动物营养学报 >

2026 , Vol. 38 >Issue 3: 2306 - 2317

DOI: https://doi.org/10.12418/CJAN2026.184

研究论文

不同添加剂对水稻秸秆微贮饲料发酵品质和体外瘤胃发酵特性的影响

  • 郑春宾 , 1 ,
  • 霍永久 1 ,
  • 陈亚方 1 ,
  • 管彤 1 ,
  • 黄倩倩 1 ,
  • 赵国琦 1, 2 ,
  • 林淼 , 1, * ,
  • 王琳 , 1, 2, *
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  • 1 扬州大学动物科学与技术学院, 扬州 225009
  • 2 扬州大学农业科技发展研究院(国际联合实验室), 扬州 225009
*林 淼,教授,硕士生导师,E-mail: ;
王 琳,助理研究员,硕士生导师,E-mail:

郑春宾(2000—),男,黑龙江佳木斯人,硕士研究生,动物营养与饲料科学专业。E-mail:

收稿日期: 2025-07-31

  网络出版日期: 2026-03-16

基金资助

现代农业产业技术体系资助项目(CARS-36)

收起

Effects of Different Additives on Fermentation Quality and in Vitro Rumen Fermentation Characteristics of Rice Straw Microbial Silage

  • ZHENG Chunbin , 1 ,
  • HUO Yongjiu 1 ,
  • CHEN Yafang 1 ,
  • GUAN Tong 1 ,
  • HUANG Qianqian 1 ,
  • ZHAO Guoqi 1, 2 ,
  • LIN Miao , 1, * ,
  • WANG Lin , 1, 2, *
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  • 1 College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China
  • 2 Institute of Agricultural Science and Technology Development (International Joint Laboratory), Yangzhou University, Yangzhou 225009, China
*LIN Miao, professor, E-mail: ;
WANG Lin, assistant professor, E-mail:

Received date: 2025-07-31

  Online published: 2026-03-16

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摘要

本试验旨在探究不同添加剂对水稻秸秆微贮饲料发酵品质和体外瘤胃发酵特性的影响。试验1设置8个组,其中对照组(CK组)不添加任何添加剂;试验组分别添加糖蜜(M组)、巨大芽孢杆菌(BM组)、嗜酸性乳杆菌(LB组)、糖蜜+巨大芽孢杆菌(MBM组)、糖蜜+嗜酸性乳杆菌(MLB组)、巨大芽孢杆菌+嗜酸性乳杆菌(BMLB组)以及糖蜜+巨大芽孢杆菌+嗜酸性乳杆菌(MBMLB组),每组3个重复,微贮45 d后进行开袋取样,测定营养成分含量和发酵品质。试验2以试验1中未经发酵的水稻秸秆原料为FS组以及CK组和BMLB组进行体外瘤胃发酵,测定产气、营养物质降解率和发酵参数。结果表明:1)LB组水稻秸秆微贮饲料干物质(DM)含量最高;BMLB组粗蛋白质(CP)和粗脂肪含量较高,中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)含量较低。2)BMLB组水稻秸秆微贮饲料pH最低,乳酸含量最高。3)与FS组相比,BMLB组体外瘤胃发酵48和72 h产气量均显著提高(P<0.05),DM、CP、NDF和ADF降解率均显著提高(P<0.05);同时,BMLB组体外瘤胃发酵pH和乙酸/丙酸值显著降低(P<0.05),总挥发性脂肪酸、乙酸、丙酸和氨态氮含量均显著提高(P<0.05)。综上所述,巨大芽孢杆菌与乳酸菌复合添加可改善水稻秸秆微贮饲料营养价值、发酵品质及体外瘤胃发酵效率。

关键词： 巨大芽孢杆菌; 水稻秸秆; 微贮; 发酵品质; 体外发酵特性

本文引用格式

郑春宾 , 霍永久 , 陈亚方 , 管彤 , 黄倩倩 , 赵国琦 , 林淼 , 王琳 . 不同添加剂对水稻秸秆微贮饲料发酵品质和体外瘤胃发酵特性的影响[J]. 动物营养学报, 2026 , 38(3) : 2306 -2317 . DOI: 10.12418/CJAN2026.184

