衍生化气相色谱法检测瘤胃液和青贮液中乳酸对映体和挥发酸

利用手性衍生化试剂L-薄荷醇和气相色谱柱前衍生化,将瘤胃液和青贮液中L-和D-乳酸对映体转化为酯类非对映体对,建立同时分析发酵液中乳酸对映体和挥发酸的气相色谱检测方法。样品乳酸对映体的质量浓度在0.05~10.00 mg/mL范围内时,标准曲线的决定系数分别达到R2=0.999 5和R2=0.999 6。乳酸对映体的回收率为96.15%~108.24%,且相对标准偏差小于5%。采用本方法检测瘤胃液和青贮液样品的L-和D-乳酸对映体含量,并且与酶学检测法进行比较,2种方法的测定结果没有显著性差异(P>0.05)。发酵液中挥发酸经衍生化处理,可以与L-和D-乳酸对映体在同一色谱条件下检测,各组分在0.05~5.00 mg/mL范围内线性良好,R2=0.998 2~0.999 4。乳酸对映体和挥发酸的色谱分析在6 min完成。     

不同精粗比饲粮中添加壳聚糖对体外瘤胃发酵甲烷产量和发酵特性的影响

本试验旨在利用长期人工瘤胃系统——Rusitec系统研究不同精粗比饲粮中添加壳聚糖对体外瘤胃发酵甲烷产量和发酵特性的影响。采用2×2两因子试验设计,2个因子分别为饲粮精粗比[30∶70(低精料饲粮)和70∶30(高精料饲粮)]和是否添加壳聚糖(0和1 500 mg/罐),共构成4组,每组4个重复。结果表明:与低精料饲粮相比,高精料饲粮显著降低了甲烷产量,中性洗涤纤维和粗蛋白质的降解率,发酵液乙酸和丁酸含量、乙酸/丙酸、氨氮浓度(P<0.05),显著提高了总产气量和丙酸含量(P<0.05);添加壳聚糖显著降低了甲烷产量和总产气量、粗蛋白质的降解率、发酵液乙酸和丁酸含量、乙酸/丙酸、氨氮浓度(P<0.05),显著提高了丙酸含量(P<0.05);在甲烷产量和总产气量,粗蛋白质的降解率,0、6 h发酵液氨氮浓度,0、12 h发酵液乙酸/丙酸上,2个因子存在互作(P<0.05),高精料饲粮中添加壳聚糖对甲烷产量和总产气量、粗蛋白质降解率、发酵液氨氮浓度的降低幅度大于低精料饲粮,对总挥发性脂肪酸浓度和乙酸/丙酸的降低幅度小于低精料饲粮。以上结果表明,改变饲粮的精粗比和添加壳聚糖均能够改变瘤胃发酵模式,且二者之间存在互作,壳聚糖在精粗比为70∶30的饲粮中添加效果优于精粗比为30∶70的饲粮。     

断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响

为了研究断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响,选取24只体况良好、胎次一致、体重相近的10日龄羔羊(湖羊),将其随机分为3组,在饲喂相同母乳的基础上,分别补饲以纯木薯(CS)、玉米(MS)和豌豆淀粉(PS)(直/支链淀粉比分别约为0.11,0.27和0.44)为唯一淀粉来源的开食料。试验期间,每天4:00-19:00将羔羊抱入补饲栏补饲不同直/支链淀粉比的开食料,并且在6:30, 10:30和15:30将羔羊抱回母羊舍哺乳1 h。羔羊自由饮水,单栏饲喂,自由采食燕麦干草。56日龄时屠宰采样,采集瘤胃组织样品制作石蜡切片进行瘤胃乳头形态测定,采集瘤胃上皮样品提取RNA测定相关基因表达。结果表明:CS组羔羊瘤胃乳头长度和表面积显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃乳头宽度显著(P=0.001)高于PS组。对瘤胃上皮各层厚度统计显示,CS和PS组羔羊瘤胃上皮角质层厚度显著(P=0.001)高于MS组;CS和MS组羔羊瘤胃上皮颗粒层厚度显著高于(P<0.001)PS组;CS组羔羊瘤胃上皮棘基层厚度和总厚度显著(P<0.001)高于MS和PS组。qRT-PCR结果显示:不同直/支链淀粉比开食料显著影响羔羊瘤胃上皮CDK2和CDK6的mRNA表达量(CS>MS>PS, P<0.001);CS组羔羊瘤胃上皮细胞cyclin A和CDK4的mRNA表达量显著(P<0.05)高于PS组,但与MS组无显著差异;CS组羔羊瘤胃上皮cyclin D1的mRNA表达量显著(P=0.012)低于PS组,但与MS组无显著差异。CS组羔羊瘤胃上皮类胰岛素生长因子-1(insulin like growth factor,IGF-1)的mRNA表达量显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃上皮IGF-1R的mRNA表达量显著(P=0.001)高于PS组。上述结果表明:较高支链淀粉比开食料与较高直链淀粉比开食料相比,在瘤胃中较易降解生成挥发性脂肪酸,促进瘤胃上皮IGF-1及细胞周期蛋白mRNA的表达,进而有利于断奶前羔羊瘤胃上皮的发育。研究结果对制订羔羊营养方案具有重要指导意义。     

