirectly by measuring removal of 15N. There were no differences (P > 0.05) among detachment procedures, neither in the detaching efficiency (mean values of 79.7 and 88.1% for C20 and C80 diets, respectively) nor in the total recovery of PAM (54.9 and 34.9% for C20 and C80, respectively). There were no differences (P > 0.05) among PAM pellets obtained by the different detachment procedures in their N content, purine bases (PB) concentration, or PB:N ratio. For the C80 diet, 15N enrichment was greater (P < 0.05) in PAM pellets obtained with methylcellulose than in those obtained by the other methods. However, there were no differences (P > 0.05) due to the detachment procedure in the values of daily microbial growth estimated using as reference the different PAM pellets. The PAM pellets for diet C20 presented greater (P < 0.01) N content and lower (P < 0.01) PB concentration than those for diet C80 (mean values of 74.3 vs 49.1 mg of N/g of dry matter, and 22.8 vs 26.0 micromol PB/mg of dry matter, respectively). Daily microbial growth was greater (P < 0.05) for the C80 diet than for the C20 diet (121 vs 114 mg of microbial N, respectively). Results suggest that the treatment of ruminal digesta with a saline solution with 0.1% methylcellulose at 38 degrees C for 15 min combined with homogenizing and chilling at 4 degrees C for 24 h removed a major proportion of PAM, although further research is needed to decrease microbial losses during the isolation process.     

Effects of Saccharomyces cerevisiae on ruminal pH and microbial fermentation in dairy cows: Yeast supplementation on rumen fermentation

An experiment was conducted with eight ruminally-cannulated cows using a crossover design with 2 periods to determine the effects of yeast supplementation on rumen fermentation. Holstein dairy cows in late lactation were either supplemented with 0.5 g/hd/d of Saccharomyces cerevisiae, an active dry yeast (CNCM-1077, Levucell SC20 (r) SC, Lallemand Animal Nutrition) or not supplemented (control). A basal diet consisting of 60% forage and 40% concentrate (DM basis) was fed once daily to both groups of cows throughout the entire experiment. Ruminal pH was measured continuously every 22 min using a pH probe that was placed in the ventral rumen sac for 6 consecutive days. Volatile fatty acid and ammonia N concentrations in the rumen were measured on days 5 or 6 of the 12-d period for each cow and DM intake was monitored throughout the experiment. Data were analyzed using a mixed-effects model with repeated measures. There were no differences in dry matter intake between treatments. Mean ruminal pH was greater (P < 0.05) when yeast was supplemented (6.53 ± 0.07) compared with the control (6.32 ± 0.07). Average maximum and minimum ruminal pH were also greater (P < 0.05) when yeast was supplemented (7.01 ± 0.09 and 5.97 ± 0.08, respectively) compared with the control (6.80 ± 0.09 and 5.69 ± 0.09, respectively). Time spent under the subacute acidosis threshold, pH less than 5.6, was lower (P < 0.05) with yeast supplementation compared with control cows. No difference was observed for ruminal ammonia N concentrations (mean = 14.0 ± 1.2 mg/dL) between treatments. Total VFA concentration (mM) in the rumen tended to be lower (P = 0.10) in the yeast-supplemented cows (107.3 ± 6.35) than in the control cows (122.4 ± 6.35), which could be related to the greater pH observed with yeast supplementation. Supplementing dairy cows with active dry yeast in the current experiment increased the mean, minimum and maximum ruminal pH; decreased time spent in subacute rumen acidosis, and tended to decrease total VFA concentration in the rumen compared with control cows.     

Effects of tallow supplementation and protein withdrawal on ruminal fermentation, microbial synthesis and site of digestion