Abstract

This experiment was conducted to explore the effects of different additives on the fermentation quality and in vitro rumen fermentation characteristics of rice straw microbial silage. In experiment 1, eight groups were set, among which the control group (CK group) was not added with any additives, and the experimental groups were added with molasses (M group), Bacillus megaterium (BM group), Lactobacillus acidophilus (LB group), molasses+Bacillus megaterium (MBM group), molasses+Lactobacillus acidophilus (MLB group), Bacillus megaterium+Lactobacillus acidophilus (BMLB group), and molasses+Bacillus megaterium+Lactobacillus acidophilus (MBMLB group), respectively, with three replicates in each group. After 45 days of microbial fermentation, the bags were opened for sampling to determine the nutrient contents and fermentation quality. In experiment 2, taking the unfermented rice straw raw materials as FS group, as well as CK group and BMLB group in experiment 1 were used for in vitro rumen fermentation, and the gas production, nutrient degradation rates and fermentation parameters were determined. The results showed as follows: 1) the dry matter (DM) content in rice straw microbial silage in LB group was the highest; the contents of crude protein (CP) and ether extract in BMLB group were higher, while the contents of neutral detergent fiber (NDF) and acid detergent fiber (ADF) were lower. 2) The pH of rice straw microbial silage in BMLB group was the lowest, and the lactic acid content was the highest. 3) Compared with FS group, the gas production after in vitro rumen fermentation for 48 and 72 hours in BMLB group was significantly increased (P<0.05), and the degradation rates of DM, CP, NDF and ADF were also significantly increased (P<0.05). Meanwhile, the pH and acetate to propionate ratio during in vitro rumen fermentation in BMLB group were significantly decreased (P<0.05), while the contents of total volatile fatty acids, acetate, propionate and ammonia nitrogen were significantly increased (P<0.05). In conclusion, the combined supplementation of Bacillus megaterium and lactic acid bacteria can improve the nutritional value, fermentation quality and in vitro rumen fermentation efficiency of rice straw microbial silage.

Key words： Bacillus megaterium; rice straw; microbial silage; fermentation quality; in vitro fermentation characteristics

我国作为全球最大的水稻生产国,年均水稻秸秆产量超过2亿t,但综合利用率不足60%,大量秸秆被直接焚烧或废弃,导致严重的环境污染和资源浪费[1]。我国畜牧业的快速发展导致粗饲料日益短缺,制约了行业的可持续发展;合理利用秸秆饲料是缓解环境污染并补充饲料供给的有效途径[2]。水稻秸秆富含纤维素和半纤维素,但木质素含量高、可消化养分低且适口性差,难以满足畜禽生产的营养需求[3]。因此,优化水稻秸秆饲料品质,对促进水稻秸秆饲料化利用具有十分重要的现实意义[4-5]。
尽管通过微贮的方式能优化水稻秸秆的营养与适口性[6-7],但其低含量的水溶性碳水化合物及匮乏的附生乳酸菌限制了发酵品质[8-10]。研究表明,适量添加剂的引入可有效提升微贮效果,例如发酵促进剂(乳酸菌)、不良发酵抑制剂(丙酸、甲酸)、营养性添加剂(糖蜜、尿素)及吸附剂(秸秆、麸皮),可以明显改善微贮的养分含量及发酵品质,提高水稻秸秆微贮的成功率[11-14]。Fang等[15]研究发现,添加糖蜜可以通过促进微贮乳酸的生成来增强乳酸发酵模式,降低微贮饲料pH,减少有机物损失,在用于低发酵碳水化合物饲料作物时特别有益[16]。贾冬文等[17]研究表明,乳酸菌利用可发酵碳水化合物产生大量乳酸,能够快速降低pH,抑制腐败菌(如梭菌属、肠杆菌科)增殖,从而减少好氧降解造成的营养损失。
巨大芽孢杆菌(Bacillus megaterium)是产芽孢杆菌,为革兰氏阳性菌,具有降解纤维素的能力[18-19]。Deng等[20]研究发现,巨大芽孢杆菌不仅能促进动物对蛋白质等营养物质的消化、调节肠道pH、增强机体抗氧化能力并提升产品品质,还能显著提升奶牛对粗蛋白质(crude protein,CP)和中性洗涤纤维(neutral detergent fiber,NDF)的消化率,优化瘤胃发酵特性,进而提高饲料利用率,降低粪污中氮排放,减轻环境污染[21]。不过,现阶段巨大芽孢杆菌作为发酵饲料添加剂在优化秸秆饲料发酵品质中的应用鲜有报道。因此,本试验旨在探究添加巨大芽孢杆菌对水稻秸秆微贮饲料发酵品质及体外瘤胃发酵特性的影响,并通过不同添加剂单独添加或复合添加来寻找理想的添加剂类型,为水稻秸秆的饲料化应用奠定科学基础。

