Identification of bacteria in total mixed ration silage produced with and without crop silage as an ingredient

As a forage source for total mixed ration (TMR) silage production, locally produced crop silage is now used in addition to imported hay. This type of TMR ensiling is regarded as a two‐step fermentation process; hence, a survey was carried out to determine whether the bacteria in crop silage affect the subsequent TMR ensiling. Fermentation product contents and bacterial community were determined for TMR silage and its ingredient silages collected in August, October and November. August product contained corn, sorghum and Italian ryegrass silages, October product had wheat silage exclusively and November product did not include any crop silages. Acetic acid, lactic acid, 2,3‐butanediol and ethanol were predominant fermentation products in corn, sorghum, Italian ryegrass and wheat silages, respectively. Robust lactic acid fermentation was seen in TMR silage, even if acetate‐type and alcohol‐type silages were mixed as ingredients. The finding that bacterial community of the TMR silage appeared unrelated to those of ingredient silage supported this. Silages of various fermentation types can therefore be formulated without interfering with lactate‐type fermentation in TMR silage.     

乳酸菌青贮技术研究进展

本文综述了国内外乳酸菌青贮技术的内容、应用及效果, 总结了添加乳酸菌对青贮饲料发酵品质和有氧稳定性的影响, 以及不同乳酸菌菌株的筛选和鉴定及其对动物生产性能的影响。通过分析以上内容, 对乳酸菌青贮技术今后的发展方向, 特别是分子生物学在乳酸菌青贮技术研究中应用的广阔前景做了展望。     

乳酸菌青贮添加剂的研究与应用

很多添加剂被用来促进青贮发酵过程,其中研究最广泛的就是乳酸菌青贮添加剂。本文主要阐述了在青贮中加入乳酸菌添加剂可以促进作物的发酵速度,并且改善家畜的生产性能。     

同/异型发酵乳酸菌对全株玉米青贮营养成分和瘤胃降解特征的影响

以新饲玉10号全株青贮玉米为研究材料,旨在探究同/异型发酵乳酸菌对其主要营养成分和瘤胃降解率特征的影响。采用真空袋法调制青贮,共设计4个发酵处理,分别为不添加任何菌剂(CK);复合同型发酵乳酸菌植物乳杆菌+戊糖片球菌(T),添加量为1∶1,1×10^5 cfu·g^-1;异型发酵乳酸菌布氏乳杆菌(Y),添加量为1×10^5 cfu·g^-1;复合同、异型发酵乳酸菌(TY),添加量为1∶1∶1,1×10^5 cfu·g^-1。通过对模拟开窖60 d主要营养成分和绵羊瘤胃降解特征参数的分析。结果表明:干物质(DM)、中/酸性洗涤纤维(N/ADF)、可溶性碳水化合物和粗脂肪含量最优均为Y处理,粗蛋白和淀粉含量各试验处理均极显著高于CK处理(P<0.01),粗灰分含量最高为CK处理。DM和NDF有效降解率按高低排序均为T和Y处理>CK和TY处理(P<0.01)。有机物(OM)有效降解率最高为Y处理(P=0.003)。各处理ADF有效降解率间差异不显著(P>0.05)。说明玉米青贮中Y处理可显著增加开窖60 d时各营养成分含量(粗灰分除外),且显著提高DM和OM有效降解率,TY处理可显著提高NDF有效降解率。综合营养成分和有效降解率12项指标进行隶属函数评价,各处理按优劣排序为:Y处理>T处理>TY处理>CK处理。     

Characterization and application of lactic acid bacteria for tropical silage preparation

Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid‐producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA‐DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small‐scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science     