The effects of tallow supplementation [0% (NT) vs 7.5% (T)] and crude protein level [8.5% (LP) vs 12.0% (HP)] on ruminal fermentation, microbial protein (MCP) synthesis, digesta passage and site of digestion were estimated using yearling Angus X Simmental steers (390 kg) fitted with ruminal and T-type duodenal cannulae. Chromium-EDTA and ytterbium (Yb) chloride were used as markers of the liquid and solid phases. Passage and site of digestion data were estimated from the concentrations of Yb in the duodenal digesta and feces. Dry matter (DM) intakes were 6.8, 6.5, 6.3 and 6.6 kg/d for the NT-LP, NT-HP and T-HP diets, respectively. Ruminal ammonia concentrations (mg/100 ml) for the NT-LP, NT-HP, T-LP and T-HP diets were 1.22, 4.75, 1.05 and 3.41, respectively. Tallow decreased (P less than .05) acetate (mol/100 mol), increased (P less than .01) propionate (mol/100 mol) and decreased the total volatile fatty acid concentration. Tallow depressed apparent ruminal DM and organic matter (OM) digestibilities only on the HP diet. High protein increased ruminal DM, OM and fiber digestibilities. Tallow and LP tended to shift the site of OM digestion to the lower tract. The liquid and solid dilution rates for the NT-LP, NT-HP, T-LP and T-HP diets were 9.53, 3.37; 5.63, 3.28; 6.66, 5.10 and 6.79, 5.34%/h, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of Lonicera japonica extract supplementation on in vitro ruminal fermentation,methane emission,and microbial population

Lonicera japonica (LJ; honeysuckle) is used in traditional folk medicine in Korea and is a rich source of ascorbic acid and phenolic components that are reported to have antioxidant and antibiotic properties. We performed an in vitro experiment to assess the effects of LJ extracts (LJE) on ruminal fermentation. Timothy hay (0.3 g dry matter [DM]) was incubated with buffer, ruminal fluid, and 0%, 3%, 5%, 7%, and 9% LJE. Batch culture fermentation was conducted separately for 12, 24, and 48 hr to determine gas production (GP), ruminal fermentation characteristics, and microbial population characteristics. The effects on GP were generally similar to those on DM degradability, with a linear decrease observed at 9% extract at 24 hr. NH₃‐N showed a linear increase with increasing extract concentrations at 12 hr, whereas a decrease was observed at 24 hr. Extract supplementation decreased methane (CH₄) production at 12, 24, and 48 hr. In addition, the abundance of fibrolytic bacteria and ciliate‐associated methanogen was reduced at all concentrations of extracts. These results indicate that LJE have the potential to serve as a ruminal fermentation modifier to suppress CH₄ production with minimal effects on nutrient digestion in the rumen.     

日粮补充异丁酸对犊牛生长性能、瘤胃发酵和纤维分解菌菌群的影响

为探索异丁酸对犊牛生长性能、瘤胃发酵、纤维分解菌菌群和酶活性的影响,试验选用15日龄荷斯坦犊牛36头,随机分成4组,对照组、低水平异丁酸组(LIB)、中水平异丁酸组(MIB)和高水平异丁酸组(HIB)分别饲喂异丁酸0、3、6和9g·d^-1,预饲15d,分别于60日龄(断奶)和90日龄测定体重和采集瘤胃液进行分析。结果表明,MIB组和HIB组犊牛干物质采食量、日增重和经济效益显著高于对照组(P<0.05)。90日龄时HIB组犊牛瘤胃pH值显著低于对照组和LIB组(P<0.05)。MIB和HIB组瘤胃总挥发性脂肪酸和乙酸浓度显著高于对照组和LIB组(P<0.05);MIB和HIB组犊牛60日龄时瘤胃乙酸/丙酸显著高于对照组(P<0.05),90日龄时显著高于对照组和LIB组(P<0.05)。60和90日龄时MIB和HIB组瘤胃异丁酸均显著高于对照组(P<0.05)。60日龄时MIB和HIB组木聚糖酶和α-淀粉酶显著高于对照组(P<0.05),MIB和HIB组纤维二糖酶、果胶酶、溶纤维丁酸弧菌、黄色瘤胃球菌和产琥珀酸丝状杆菌显著高于对照组和LIB组(P<0.05);90日龄时MIB和HIB组各种瘤胃酶活力和纤维分解菌均显著高于对照组和LIB组(P<0.05)。结果显示,日粮补充异丁酸促进了犊牛瘤胃纤维素分解菌的生长,提高了纤维素分解酶的活性,进而促进了犊牛瘤胃发酵和生长发育。由于MIB与HIB之间各指标差异不显著,兼顾经济效益,在本试验条件下异丁酸的最佳添加量为6.0g·d^-1。     

Ruminal fermentation,microbial growth and amino acid flow in single‐flow continuous culture fermenters fed a diet containing olive leaves