1 材料与方法

1.1 试验材料

本试验所用水稻秸秆采自扬州大学水稻试验田(10月收获),留茬7 cm,铡成2~3 cm备用。水稻秸秆营养成分含量分别为:66.25%干物质(dry matter,DM),4.04% DM CP,2.43% DM粗脂肪(ether extract,EE),20.47% DM粗灰分(crude ash,Ash),70.69% DM NDF,54.38% DM酸性洗涤纤维(acid detergent fiber,ADF)。水稻秸秆营养成分含量均为实测值,测定方法见1.3。
巨大芽孢杆菌由扬州大学动物营养与饲料工程技术研究中心提供,菌粉活菌数为4×1010 CFU/g;乳酸菌为嗜酸性乳杆菌(Lactobacillus acidophilus),菌粉活菌数为1×1010 CFU/g;糖蜜为市购产品。

1.2 微贮制作

试验1:将10 g巨大芽孢杆菌菌粉溶于1 L的去离子水中,制成浓度为4×108 CFU/mL的巨大芽孢杆菌稀释液;将4 g的嗜酸性乳杆菌菌粉溶于1 L去离子水中,制成浓度为4×107 CFU/mL的嗜酸性乳杆菌稀释液;将10 mL糖蜜溶于490 mL去离子水中,制备成2%糖蜜。试验共设计8个组,其中对照组(CK组)不添加任何添加剂,只添加去离子水(100 mL);试验组分别添加5 mL 2%糖蜜+95 mL去离子水(M组)、5 mL菌体浓度为4×108 CFU/mL的巨大芽孢杆菌+95 mL去离子水(BM组)、5 mL菌体浓度为4×107 CFU/mL的嗜酸性乳杆菌+95 mL去离子水(LB组)、5 mL 2%糖蜜+5 mL菌体浓度为4×108 CFU/mL的巨大芽孢杆菌+90 mL去离子水(MBM组)、5 mL 2%糖蜜+5 mL菌体浓度为4×107 CFU/mL的嗜酸性乳杆菌+90 mL去离子水(MLB组)、5 mL菌体浓度为4×108 CFU/mL的巨大芽孢杆菌+5 mL菌体浓度为4×107 CFU/mL的嗜酸性乳杆菌+90 mL去离子水(BMLB组)以及5 mL 2%糖蜜+5 mL菌体浓度为4×108 CFU/mL的巨大芽孢杆菌+5 mL菌体浓度为4×107 CFU/mL的嗜酸性乳杆菌+85 mL去离子水(MBMLB组)。原料经喷雾器均匀喷洒、混匀后,装入聚乙烯袋(30 cm×50 cm)真空封口,每袋100 g。每组3个重复,共24袋,于避光室温(23~30 ℃)贮藏45 d后开袋分析。

1.3 水稻秸秆微贮饲料营养成分含量测定

各组水稻秸秆微贮开袋混匀后,采用“四分法”取90 g代表性样品,经烘干、粉碎过40目筛后,测定其营养成分含量。其中,DM含量参照GB/T 6435—2014中方法测定,CP含量参照GB/T 6432—2018中方法测定,EE含量参照GB/T 6433—2006中方法测定,Ash含量参照GB/T 6438—2007中方法测定,NDF含量参照GB/T 20806—2022中方法测定,ADF含量参照NY/T 1459—2022中方法测定。

