Chinese Journal of Animal Nutrition >

2026 , Vol. 38 >Issue 8: 6276 - 6288

DOI: https://doi.org/10.12418/CJAN2026.501

RESEARCH PAPER

Effects of Co-Fermentation with Lactobacillus buchneri and Cellulase on Nutrient Composition, Fermentation Quality and Microbial Diversity of Cenchrus fungigraminus Fermented Total Mixed Ration

  • WANG Haimei , 1, 2, 3 ,
  • LIU Hui 2 ,
  • YANG Jie 1, 2, 3 ,
  • GUO Xin 1, 2, 3 ,
  • HAN Wei 2, 3 ,
  • XIN Guosheng , 1, 2, 3, *
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  • 1 School of Life Sciences, Ningxia University, Yinchuan 750021, China
  • 2 Feed Engineering Technology Research Center, Ningxia University, Yinchuan 750021, China
  • 3 Key Laboratory of Western Characteristic Biological Resources Protection and Utilization, Ministry of Education, Ningxia University, Yinchuan 750021, China
*professor, E-mail:

Received date: 2025-12-19

  Online published: 2026-08-13

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Abstract

This study aimed to investigate the effects of co-fermentation with Lactobacillus buchneri and cellulase on the nutrient composition, fermentation quality and microbial diversity of Cenchrus fungigraminus fermented total mixed ration (FTMR). The total mixed ration (TMR) of Cenchrus fungigraminus was used as the fermentation substrate, and sterile water (NC group), Lactobacillus buchneri at 1.0×106 CFU/g (LAB group), cellulase at 10 U/g (CE group), and cellulase at 10 U/g combined with Lactobacillus buchneri at 1.0×106 CFU/g (CE+LAB group) were added for anaerobic fermentation, respectively. The addition amounts of both enzyme and bacteria were based on the fresh weight of Cenchrus fungigraminus TMR, and each group had 8 replicates. After 45 days of fermentation, samples were collected, and the sensory quality of Cenchrus fungigraminus FTMR was evaluated, and its nutrient composition, fermentation parameters, mycotoxin contents and microbial diversity were determined. The results showed as follows: 1) the sensory quality of Cenchrus fungigraminus FTMR in all groups was good, with no mold contamination. The sensory evaluation grades of the three experimental groups (LAB group, CE group and CE+LAB group) were all grade Ⅰ (excellent), among which the CE+LAB group had the highest sensory evaluation score (20 points). 2) The contents of neutral detergent fiber (NDF) and hemicellulose (HC) in the three experimental groups were significantly lower than those in the NC group (P<0.05), and the acid detergent fiber (ADF) content in the CE+LAB group was significantly lower than that in the other groups (P<0.05). 3) Compared with the NC group, the pH and zearalenone (ZEN) content in the CE+LAB group and LAB group were significantly reduced (P<0.05), while the propionic acid and lactic acid contents were significantly increased (P<0.05); the acetic acid content in the three experimental groups was significantly increased (P<0.05), and the deoxynivalenol (DON) content was significantly reduced (P<0.05). The ammonia nitrogen (NH3-N) content in the CE+LAB group and CE group was significantly lower than that in the NC group and LAB group (P<0.05). 4) At the phylum level, the relative abundance of Firmicutes in the CE+LAB group and LAB group was significantly higher than that in the CE group (P<0.05). At the genus level, the relative abundance of Lentilactobacillus in the CE+LAB group and LAB group was significantly higher than that in the NC group (P<0.05). 5) Correlation analysis showed that the relative abundance of Lentilactobacillus was extremely significantly positively correlated with acetic acid content (r=0.55, P<0.01), and significantly negatively correlated with NDF content (r=-0.35, P=0.04). In conclusion, the addition of Lactobacillus buchneri alone or in combination with cellulase can reduce the fiber content, maintain nutrient composition, increase organic acid contents, and enhance the relative abundance of Lentilactobacillus in Cenchrus fungigraminus FTMR. Moreover, the combined addition of the two shows better effects in fiber degradation and fermentation quality improvement.

