应用PCR-DGGE技术对发酵全混合日粮中可培养乳酸菌的调查

为了了解发酵全混合日粮(TMR silage)发酵过程中的乳酸菌的菌种变化,应用MRS琼脂培养基对新鲜啤酒糟和豆腐渣以及它们的发酵TMR(贮存0、14和56 d)的乳酸菌进行培养,所得的可培养乳酸菌应用PCR-DGGE进行筛选,对菌体DNA的碱基序列进行测定,在GenBank上检索相近的16S rDNA序列对其鉴定.结果表明:2种原料中的乳酸菌种类完全不同;加入相同的其他材料变成TMR后,优势乳酸菌发生改变;发酵初期(14 d)的2种发酵TMR中都有2种乳酸菌,并且有1种乳酸菌相同;发酵后期(56 d)的2种发酵TMR中都有5种乳酸菌,而且有4种乳酸菌相同.由此可知,采用不同食品废弃物所制作的发酵TMR,在发酵后期所含菌种趋于相同、种类增加.     

燕麦和全株玉米不同配比对TMR发酵品质及有氧稳定性的影响

文章旨在探究全株燕麦和全株玉米的不同配合比例对TMR发酵品质及有氧稳定性的影响。将新鲜全株燕麦和/或全株玉米与玉米粉、豆粕、豆腐渣、苜蓿干草、麸皮、维生素-矿物质预混料、食盐,按照干物质30:34.5:10.5:10.5:10:3:1:0.5的比例混合调制TMR,其中,全株鲜燕麦和全株玉米的比例分别为0:30、10:20、20:10和30:0。分别于厌氧发酵后7、14、28 d和56 d开封,测定发酵品质、微生物组成、化学成分以及发酵56 d的TMR有氧稳定性。结果表明,所有处理组的TMR均发酵良好,具有较高的乳酸含量、较低的p H值和氨态氮含量,并具有良好的有氧稳定性。随着发酵时间的延长,p H值和乙酸含量显著降低、乳酸含量极显著升高(P<0.01);干物质(DM)、中性洗涤纤维(NDF)和可溶性碳水化合物(WSC)含量极显著降低(P<0.01)。随着全株燕麦配合比例的增加,DM、CP、NDF和酸性洗涤纤维(ADF)含量极显著升高(P<0.01),WSC含量极显著降低(P<0.01);p H值和乳酸含量极显著增加(P<0.01)。全株燕麦调制的TMR均发酵良好,但有氧稳定性随着全株燕麦比例的增加显著降低。     

应用体外产气法评定广西区内豆腐渣、木薯渣、啤酒糟的营养价值

试验应用注射器式体外产气法评定来源于广西的豆腐渣、木薯渣、啤酒糟的营养价值。瘤胃液来源于2头健康状况良好的带有瘤胃瘘管的水牛,记录了2、4、6、9、12、24、36、48、72、96 h的累积产气量,并取发酵了96 h的底物作为测定乙酸、丙酸和丁酸以及氨态氮的含量。试验结果显示,从各时间点累计产气量来看,豆腐渣和木薯渣的产气量相近,啤酒糟较低。潜在产气量最高的为豆腐渣,其次为木薯渣,啤酒糟最低。总挥发性脂肪酸最高的是木薯渣,其次为啤酒糟,豆腐渣最低,乙酸比例最高的是木薯渣,啤酒糟豆腐渣的丙酸比例相近,皆高于木薯渣的丙酸比例。根据饲料相对评定指数(RFV)计算公式,得到豆腐渣、木薯渣、啤酒糟的RFV值,从大到小排列顺序为:木薯渣(164.69)>豆腐渣(155.96)>啤酒糟(143.60)。由24 h体外产气量计算的可消化有机物质含量和代谢能从大到小的顺序为:木薯渣(762.12 g/kg,10.81 MJ/kg DM)、豆腐渣(742.77 g/kg,10.54 MJ/kg DM)、啤酒糟(513.03 g/kg,7.33 MJ/kg DM)。     

不同精粗比和含水量裹包TMR贮存效果研究

本试验是研究不同精粗比(60:40、50:50、40:60)和不同含水量(40%、50%和60%)全混合日粮(TMR)经过裹包处理后的贮存效果.试验对贮存15d的裹包TMR进行了感官评价并研究了干物质(DM)、粗蛋白(CP)、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)、灰分(Ash)、pH值、氨态氮(NH3-N)等成分变化情况.结果表明,在现试验条件下,各处理TMR都可以通过裹包的方式贮存,精粗比40:60或含水量40%、50%的TMR贮存效果优于其他处理组.     

