Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 5: 3180 - 3193

DOI: https://doi.org/10.12418/CJAN2025.263

RESEARCH PAPER

Effects of Complex Probiotics on Nutrient Apparent Digestibility, Rumen Fermentation Parameters and Microbiota of Lambs

  • WANG Haiyu , 1 ,
  • ZHAO Miaomiao 1 ,
  • ZUO Yanhua 2 ,
  • GUO Yunxia 1 ,
  • WEI Ze 1 ,
  • DING Yawei 1 ,
  • DUAN Chunhui , 1, * ,
  • ZHANG Yingjie 1 ,
  • LIU Yueqin 1
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  • 1 College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China
  • 2 Agricultural and Rural Affairs Bureau of Qinglong Manzu Autonomous County, Qinhuangdao 066000, China
* associate professor, E-mail:

Received date: 2024-09-25

  Online published: 2025-05-14

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Abstract

This study was conducted to investigate the effects of dietary complex probiotics on nutrient apparent digestibility, rumen fermentation parameters and microbiota of lambs. Forty-eight 36-day-old Hu sheep with an initial body weight of (12.43±0.27) kg were randomly divided into 4 groups with 12 lambs (half male and half female) in each group. Lambs in the control group (group Ⅰ) were fed a basal diet without complex probiotic powder, and those in the experimental groups were fed the basal diet supplemented with 0.5% (group Ⅱ), 1.0% (group Ⅲ) and 1.5% (group Ⅳ) complex probiotic powder (Lactobacillus RSG-1∶Bacillus subtilis B-1∶Bacillus licheniformis Y5-39=1∶1∶4, total viable bacteria number≥1.0×108 CFU/g), respectively. The pre-trial period lasted for 10 days, and the experimental period lasted for 29 days. Lambs were weaned at 60 days of age, and the rumen fluid was collected on day 14 before weaning, day 7 before weaning, weaning day, day 3 after weaning and day 7 after weaning, respectively, to determine the rumen fermentation parameters and microbiota. The results showed as follows: 1) compared with group Ⅰ, the apparent digestibility of dry matter, crude protein, ether extract and gross energy in group Ⅲ was significantly increased (P<0.05). 2) On days 3 and 7 after weaning, the rumen ammonia nitrogen (NH3-N) concentration in groups Ⅱ, Ⅲ and Ⅳ was significantly higher than that in group Ⅰ (P<0.05); on weaning day, the rumen total volatile fatty acid (TVFA) concentration in groups Ⅱ, Ⅲ and Ⅳ was significantly higher than that in group Ⅰ (P<0.05); on day 3 after weaning, the rumen butyrate concentration in groups Ⅱ, Ⅲ and Ⅳ was significantly higher than that in group Ⅰ (P<0.05); on day 7 after weaning, the rumen butyrate concentration in group Ⅲ was significantly higher than that in group Ⅰ (P<0.05). 3) On weaning day and days 3 and 7 after weaning, the Chao1 index and Observed_species index of rumen microbiota in group Ⅲ were significantly higher than those in group Ⅰ (P<0.05). The relative abundances of Bacteroidota and Firmicutes in rumen in group Ⅰ on day 3 after weaning were lower than those on day 7 before weaning and weaning day. The abundances of genes related to carbohydrate metabolism function of rumen microbiota in group Ⅲ were significantly higher than those in group Ⅰ on day 3 after weaning (P<0.05). In conclusion, dietary complex probiotics can improve the nutrient apparent digestibility of lambs and promote the rumen fermentation to produce volatile fatty acids by regulating rumen microbiota, thus alleviating the effects of weaning stress on lambs. Under the experimental conditions, the effect of 1.0% complex probiotics is better.