Six single‐flow continuous culture fermenters were used to determine fermentation profile, microbial growth and amino acid (AA) flow promoted by olive leaves supplemented with barley grains and faba beans (OLSUP), and alfalfa hay (AH). Two incubation runs were carried out with three fermenters inoculated with ruminal fluid from wethers and three from goats. The inoculum source did not affect (p = 0.059 to 0.980) any of the parameters. Daily volatile fatty acid (VFA) production and carbohydrate digestibility were greater (p = 0.009 and 0.024, respectively) for AH, therefore the pH values were lower (p = 0.015) than for OLSUP. Acetate was greater (p < 0.001) and isobutyrate, isovalerate and caproate lower (p < 0.001 to 0.006) for AH with greater acetate/propionate (p = 0.014) and ‘VFA/digested carbohydrate’ (p = 0.026) ratios. Daily microbial N flow and efficiency were greater (p = 0.016 and p = 0.041) for diet AH. Individual AA flows were greater (p < 0.001 to 0.016) for AH, but microbial essential AA proportion was greater for OLSUP (p = 0.015). The results indicate that OLSUP promoted lower bacterial growth and AA flow than AH, which could have been partially due to a limitation of N availability to ruminal microbes.     

Influence of DL-methionine supplementation on growth, ruminal fermentation and dilution rates in heifers

A 3 X 3 replicated Latin square design was used to evaluate three isonitrogenous supplements designed to supply 250 g crude protein (CP) daily. Measurements included in situ dry matter (DM) and neutral detergent fiber (NDF) fermentation and ruminal dilution rates. Supplements contained beet pulp plus DL-methionine and urea (MET), ammonium sulfate and urea (U) or soybean meal (SBM). Six mature, ruminally cannulated crossbred beef cows were individually fed supplement and a mixture (63% NDF and 6.1% CP) of chopped 75% grass hay and 25% barley straw in ad libitum. Fermentation rate of DM was increased (P less than .05) by 30% with MET in comparison to SBM or U (9.54 vs 7.28% and 7.74%/h for MET, SBM and U, respectively). Even though MET improved fermentation rate by 30%, particle dilution rate was more important in affecting ruminal digestibility than fermentation rate. Two 90-d heifer growth trials were conducted to evaluate similar supplements. Supplements similar to those used in the in situ trial were mixed with roughage to provide a complete diet balanced for .3 kg daily gain. Heifers consumed 112% of the National Research Council CP requirement. Weight gain, intake and feed conversion were similar (P greater than .10) for all treatments. In heifer trial 2, 90% of the National Research Council CP requirement was fed. The heifers supplemented with MET and SBM had faster (P less than .05) weight gains than heifers receiving U. These studies show that feeding DL-methionine with urea, as compared with feeding an isonitrogenous supplement containing SBM, increased the fermentation rate of DM.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of tributyrin supplementation on ruminal microbial protein yield,fermentation characteristics and nutrients degradability in adult Small Tail ewes

Two trials were conducted to assess the effects of tributyrin (TB) supplementation on ruminal microbial protein yield and fermentation characteristics in adult sheep. In an in vitro trial, substrate was made to offer TB at 0, 2, 4, 6, and 8 g/kg on a dry matter (DM) basis and incubated for 48 hr. In an in vivo trial, 45 adult ewes were randomly assigned by initial body weight (55 ± 5 kg) to five treatments of nine animals over an 18‐day period. Total mixed ration was made to offer TB to ewes at 0, 2, 4, 6, and 8 g/kg on a DM basis. The in vitro trial showed that TB enhanced apparent degradation of DM (p = .009), crude protein (p < .001), neutral detergent fiber (p = .007) and acid detergent fiber (p = .010) and increased methanogenesis (p < .001), respectively. The in vivo trial showed that TB decreased DM intake (p < .001) and enhanced rumen microbial N synthesis (p < .001), respectively. Both in vitro and in vivo trials showed that TB increased total volatile fatty acid concentration and enhanced fibrolytic enzyme activity. The results indicated that TB might exert positive effects on microbial protein yield and fermentation in the rumen.     