1.4 水稻秸秆微贮饲料发酵品质测定

开袋后,取新鲜水稻秸秆微贮饲料样品置于酒精消毒塑料容器内,混匀后取代表性样品5 g,加入45 mL去离子水,于4 ℃冰箱浸提24 h,用榨汁机粉碎,4层纱布过滤,浸提液用于测定pH、乳酸、乙酸、丙酸、丁酸和氨态氮(ammoniacal nitrogen,NH3-N)含量。采用pH计测定pH,采用苯酚次氯酸钠比色法测定NH3-N含量[22],采用气相色谱法测定有机酸组成[23]。

1.5 体外瘤胃发酵

试验2:选用试验1中未经发酵的水稻秸秆原料为FS组以及CK组和BMLB组进行体外发酵试验。瘤胃液供体动物为3头安装永久性瘤胃瘘管的健康干奶期奶牛,体重(650.0±30.0) kg。每日分别于07:30、14:30和21:00定时饲喂,自由饮水。饲粮组成及营养水平见表1。饲粮产奶净能参照冯仰廉等[24]的方法计算;钙含量采用原子吸收光谱法测定,磷含量采用钒钼酸铵试剂显色法测定,其余营养水平的测定方法同1.3。
表1 饲粮组成及营养水平(干物质基础)

Table 1 Composition and nutrient levels of the diet (DM basis)

项目 Items 含量 Content
原料 Ingredients
苜蓿干草 Alfalfa hay 25.30
玉米青贮 Corn silage 28.51
玉米 Corn 17.48
燕麦草 Oat grass 6.16
大麦 Barley 5.17
棉籽粕 Cottonseed meal 4.06
酒糟 Distillers grains 5.31
豆粕 Soybean meal (42% CP) 5.26
石粉 Limestone 0.32
氯化钠 NaCl 0.31
磷酸氢钙 CaHPO4 0.56
碳酸氢钠 NaHCO3 0.36
预混料 Premix1) 1.20
合计 Total 100.00
营养水平 Nutrient levels2)
产奶净能 NEL/(MJ/kg) 6.29
粗蛋白质 CP 15.02
粗脂肪 EE 3.96
中性洗涤纤维 NDF 41.11
酸性洗涤纤维 ADF 22.04
钙 Ca 0.82
磷 P 0.42

1)每千克预混料含 One kilogram of the premix contained the following:VA 300 000 IU,VD 385 000 IU,VE 1 455 IU,烟酸 niacin 550 mg,Cu 770 mg,Mn 930 mg,Fe 1 200 mg,Zn 3 600 mg,As 21 mg,I 50 mg,Co 12 mg。

2)产奶净能为计算值,其他营养水平为实测值。NEL was a calculated value, while the other nutrient levels were measured values.

1.5.1 人工瘤胃缓冲液和培养液配制

人工瘤胃缓冲液参照Menke等[25]按以下比例和顺序配制:400 mL蒸馏水+0.1 mL A液+200 mL B液+200 mL C液+1 mL刃天青溶液+40 mL还原剂溶液,各溶液即配即用,通入二氧化碳(CO2)至饱和,预热至39 ℃备用。晨饲前采集供体牛瘤胃液,参照邹诗雨等[26]的方法保存,迅速带回实验室,经4层纱布过滤得新鲜滤液,将滤液与人工瘤胃缓冲液以体积比1∶2配制成混合培养液,持续通入CO2并磁力搅拌。

1.5.2 体外发酵

采用Menke等[25]的体外产气法,取0.5 g风干样品置于培养瓶,加入75 mL人工瘤胃培养液;通入CO2约10 s维持厌氧环境,随即加盖密封(胶盖+铝盖封口钳压紧)。将3组培养管迅速放入已预热[(39.5±0.5) ℃]的水浴箱中恒温振荡72 h,每个样品设5个重复。

1.5.3 产气量及气体成分分析

从放进恒温摇床的时间开始计算,本试验共持续72 h,并分别在0、2、4、8、12、24、48和72 h使用数字压力传感器(DPG1000B15PSIG-5,Cecomp Electronics,美国)读取培养瓶中气体压力,参考Datsomor等[27]的公式计算产气量。计算公式为:
Vgas=Vj×Ppsi×0.068 004 084。
式中:Vgas为39 ℃校正气体体积(mL);Vj为各时间点发酵液面至顶空体积(mL);Ppsi为培养瓶压力(psi);实测值用空白培养产气量进行校正。
分别收集48和72 h的培养气体10 mL,用集气袋保存,采用气相色谱仪(GC9800,上海科创色谱仪器有限公司)测定气体成分。