Key words： Cenchrus fungigraminus; cellulase; Lactobacillus buchneri; fermentation quality; microbial diversity

Cite this article

WANG Haimei , LIU Hui , YANG Jie , GUO Xin , HAN Wei , XIN Guosheng . Effects of Co-Fermentation with Lactobacillus buchneri and Cellulase on Nutrient Composition, Fermentation Quality and Microbial Diversity of Cenchrus fungigraminus Fermented Total Mixed Ration[J]. Chinese Journal of Animal Nutrition, 2026 , 38(8) : 6276 -6288 . DOI: 10.12418/CJAN2026.501

近年来,随着居民消费水平提升,畜牧业规模化发展迅速,对优质粗饲料的需求日益增加。然而,受耕地资源刚性约束和“粮改饲”政策影响,传统饲草种植面积有限,优质草料供需矛盾日益突出。因此,挖掘新型、非竞争性饲草资源已成为保障畜牧业可持续发展的关键途径[1]。巨菌草(Cenchrus fungigraminus)是一种禾本科植物,具有生物产量高、分蘖能力强及营养品质优良等特性,被视为极具开发潜力的优质饲草资源[2]。然而,巨菌草因木质素与纤维素含量较高,直接饲喂时适口性差,易导致采食量降低与消化率下降,从而制约动物生产性能的发挥[3]。青贮处理虽能在一定程度上改善适口性与消化率,但巨菌草中可溶性碳水化合物(WSC)含量偏低,单独青贮时发酵品质往往不佳,限制了其作为优质粗饲料的利用潜力[4]。为破解这一矛盾,而根据反刍动物在特定生理阶段的生产目标与营养需求,将粗饲料、精饲料及矿物质等按科学配比调制的全混合日粮(TMR)则可有效的解决这一问题[5]。经厌氧发酵制备的发酵全混合日粮(FTMR),不仅能弥补单一原料青贮的营养失衡问题,还能有效保存养分并提升发酵品质[6],这为巨菌草的高效饲用提供了可行路径。
菌酶复合技术通过将特定益生菌与外源酶制剂科学配伍,已成为提升饲料发酵品质的有效策略。其中,纤维素酶能够破坏植物细胞壁结构,将纤维素降解为葡萄糖,从而为乳酸菌等微生物提供发酵底物[7]。布氏乳杆菌(Lactobacillus buchneri)则可将乳酸进一步转化为乙酸和1,2-丙二醇,有效抑制霉菌生长并降低二次发酵风险[8-10]。酶菌协同作用下,菌酶复合制剂不仅能直接优化发酵参数,还可通过调控发酵体系中的微生物群落结构与多样性,提升发酵饲料的营养品质与发酵品质,进一步提升发酵饲料的质量[11]。
近年来,虽然针对菌酶复合制剂对发酵饲料品质影响的研究已有报道[12-13],但针对巨菌草TMR中添加布氏乳杆菌与纤维素酶进行协同发酵的系统性研究尚属空白。基于此,本试验旨在探究布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR营养成分、发酵品质及微生物多样性的影响,以期为反刍动物饲草资源的高效开发与利用提供理论依据和技术支撑。

1 材料与方法

1.1 试验材料

试验所用巨菌草于2024年10月采自宁夏回族自治区石嘴山市平罗县宝丰村,收割时处于生长后期,株高200~300 cm;布氏乳杆菌(活菌数为1.0×1011 CFU/g)与纤维素酶(活性约20 000 U/g)均为市售产品。

1.2 试验设计

试验采用单因素完全随机设计,以巨菌草TMR为发酵底物,在发酵制备巨菌草FTMR时分别进行如下处理:NC组,添加无菌水;LAB组,接种布氏乳杆菌(接种量为1.0×106 CFU/g鲜重);CE组,添加纤维素酶(添加量为10 U/g鲜重);CE+LAB组,添加纤维素酶(添加量为10 U/g鲜重)+接种布氏乳杆菌(接种量为1.0×106 CFU/g鲜重)。每组设8个重复。

1.3 巨菌草FTMR的制备

巨菌草TMR组成及营养水平见表1,精粗比为7∶3(干物质基础)。将收割的巨菌草切割至1.5~2.5 cm长度后,按照试验设计将各组所需的布氏乳杆菌和/或纤维素酶溶解于100 mL无菌水中,NC组则以等量无菌水替代,均匀喷洒于巨菌草表面并充分混合后,配制成巨菌草TMR,含水率控制在50%左右。随后,将混合好的巨菌草TMR装入1.5 L发酵瓶中,压实并密封,每个组制备8瓶,每瓶装填约1.5 kg(鲜重),置于25 ℃左右的环境中厌氧发酵45 d后开瓶取样分析。
表1 巨菌草TMR组成及营养水平(干物质基础)