Key words： complex probiotics; lambs; nutrient apparent digestibility; rumen fermentation; microbiota

Cite this article

WANG Haiyu , ZHAO Miaomiao , ZUO Yanhua , GUO Yunxia , WEI Ze , DING Yawei , DUAN Chunhui , ZHANG Yingjie , LIU Yueqin . Effects of Complex Probiotics on Nutrient Apparent Digestibility, Rumen Fermentation Parameters and Microbiota of Lambs[J]. Chinese Journal of Animal Nutrition, 2025 , 37(5) : 3180 -3193 . DOI: 10.12418/CJAN2025.263

羔羊断奶时由于母子分离、饲养方式转变等会产生断奶应激,且断奶时羔羊胃肠道功能发育尚不完全,饲养管理不当极易导致饲粮消化利用率低、生长发育缓慢等问题,致使发病死亡率升高,严重影响养殖场经济效益[1]。研究表明,益生菌具有调节动物机体胃肠道微生物平衡、提高营养物质消化利用率的作用[2-3]。饲粮添加益生菌有助于丰富反刍动物瘤胃微生物多样性,促进幼畜瘤胃功能发育,提高营养物质消化率[4-6]。断奶应激会导致羔羊营养物质的消化利用率降低,影响瘤胃液挥发性脂肪酸(VFA)和氮含量以及消化酶活性[2]。研究发现,犊牛断奶前后瘤胃和粪便中微生物群落β多样性有显著差异,断奶应激缓解后肠道菌群α多样性随日龄增长显著提高,纤维素及碳水化合物降解菌数量快速增多[7-8]。益生菌在抑制胃肠道致病菌生长,促进有益菌繁殖,提高营养物质消化利用率方面有显著效果[4]。多种益生菌单独或复合添加均表现出提高断奶应激期奶牛平均日增重和饲料利用率,以及缓解断奶应激的效果[9]。益生菌在增殖过程中可分泌大量分解蛋白质、脂肪和纤维素的酶,直接参与饲粮中营养物质的消化和利用,提升饲粮适口性[10]。不过,关于益生菌对断奶应激的影响研究多集中于仔猪和犊牛上,在羔羊上的研究鲜见报道。研究表明,幼畜断奶应激与瘤胃功能和微生物区系的变化有密切关系[11]。因此,本试验比较了饲粮添加不同比例复合益生菌对羔羊营养物质表观消化率、瘤胃发酵参数和微生物区系的影响,旨在为复合益生菌在羔羊生产中的应用提供参考。

1 材料与方法

1.1 试验材料

复合益生菌菌粉由河北农业大学生命科学学院制药工程系实验室配制,其中乳酸杆菌RSG-1∶枯草芽孢杆菌B-1∶地衣芽孢杆菌Y5-39=1∶1∶4,总活菌数≥1.0×108 CFU/g。

1.2 试验设计

本试验于2021年8—10月在河北省衡水志豪畜牧科技有限公司进行,试验经河北农业大学实验动物伦理委员会批准(批准编号:No.2021083)。试验选择48只初始体重为(12.43±0.27) kg的36日龄湖羊羔羊,随机分为4组,每组12只(公母各占1/2)。每组羔羊在同一个圈舍,混群饲养。其中,对照组(Ⅰ组)饲喂基础饲粮,不添加复合益生菌菌粉;试验组分别饲喂在基础饲粮的基础上添加0.5%(Ⅱ组)、1.0%(Ⅲ组)和1.5%(Ⅳ组)复合益生菌菌粉的饲粮。预试期10 d,正试期29 d。羔羊60日龄断奶,记为第0天(0D)。
基础饲粮参照《肉羊营养需要量》(NY/T 816—2021)[12]配制,其组成及营养水平见表1。
表1 基础饲粮组成及营养水平(干物质基础)

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

原料Ingredients 含量Content 营养水平Nutrient levels2) 含量Content
玉米Corn 41.50 干物质(风干基础) DM (air-dry basis) 87.32
麦麸Wheat bran 8.00 代谢能ME/(MJ/kg) 9.30
次粉Wheat middling 10.00 粗蛋白质CP 18.35
玉米胚芽粕Corn germ meal 7.00 粗脂肪EE 7.49
豆粕Soybean meal 16.00 粗灰分Ash 9.24
干酒糟及其可溶物DDGS 8.50 中性洗涤纤维NDF 19.50
乳清粉Whey powder 2.00 酸性洗涤纤维ADF 11.40
甜菜粕Sugar beet pulp 3.00 钙Ca 0.72
预混料Premix1) 4.00 总磷TP 0.38
合计Total 100.00

1)预混料为每千克饲粮提供 The premix provided the following per kilogram of the diet:VA 8 000 IU,VD 800 IU,VE 16 mg,Cu 4 mg,Fe 20 mg,Mn 30 mg,Zn 30 mg,I 0.6 mg,Se 0.4 mg,Co 0.2 mg,Ca 0.8 g,P 0.08 g。

2)代谢能为计算值(参照NY/T 816—2021计算),其他均为实测值。ME was a calculated value referred to NY/T 816—2021, while the others were measured values.