TMR饲料对牛育成期瘤胃发酵特性及微生物生态的影响

对去势的黑白花奶牛饲喂精粗分离饲料和TMR饲料后,瘤胃pH在两处理间无显著差异,采食后9 h的平均NH3-N浓度是试验组极显著高于对照组(P<0.01).丙酸浓度在采食后3 h时试验组显著高于对照组(P<0.05),丁酸浓度在6 h和9 h时试验组显著低于对照组(P<0.05),而乙酸浓度无差异.乙酸/丙酸比是试验组极显著低于对照组(P<0.01).CMCase活力试验组表现出高于对照组的倾向,但木聚糖酶活力试验组极显著高于对照组(P<0.01).另外,采食后瘤胃内总细菌数是试验组比对照组有增加趋势,但相反地,总真菌数表现出减少的倾向.瘤胃原虫的数量是采食0 h和9 h时试验组显著高于对照组(P<0.05).     

绞股蓝皂甙对体外瘤胃微生物甲烷产量及发酵特性的影响

利用体外产气量法研究绞股蓝皂甙对山羊瘤胃微生物体外甲烷产量及发酵特性的影响。试验包括2个部分,试验一研究了绞股蓝皂甙对瘤胃微生物甲烷产量及发酵特性的影响,试验二分析绞股蓝皂甙对瘤胃微生物发酵动力学参数的影响。试验以0.42 g羊草＋0.126 g玉米＋0.054 g豆粕为发酵底物,60 mL培养基中的绞股蓝皂甙添加量分别为0(对照),5,10,20和40 mg,发酵24 h。与对照组比较,发酵8 h,各处理组甲烷浓度显著下降(P<0.05),分别下降30.20％,43.49％,44.67％和75.8％;12 h,20 mg组显著下降(P<0.05),40 mg组极显著下降(P<0.01),处理组甲烷浓度分别下降6.97％,9.63％,18.90％和61.82％;24 h,10 mg组显著下降(P<0.05),40 mg组极显著下降(P<0.01),处理组甲烷浓度分别下降2.34％,9.39％,6.90％和20.73％,甲烷浓度与皂甙剂量之间有极显著的线性效应(P<0.01)。10 mg组的氢利用率极显著低于对照组,其他试验组无显著变化。10 mg组显著提高了TVFA及乙酸、丙酸、异丁酸、戊酸、异戊酸和支链脂肪酸浓度(P<0.05),40 mg组丁酸的浓度极显著下降(P<0.01)。10 mg组和20 mg组乙丙比显著高于对照组(P<0.05)。随着皂甙剂量增加,乙酸、丁酸、异丁酸、戊酸、异戊酸、支链脂肪酸、总挥发性脂肪酸浓度及乙丙比呈显著或极显著的二次方效应,丁酸同时具有极显著的线性效应。处理组原虫数量显著(P<0.05)或极显著下降(P<0.01)(40 mg组),且与皂甙剂量间存在极显著的线性和二次方效应(P<0.01)。微生物蛋白含量没有显著变化,但呈上升趋势。10 mg组和40 mg组的氨态氮浓度显著升高(P<0.05),氨态氮浓度与皂甙剂量之间有显著的线性效应(P<0.05)。高剂量绞股蓝皂甙降低了微生物发酵的理论与实际产气量,并呈显著的线性和二次方效应,产气速率与皂甙剂量之间有着显著的线性效应。以上结果表明绞股蓝皂甙能改变瘤胃微生物发酵模式,降低瘤胃微生物的甲烷产量,提高VFA的产量,有利于饲料能量的利用,同时缓解甲烷对大气环境的污染。     

Inhibitory effects of isoleucine and antagonism of the other branched-chain amino acids on fermentation parameters by mixed ruminal microbes in batch cultures and rumen simulating fermenters (Rusitec)

The inhibitory effect of isoleucine (Ile) and antagonistic effect of leucine (Leu) and valine (Val) against the inhibition by Ile were investigated using simple batch and semicontinuous cultures of mixed ruminal microbes under feeding conditions using ordinary feed (timothy hay and corn grain). In the batch culture experiment, supplementation with Ile inhibited digestion and fermentation at 8–24 h of incubation as the concentration of Ile increased up to 5 mmol/L, but these inhibitions diminished at 48 h. The inhibition caused by Ile was mitigated by the addition of Leu and Val. In a semi‐continuous culture experiment using Rusitec, digestion and fermentation of the diet was suppressed by the administration of 1 and 5 mmol/L of Ile. The daily yield of microbial‐N was also reduced by Ile, but the density of the protozoa was not. The presence of Leu and Val alleviated the inhibition by Ile. These results suggest that the dietary inclusion of Ile possibly represses the bacterial growth, feed digestion and fermentation in the rumen in ordinary feeding situations by feedback inhibition, which could be diminished in the presence of other branched‐chain amino acids. The influences of the passage and level of supplementation on inhibition by amino acids are discussed.     