Use of NaHCO3 and MgO as additives for sheep fed only pasture for a restricted period of time per day: effects on intake,digestion and the rumen environment

Effects of NaHCO₃ and MgO buffer addition on intake and digestive utilization of a pasture were studied in wethers allowed a restricted time of access to forage. Twelve wethers housed in metabolic cages and fed fresh forage (predominantly Lotus corniculatus) ad libitum for 6 h/d were randomly assigned to one of the following treatments: a control forage without buffer (C) or a forage plus buffer composed of a mixture of 750 g/kg NaHCO₃ and 250 g/kg MgO at 20 g/kg dry matter intake (B). Feeding behaviour, feed and water intake and digestibility, urine output, Na urine elimination, kinetics of passage, ruminal pH and ammonia concentration, N balance and ruminal microbial N synthesis were determined in vivo, and the ruminal liquor activity was evaluated in vitro by fermentation of wheat straw. Addition of buffer increased total water intake (p = 0.05), Na urinary output (p = 0.01), purine derivative excretion in urine (p = 0.05) and tended to decrease mean total retention time in the digestive tract (p = 0.09). However, buffer addition increased ruminal pH (p < 0.001) and tended to decrease the ammonia concentration (p = 0.05). That use of buffer decreased ruminal activity was evidenced by a lower volume of gas produced in vitro (p = 0.01) possibly due to a lower microbial concentration in rumen liquor. The higher rumen dilution rate, likely due to a higher water intake, seems to have been the key driver of the actions of buffer supplementation on the rumen environment. Moreover, addition of NaHCO₃ led to an increased urinary Na excretion, which should be considered due to its likely negative environmental impacts.     

Effect of monensin withdrawal on rumen fermentation,methanogenesis and microbial populations in cattle

This study was designed to obtain information on the residual influence of dietary monensin on ruminant fermentation, methanogenesis and bacterial population. Three ruminally cannulated crossbreed heifers (14 months old, 363 ± 11 kg) were fed Italian ryegrass straw and concentrate supplemented with monensin for 21 days before sampling. Rumen fluid samples were collected for analysis of short chain fatty acid (SCFA) profiles, monensin concentration, methanogens and rumen bacterial density. Post‐feeding rumen fluid was also collected to determine in vitro gas production. Monensin was eliminated from the rumen fluid within 3 days. The composition of SCFA varied after elimination of monensin, while total production of SCFA was 1.78 times higher than on the first day. Methane production increased 7 days after monensin administration ceased, whereas hydrogen production decreased. The methanogens and rumen bacterial copy numbers were unaffected by the withdrawal of monensin.     

Buffer nitrogen solubility,in vitro ruminal partitioning of nitrogen and in vitro ruminal biological activity of tannins in leaves of four fodder tree species

This study explores the chemical composition, buffer N solubility, in vitro ruminal N degradability and in vitro ruminal biological activity of tannins in leaves from Gliricidia sepium, Leucaena leucocephala, Morus alba and Trichanthera gigantea trees. These tree leaves are a potential protein source for ruminants, but their site‐influenced nutritive value is largely unknown. Leucaena leucocephala leaves had the highest N content (42.1 g/kg DM), while T. gigantea leaves had the least (26.1 g/kg DM). Leucaena leucocephala had the highest buffer solubility index (20%), while 10% of the total N in leaves of the other three species was soluble. The rapidly fermentable N fraction ‘a’ was highest in M. alba leaves (734.9 g/kg DM) and least in T. gigantea leaves (139.5 g/kg DM). The rate of fermentation (c) was highest for M. alba (7%/hours) leaves. No significant correlations were recorded between buffer solubility index of N and in vitro ruminal N degradability parameters: a, b, and c. The highest response to tannin inactivation using polyethylene glycol, in terms of percentage increase in 36‐hours cumulative gas production, was recorded in M. alba (39%) and T. gigantea (38%) leaves. It was concluded that buffer solubility of N is not a good indicator of ruminal N degradation in the leaves of these tree species. Leaves of M. alba could be more valuable as a source of rapidly fermentable N when animals are offered low‐protein, high‐fibre diets compared with other tree species evaluated in the current study. However, when feeding M. alba leaves, the role of tannins must be considered because these secondary plant compounds showed significant in vitro ruminal biological activity.     