1.5.4 发酵参数测定

体外发酵后,测定发酵液pH;采用气相色谱仪(GC9800,上海科创色谱仪器有限公司)测定乙酸、丙酸、丁酸和总挥发性脂肪酸(TVFA)含量,并计算乙酸/丙酸值;NH3-N含量参照冯宗慈等[22]的比色法进行测定。

1.5.5 营养物质降解率

按下列公式计算体外瘤胃发酵营养物质降解率:
体外瘤胃发酵某营养物质降解率(%)=100×[发酵前该营养物质含量(g)-发酵后该营养物质含量(g)]/发酵前该营养物质含量(g)。

1.6 数据统计和分析

试验数据经Excel 2021整理后,采用SPSS 25.0软件进行单因素方差分析(one-way ANOVA),差异显著时采用Duncan氏法进行多重比较,结果数据采用平均值和均值标准误(SEM)表示,P<0.05为差异显著。

2 结果与分析

2.1 不同添加剂对水稻秸秆微贮饲料营养成分含量的影响

由表2可知,LB组水稻秸秆微贮饲料DM含量最高,显著高于BM组、MBM组、MLB组和MBMLB组(P<0.05);BMLB组和MBM组CP含量较高,显著高于CK组和LB组(P<0.05);BMLB组EE含量显著高于其他组(P<0.05),同时NDF和Ash含量显著低于CK组(P<0.05);BMLB组和MBMLB组ADF含量显著低于MBM组(P<0.05)。
表2 不同添加剂对水稻秸秆微贮饲料营养成分含量的影响

Table 2 Effects of different additives on nutrient contents of rice straw microbial silage

项目
Items		 组别 Groups 均值
标准误
SEM		 P值
P-value
CK BM LB BMLB M MBM MLB MBMLB
干物质 DM/% 56.52ab 53.66b 61.45a 53.77ab 57.64ab 52.69b 52.34b 53.53b 0.822 0.048
粗蛋白质 CP/% DM 4.14b 4.19ab 4.15b 4.39a 4.19ab 4.32a 4.22ab 4.20ab 0.741 0.047
粗脂肪 EE/% DM 5.46d 6.35c 6.22c 6.91a 6.62b 6.59b 6.29c 6.35c 0.011 <0.001
中性洗涤纤维
NDF/% DM		 73.61a 67.54b 70.98ab 67.01b 67.77b 69.69ab 70.86ab 70.63ab 0.154 <0.001
酸性洗涤纤维
ADF/% DM		 59.33ab 58.31ab 58.45ab 56.51b 58.39ab 60.12a 58.41ab 55.19b 0.053 0.024
粗灰分
Ash/% DM		 21.89a 19.57bc 18.71bc 18.01c 20.76ab 18.75bc 20.04abc 18.89bc 0.310 0.016

同行数据肩标无字母或相同字母表示差异不显著(P>0.05),不同字母表示差异显著(P<0.05)。下表同。

In the data within the same row, the absence of superscript letters or the presence of the same superscript letters indicated no significant difference (P>0.05), while different superscript letters indicated significant difference (P<0.05). The same applied to the following tables.

2.2 不同添加剂对水稻秸秆微贮饲料发酵品质的影响

由表3可知,BMLB组水稻秸秆微贮饲料pH显著低于除MBMLB组外的其他组(P<0.05);试验组NH3-N含量均显著低于CK组(P<0.05),MLB组和MBMLB组NH3-N含量显著低于CK组、M组和MBM组(P<0.05);CK组和MBM组乙酸含量显著高于其他组(P<0.05),BMLB组乙酸含量最低;LB组、BMLB组和M组丙酸含量显著低于CK组、MBM组、MLB组和MBMLB组(P<0.05);CK组丁酸含量最高,显著高于其他组(P<0.05);此外,BMLB组乳酸含量最高,显著高于CK组、BM组、M组和MBM组(P<0.05)。
表3 不同添加剂对水稻秸秆微贮饲料发酵品质的影响

Table 3 Effects of different additives on fermentation quality of rice straw microbial silage