Table 1 Composition and nutrient levels of Cenchrus fungigraminus TMR (DM basis)

项目 Items 含量 Content
原料 Ingredients
玉米 Corn 40.00
豆粕 Soybean meal 18.00
麸皮 Wheat bran 5.00
干酒糟及其可溶物 DDGS 3.00
巨菌草 Cenchrus fungigraminus 30.00
预混料 Premix1) 2.00
小苏打 NaHCO3 1.00
食盐 NaCl 0.50
磷酸氢钙 CaHPO4 0.30
石粉 Limestone 0.20
合计 Total 100.00
营养水平 Nutrient levels2)
消化能 DE/(MJ/kg) 13.14
粗蛋白质 CP 13.83
粗纤维 CF 13.49
粗灰分 Ash 3.87
钙 Ca 0.62
磷 P 0.35

1)每千克预混料提供 Per kilogram of premix provided the following:维生素A乙酸酯VA acetate 6.33×105 IU,VD3 2.51×105 IU,DL-α-生育酚乙酸酯 DL-α-tocopherol acetate 3.69×103 IU,烟酰胺 nicotinamide 1.71×103 mg,Fe 1.05×103 mg,Cu 640 mg,Zn 3.90×103 mg,Mn 5.80×103 mg,I 58.80 mg,Se 13.49 mg,Co 71.2 mg。

2)消化能依据《肉羊营养需要量》(NY/T 816—2021)计算得出,其余为实测值。DE was calculated according to the Nutrient Requirements of Meat-Type Sheep and Goat (NY/T 816—2021), while the others were measured values.

1.4 测定指标及方法

1.4.1 感官品质的评价

感官品质的评分标准参照德国DLG青贮感官评分方法[14]。根据巨菌草FTMR的颜色、气味及色泽情况,由5名专业人员同时进行评价并打分,去除最高与最低分后求平均值。

1.4.2 营养成分的测定

将样品在105 ℃烘箱中烘30 min后,将烘箱温度调至85 ℃继续干燥48 h直至样品衡重,测得干物质(DM)含量,将干燥后的样品经粉碎机粉碎过40目筛后进行营养成分测定。其中,粗蛋白质(CP)含量参照GB/T 6432—2018测定,粗灰分(Ash)含量参照GB/T 6438—2007测定,粗脂肪含量参照GB/T 6433—2006测定,中性洗涤纤维(NDF)含量参照GB/T 20806—2006测定,酸性洗涤纤维(ADF)含量参照NY/T 1459—2022测定,半纤维素(HC)含量为NDF和ADF含量的差值(HC含量=NDF含量-ADF含量),钙(Ca)含量参照GB/T 6436—2018测定,磷(P)含量参照GB/T 6437—2018测定。

1.4.3 发酵参数的测定

称取20.00 g样品,加180 mL蒸馏水后振荡10 min,4 ℃冰箱置放24 h,4层纱布和定性滤纸过滤得浸出液。所得浸出液一部分测定pH;另一部分采用气相色谱仪(7820A,Agilent Technologies,美国)测定挥发性脂肪酸(VFAs)含量,采用苯酚-次氯酸钠比色法[15]测定氨态氮(NH3-N)含量。
称取样品1.00~5.00 g,置于100 mL容量瓶中,加入60 mL蒸馏水,在50 ℃水浴中超声提取20 min,冷却至室温,用蒸馏水定容至刻度,摇匀后过滤,收集滤液,采用高效液相色谱仪(L-2000,Agilent Technologies,美国)测定乳酸(LA)含量。

1.4.4 霉菌毒素含量的测定

准确称取样品5.00 g,置于50 mL离心管中,加25 mL蒸馏水振荡提取3 min,静置数秒后取1 mL于离心管中,1 789×g离心2 min,采用荧光定量快速检测仪(FD-500,上海飞测生物科技有限公司)检测玉米赤霉烯酮(ZEN)、呕吐毒素(DON)及黄曲霉毒素B1(AFB1)含量。ZEN含量检测范围为20~200 μg/kg,DON含量检测范围为200~1 000 μg/kg,AFB1含量检测范围为1~10 μg/kg。