添加乳酸菌和纤维素酶对苜蓿青贮品质的影响

初花期收获的苜蓿(Medicago sativa)经过0、8、32 h的晾晒,添加乳酸菌和纤维素酶进行青贮。结果表明,苜蓿凋萎（干物质含量为38.45%）青贮可以使青贮料的氨态氮含量显著降低,并保存有更多的粗蛋白质,生成更多的乳酸。苜蓿青贮时添加乳酸菌和纤维素酶能明显改善苜蓿青贮料的发酵品质,即降低青贮料的氨态氮含量和保存更多的粗蛋白质以及生成更多的乳酸。晾晒8 h的苜蓿添加乳酸菌（106 cfu/g）和纤维素酶（0.05 g/kg）的青贮效果最好;而晾晒32 h的苜蓿直接青贮的效果最好。     

青贮饲料乳酸菌添加剂的应用现状

青贮饲料乳酸菌添加剂对改善青贮饲料品质具有显著作用。为了明确目前国内外对乳酸菌饲料添加剂的研究及应用情况,本文对近年来青贮饲料乳酸菌添加剂的发酵特性,筛选鉴定及应用等方面的研究做了系统的归纳和阐述。目前研究已明确了青贮饲料乳酸菌添加剂的发酵原理和青贮的过程,研究内容主要包括以下几个方面:1)乳酸菌添加剂本身、环境因素及饲料作物特性这3个方面对发酵产生的影响;2)青贮饲料乳酸菌添加剂的筛选和优良乳酸菌添加至青贮饲料的研究;3)乳酸菌菌株单独添加、与酶制剂或其他功能性微生物混合添加应用研究。综合以上研究,认为未来我国可以对不同生态环境中的乳酸菌资源进行发掘、筛选,并根据草牧业发展的实际需求以及我国现有的农艺,加强乳酸菌加工利用工艺的研发。     

添加乳酸菌和纤维素酶对苜蓿青贮品质的影响(简报)

初花期收获的苜蓿经过0,8和32 h的晾晒,添加乳酸菌和纤维素酶进行青贮。结果表明,不同干物质含量的苜蓿青贮时,乳酸菌 纤维素酶的添加均能使青贮pH值快速下降;苜蓿半干青贮(干物质含量为38.45%)可使青贮氨态氮含量显著降低,并保存有较多的粗蛋白质,生成较多乳酸,其中以添加乳酸菌106cfu/g 纤维素酶0.05g/kg的青贮效果最好,其青贮综合评定得分最高,达93分。     

玉米秸秆与废弃白菜的混贮品质及乳酸菌多样性研究

为研究玉米秸秆和废弃白菜混合青贮可行性,考查二者在不同质量比时的混贮品质,设计了6个不同的混贮比例,分别为29∶19,27∶21,25∶23,23∶25,21∶27和19∶29。混合青贮30 d后对其化学组分和发酵品质进行分析,筛选品质最佳的混贮比例,并进一步研究了品质最佳混贮组的乳酸菌多样性。结果表明,质量比为21∶27的混贮5(ME₅)组的pH和氨态氮/总氮显著低于其余混贮组(P<0.05),乳酸含量显著高于其余混贮组(P<0.05)。ME₅组的干物质和能源组分综纤维素含量较高,而酸性洗涤木质素含量较低,综合判定该组的混贮品质优于其他5组。乳酸菌多样性结果显示,从ME₅组中共分离出10株乳酸菌,分属于3个属,4个种。3个属分别是乳杆菌属、肠球菌属和明串珠菌属。4个种分别是2株短乳杆菌、1株屎肠球菌、5株肠膜明串珠菌肠膜亚种和2株植物乳杆菌,其中同型发酵乳酸菌乳杆菌属和肠球菌属为该青贮体系的关键乳酸菌。     

Identification and characterization of lactic acid bacteria isolated from mixed pasture of timothy and orchardgrass,and its badly preserved silage