项目
Items		 组别 Groups 均值
标准误
SEM		 P值
P-value
CK BM LB BMLB M MBM MLB MBMLB
pH 6.63a 6.52a 6.08bc 5.80d 6.19bc 6.36ab 6.17bc 5.95cd 0.642 0.032
氨态氮
NH3-N/(mg/dL)		 2.35a 1.65bc 1.67bc 1.53bc 1.91b 1.75b 1.23c 1.26c 0.680 <0.001
乙酸
Acetate/(mmol/L)		 8.75a 5.19c 5.19c 3.89c 5.04c 8.67a 4.57c 6.88b 0.793 <0.001
丙酸
Propionate/(mmol/L)		 0.36bc 0.20cd 0.17d 0.16d 0.13d 0.43b 0.65a 0.39bc 0.331 <0.001
丁酸
Butyrate/(mmol/L)		 1.28a 0.97b 0.44d 0.33d 0.36d 0.70c 0.26d 0.32d 0.424 <0.001
乳酸
Lactic acid/(mmol/L)		 3.21c 3.94bc 4.19ab 4.98a 3.76c 4.03bc 4.76a 4.65ab 0.972 0.046

2.3 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵产气的影响

由表4可知,CK组和BMLB组水稻秸秆微贮饲料体外瘤胃发酵48h产气量显著高于FS组(P<0.05),同时BMLB组72 h产气量显著高于FS组和CK组(P<0.05);与FS组相比,CK组48 h产气中甲烷比例显著降低(P<0.05)。
表4 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵产气的影响

Table 4 Effects of combined supplementation of Bacillus megaterium and lactic acid bacteria on gas production in rumen fermentation of rice straw microbial silage in vitro

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
FS CK BMLB
48 h产气量 GP48 h/(mL/g DM) 150.15b 189.45a 208.60a 0.141 0.013
72 h产气量 GP72 h/(mL/g DM) 210.70b 222.05b 242.35a 0.242 0.024
48 h气体成分 Gas composition at 48 h/%
氢气 H2 0.02 0.01 0.01 0.013 0.230
甲烷 CH4 7.32a 4.93b 5.59ab 0.340 0.049
二氧化碳 CO2 88.47 88.30 88.81 0.631 0.421
72 h气体成分 Gas composition at 72 h/%
氢气 H2 0.22 0.19 0.11 0.011 0.643
甲烷 CH4 6.32 5.44 6.53 0.933 0.352
二氧化碳 CO2 73.56 79.76 76.27 0.151 0.192

2.4 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵营养物质降解率的影响

由表5可知,CK组和BMLB组水稻秸秆微贮饲料体外瘤胃发酵DM降解率显著高于FS组(P<0.05),同时BMLB组CP、NDF和ADF降解率均显著高于CK组和FS组(P<0.05)。
表5 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵营养物质降解率的影响

Table 5 Effects of combined supplementation of Bacillus megaterium and lactic acid bacteria on nutrient degradation rates in rumen fermentation of rice straw microbial silage in vitro

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
FS CK BMLB
干物质 DM 52.84b 66.69a 73.43a 0.810 0.025
粗蛋白质 CP 41.90b 42.21b 45.52a 3.398 0.031
中性洗涤纤维 NDF 38.70c 40.63b 43.77a 2.313 <0.001
酸性洗涤纤维 ADF 62.81b 62.93b 64.96a 0.424 0.028

2.5 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵参数的影响

由表6可知,与FS组相比,CK组和BMLB组水稻秸秆微贮饲料体外瘤胃发酵pH和乙酸/丙酸值显著降低(P<0.05),TVFA、乙酸、丙酸和丁酸含量显著提高(P<0.05);与CK组相比,BMLB组TVFA、乙酸和丙酸含量显著提高(P<0.05);此外,BMLB组NH3-N含量显著高于FS组和CK组(P<0.05)。
表6 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵参数的影响

Table 6 Effects of combined supplementation of Bacillus megaterium and lactic acid bacteria on rumen fermentation parameters of rice straw microbial silage in vitro