1.4.5 微生物多样性的检测

取250~500 mg样品置于2.0 mL离心管中,采用细菌基因组DNA提取试剂盒提取总DNA。以提取的DNA为模板,采用引物对341F(5'-CCTAYGGGRBGCASCAG-3')和806R(5'-GGACTACNNGGGTATCTAAT-3'),使用PCR仪(T100,Bio-Rad Laboratories,美国)扩增细菌16S rRNA基因V3~V4区。PCR扩增程序:95 ℃预变性5 min;95 ℃变性30 s,55 ℃退火30 s,72 ℃延伸45 s,共25个循环;最后72 ℃延伸7 min。每个样品设3个重复。扩增产物经纯化后,采用QuantiFluorTM荧光计(Promega,美国)进行定量,将纯化后的扩增产物等量混合,连接测序接头,构建测序文库。原始测序数据进行拼接、过滤后得到有效数据(clean reads),用于后续生物信息学分析。

1.5 数据统计与分析

使用Excel 2016软件对数据进行整理后,采用Shapiro-Wilk法对数据进行正态性检验。若数据符合正态分布,则采用单因素方差分析(one-way ANOVA)进行组间比较,并采用Duncan氏法进行多重比较;若数据不满足正态分布假设,则改用Kruskal-Wallis非参数检验。结果以“平均值±标准误”表示。P<0.01为差异极显著,P<0.05为差异显著,P>0.05为差异不显著。微生物测序数据由诺禾云平台(https://magic-plus.novogene.com)进行分析与制图;差异菌群分析采用Wilcoxon秩和检验。采用Spearman相关分析法,对属水平微生物相对丰度与部分发酵参数、营养成分及霉菌毒素含量进行相关性分析。

2 结果与分析

2.1 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR感官品质的影响

由表2可以看出,NC组感官评价总分小于16分,评定等级为Ⅱ级(尚好),3个试验组(LAB组、CE组和CE+LAB组)感官评价总分在16分及以上,评价等级均为Ⅰ级(优良);巨菌草FTMR感官评价总分排序为CE+LAB组(20分)>LAB组(19分)>CE组(16分)>NC组(15分),说明CE+LAB组巨菌草FTMR的感官品质最好。
表2 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR感官品质的影响

Table 2 Effects of co-fermentation with Lactobacillus buchneri and cellulase on sensory quality of Cenchrus fungigraminus FTMR

项目
Items		 组别 Groups
NC LAB CE CE+LAB
色泽 Color/分 1 2 1 2
气味 Odor/分 10 13 11 14
质地 Texture/分 4 4 4 4
总分 Total score/分 15 19 16 20
等级 Grade Ⅱ级 Ⅰ级 Ⅰ级 Ⅰ级
评语 Comment 尚好 优良 优良 优良

2.2 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR营养成分的影响

由表3可以看出,CE+LAB组的ADF含量显著低于其他组(P<0.05);3个试验组的NDF、HC含量均显著低于NC组(P<0.05);3个试验组的DM、CP、EE、Ash、Ca和P含量相比于NC组无显著差异(P>0.05)。
表3 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR营养成分的影响

Table 3 Effects of co-fermentation with Lactobacillus buchneri and cellulase on nutrient composition of Cenchrus fungigraminus FTMR

项目
Items		 组别 Groups P值
P-value
NC LAB CE CE+LAB
干物质 DM/% FM 42.01±0.51 42.26±0.53 42.08±0.29 41.72±0.45 0.804
粗蛋白质 CP/% DM 18.22±0.28 19.46±0.26 19.07±0.21 20.16±0.25 0.110
粗脂肪 EE/% DM 2.27±0.19 2.36±0.22 2.52±0.25 2.78±0.24 0.232
中性洗涤纤维 NDF/% DM 47.40±0.41a 41.73±0.41b 41.87±0.43b 42.04±0.29b 0.001
酸性洗涤纤维 ADF/% DM 22.91±0.56a 22.97±0.73a 21.23±0.32a 18.61±0.28b 0.002
半纤维素 HC/% DM 26.03±0.15a 19.60±0.22b 19.01±0.32b 22.38±0.24b 0.001
粗灰分 Ash/% DM 9.53±0.09 9.61±0.08 9.59±0.09 9.51±0.09 0.067
钙 Ca/% DM 0.81±0.01 0.82±0.01 0.83±0.01 0.82±0.01 0.064
磷 P/% DM 0.30±0.02 0.34±0.03 0.36±0.01 0.33±0.01 0.102

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

In the same row, values with no letter or the same letter superscripts mean no significant difference (P>0.05), while with different small letter superscripts mean significant difference (P<0.05). The same as below.