1.3 饲养管理

试验开始前,所有试验羊均登记耳号并按照羊场正常免疫程序进行免疫和驱虫。羔羊断奶前随母羊哺乳并补饲羔羊料,断奶后饲喂羔羊料。在母子舍中母羊栏相邻的圈舍设置羔羊补饲栏,仅羔羊可以自由进出补饲栏。羔羊每天饲喂2次(06:30和16:30各1次),自由采食和饮水,每次饲喂前将菌粉与饲粮充分混匀,保证菌粉被全部采食完。羔羊60日龄断奶,转移母羊,羔羊原圈饲养。

1.4 样品采集

正试期间,分别于断奶前第14天(-14D)、断奶前第7天(-7D)、断奶当天(0D)、断奶后第3天(3D)和断奶后第7天(7D)晨饲前通过胃管采集羔羊瘤胃液。断奶后第7天每组随机选择3只羊(1母,2公)称重进行消化试验,期间羔羊料及羊草充足供应,羔羊自由饮水,第7~9天为适应期,记录第10~14天每只羊的采食量,收集剩料及全部粪便称重。取饲粮样及每只羊全部粪便的20%,分别标记好,密封保存于-20 ℃冰箱。

1.5 测定指标及方法

1.5.1 饲粮营养水平

饲粮干物质(DM)、粗蛋白质(CP)、粗脂肪(EE)、粗灰分(Ash)、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)、钙和磷含量分别参照GB/T 6435—2014、GB/T 6432—2018、GB/T 6433—2006、GB/T 6438—2007、GB/T 20806—2022、NY/T 1459—2022、GB/T 6436—2018和GB/T 6437—2018中方法进行测定。

1.5.2 营养物质表观消化率

消化试验期间,每天记录代谢笼内羔羊的采食量和粪便量。将各组饲粮和粪便样品放置于65 ℃烘干箱内脱水烘干48 h,室温静置24 h。烘干样品粉碎后分别粉碎过10和40目筛备用,用于后续营养物质表观消化率的测定。饲粮和粪便中DM、CP、EE、NDF和ADF含量分别参照GB/T 6435—2014、GB/T 6432—2018、GB/T 6433—2006、GB/T 20806—2022和NY/T 1459—2022中方法进行测定,总能(GE)参照ISO 9831:1998进行测定。营养物质表观消化率计算公式如下:
$ \text { 某营养物质表观消化率 }(\%)=100 \times \text {（该营养物质摄入量－粪中该营养物质含量）／该营养物质摄入量。 }$

1.5.3 瘤胃发酵参数

分别于断奶前第14天、断奶前第7天、断奶当天、断奶后第3天和断奶后第7天晨饲前每组选取6只羔羊利用瘤胃管采集瘤胃液,随即使用便携式pH检测仪测定瘤胃液pH[13];然后经4层纱布过滤,取上清液分装入2个15 mL离心管于-20 ℃保存,并分装4个冻存管存入液氮中。采用靛酚比色法测定氨态氮(NH3-N)浓度[14],并采用气相色谱分析法[14]测定总挥发性脂肪酸(TVFA)和短链脂肪酸(乙酸、丙酸、丁酸)浓度。

1.5.4 瘤胃微生物区系

将液氮保存的瘤胃液送至南京奥维森基因科技有限公司完成DNA抽提、PCR扩增、MiSeq文库构建以及MiSeq上机测序。
α和β多样性分析:基于操作分类单元(OTU)及其丰度结果,使用QIIME(v1.8.0)软件计算α多样性指数和β多样性距离矩阵,并使用R(v3.6.0)软件绘图,建立微生物相对丰度与样品类别之间的关系模型,来实现对样品类别的预测。
物种注释:将OTU代表序列使用BLAST算法与Silva138数据库进行比对,E值(E-value)阈值设置为e-5,得到每个OTU对应的物种分类信息。
基于物种注释及相对丰度结果,通过Spearman检验方法进行瘤胃微生物与发酵参数的相关性分析,并通过PICRUSt 2进行功能预测。