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
FS CK BMLB
pH 6.97a 6.43b 6.02b 0.112 0.016
总挥发性脂肪酸 TVFA/(mmol/L) 87.60c 95.32b 104.88a 2.570 <0.001
乙酸 Acetate/(mmol/L) 62.10c 64.37b 69.62a 1.144 <0.001
丙酸 Propionate/(mmol/L) 19.24c 22.85b 27.05a 1.223 <0.001
乙酸/丙酸 Acetate/propionate 3.24a 2.82b 2.59b 0.114 0.018
丁酸 Butyrate/(mmol/L) 6.10b 7.86a 8.01a 0.993 0.032
氨态氮 NH3-N/(mg/dL) 15.51b 16.81b 19.34a 0.614 <0.001

3 讨论

3.1 不同添加剂对水稻秸秆微贮饲料营养成分含量和发酵品质的影响

研究表明,在微贮过程中添加乳酸菌、巨大芽孢杆菌和糖蜜可以提高发酵品质和营养成分含量[28-31]。本研究通过乳酸菌、巨大芽孢杆菌和糖蜜单一或组合添加来分析水稻秸秆微贮营养成分含量的变化,结果发现LB组DM含量最高,这可能源于乳酸菌快速产酸抑制了好氧腐败微生物的生长,减少了有机物损失;CK组因发酵活动弱,基质消耗少且水分逸出有限,DM保留较多但发酵品质较差;而BM组、MBM组、MLB组和MBMLB组DM含量降低是有益发酵活动增强的体现,部分基质转化为挥发性脂肪酸(VFA)、CO2等挥发性产物而损失,但实质上提升了纤维降解效率和营养价值[32]。此外,与CK相比,BMLB组CP和EE含量显著提高,表明巨大芽孢杆菌与乳酸菌两者协同作用可促进纤维素的分解,由此产生更多的可溶性糖,抑制不良微生物的繁殖,促进有益微生物的繁殖,积累大量蛋白质产物,且减少对蛋白质和氨基酸的分解,这与陈亚方等[31]、陈鑫珠等[33]的研究结果类似。NDF和ADF作为反映饲料纤维质量和适口性的关键指标,其含量降低可改善适口性,提升采食量及营养物质消化率[34]。本研究发现,BMLB组NDF和ADF含量较低,可能是由于巨大芽孢杆菌与乳酸菌复合添加可以促进酶的分泌,有效降解秸秆中的结构性碳水化合物[35]。
适宜的pH可稳定瘤胃内环境,促进微生物代谢及粗纤维降解消化,改善发酵功能。本试验中,各组水稻秸秆微贮饲料NH3-N含量均处于正常范围(0.8~56.1 mg/dL)[36],且试验组NH3-N含量均显著低于CK组,说明本试验的不同添加剂处理有效抑制了蛋白质水解,此作用机制源于微贮发酵的酸性环境对蛋白质水解酶活性的抑制,从而降低了NH3-N含量。从发酵品质结果来看,与CK组相比,各试验组水稻秸秆微贮饲料pH以及NH3-N和丁酸含量降低,乳酸含量升高,并且添加剂组合添加效果优于单一添加,其中巨大芽孢杆菌与乳酸菌复合添加效果最为显著,表明水稻秸秆经巨大芽孢杆菌处理后,更有利于改善饲料的发酵品质,进而为机体提供更多的能量[37]。研究表明,在水稻秸秆微贮过程中,添加发酵促进剂、营养性添加剂和微生物菌剂等添加剂,可通过调控发酵代谢路径实现发酵品质的提升[37-38]。具体而言,适宜剂量的发酵促进剂、营养性添加剂与菌制剂协同作用能够显著促进乳酸菌等有益微生物的增殖,使其在发酵过程中产生大量乳酸,随着乳酸的积累,使得微贮过程中的pH得以有效降低[39-40]。同时,适宜的酸性发酵环境可增强微生物对NH3-N的利用效率,促使菌体蛋白合成代谢,从而降低发酵过程中NH3-N含量,提高水稻秸秆微贮的发酵品质[40-41]。值得注意的是,MBMLB组在某些营养成分含量和发酵品质指标上与BMLB组无显著差异,但BMLB组在无需外加碳源的条件下仍能达到同等优良效果,且其发酵品质总体更为稳定优异。从实际应用角度,BMLB组处理更具经济性和操作便利性,避免了糖蜜添加所带来的成本提高和工艺复杂性。