杏饮料残渣青贮饲料的发酵品质研究

为了开发新的饲料资源,试验对杏饮料残渣青贮饲料的营养成分和发酵品质进行了研究。试验分为对照组(Ⅰ)、乳酸菌处理组(Ⅱ)、干啤酒糟处理组(Ⅲ)、干啤酒糟+乳酸菌处理组(Ⅳ),分别于青贮后第0、7、14、28和40d开封分析其营养成分和发酵品质。结果表明,不同的处理方法和贮藏时间以及处理方法和时间的交互作用对青贮的发酵品质和营养成分有显著影响;添加乳酸菌和干啤酒糟能够改善杏饮料残渣的发酵品质和营养价值。其中,以同时添加乳酸菌和干啤酒糟混合青贮效果最佳。     

蘑菇下脚料中提取木霉分解啤酒糟中纤维素的试验研究

为提高啤酒糟中纤维素的利用率,以啤酒糟作为原料,利用从蘑菇下脚料中提取的木霉在不同添加量、发酵时间、温度及p H条件下分解纤维素单因素试验,在此基础上进行L9(34)正交试验优化。结果表明:在单因素试验中,木霉分别在27℃,p H 5.0,发酵时间2 d,木霉添加量0.030 g(每5 g干啤酒糟)时分解率最高。正交试验的最佳条件是:在发酵温度27℃、发酵时间3 d、发酵p H 5.0和木霉添加量为0.005 g(每5 g干啤酒糟)的组合条件下,分解纤维素的量为7.13%,影响顺序为发酵p H发酵时间发酵温度木霉添加量。试验说明,从蘑菇下脚料中提取的木霉能够较好地分解啤酒糟中的纤维素。     

TMR制作粒度对肉牛增重与瘤胃微生物发酵的影响

为研究不同粒度全混合日粮(TMR)对肉牛增重和瘤胃发酵的影响,试验选取新疆褐牛(♂)与哈萨克牛(♀)杂交一代、体重为(305.6±5.6)kg的公牛70头,随机分为2组,预试期14 d,正试期44 d。日粮为不同搅拌时间的TMR,使用宾州筛进行粒度分布测定。对照组TMR搅拌时间为30 min,平均粒度为7.1 mm;试验组TMR搅拌时间为40 min,平均粒度为6.1 mm。结果表明:与对照组相比,试验组第1层(>19 mm)比例下降了6.3个百分点,第2层(>8 mm)和底层(≤1.18 mm)比例下降了3个百分点(P<0.01);结合剩料粒度分析结果研究发现,虽然延长搅拌时间后没有提高TMR总采食量,但提高了肉牛对TMR中粒度大于8 mm长纤维饲料的进食量,并提高了肉牛育肥全期平均日增重(P<0.05),饲料转化效率提高了12.7%(P=0.01);对TMR及宾州筛各层样品采用体外瘤胃发酵测试结果表明,延长TMR搅拌时间对体外干物质消化率、发酵产气量与产气速率、氨氮和挥发性脂肪酸(VFA)生成量无影响;但随着筛分粒度的下降,总VFA生成量升高(P=0.02),其中乙酸比例下降而丁酸比例升高(P<0.05)。综上所述,通过延长搅拌时间降低TMR粒度虽未能提高采食量,但可促进肉牛生长继而显著提高饲料转化效率,并有利于维持瘤胃微生物的正常发酵功能。     