In order to understand the relationship between lactic acid bacteria (LAB) species and silage fermentation, a total of 65 LAB strains isolated from mixed pasture of timothy (Phleum pratense L.) and orchardgrass (Dactylis glomerata L.), and its badly preserved silages were subjected to phenotypic and genetic analysis. According to these analyses, the isolates were divided into 13 groups, including Enterococcus gallinarum, Lactobacillus acidipiscis, L. coryniformis subsp. coryniformis, L. coryniformis subsp. torquens, L. curvatus, L. paraplantarum, L. plantarum subsp. argentoratensis, L. plantarum subsp. plantarum, L. sakei subsp. carnosus, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Leuconostoc pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Weissella hellenica, Weissella paramesenteroides and Carnobacterium divergens. This is the first report to document that C. divergens, L. acidipiscis, L. sakei subsp. carnosus, L. garvieae, phenotypically novel L. lactis subsp. cremoris, E. gallinarum and W. hellenica are present in vegetative forage crops. L. plantarum group strains were most frequently isolated from the badly preserved silages. Some isolates showed a wide range of growth preferences for carbohydrate utilization, optimal growth pH and temperature in vitro, indicating that they have a high growth potential. These results are useful in understanding the diversity of LAB associated with decayed silage of timothy and orchardgrass.     

苜蓿青贮乳酸菌的筛选及其对苜蓿青贮发酵的影响

为获得苜蓿青贮用乳酸菌,本研究从苜蓿附生乳酸菌中通过抑菌活性、酸化苜蓿粉及结构性糖代谢中间产物的性能指标筛选优良菌株,并研究其对苜蓿(Medicago sativa)青贮发酵的影响。结果表明,1)筛选到抑菌活性较高、酸化苜蓿粉及结构性糖代谢中间产物综合性能较好的菌株ZZU A341,被鉴定为植物乳杆菌(Lactobacillus plantarum);2)青贮90 d,添加该菌株的处理组pH、氨态氮含量低于添加商业菌株(YX)的处理组,显著低于自然青贮(P <0.05)。综上所述,使用该筛选方法获得的优良乳酸菌可有效改善苜蓿发酵品质。     

Fermentation quality and aerobic stability of mulberry silage prepared with lactic acid bacteria and propionic acid

The objective of this study was to examine the fermentation quality and aerobic stability of mulberry (Morus alba L.) silage prepared with lactic acid bacteria (LAB) and propionic acid (PA). The selected LAB strains Lactobacillus (L.) plantarum LC365281 (L1) and L. brevis LC365282 (L2), and commercial inoculant strains L. plantarum Gaofuji (GF) and L. buchneri Fresh (FR), and PA were used as additives for silage preparation. Silage treatments were designed as control, L1, L2, GF, FR, PA, PA + L1, PA + L2, PA + GF, or PA + FR. After 30 days of ensiling, the fermentation quality of silages treated with PA + L1 was improved, with a lower (p < 0.05) pH and NH₃‐N content than those of other treatments. During the aerobic exposure, the PA + LAB‐treated silages displayed an aerobic stability with stable pH value and lactic acid content. The results confirm that L. plantarum L1 and PA were the best additive combination for ensiling mulberry.     

乳酸菌对苜蓿青贮发酵品质和细菌菌群的影响

为探究乳酸菌作为青贮添加剂对苜蓿青贮发酵品质和细菌菌群的影响,试验选取萎焉苜蓿为原料分为三个处理组,分别为无添加剂对照组(CK组)和分别添加1×106 cfu/g乳酸菌YX(LPI组)、乳酸菌ZZU A341(LPA组).结果表明:LPI或LPA组乳酸积累量显著高于CK组(P<0.05),pH和氨态氮含量显著低于CK组...     