阴离子饲粮对活体外瘤胃发酵特性和营养物质消化的影响

应用双外流连续培养系统模拟瘤胃发酵,研究不同阴阳离子差(DCAD)水平的阴离子饲粮对活体外瘤胃发酵和营养物质消化率的影响。试验分2期进行,每期12个发酵罐,3个1组,分别投入4种不同DCAD水平的饲粮:+120-、9-、77-、145 mEq/kgDM,后3种饲粮通过添加阴离子添加剂降低DCAD。结果表明:①阴离子饲粮对瘤胃发酵pH值、氨态氮浓度、总挥发性脂肪酸浓度、主要挥发酸的摩尔比例均没有显著影响(P>0.2)。②对饲粮干物质、有机质、中性洗涤纤维、粗蛋白消化率没有显著影响(P>0.2)。③对瘤胃微生物合成效率没有显著影响(P>0.2)。试验结论为阴离子饲粮对活体外瘤胃发酵和饲粮营养物质消化率没有显著影响。     

不同RDP/RUP比例日粮对鲁西黄牛瘤胃发酵的影响

试验选用3头体况健康,体重350 kg左右,并安装有瘤胃瘘管的鲁西黄牛,采用3×3拉丁方试验设计,饲喂3种RDP/RUP比例分别为1.00∶1.00(A组)、1.22∶1.00(B组)、1.50∶1.00(C组)的等能等氮日粮,研究其对瘤胃内环境参数的昼夜动态变化规律的影响。结果表明:瘤胃pH受日粮RDP/RUP比值差异的影响不显著(P>0.05),A、B和C组pH变化范围分别为6.53~6.94、6.44~6.84和6.45~7.03。各组瘤胃pH均在饲喂后2~4 h达到最低值,随后逐渐上升;瘤胃液尿素氮以C组最高,A、B 2组差异不显著(P>0.05)。各组瘤胃液尿素氮在08:00饲喂后2 h达到最高值,但20:00饲喂后各组变化趋势不一致。A、B 2组瘤胃液尿素氮浓度白天较低,夜晚较高,而C组正好相反;各组瘤胃液氨氮平均浓度差异不显著(P>0.05),动态变化趋势基本相同,均在饲喂后2 h达到最高值;瘤胃微生物蛋白(MCP)平均浓度随RDP/RUP比值升高而显著提高,C组显著高于A、B 2组(P<0.05)。各组瘤胃液MCP浓度均在饲喂后2 h达到最低值。结果显示,以瘤胃液MCP浓度为衡量指标,日粮适宜的RDP/RUP比例为1.50∶1.00。     

Chemical Composition and Ruminal Dry Matter and Crude Protein Degradability of Spineless Cactus

The objective of this study was to determine ruminal degradability of the dry matter (DM) and crude protein (CP) of 10 varieties of spineless cactus (Opuntia spp.) grown in north‐eastern Brazil. Two ruminally fistulated steers were used in a randomized complete block design. Ash, CP, acid detergent fiber, and acid detergent lignin levels ranged from 10.4 to 13.3 %, 6.2–7.7 %, 19.8–24.8 % and 3.4–5.4 %, respectively. Relative to the other cactus varieties, Redonda had the highest (P < 0.05) in situ soluble DM fraction and effective DM degradability. No differences in effective DM degradability were observed between the other cactus varieties. In situ soluble CP fraction ranged from 1.7 % of CP for the 69 IPA/UFRPE variety to 11.1 % of CP for the Gigante variety. Slowly degradable CP fraction and its rate of degradation were similar among the cactus varieties (average 90 % of CP and 9 % h^?1, respectively). The cactus variety 69 IP/UFRPE had a lower (P < 0.05) effective ruminal CP degradability (ECPD) than the other varieties, which had a similar ECPD (average 66 % of CP). It was concluded that differences in ruminal degradability exist between cactus varieties, with more variations observed for ruminal DM than for CP degradability.     

日粮精粗比对泌乳奶牛瘤胃纤维降解酶活性及血液指标的影响

选择4头安装有瘤胃瘘管的泌乳奶牛,按4×4拉丁方设计,研究日粮精粗质量比约为30∶70的“高低质粗料型”日粮、30∶70的混合型高青贮日粮、50∶50的精粗料比例相当的日粮及65∶35的高精料日粮等4种不同精粗比日粮对瘤胃纤维降解酶活性及血液指标的影响。结果显示,奶牛采食4种精粗比不同的日粮,均能达到满足纤维降解及纤维分解菌生长要求的瘤胃pH值;日粮精粗比对微晶纤维素、羧甲基纤维素酶、纤维二糖酶和木聚糖酶等瘤胃纤维降解酶活性的影响差异不显著(P>0.05),对奶牛颈静脉血中血浆葡萄糖浓度和血清生长激素、胰岛素、胰高血糖素浓度亦无显著(P>0.05)影响。说明4种精粗比日粮对奶牛瘤胃纤维降解酶活性及上述血液指标没有影响。