添加剂对西藏啤酒糟全混合日粮青贮发酵品质及有氧稳定性的影响

本试验旨在评价布氏乳杆菌和双乙酸钠对西藏啤酒糟全混合日粮发酵品质和有氧稳定性的影响。试验设对照组(C)、布氏乳杆菌组(LB,106 cfu/g)和双乙酸钠组(SDA,0.5%)3个处理。青贮70d后全部开窖,取样分析发酵品质、营养和微生物成分,同时将剩余的发酵全混合日粮(TMR)暴露到空气中,用多通道温度记录仪记录温度变化,并分别在有氧暴露第6,9和14天取样分析,评定其有氧稳定性。结果表明,各组具有较高的乳酸含量,较低的氨态氮/总氮和微量的丁酸含量,发酵品质良好,添加双乙酸钠提高了乳酸含量,显著(P0.05)降低了pH、氨态氮/总氮和丁酸含量,同时显著(P0.05)提高了干物质回收率,但粗蛋白、粗灰分、粗脂肪、中性洗涤纤维和酸性洗涤纤维含量各组无显著差异;布氏乳杆菌处理组显著(P0.05)降低了乳酸、粗蛋白含量和干物质回收率,显著(P0.05)提高了pH和氨态氮/总氮值。在有氧暴露14d内,各组pH缓慢上升,且温度未超过环境温度2℃,显示有较高的有氧稳定性。添加布氏乳杆菌和双乙酸钠均在不同程度上降低了霉菌和酵母菌数量,对啤酒糟TMR有氧稳定性有积极的改善效果,其中双乙酸钠的效果优于布氏乳杆菌。综合考虑,在西藏地区生产啤酒糟发酵TMR时可添加0.5%的双乙酸钠,既能改善发酵品质,又能提高其有氧稳定性。     

添加中草药对发酵TMR化学成分、微生物组成及发酵品质的影响

文章旨在探究添加中草药对发酵全混合日粮(TMR)化学成分、微生物组成及发酵品质的影响。将苜蓿干草、玉米粉、豆浆渣、豆粕、预混料、食盐按照干物质(DM)36∶25.5∶21∶12∶5∶0.5的比例混合调制TMR,分为4组,分别加入2%无菌水、2%当归、2%黄芩、2%茯苓,室温条件下厌氧发酵,并于发酵7、14、28、56 d开封,测定化学成分、微生物组成及发酵品质。结果表明,发酵56 d后,所有组别的TMR发酵品质良好,具有较高的乳酸(LA)含量(7.31%DM～8.30%DM)、较低的p H(4.56～4.63)和氨态氮(NH3-N)含量(3.13%TN～3.75%TN)。随着发酵时间的延长,所有组别的p H和好氧细菌(AB)数量极显著降低(P0.01)、乳酸(LA)含量极显著升高(P0.01);DM和可溶性碳水化合物(WSC)含量极显著降低(P0.01)。添加中草药能够显著降低TMR发酵56 d后的p H(P0.05),提高LA含量(P0.05)。其中,当归组发酵56 d后LA含量最高,p H和NH3-N含量最低。综上所述,添加中草药可以有效抑制不良微生物的生长,改善TMR的发酵品质,其中以添加2%的当归效果最佳。     

裹包青贮技术在产奶牛用全混合日粮贮存中的应用研究

为使中小奶牛养殖场和奶牛养殖小区的奶牛养殖户用上营养平衡的全混合日粮（TMR）,研究利用拉伸膜包裹TMR饲料成品后贮存的效果。该研究对高产奶牛和中产奶牛用两种TMR进行裹包贮存,并于0d、3d、7d、15d、30d和60d分别取样进行裹包贮存外观评定、感官评定及营养物质含量和发酵指标的测定。试验结果表明,两种TMR裹包贮存后外观质量和感官质量良好;随贮存时间的延长,干物质含量有所降低,而粗蛋白、粗脂肪、粗纤维、粗灰分等营养指标有所增加;随贮存时间的延长裹包TMR的pH值显著降低（P〈0．05）,氨态氮含量显著上升（P〈0．05）。综合外观和感官评定及各项营养指标、发酵指标的变化,TMR裹包贮存技术可以为TMR饲喂技术在中小奶牛养殖场和奶牛养殖小区中应用提供技术支持。     