2.3 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR发酵参数和霉菌毒素含量的影响

由表4可以看出,与NC组、CE组相比,CE+LAB组和LAB组的pH显著降低(P<0.05), 丙酸含量显著升高(P<0.05);与NC组相比,CE+LAB组和LAB组的乳酸含量显著升高(P<0.05),3个试验组的乙酸含量均显著升高(P<0.05);CE+LAB组和CE组的氨态氮含量显著低于NC组和LAB组(P<0.05);3个试验组的丁酸含量与NC组无显著差异(P>0.05)。
表4 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR发酵参数的影响

Table 4 Effects of co-fermentation with Lactobacillus buchneri and cellulase on fermentation parameters of Cenchrus fungigraminus FTMR

项目
Items		 组别 Groups P值
P-value
NC LAB CE CE+LAB
pH 4.35±0.01a 4.19±0.00c 4.30±0.01b 4.20±0.00c <0.001
乳酸 LA/% DM 2.70±0.08b 3.32±0.06a 3.03±0.07ab 3.33±0.08a 0.004
乙酸 AA/% DM 0.65±0.02c 0.84±0.02b 0.78±0.02b 0.97±0.03a <0.001
丙酸 PA/% DM 0.05±0.01b 0.10±0.01a 0.05±0.01b 0.08±0.01a <0.008
丁酸 BA/% DM 0.02±0.01 0.02±0.00 0.01±0.00 0.01±0.00 0.144
氨态氮 NH3-N/% DM 0.24±0.02a 0.24±0.03a 0.13±0.01c 0.18±0.01b <0.001
由表5可以看出,各组的AFB1、ZEN及DON含量均未超过《饲料卫生标准》(GB 13078—2017)规定的限量标准。LAB组的AFB1含量显著低于NC组和CE+LAB组(P<0.05);与NC组相比,CE+LAB组和LAB组的ZEN含量显著降低(P<0.05),3个试验组的DON含量均显著降低(P<0.05)。
表5 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR霉菌毒素含量的影响

Table 5 Effects of co-fermentation with Lactobacillus buchneri and cellulase on mycotoxin contents of Cenchrus fungigraminus FTMRμg/kg FM

项目
Items		 组别 Groups P值
P-value
NC LAB CE CE+LAB
黄曲霉毒素B1 AFB1 1.25±0.08a 1.00±0.01b 1.13±0.04ab 1.22±0.04a 0.013
玉米赤霉烯酮 ZEN 36.86±2.26a 29.50±1.02b 32.21±2.41ab 26.42±1.05b 0.016
呕吐毒素 DON 266.16±12.33a 221.37±7.85b 220.27±6.15b 227.48±3.86b 0.036

根据《饲料卫生标准》(GB 13078—2017),黄曲霉毒素B1含量限量标准为≤20 μg/kg,玉米赤霉烯酮含量限量标准为≤0.5 mg/kg,呕吐毒素含量限量标准为≤3 mg/kg。

According to the Hygienic Standard for Feeds (GB 13078—2017), the maximum allowable limits for AFB1, ZEN and DON are ≤20 μg/kg, ≤0.5 mg/kg and≤3 mg/kg, respectively.

2.4 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR微生物多样性的影响

巨菌草FTMR物种稀释曲线如图1-A所示。随着测序深度的增加,各样本的稀释曲线逐渐趋于平缓,表明测序数据量已基本覆盖样品中的大部分微生物类群,继续增加测序深度仅能发现少量新的物种。基于操作分类单元(OTU)的韦恩图(图1-B)显示,4组共有的OTU数为270个,各组特有的OTU数NC组为50个,CE组为43个,CE+LAB组为28个,LAB组为25个。
图1 巨菌草FTMR物种稀释曲线(A)和OTU韦恩图(B)

NC、LAB、CE、CE+LAB分别代表NC、LAB、CE、CE+LAB组。下图同。

Fig.1 Species dilution curves (A) and OTU Venn diagram (B) of Cenchrus fungigraminus FTMR

NC, LAB, CE and CE+LAB represented the NC, LAB, CE and CE+LAB groups, respectively. The same as below.