1.6 数据统计分析

营养物质表观消化率采用SPSS 22.0软件进行单因素方差分析(one-way ANOVA)和LSD多重比较,瘤胃发酵参数采用SAS 9.2软件的MIXED模型进行数据分析,P<0.05为差异显著,结果数据采用“平均值±标准误(mean±SE)”形式表示。

2 结果与分析

2.1 复合益生菌对羔羊营养物质表观消化率的影响

由表2可知,饲粮添加复合益生菌对羔羊DM、GE、CP和EE表观消化率有显著影响(P<0.05)。其中,Ⅱ组和Ⅲ组DM表观消化率显著高于Ⅰ组(P<0.05),Ⅳ组DM表观消化率有高于Ⅰ组的趋势(P>0.05);Ⅱ组和Ⅲ组GE表观消化率显著高于Ⅰ组和Ⅳ组(P<0.05);Ⅲ组CP和EE表观消化率显著高于Ⅰ组和Ⅱ组(P<0.05),Ⅱ组和Ⅳ组EE表观消化率显著高于Ⅰ组(P<0.05)。各组间NDF和ADF表观消化率无显著差异(P>0.05),不过Ⅲ组NDF和ADF表观消化率较Ⅰ组分别提高了17.04%和5.71%。
表2 复合益生菌对羔羊营养物质表观消化率的影响

Table 2 Effects of complex probiotics on nutrient apparent digestibility of lambs %

项目Items 组别Groups P值P-value
Ⅰ Ⅱ Ⅲ Ⅳ
干物质DM 71.24±2.67b 77.58±2.55a 79.98±1.66a 74.54±1.15ab 0.03
总能GE 69.24±2.36b 77.38±1.92a 78.93±1.29a 72.64±0.90b 0.01
粗蛋白质CP 64.54±3.27b 65.15±2.09b 76.80±2.61a 68.36±4.48ab 0.04
粗脂肪EE 60.31±2.65c 68.22±2.68b 75.07±1.54a 70.40±1.71ab 0.01
中性洗涤纤维NDF 45.67±6.07 54.98±3.73 53.45±3.59 52.50±4.28 0.24
酸性洗涤纤维ADF 39.76±5.34 34.65±5.47 42.03±5.03 32.88±4.62 0.55

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

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

2.2 复合益生菌对羔羊瘤胃发酵参数的影响

由表3可知,采样时间显著影响羔羊瘤胃pH以及NH3-N、TVFA、乙酸、丙酸和丁酸浓度(P<0.05),且上述指标随着采样时间的变化整体呈升高趋势。饲粮添加复合益生菌对瘤胃NH3-N、TVFA和丁酸浓度有显著影响(P<0.05)。其中,断奶后第3天和第7天,Ⅱ组、Ⅲ组和Ⅳ组瘤胃NH3-N浓度显著高于Ⅰ组(P<0.05);断奶当天,Ⅱ组、Ⅲ组和Ⅳ组瘤胃TVFA浓度显著高于Ⅰ组(P<0.05);断奶后第7天,Ⅳ组瘤胃TVFA浓度显著高于Ⅰ组(P<0.05);断奶前第7天和断奶当天,Ⅳ组瘤胃丁酸浓度显著高于Ⅰ组(P<0.05);断奶后第3天,Ⅱ组、Ⅲ组和Ⅳ组瘤胃丁酸浓度显著高于Ⅰ组(P<0.05);断奶后第7天,Ⅲ组瘤胃丁酸浓度显著高于Ⅰ组(P<0.05)。采样时间与饲粮添加复合益生菌对瘤胃丁酸浓度有显著交互作用(P<0.05)。
表3 复合益生菌对羔羊瘤胃发酵参数的影响