Milk production,nutrient digestibility and nitrogen balance in lactating cows fed total mixed ration silages containing steam‐flaked brown rice as substitute for steam‐flaked corn,and wet food by‐products

The objective of this study was to evaluate the effect of substituting brown rice grain for corn grain in total mixed ration (TMR) silage containing food by‐products on the milk production, whole‐tract nutrient digestibility and nitrogen balance in dairy cows. Six multiparous Holstein cows were used in a crossover design with two dietary treatments: a diet containing 30.9% steam‐flaked corn (corn TMR) or 30.9% steam‐flaked brown rice (rice TMR) with wet soybean curd residue and wet soy sauce cake. Dietary treatment did not affect the dry matter intake, milk yield and compositions in dairy cows. The dry matter and starch digestibility were higher, and the neutral detergent fiber digestibility was lower for rice TMR than for corn TMR. The urinary nitrogen (N) excretion as a proportion of the N intake was lower for rice TMR than for corn TMR with no dietary effect on N secretion in milk and fecal N excretion. These results indicated that the replacement of corn with brown rice in TMR silage relatively reduced urinary N loss without adverse effects on feed intake and milk production, when food by‐products such as soybean curd residue were included in the TMR silage as dietary crude protein sources.     

Effects of utilization of local food by‐products as total mixed ration silage materials on fermentation quality and intake,digestibility, rumen condition and nitrogen availability in sheep

Four wethers were used in a 4 × 4 Latin square design experiment to evaluate in vivo digestibility of total mixed ration (TMR) silage with food by‐products for dairy cows, and the ruminal condition and nitrogen (N) balance were examined. Five by‐products (i.e. potato waste, noodle waste, soybean curd residue, soy sauce cake and green tea waste) were obtained. Four types of TMR silage were used: control (C) containing roughage and commercial concentrate, T1:20% and T1:40% containing the five by‐products replacing 20% and 40% of the commercial concentrate on a dry matter (DM) basis, respectively, and T2:40% containing three by‐products (potato waste, noodle waste and soybean curd residue) replacing 40% of the commercial concentrate on a DM basis. The ingredients were mixed and preserved in oil drum silos for 4 months. The TMR silages showed 4.02–4.44% and 1.75–2.19% for pH and lactic acid contents, respectively. The digestibility of DM and neutral detergent fiber, and total digestible nutrient content were higher (P < 0.05) for T2:40% feeding than for C feeding. Urinary nitrogen excretion tended to be lower (P = 0.07) for T2:40% than for C. The results suggested 40% replacing of commercial concentrate by using the three food by‐products can be most suitable for TMR silage.     

不同水分吸附材料对象草青贮发酵品质及好氧稳定性的影响

象草收获时往往具有较高水分，单独青贮发酵品质不佳。为提高象草青贮品质，研究了不同水分吸附材料对象草青贮发酵品质和好氧稳定性的影响。处理设为对照、分别添加象草鲜重10%的米糠、小麦粉、干豆腐渣和麦麸。青贮60 d后分析发酵品质和好氧稳定性。结果表明：所有添加处理均比对照显著增加了青贮原材料的干物质（DM， P<0.01），降低了缓冲能（P<0.05）；青贮后，所有添加处理均显著提高了乳酸含量（P<0.01）。添加小麦粉显著增加了丁酸（0.81% DM）和氨态氮（NH₃-N， 14.40% TN）含量（P<0.05），而其他添加处理无显著影响（P>0.05），添加干豆腐渣的处理丁酸和氨态氮含量最低，分别只有0.04% DM和9.75% TN。添加小麦粉和干豆腐渣对青贮饲料好氧稳定性改善效果最佳。综合比较分析营养特性、青贮发酵品质和好氧稳定性，添加干豆腐渣的处理青贮效果最优。     