3.2 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵产气的影响

产气量是评估微贮饲料发酵程度和瘤胃饲料利用率的关键指标[42],其气体源于微生物分解碳水化合物和蛋白质的过程。产气量与瘤胃微生物活性呈正相关,产气量越高表明发酵活动越强[43-45]。本试验中,BMLB组48和72 h累计产气量均高于FS组和CK组,说明添加剂提高了底物的可发酵性,减少了营养物质损失并增强了瘤胃微生物活性,从而提高了体外产气量,这与Li等[46]的研究结果一致。甲烷作为瘤胃发酵产物的一种,是造成温室效应加重的原因之一[47]。因此,减少反刍动物甲烷排放可促进生态养殖。同时,反刍动物甲烷排放量的减少,主要可通过减少氢的生成、降低产甲烷菌数量以及改善瘤胃发酵性能等方式实现[48]。本试验中,与FS组相比,CK组48 h产气中甲烷比例显著降低,其主要原因可能是水稻秸秆经微贮发酵后,能改善在瘤胃内的发酵性能,促进丙酸合成,进而减少甲烷生成量,这与曾旭等[49]的研究结果相一致。但与CK组相比,BMLB组甲烷比例并无显著差异,这仍需进一步进行动物试验验证。

3.3 巨大芽孢杆菌与乳酸菌复合添加对水稻秸秆微贮饲料体外瘤胃发酵营养物质降解率和发酵参数的影响

本试验中,与FS组相比,CK组和BMLB组DM、CP、NDF和ADF降解率均有不同程度的提高,且巨大芽孢杆菌与乳酸菌复合添加效果更为显著,表明微生物菌剂复合发酵可提升水稻秸秆营养价值,促进反刍动物利用,这与邱亚兰等[35]的研究结果相一致。
pH是瘤胃微生物生长繁殖的重要条件,保持瘤胃内环境稳定的适宜pH为5.6~7.5[50]。本试验中,各组体外发酵瘤胃液pH均在适宜范围内,BMLB组pH虽显著低于FS组,但仍处于正常范围内,说明体外瘤胃发酵环境处于稳定状态,能促进微生物的生长和代谢,这与赵超等[51]的试验结果相符。反刍动物70%~80%的能量源自瘤胃微生物代谢产生的VFA,其主要成分为乙酸、丙酸和丁酸,三者占比约达95%[35,52]。VFA不仅是反映动物对饲粮利用情况的重要指标,还对胃肠道激素分泌和肠道上皮细胞生长增殖具有调节作用[53]。其中,乙酸能够合成乳脂,丙酸在动物体内代谢可产生葡萄糖,对动物育肥有积极作用[54]。VFA含量受饲粮组成、添加剂和内环境等多种因素调控,其中饲粮组成最为关键,优化饲粮搭配可显著提升其产量。作为维持牛等反刍动物生命的重要能量来源,VFA参与瘤胃代谢过程,其生成与含量调控对反刍动物的能量平衡和生长发育至关重要[43,55]。本试验中,BMLB组TVFA、乙酸和丙酸含量均显著高于其他组,说明巨大芽孢杆菌与乳酸菌复合添加能够改善微贮效果,这与吴鹏辉等[56]的研究结果一致。瘤胃微生物区系适宜NH3-N含量为0.35~29.00 mg/dL[57-58],本试验各组体外瘤胃发酵NH3-N含量均在此范围内,并且BMLB组NH3-N含量显著高于其他组,表明巨大芽孢杆菌与乳酸菌复合添加有助于促进发酵底物中含氮化合物的分解。

4 结论

① 单独添加乳酸菌能有效保存水稻秸秆微贮饲料DM,巨大芽孢杆菌与乳酸菌复合添加在提高CP和EE含量、降低纤维组分含量方面效果较为均衡和突出。
② 在发酵品质上,巨大芽孢杆菌与乳酸菌复合添加能有效降低水稻秸秆微贮饲料pH,促进乳酸发酵,其发酵效果最佳。
③ 在体外瘤胃发酵中,与水稻秸秆原料相比,巨大芽孢杆菌与乳酸菌复合添加能显著提高水稻秸秆微贮饲料产气量、营养物质降解率和VFA产量。

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