Table 3 Effects of complex probiotics on rumen fermentation parameters of lambs

项目Items 时间Time 组别Groups P值P-value
Ⅰ Ⅱ Ⅲ Ⅳ 时间Time 处理Treatment 时间×处理Time×treatment
pH -14D 5.84±0.07B 5.82±0.18C 5.87±0.19B 5.87±0.10B <0.01 0.49 0.77
-7D 6.39±0.21A 6.42±0.18B 6.48±0.23AB 6.57±0.16A
0D 6.47±0.22A 6.52±0.10AB 6.67±0.31A 6.85±0.15A
3D 6.41±0.13A 6.42±0.09B 6.57±0.19AB 6.65±0.30A
7D 6.69±0.18A 6.93±0.20A 6.97±0.19A 7.02±0.12A
氨态氮NH3-N/(mg/dL) -14D 9.78±0.25B 8.74±0.95C 8.47±1.04C 8.18±0.97C <0.01 0.02 0.62
-7D 15.20±1.25A 16.97±2.64B 17.21±1.69AB 16.02±1.56B
0D 10.02±1.09B 14.16±4.13BC 13.40±3.41BC 15.81±2.29B
3D 11.15±0.38Bb 17.99±1.34Ba 17.50±2.30ABa 18.71±3.87Ba
7D 17.39±1.28Ab 25.82±2.18Aa 24.57±1.98Aa 26.10±2.34Aa
总挥发性脂肪酸TVFA/(mmol/L) -14D 88.89±1.33 97.14±5.01B 89.43±5.86B 86.53±3.86B <0.01 <0.01 0.86
-7D 96.19±6.19 108.61±0.55AB 114.69±9.66AB 112.45±2.30A
0D 92.48±2.33b 113.10±4.06Aa 109.45±7.61ABa 110.68±4.19Aa
3D 90.07±5.37 112.51±7.79A 114.39±8.35AB 110.89±8.23A
7D 98.26±6.46b 118.68±1.99Aab 122.24±11.08Aab 127.66±8.68Aa
乙酸Acetate/(mmol/L) -14D 55.41±0.36AB 55.39±0.85BC 53.45±0.44BC 53.57±0.71B <0.01 0.47 0.49
-7D 58.44±1.32A 58.48±0.66AB 57.12±0.66B 57.74±0.33A
0D 46.36±0.29C 45.07±2.80D 48.78±1.10C 45.53±0.56D
3D 50.24±0.79BC 52.33±1.07C 49.09±0.47C 50.12±0.99C
7D 58.99±3.93A 60.43±0.61A 62.24±3.13A 57.90±0.79A
丙酸Propionate/(mmol/L) -14D 12.92±0.70B 13.43±0.81B 11.14±0.23B 11.91±0.55C <0.01 0.07 0.11
-7D 14.09±0.78AB 14.80±1.33B 14.24±0.48B 15.96±1.40B
0D 13.01±0.92B 13.35±0.69B 15.53±0.87B 14.21±0.59B
3D 15.36±0.78AB 15.39±0.44B 20.33±0.56A 14.90±0.22B
7D 16.40±0.91A 20.52±2.67A 23.10±3.33A 19.62±0.52A
丁酸Butyrate/(mmol/L) -14D 6.68±0.58B 7.81±0.29C 6.75±0.24D 6.93±0.54B <0.01 0.01 0.01
-7D 8.60±0.17ABb 9.62±0.28Bab 9.53±0.97BCab 10.67±0.39Aa
0D 8.27±0.37ABb 8.31±0.36Cb 8.27±0.37CDb 9.75±0.43Aa
3D 8.36±0.64ABb 10.83±0.28Aa 11.22±0.79ABa 10.22±0.49Aa
7D 9.53±0.97Ab 10.11±0.29ABb 12.46±0.36Aa 11.05±0.43Aab

-14D、-7D、0D、3D和7D分别表示断奶前第14天、断奶前第7天、断奶当天、断奶后第3天和断奶后第7天。同列数据肩标无字母或相同大写字母表示不同时间之间差异不显著(P>0.05),不同大写字母表示不同时间之间差异显著(P<0.05)。表4同。

-14D, -7D, 0D, 3D and 7D represented day 14 before weaning, day 7 before weaning, weaning day, day 3 after weaning, and day 7 after weaning, respectively. In the same column, values with no letter or the same capital letter superscripts mean no significant difference between different time (P>0.05), while with different capital letter superscripts mean significant difference between different time (P<0.05). The same as Table 4.