由图2可知,各组间ACE指数、Chao1指数、Shannon指数及Simpson指数均无显著差异(P>0.05)。
图2 巨菌草FTMR微生物α多样性指数

Fig.2 Microbial α diversity indexes of Cenchrus fungigraminus FTMR

图3为巨菌草FTMR微生物在门水平上相对丰度排名前5的物种(A)及差异菌群分析(B),各组优势菌门为变形菌门(Proteobacteria)和厚壁菌门(Firmicutes)。其中,CE+LAB组拟杆菌门(Bacteroidota)相对丰度显著低于NC组(P<0.05);CE+LAB组和LAB组厚壁菌门相对丰度显著高于CE组(P<0.05),变形菌门相对丰度显著低于CE组(P<0.05)。
图3 巨菌草FTMR微生物门水平上菌群组成(A)和差异菌门分析(B)

Fig.3 Microbial community composition at phylum level (A) and differential phyla analysis (B) of Cenchrus fungigraminus FTMR

图4为巨菌草FTMR微生物在属水平上相对丰度排名前10的物种(A)及差异菌属分析(B),各组优势菌属为不动杆菌属(Acinetobacter)、乳植杆菌属(Lactiplantibacillus)、肠杆菌属(Enterobacter)、促生乳杆菌属(Levilactobacillus)、魏斯氏菌属(Weissella)、伴生乳杆菌属(Companilactobacillus)和慢生乳杆菌属(Lentilactobacillus)。其中,CE+LAB组和LAB组慢生乳杆菌属相对丰度显著高于NC组(P<0.05),CE+LAB组伴生乳杆菌属相对丰度显著低于NC组(P<0.05)。
图4 巨菌草FTMR微生物属水平上菌群组成(A)和差异菌属分析(B)

Fig.4 Microbial community composition at genus level (A) and differential genera analysis (B) of Cenchrus fungigraminus FTMR

2.5 巨菌草FTMR中微生物与发酵品质、营养成分的相关性

为探究巨菌草FTMR中微生物与发酵品质、营养成分的相关性,对其属水平微生物相对丰度与部分发酵参数、营养成分及霉菌毒素含量进行Spearman相关性分析。相关性分析结果(图5)显示,不动杆菌属相对丰度与丙酸(r=0.46,P<0.01)和乙酸含量(r=0.36,P=0.04)分别呈极显著和显著正相关,与pH呈极显著负相关(r=-0.63,P<0.01);慢生乳杆菌属相对丰度与乙酸含量呈极显著正相关(r=0.55,P<0.01),与NDF含量呈显著负相关(r=-0.35,P=0.04);伴生乳杆菌属相对丰度与pH(r=0.72,P<0.01)、ZEN含量(r=0.38,P=0.03)分别呈极显著和显著正相关,与丙酸含量呈极显著负相关(r=-0.50,P<0.01);促生乳杆菌属相对丰度与ZEN含量呈极显著正相关(r=0.47,P<0.01)。
图5 巨菌草FTMR属水平微生物相对丰度与部分发酵参数、营养成分及霉菌毒素含量的相关性热图

*表示显著相关(P<0.05),**表示极显著相关(P<0.01)。

Fig.5 Correlation heatmap of genus-level microbial relative abundances with selected fermentation parameters, nutrient composition and mycotoxin contents of Cenchrus fungigraminus FTMR

* indicates significant correlation (P<0.05), and ** indicates extremely significant correlation (P<0.01).