饲粮中添加燕麦干草对绵羊体外发酵的影响

为明确燕麦干草不同添加量对绵羊瘤胃体外发酵的影响,选用6头体况相似、体重 (70.24±2.03) kg且装有永久性瘤胃瘘管的杂种(美利奴♂×蒙古羊♀)公羊作为瘤胃液供体动物进行体外发酵试验,按粗饲料中燕麦干草和青贮玉米比例(干物质基础)将饲粮分为C₁组(精料+100%玉米青贮),C₂组(精料+50%玉米青贮+50%燕麦干草)和C₃组(精料+100%燕麦干草)。饲粮精粗比为35∶65。结果表明:1)饲粮中加入燕麦干草显著提高了体外发酵的产气量和甲烷产量(P<0.05);2)饲粮中加入燕麦干草对体外培养液pH没有明显影响,但显著提高了培养液NH₃-N浓度(P<0.05),未明显改变试羊瘤胃丙酸摩尔比,但TVFA浓度、乙酸、丁酸摩尔比以及乙酸/丙酸上升;3)燕麦干草的加入显著提高了绵羊瘤胃体外发酵的IVDMD和IVCPD,对饲粮中营养物质的降解有促进作用;4)多项指标综合指数以C₂组最高,C₁组最低。由此可知,在精料水平相同的条件下,燕麦干草与青贮玉米1∶1混合作为粗饲料能改善绵羊的瘤胃内环境,提高瘤胃发酵能力,得到最大组合效应。     

Effects of urea infusion and ruminal degradable protein concentration on microbial growth,digestibility, and fermentation in continuous culture

The effects of urea and rumen-degradable protein (RDP) on microbial growth, digestibility, and fermentation were examined using dual-flow continuous culture. The experimental design was a 4 x 4 Latin square with a 2 x 2 factorial arrangement of treatments. Factors were urea infusion (0.4 g/L of artificial saliva) and RDP concentration, and the treatments were as follows: 1) low RDP (8% of dietary dry matter) without urea (LDNU), 2) high RDP (11% of dietary dry matter) without urea (HDNU), 3) low RDP (8% of dietary dry matter) with urea (LDU), and 4) high RDP (11% of dietary dry matter) with urea (HDU). The LDNU (i.e., negative control) and HDNU treatments were formulated to be nitrogen limiting. Results indicated that infusion of urea increased all digestibility measurements (P < 0.05), which in turn increased (P < 0.05) volatile fatty acid, NH3 nitrogen, trichloroacetic acid-soluble nitrogen, and soluble protein concentrations. Increasing dietary RDP improved dry matter and organic matter digestibility (P < 0.05) but did not alter acid detergent fiber or nonfiber carbohydrate digestibilities (P > 0.05). Isobutyrate concentration decreased (P = 0.05) with increased RDP. Increased dietary RDP increased crude protein degradation and soluble protein concentration (P < 0.05), but NH3 nitrogen, trichloroacetic acid-soluble nitrogen, and peptide nitrogen were unaffected by changing RDP levels. Microbial growth efficiency was 19.9, 24.9, 28.0, and 32.2 g N/g organic matter truly digested for LDNU, HDNU, LDU, and HDU, respectively, and was significantly improved both by urea infusion (P = 0.002) and increased RDP concentration (P = 0.021). The interactions of urea and RDP (P < 0.05) were explained by the high digestibility of neutral detergent fiber, nonstructural carbohydrate, and especially hemicellulose, with the HDNU treatment. The results of this study indicated that hemicellulose-degrading bacteria were able to effectively compete with nonstructural carbohydrate-degrading bacteria for available peptide and amino acid nitrogen. Further, the extent of protein degradation was dependent on the availability of NH3 nitrogen in the system.     