2.3 复合益生菌对羔羊瘤胃微生物区系的影响

2.3.1 瘤胃微生物α多样性分析

如图1所示,采样时间对羔羊瘤胃微生物Chao1指数有显著影响(P<0.05)。饲粮添加复合益生菌对瘤胃微生物Chao1指数和Observed_species指数有显著影响(P<0.05)。其中,断奶当天以及断奶后第3天和第7天,Ⅲ组Chao1指数和Observed_species指数显著高于Ⅰ组(P<0.05)。采样时间与饲粮添加复合益生菌对瘤胃微生物Chao1指数有显著交互作用(P<0.05)。采样时间和饲粮添加复合益生菌以及两者的交互作用对瘤胃微生物Shannon指数和Simpson指数均无显著影响(P>0.05)。
图1 复合益生菌对羔羊瘤胃微生物α多样性的影响

-14D、-7D、0D、3D和7D分别表示断奶前第14天、断奶前第7天、断奶当天、断奶后第3天和断奶后第7天。-14D, -7D, 0D, 3D and 7D represented day 14 before weaning, day 7 before weaning, weaning day, day 3 after weaning, and day 7 after weaning, respectively.

数据柱标注不同字母表示不同组间差异显著(P<0.05)。**表示P<0.01,ns表示P>0.05。Value columns with different letters mean significant difference between different groups (P<0.05). ** mean P<0.01, and ns mean P<0.05.

Fig.1 Effects of complex probiotics on α diversity of rumen microbiota of lambs

2.3.2 瘤胃微生物β多样性分析

如图2所述,基于OTU及其丰度结果,使用QIIME(v1.8.0)软件计算β多样性距离矩阵,采用偏最小二乘法判别分析(PLS-DA)分析发现,各组羔羊瘤胃微生物结构在断奶前第14天、断奶前第7天、断奶当天、断奶后第3天和断奶后第7天同时具有相似性和特异性。
图2 复合益生菌对羔羊瘤胃微生物β多样性的影响

Ⅰ,7~Ⅳ,7分别表示Ⅰ组~Ⅳ组断奶后第7天样本,Ⅰ,3~Ⅳ,3分别表示Ⅰ组~Ⅳ组断奶后第3天样本,Ⅰ,0~Ⅳ,0分别表示Ⅰ组~Ⅳ组断奶当天样本,Ⅰ,-7~Ⅳ,-7分别表示Ⅰ组~Ⅳ组断奶前第7天样本,Ⅰ,-14~Ⅳ,-14分别表示Ⅰ组~Ⅳ组断奶前第14天样本。下图同。

Fig.2 Effects of complex probiotics on β diversity of rumen microbiota of lambs

Ⅰ,7 to Ⅳ,7 represented samples in groups Ⅰ to Ⅳ on day 7 after weaning, Ⅰ,3 to Ⅳ,3 represented samples in groups Ⅰ to Ⅳ on day 3 after weaning, Ⅰ,0 to Ⅳ,0 represented samples in groups Ⅰ to Ⅳ on weaning day, Ⅰ,-7 to Ⅳ,-7 represented samples in groups Ⅰ to Ⅳ on day 7 before weaning, and Ⅰ,-14 to Ⅳ,-14 represented samples in groups Ⅰ to Ⅳ on day 14 before weaning, respectively. The same as below.

2.3.3 瘤胃微生物物种注释及群落组成差异分析

如图3所示,拟杆菌门(Bacteroidota)、厚壁菌门(Firmicutes)、髌骨菌门(Patescibacteria)和放线菌门(Actinobacteriota)是羔羊瘤胃中相对丰度较高的菌门。其中,Ⅰ组羔羊断奶后第3天瘤胃拟杆菌门和厚壁菌门相对丰度与断奶前第7天和断奶当天相比有所降低。Ⅱ组、Ⅲ组和Ⅳ组瘤胃厚壁菌门、拟杆菌门和髌骨菌门相对丰度高于Ⅰ组。Ⅱ组、Ⅲ组和Ⅳ组断奶当天以及断奶后第3天和第7天瘤胃放线菌门相对丰度高于Ⅰ组,并在Ⅲ组富集。同时,断奶当天和断奶后第3天,瘤胃变形菌门(Proteobacteria)在Ⅲ组富集。此外,瘤胃互养菌门(Synergistota)相对丰度在Ⅰ组断奶后第7天以及Ⅳ组断奶后第3天明显降低,瘤胃疣微菌门(Verrucomicrobiota)相对丰度在Ⅳ组断奶后第3天明显降低,WPS-2相对丰度在Ⅰ组(断奶当天以及断奶后第3天和第7天)、Ⅱ组(断奶后第3天和第7天)和Ⅳ组(断奶当天和断奶后第3天)明显降低,迷踪菌门(Elusimicrobiota)相对丰度在Ⅰ组和Ⅳ组断奶后第3天和第7天明显降低。
图3 复合益生菌对羔羊瘤胃微生物在门水平上组成的影响