添加不同防霉剂对全混合日粮存放的影响

选择山梨酸钾、脱氢醋酸钠、苯甲酸钠、丙酸和丙酸钙5种防霉剂,分别以0.15％和0.3％的剂量添加到奶牛全混合日粮（total mixed ration,TMR）中进行7 d的贮存试验,测定防霉剂对TMR饲料贮存效果的影响。试验结果表明,添加脱氢醋酸钠和0.3％山梨酸钾组的TMR饲料在贮存期内没有发热产生,也未发生霉变,其它各防霉剂添加组从贮存第2天开始逐渐出现升温发霉现象。贮存第2天,添加丙酸、丙酸钙及0.15％苯甲酸钠组TMR饲料的含水量极显著低于对照组（P＜0.01）;贮存第3天和第5天,添加脱氢醋酸钠和添加0.3％山梨酸钾组TMR饲料的含水量高于对照组（P＜0.05）。贮存过程中TMR饲料的粗蛋白质和粗脂肪均无明显变化。综合各项指标,TMR饲料中添加脱氢醋酸钠和山梨酸钾比苯甲酸钠、丙酸和丙酸钙有较好的防霉效果。     

Effects of ethanol,molasses and Lactobacillus plantarum on the fermentation quality,in vitro digestibility and aerobic stability of total mixed ration silages in the Tibetan plateau of China

In Tibet, it is common practice to make and relocate total mixed ration (TMR) silages before feeding due to the uneven distribution of forages temporally and spatially. This study was conducted to investigate the effects of Lactobacillus plantarum (L), molasses (M) or ethanol (E) on the fermentation quality and aerobic stability of local adaptive TMR silage. After 45 days of ensiling, pH and ammonia nitrogen in inoculated TMR silages were significantly lower than control. During the first 6 days of the aerobic exposure test, a small fluctuation in lactic acid concentration for all TMR silages was observed, and then silages with ethanol continued this trend, while lactic acid in silage without ethanol sharply decreased until the end of the aerobic exposure period. Meanwhile, pH gradually increased along the aerobic exposure; silages treated with ethanol showed lower pH after 9 days of aerobic exposure. The population of yeast gradually increased during 6 days of aerobic exposure, after that an accelerated rise was observed in TMR silages without ethanol. The combinational beneficial effect of L. plantarum and ethanol was found in combined addition of ethanol and Lactobacillus plantarum silages (EL), indicated by intermediate fermentation quality and higher aerobic stability.     

Yeast population dynamics on air exposure in total mixed ration silage with sweet potato residue

To investigate the yeast population dynamics during air exposure in total mixed ration (TMR) silage containing sweet potato residue. TMR were ensiled in laboratory silos (1 kg) with or without two lactic acid bacteria strains, Lactobacillus plantarum (LP), and Lactobacillus amylovorus (LA). Fermentation characteristics were measured and yeast population was investigated by ITS1 region gene sequencing using Illumina MiSeq platform. All treatments were well ensiled, and L. amylovorus improved aerobic stability. During aerobic exposure, Pichia kudriavzevii was detected with increased relative abundance in all treatments and more relative abundant in LP. Pichia fermentans was more relative abundant in control. Higher relative abundance of Pichia anomala was detected in deteriorating LP. The relative abundance of Pichia ohmeri increased during later aerobic exposure in the control and LA, with a significant increase in the count of yeast population. Despite Cryptococcus was detected more relative abundant during early stage of aerobic exposure, the yeast population was below the detection limit. Aerobic deterioration was characterized by an increase in operational taxonomic units of Pichia. High relative abundance of P. anomala and P. kudriavzevii made aerobic deterioration easier. Inhibition of P. fermentans might be an effective strategy for improving the aerobic stability to some instance.     