3 讨论

3.1 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR感官品质的影响

感官品质是评价饲料发酵质量的重要指标。本研究中,各组的巨菌草FTMR未出现霉变,质地松散不粘手,叶茎结构保持完整且具有芳香味,综合评级由“尚好”提升至“优良”,以菌酶协同发酵组(CE+LAB组)感官评价总分最高,这与已有研究结论[16-17]一致。这一改善主要源于纤维素酶对高纤维草料中纤维结构的破坏作用,使草料质地更为柔软[18];除此之外,布氏乳杆菌的添加显著降低了霉菌毒素含量,使得发酵饲料无霉变和刺鼻味,色泽更鲜绿,这也是提升发酵饲料感官品质的一大原因。廖隽锐等[19]以乳酸菌为添加剂发酵蚕豆秸秆饲料的研究结果也表明,添加剂的使用可保持发酵原料的颜色,减少刺鼻酸味,与本研究结果相互印证。

3.2 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR营养成分的影响

营养成分是衡量发酵饲料质量的基础指标,其中,NDF和ADF含量是评价饲料纤维质量的重要参数。已有研究表明,NDF和ADF含量与动物采食量及消化率呈负相关,二者含量的降低通常表明饲料品质的提高[20]。添加菌酶制剂能改善发酵饲料的营养价值,尤其在纤维降解方面具有重要作用[21]。王红梅等[22]以植物乳杆菌和纤维素酶作为添加剂,显著降低了膨化预处理玉米秸秆的NDF和ADF含量。本研究中,各试验组尤其是CE+LAB组巨菌草FTMR的NDF和ADF含量较NC组显著降低,说明菌酶协同发酵在降解纤维方面效果显著。这可能与外源添加的纤维素酶或发酵过程中微生物产生的水解酶对纤维成分的降解有关[11]。

3.3 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR发酵品质的影响

pH能直接反映饲料发酵进程,通常pH低于4.2表明饲料发酵充分。乳酸是评价发酵饲料的可靠指标,是发酵饲料中的有益酸,其积累会导致pH的下降[23]。此外,发酵饲料中挥发性脂肪酸如乙酸、丙酸、丁酸等也是衡量发酵品质的关键指标[24-25]。赵超等[26]研究表明,添加乳酸菌和纤维素酶可显著提升豆渣与桑叶混贮中乳酸和挥发性脂肪酸含量,降低pH。本试验中,CE+LAB组和LAB组巨菌草FTMR的pH均降至4.2及以下,且CE+LAB组较NC组氨态氮含量显著降低,乳酸、乙酸及丙酸含量显著升高。其原因可能在于纤维素酶或布氏乳杆菌的添加增加了乳酸菌等有益菌的相对丰度,快速产酸并降低pH,从而促进有机酸的积累[27]。此外,严格的厌氧环境不仅有利于有机酸的积累,还能抑制有害微生物对蛋白质的水解,从而降低氨态氮含量[28-29]。

3.4 布氏乳杆菌与纤维素酶协同发酵对巨菌草FTMR微生物多样性的影响

发酵是由多种微生物共同参与的复杂过程,而添加剂的使用会改变已有的微生物群落结构[30]。杨琪等[10]在全株玉米中添加乳酸菌制剂后,随着青贮时间的延长,优势菌群由变形菌门转变为厚壁菌门。本试验中,相较于NC组,各试验组的Chao1指数和ACE指数虽有所降低但差异不显著,其中厚壁菌门和变形菌门为优势菌门,且布氏乳杆菌的添加降低了变形菌门的相对丰度,提高了厚壁菌门的相对丰度。这种变化可能是由于布氏乳杆菌加速了乳酸的积累,适宜的酸性环境和充足的底物有利于厚壁菌门增殖,从而使厚壁菌门相对丰度增加[31-32]。乳植杆菌属、伴生乳杆菌属、慢生乳杆菌属等都是与饲料发酵密切相关的乳酸菌,其均可利用碳水化合物产生大量乳酸[33-34]。本试验中,CE+LAB组慢生乳杆菌属相对丰度显著高于其他组,且相关性分析显示,慢生乳杆菌属的相对丰度与乙酸含量呈极显著正相关,主要原因可能是在发酵过程中,纤维素酶和布氏乳杆菌的联合使用增强了慢生乳杆菌利用底物的能力,从而增强其增殖代谢能力[12]。

4 结论

本试验条件下,布氏乳杆菌单独或与纤维素酶复合添加均能降低巨菌草FTMR的纤维含量,保持营养成分,提升有机酸含量,并提高慢生乳杆菌属的相对丰度,且二者复合添加在降解纤维和改善发酵品质方面效果更优,表明菌酶协同在巨菌草FTMR调制中具有良好的应用潜力。

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