添加乳酸菌和菠萝皮对柱花草青贮品质的影响

柱花草是热带、亚热带的优质豆科牧草,蛋白含量高、营养品质好,为促进柱花草的加工利用,本试验研究了添加不同乳酸菌和菠萝皮对其青贮发酵品质和有氧稳定性的影响。青贮处理为CK(对照)、LQ(青贮宝)、LF(发酵乳杆菌)、LP(植物乳杆菌)、LS(鼠李糖乳杆菌)、B(20%菠萝皮)、BLQ(B+LQ)、BLF(B+LF)、BLP(B+LP)、BLS(B+LS)。添加处理后青贮60 d进行分析。结果表明,柱花草含有较少的可溶性碳水化合物和乳酸菌,自然青贮其pH超过5.0,发酵品质差;所有添加物都显著降低pH、增加乳酸含量(P＜0.05),明显改善了柱花草青贮料的发酵品质。单独添加菠萝皮的丁酸和NH₃-N含量显著高于所有单独添加乳酸菌(P＜0.05),pH与LP以外的其他3种乳酸菌差异不显著(P＞0.05)。除LP的乙酸含量较高,丁酸含量较低外,4种乳酸菌对其他各个发酵指标的影响没有显著差异(P＞0.05)。本试验所用4种乳酸菌与菠萝皮混合添加,都进一步改善了柱花草青贮料的发酵品质,特别是BLP的青贮效果最佳,其pH值、乙酸和NH₃-N含量低,乳酸含量及乳酸与乙酸比高。青贮袋开封后,包括对照在内的所有青贮料的有氧稳定性均较佳。     

含水量和乳酸菌添加剂对多花黑麦草青贮品质的影响

本试验旨在研究含水量和乳酸菌添加剂对多花黑麦草青贮品质的影响,探究优质多花黑麦草青贮饲料的调制方法。试验选用孕穗期的多花黑麦草为原料,原料含水量设85.23%(W₁),74.70%(W₂)和65.70%(W₃)3个水平,乳酸菌添加浓度设置对照组0 cfu/g鲜样(L₀),10⁵ cfu/g鲜样(L₁),10⁶ cfu/g鲜样(L₂),10⁷ cfu/g鲜样(L₃)4个水平。桶装青贮42 d后开盖,取样测定青贮饲料的青贮品质。结果表明,在高含水量(85.23%)的条件下,无论是否添加乳酸菌,丁酸的含量和氨态氮/总氮均未能达到优质青贮的条件,青贮失败;当含水量为74.70%时,添加乳酸菌组的青贮品质优于未添加组,且随着添加浓度的增加青贮品质更佳;在含水量为65.70%的条件下,无论是否添加乳酸菌添加剂,青贮饲料均能获得较好的青贮品质,但添加乳酸菌后效果更佳。因此,W₃L₃(含水量65.70%+10⁷ cfu/g鲜样)处理的青贮效果最佳。     

短芒大麦草青贮微生物特性研究及优良乳酸菌筛选

以短芒大麦草为研究对象,利用传统培养法从叶围和青贮发酵体系中分离出乳酸菌、大肠杆菌、好氧细菌、酵母菌和霉菌,并计数;结合细菌形态学、生理生化特征及16S rDNA序列分析鉴定分离出的乳酸菌菌株;通过研究乳酸菌的生长曲线、产酸特性及耐酸性,筛选优质乳酸菌。以期探明短芒大麦草叶围及青贮发酵体系中微生物菌群特性及青贮料中乳酸菌多样性,筛选出具有促发酵效果的乳酸菌菌株,为有益微生物饲料研发奠定基础。试验结果表明,短芒大麦草经青贮发酵后各微生物菌群数量发生不同程度变化,乳酸菌数量由0 cfu/g FM增加到4.00×10⁸ cfu/g FM,酵母菌数量由8.50×10⁵ cfu/g FM增加到1.02×10⁸ cfu/g FM,而好氧细菌、大肠杆菌和霉菌数量变化不明显;从短芒大麦草青贮发酵体系分离得到4株乳酸菌,经鉴定Lx36为Lactobacillus pentosus,Lx37为Lactobacillus brevis,Lx53为Pediococcus pentosaceus,Lx54为Lactobacillus parabuchneri;筛选得到1株益于青贮的乳酸菌株Lx36,约在20 h后进入稳定生长期,OD值达到4.21,且发酵12 h的pH仅为4.08,并可以在pH=3.0环境条件下生长。综上所述,青贮发酵是体系中各种微生物相互作用的过程,微生物菌群的数量及变化直接影响青贮饲料发酵品质。短芒大麦草青贮饲料中乳酸菌种类较丰富,筛选得到的戊糖乳杆菌繁殖速度快、产酸能力强同时表现出了较强的耐酸性,具有潜在的生产应用价值,适宜用作促发酵的青贮添加剂菌种。     