Effects of forage:concentrate ratio and forage type on apparent digestibility, ruminal fermentation, and microbial growth in goats

The effects of forage type and forage:concentrate ratio (F:C) on apparent nutrient digestibility, ruminal fermentation, and microbial growth were investigated in goats. A comparison between liquid (LAB) and solid (SAB)-associated bacteria to estimate microbial N flow (MNF) from urinary purine derivative excretion was also examined. Treatments were a 2 x 2 factorial arrangement of forage type (grass hay vs. alfalfa hay) and high vs. low F:C (70:30 and 30:70, respectively). Four ruminally cannulated goats were fed, at maintenance intake, 4 experimental diets according to a 4 x 4 Latin square design. High-concentrate diets resulted in greater (P < 0.001) nutrient digestibility except for ADF. However, CP digestibility increased (P < 0.001) only for the high-concentrate diets including grass hay. Likewise, N retention, ruminal NH(3)-N concentration, and urinary excretion of purine derivatives increased (P < 0.05) with increasing concentrate in animals fed diets based on grass hay (0.23 vs. 0.13 g of retained N/g of digested N, 30.1 vs. 12.9 mg of NH(3)-N/100 mL, and 11.5 vs. 8.40 mmol/d, respectively), but not (P > 0.05) when diets included alfalfa hay. Total protozoa numbers and holotricha proportion were greater and less (P < 0.001), respectively, in high- than in low-concentrate diets. The F:C affected (P < 0.001) ruminal pH but not total VFA concentration (P = 0.12). Ammonia-N concentration was similar (P = 0.13) over time, whereas pH, VFA concentration, and protozoa numbers differed (P < 0.001) among diets. Estimated MNF was strongly influenced by using either the purine bases:N ratio obtained in our experimental conditions or values reported in the literature for small ruminants. There was a F:C effect (P = 0.006) on MNF estimated from LAB but not from SAB. The effect of F:C shifting from 70:30 to 30:70 in goat diets depends on the type of forage used. The MNF measured in goats fed different diets was influenced by the bacterial pellet (LAB or SAB). In addition, the purine bases:N ratio values found were different from those reported in the literature, which underlines the need for these variables to be analyzed directly in pellets isolated from specific animals and experimental conditions.     

Effects of dietary concentrate level and chromium-methionine supplementation on ruminal fermentation,and ruminal bacterial and fatty acid composition in Tan lambs北大核心CSCD

This study evaluated the effects of dietary concentrate level and chromium methionine(Cr-Met)supplementation on ruminal fermentation and fatty acid composition, and ruminal bacteria abundance in Tan lambs. Forty male Tan lambs [(21.00±1.23) kg body weight, 5 months of age] were randomly assigned to four treatment groups with 10 lambs in each treatment. The treatments were: a low-concentrate (LC) diet (concentrate:forage, 35:65) without Cr-Met supplementation; a high-concentrate (HC) diet (concentrate:forage, 55:45) without Cr-Met supplementation; groups fed the HC diet with 0.75 or 1.50 g·d-1·lamb-1 Cr-Met (HCM and HCH, respectively). Ruminal fluid was collected on day 65 with an oral stomach tube about 3 h after the morning feed and samples used for fermentation analysis, fatty acid composition determination and bacterial DNA extraction. It was found that: 1) Ruminal pH, acetate proportion and acetate:propionate were greater (P<0.05) with the LC diet whereas microbial crude protein, propionate and valerate proportions were greater (P<0.05) with the HC diet. The acetate:propionate was greater (P<0.05) in the HCM group than in the HC group. 2) For the HC diet, the DNA abundances of Butyrivibrio fibrisolvens vaccenic acid subgroup (Butyrivibrio VA) and Ruminococcus flavefaciens were decreased (P<0.05), compared with the LC diet, whereas the DNA abundance of Anaerovibrio lipolytica was increased (P<0.05). For lambs fed the HC diet, DNA abundances of B. fibrisolvens stearic acid subgroup, Butyrivibrio VA, Butyrivibrio proteoclasticus, and A. lipolytica showed a linear decrease (P<0.05) with increasing Cr-Met supplementation level; However, the DNA abundance of R. flavefaciens showed a linear increase (P<0.05). 3) Compared with the LC group, the concentrations of t11 C18:1, trans C18:1, c9t11 CLA, t10c12 CLA, C18:2n6, and C18:3n3 were decreased by the HC diet without Cr-Met supplementation whereas the concentration of C18:0 was increased. There was a linear increase in t11 C18:1 and trans C18:1 with increasing Cr-Met supplementation level in lambs fed the HC diet. The results suggest that the HC diet inhibited the growth of ruminal bacteria involved in the ruminal biohydrogenation process, and the addition of Cr-Met had a potentially positive effect on conjugated linoleic acid (CLA) synthesis in body tissue. © 2022, Editorial Office of Acta Prataculturae Sinica. All rights reserved.     