Effects of adding food by‐products mainly including noodle waste to total mixed ration silage on fermentation quality,feed intake,digestibility, nitrogen utilization and ruminal fermentation in wethers

Four wethers were used in a 4 × 4 Latin square design experiment to evaluate the applicability of two types of total mixed ration (TMR) silage with food by‐products. Four food by‐products (i.e., potato waste, soy sauce cake, soybean curd residue and noodle waste) were obtained and mixed with commercial concentrate (CC) as TMR silage. The two types of TMR silage, T1 and T2, each contained CC, in addition to all by‐products for T1 (TRE1), and soy sauce cake and noodle waste for T2 (TRE2) on a dry matter (DM) basis. The silage was well‐fermented with low pH values and high lactic acid concentration. As the experimental treatments, T1, T2 and CC (CTL) were fed with a basal diet. The result showed that the digestibility of DM and organic matter (OM) were higher for T1 than for CC (P < 0.05), while crude protein digestibility was not significantly different among T1, T2 and CC. The retained nitrogen was not affected by inclusion of food by‐products. Ruminal pH in TRE1 and TRE2 immediately decreased compared to that in CTL. The study suggested that the two types of TMR silage, including food by‐products, might be used as a substitute for commercial concentrate.     

奶牛场饲料霉菌毒素感染现状的调查与分析

随机抽取3家不同规模奶牛场饲料原料及自配料32份,采用进口ELISA试剂盒检测黄曲霉毒素、玉米赤霉烯酮、赭曲霉毒素和呕吐毒素等4种常见霉菌毒素含量。结果表明：所检测的4种霉菌毒素在所有样品中的检出率为100％,并同时含有2～3种霉菌毒素。其中,除豆粕、苜蓿干草、稻草、豆腐渣外,几乎在所有的饲料样本中玉米赤霉稀酮含量都超标,青贮玉米最高,超标4倍,TMR饲料中玉米赤霉酮含量超出了2倍以上;玉米副产品GGDS中赭曲霉毒素、玉米赤霉酮含量超标严重,呕吐霉素存在微量或少量超标;黄曲霉毒素在青贮玉米、稻草、豆腐渣中有超标,占总样品的15．63％。     

复合微生物发酵豆腐渣对生长育肥牛生产性能的影响

[目的]为研究日粮中不同量豆腐渣和复合微生物发酵豆腐渣添加水平对生长肥育牛生产性能的影响。[方法]选用体重约190kg的西门达尔架子牛(约1岁)50头,随机分为5组,每组4个重复。[结果]结果表明:添加15%发酵豆腐渣的试验2组日增重比添加15%干豆腐渣的试验1组有显著提高(P<0.05);添加25%,35%发酵豆腐渣的试验3组和4组日增重比添加15%发酵豆腐渣的试验2组有明显提高(P<0.05),添加25%发酵豆腐渣的试验3组和添加35%发酵豆腐渣的试验4组在全期试验中差异不显著(P>0.05)。     

西藏不同饲草全混合日粮发酵品质和有氧稳定性的研究

为了充分利用西藏的农作物秸秆和饲草资源,本试验在当地奶牛场采用的以全株玉米和青稞秸秆为主要粗饲料的TMR配方基础上,用苜蓿、全株燕麦和小麦秸秆按不同比例替代全株玉米。青贮发酵45 d后评定各组发酵品质、营养特性和有氧稳定性。结果表明,发酵45 d后,TMR-2、TMR-3和TMR-4组pH值显著高于TMR-1(P<0.05),乳酸含量均低于TMR-1,但各替代组仅检测到微量丙酸和丁酸含量,且氨态氮/总氮较低,表明各组发酵品质均良好。发酵后各组水溶性碳水化合物含量均有大幅度下降,除TMR-1组外,其他3组干物质含量基本不变,粗蛋白、中性洗涤纤维、酸性洗涤纤维和粗脂肪等营养物质含量不同程度增加,其中TMR-2和 TMR-3组粗蛋白含量高于15%,能够满足改良奶牛的营养需要。在有氧暴露过程中,各组乳酸、乙酸和水溶性碳水化合物含量均出现不同程度的下降,丙酸和丁酸含量在有氧暴露第3天均有升高,之后逐渐下降。在有氧暴露9 d中,pH值均呈上升趋势,其中TMR-3组pH值上升幅度最小,其有氧稳定性最好。综上所述,虽然TMR-3发酵品质略低于TMR-1组,但其有氧稳定性有所提高,因此,TMR-3组可应用于生产实际中。