Fermentation quality and chemical composition of shrub silage treated with lactic acid bacteria inoculants and cellulase additives

Effects of lactic acid bacteria (LAB) inoculants and cellulase additives on fermentation quality and chemical compositions of shrub silages were studied by using a small‐scale fermentation system. Two LAB inoculants of Qingbao (Lactobacillus plantarum, Pediococcus acidilacticii, Lactobacillus casei and Clostridium phage) and Caihe (Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici) and a commercial cellulase made from Trichoderma reesei were used as additives for intermediate pea‐shrub, rush bushclover, arborescent ceratoides and shrubby silage preparation. The crude protein, neutral detergent fiber and water‐soluble carbohydrate contents of the four shrub materials were 10.1–14.2, 62.6–67.2 and 1.9–3.5% on a dry matter basis, respectively. All shrub silages had pH 3.40–4.43, ammonia‐N 0.1–0.2% g/kg and lactic acid 1.3–2.9% on a fresh matter basis. The silage quality of LAB‐inoculated silages did not have a greater effect than control silages, except shrubby silage preparation. Silages treated with the cellulase, the pH of rush bushclover and shrubby sweetvetch silage were significantly (P < 0.05) lower and the lactic acid content were significantly (P < 0.05) higher than the control silages. The results confirmed that shrub contained a relatively high content of crude protein; its silages can be preserved in good quality, and they are new potential resources for livestock feed.     

乳酸菌及丙酸钙对全株玉米和燕麦青贮饲料发酵品质和霉菌毒素含量的影响

为研究乳酸菌及丙酸钙对全株玉米和燕麦青贮饲料发酵品质、营养品质、微生物数量、霉菌毒素含量及有氧稳定性的影响,利用蒸馏水(CK组)、复合乳酸菌(LAB组,添加量为5×10⁵CFU·g^-1鲜样)、丙酸钙(PACA组,添加量为鲜草重的0.4%)以及乳酸菌和丙酸钙复合(LAB+PACA)分别添加进全株玉米和燕麦原料中青贮120 d。结果表明:全株玉米青贮饲料发酵品质优良而萎蔫后的燕麦青贮丁酸和氨态氮含量较高。添加剂在不同青贮饲料中表现不同。所有添加剂均显著提高了燕麦青贮饲料的乳酸和乙酸含量,降低了pH值、氨态氮、丁酸含量和酵母菌数量(P<0.05)。而PACA和LAB+PACA组则显著提高了玉米青贮饲料中的淀粉和可溶性碳水化合物含量和有氧稳定性并且降低了霉菌和酵母菌数量(P<0.05)。LAB和LAB+PACA的使用有效降低了玉米青贮饲料中的黄曲霉毒素B1含量(P<0.05)。在两种青贮饲料中丙酸含量只有PACA和LAB+PACA组能够显著增加。因此,添加剂尤其是LAB+PACA的复合添加对于提高玉米和燕麦青贮饲料的青贮品质和安全性...     

乳酸菌和纤维素酶对狼尾草和圆叶决明混合青贮品质的影响

为研究青贮添加剂对狼尾草和圆叶决明混合青贮效果的影响,利用乳酸菌、纤维素酶作为青贮添加剂对狼尾草和圆叶决明(混合比例为75∶25)进行混合青贮研究.试验设4个处理组,分别为对照组(无添加),试验Ⅰ组(添加≥1.0×105CFU·g-1乳酸菌剂),试验Ⅱ组(添加0.2g·kg-1纤维素酶),试验Ⅲ组(同时添加≥1.0×1...     

植物乳酸菌和贮藏温度对辣木叶青贮品质的影响

从辣木叶青贮中筛选分离出4个乳酸菌菌株,鉴定后分别为Lactobacillus plantarum(LP),Lactobacillus far?ciminis(LF),Weissella thailandensis(W)和Lactococcus lactis(E).试验旨在探究不同温度下添加植物乳酸菌对辣木叶青贮的青贮...