Effects of brewers' spent grain protein hydrolysates on gas production,ruminal fermentation characteristics,microbial protein synthesis and microbial community in an artificial rumen fed a high grain diet

Background: Brewers' spent grain(BSG) typically contains 20% – 29% crude protein(CP) with high concentrations of glutamine, proline and hydrophobic and non-polar amino acid, making it an ideal material for producing valueadded products like bioactive peptides which have antioxidant properties. For this study, protein was extracted from BSG, hydrolyzed with 1% alcalase and flavourzyme, with the generated protein hydrolysates(AlcH and FlaH)showing antioxidant activities. This study evaluated the effects of AlcH and FlaH on gas production, ruminal fermentation characteristics, nutrient disappearance, microbial protein synthesis and microbial community using an artificial rumen system(RUSITEC) fed a high-grain diet.Results: As compared to the control of grain only, supplementation of FlaH decreased(P < 0.01) disappearances of dry matter(DM), organic matter(OM), CP and starch, without affecting fibre disappearances;while AlcH had no effect on nutrient disappearance. Neither AlcH nor FlaH affected gas production or VFA profiles, however they increased(P < 0.01) NH_3-N and decreased(P < 0.01) H_2 production. Supplementation of FlaH decreased(P < 0.01)the percentage of CH_4 in total gas and dissolved-CH_4(dCH_4) in dissolved gas. Addition of monensin reduced(P < 0.01) disappearance of nutrients, improved fermentation efficiency and reduced CH_4 and H_2 emissions.Total microbial nitrogen production was decreased(P < 0.05) but the proportion of feed particle associated(FPA) bacteria was increased with FlaH and monensin supplementation. Numbers of OTUs and Shannon diversity indices of FPA microbial community were unaffected by AlcH and FlaH;whereas both indices were reduced(P < 0.05) by monensin. Taxonomic analysis revealed no effect of AlcH and FlaH on the relative abundance(RA) of bacteria at phylum level, whereas monensin reduced(P < 0.05) the RA of Firmicutes and Bacteroidetes and enhanced Proteobacteria. Supplementation of FlaH enhanced(P < 0.05) the RA of genus Prevotella, reduced Selenomonas, Shuttleworthia, Bifidobacterium and Dialister as compared to control;monensin reduced(P < 0.05) RA of genus Prevotella but enhaced Succinivibrio.Conclusions: The supplementation of FlaH in high-grain diets may potentially protect CP and starch from ruminal degradation, without adversely affecting fibre degradation and VFA profiles. It also showed promising effects on reducing CH_4 production by suppressing H_2 production. Protein enzymatic hydrolysates from BSG using flavourzyme showed potential application to high value-added bio-products.     

瘤胃蛋白降解率和添加频率对饲喂低质饲料的羔羊的影响研究

4头初始体重为34.5±2.0kg、插有瘤胃瘘管的萨福克阉公羊羔被用来评估瘤胃蛋白降解率和添加频率对饲喂低质饲料的羔羊瘤胃特性的影响。羔羊以成熟皇冠干草（含4.2%CP）为基础日粮,各日粮组分为4组,分别是：1）RDP-D：每天饲喂高的瘤胃降解蛋白质（RDP）;2）RDP-A：每隔1d饲喂高的RDP;3）RUP-A：每隔1d饲喂非瘤胃降解蛋白质（RUP）;4）MIX-A：每隔1d按照RDP：RUP=1：1的比例来饲喂。结果显示,各处理组的羔羊粗饲料的有机质（OM）、氮（N）、中性洗涤纤维（NDF）或酸性洗涤纤维（ADF）的采食量没有显著差异;混合组（MIX-A）的羔羊瘤胃有机质的消化率比其他组的要高（P〈0.001）;RUP-A组羔羊的真瘤胃氮消化率较其他处理组的低（P〈0.01）;混合组MIX-A瘤胃NDF和ADF的代谢较其他处理组高（P≤0.01）。当在羔羊日粮中添加RUP相对添加RDP-D可以降低其瘤胃氨气浓度,且RUP-A组的羔羊的瘤胃氨气浓度最低。各处理组的瘤胃脲酶活性差异不显著,瘤胃微生物N流量差异不显著,不过RDP-D组的羔羊的瘤胃微生物N合成效率有增加趋势（P=0.004）。隔天饲喂羔羊混合组的日粮可以改善低质粗饲料的消化率,这可能是由于内源性氮循环利用增强导致瘤胃氨气浓度有所下降。     

Influence of decoquinate on ruminal fermentation, diet digestibility and cattle performance

Three experiments utilizing three Holstein steers (235 and 299 kg avg body weight for Exp. 1 and 2, respectively) were conducted to evaluate the effects of decoquinate, a synthetic coccidiostat, on ruminal fermentation, diet digestibility and performance of steers fed a finishing diet containing monensin and tylosin. Experiment 1 utilized a 70% forage diet, whereas Exp. 2 utilized a 20% forage diet. Each experiment was a 3 X 3 Latin-square design with treatments being 0, .5 and 5 mg decoquinate/kg body weight. Ruminal fermentation characteristics, water kinetics and blood constituents were measured on d 11 of each period, and zero-time volatile fatty acid (VFA) production was measured at 3 and 6 h post-feeding on d 12 to 14. No changes were seen in plasma glucose or L-lactate, ruminal pH, NH3-N or ruminal L-lactate for either experiment. Dry matter digestibility was depressed (P less than .05) at the .5- and 5-mg levels of decoquinate in Exp. 1, but dry matter digestibility was not affected in Exp. 2. No changes were seen in ruminal volume, outflow or total VFA concentration for either experiment. Molar proportions of VFA were not affected in Exp. 1, but the proportions of isobutyrate and butyrate decreased (P less than .05) at the 5-mg level of decoquinate in Exp. 2. Volatile fatty acid production was not changed in Exp. 1, but butyrate production was decreased (P less than .05) at the 5-mg level in Exp. 2. Experiment 3 involved 135 crossbred steers (259 kg avg initial wt), which were stratified by weight into 12 pens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of a blend of garlic oil,nitrate and fumarate on in vitro ruminal fermentation and microbial population

Although garlic oil and nitrate can effectively suppress ruminal methane (CH₄) production in vitro, the application of these compounds is associated with suppressed total volatile fatty acid (VFA) concentration. On the other hand, the effectiveness of fumarate as a ruminal CH₄ mitigating agent is variable but its application increases total VFA concentration. We therefore hypothesized that the different characteristics of the compounds can compensate for the shortcomings of the other. The objective of this study was to develop an optimal blend of garlic oil, nitrate and fumarate that can suppress in vitro ruminal CH₄ without affecting total VFA concentration. Three ruminal in vitro fermentation experiments were carried out. The first one, a one factor at a time experiment was employed to investigate the effective concentration of each of the compounds on CH₄ and VFA production by ruminal bacteria. We then applied the fractional factorial design and response surface methodology in the second experiment to determine optimal concentrations of the compounds in the blend. The optimal blending of garlic oil, fumarate and nitrate was determined to be 50 mg/l, 15 mm and 20 mm , respectively. This simulated optimal blend was verified in a 48 h in vitro batch fermentation experiment. The blend achieved the intended goal of suppressing CH₄ whilst maintaining total VFA concentration. The blend and nitrate suppressed archaea populations (p < 0.001) but did not affect the total microbial population (p = 0.945). The observed results could be explained by additive effects of the agents making up the blend. Supplementing a high concentrate diet with the blend can significantly decrease ruminal CH₄ and maintain total VFAin vitro. These findings however, need to be verified in vivo using the optimized ratio of combining the three methane inhibitors as a guide.     

花生油对绵羊体外瘤胃发酵指标的影响

本研究采用体外批次培养技术,选择3只安装有永久性瘤胃瘘管的绵羊为瘤胃液供体,以微晶纤维素为培养底物,培养瓶中花生油的添加量分别为0、5、10、15 mg培和20 mg,每个水平做3个重复,在培养4、8、16、24、36 h和48 h各时间.占'测定培养液中pH值、氨氮(NH3-N)浓度、总挥发性脂肪酸(TVFA)浓度和48 h纤维素消失率,从而筛选花生油在体外培养中适宜的添加量.结果表明:随着花生油添加水平的逐渐增加,培养液中pH值呈上升趋势,NH3-N浓度、TVFA浓度和48 h纤维素消失率逐渐降低.本试验中,在不影响瘤胃发酵的情况下花生油的最适宜添加量为5 mg.