Fig.3 Effects of complex probiotics on rumen microbiota composition of lambs at phylum level

如图4所示,羔羊瘤胃中共检测到372个菌属,其中相对丰度大于1.0%的菌属有16个,合计占84.88%,其中普雷沃氏菌属(Prevotella)占比为32.26%,是断奶前后羔羊瘤胃的优势菌属。各组瘤胃普雷沃氏菌属相对丰度在断奶前第14天和第7天较高,断奶当天迅速下降,断奶后第3天降至最低,断奶后第7天又缓慢升高。同时,Ⅱ组、Ⅲ组和Ⅳ组瘤胃普雷沃氏菌属相对丰度总体上高于Ⅰ组。
图4 复合益生菌对羔羊瘤胃微生物在属水平上组成的影响

Fig.4 Effects of complex probiotics on rumen microbiota composition of lambs at genus level

2.3.4 瘤胃微生物与发酵参数相关性分析

通过Spearman相关性分析,对羔羊瘤胃微生物在属水平上相对丰度大于1.0%的菌属与瘤胃发酵参数进行关联分析,如图5所示,Shuttleworthia相对丰度与pH呈显著正相关(P<0.05);瘤胃球菌属(Ruminococcus)相对丰度与NH3-N和乙酸浓度呈极显著正相关(P<0.01),与丙酸和丁酸浓度呈显著正相关(P<0.05);韦荣氏球菌科UCG-001(Veillonellaceae_UCG-001)相对丰度与TVFA和NH3-N浓度呈显著正相关(P<0.05);罗氏菌属(Roseburia)相对丰度与乙丙比呈显著正相关(P<0.05),与NH3-N、TVFA、丙酸和丁酸浓度呈显著负相关(P<0.05);丹毒丝菌科UCG-002(Erysipelotrichaceae_UCG-002)相对丰度与NH3-N浓度呈显著负相关(P<0.05)。
图5 瘤胃微生物与发酵参数相关性分析

**表示极显著相关(P<0.01),*表示显著相关(P<0.05)。

Fig.5 Correlation analysis of rumen microbiota and fermentation parameters

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

2.3.5 瘤胃微生物功能预测

由表4可知,采样时间对羔羊瘤胃微生物脂质代谢功能有显著影响(P<0.05),饲粮添加复合益生菌对瘤胃微生物碳水化合物代谢功能有显著影响(P<0.05)。其中,Ⅰ组断奶当天瘤胃微生物脂质代谢功能相关基因丰度显著高于断奶前第7天(P<0.05),Ⅱ组和Ⅲ组断奶后第3天瘤胃微生物碳水化合物代谢功能相关基因丰度显著高于Ⅰ组(P<0.05)。采样时间和饲粮添加复合益生菌以及两者的交互作用对瘤胃微生物氨基酸代谢和能量代谢功能无显著影响(P>0.05)。
表4 复合益生菌对羔羊瘤胃微生物代谢功能相关基因丰度的影响

Table 4 Effects of complex probiotics on abundance of genes related to metabolic function in rumen microbiota of lambs %

功能Function 时间Time 组别Groups P值P-value
Ⅰ Ⅱ Ⅲ Ⅳ 时间Time 处理Treatment 时间×处理Time×treatment
碳水化合物代谢Carbohydrate metabolism -14D 14.53±0.33 14.68±0.16 14.35±0.23 14.54±0.17 0.05 0.04 0.57
-7D 14.77±0.24 14.45±0.35 14.61±0.20 14.33±0.12
0D 14.73±0.35 13.98±0.21 14.32±0.23 13.92±0.26
3D 13.87±0.21b 14.89±0.58a 14.97±0.22a 14.17±0.16ab
7D 14.90±0.19 15.00±0.36 14.88±0.46 14.70±0.33
氨基酸代谢Amino acid metabolism -14D 13.08±0.03 13.09±0.12 13.05±0.07 13.20±0.21 0.61 0.87 0.46
-7D 12.99±0.08 13.00±0.06 13.25±0.02 12.87±0.07
0D 12.86±0.12 13.16±0.11 13.11±0.19 12.98±0.08
3D 13.34±0.19 13.13±0.06 13.08±0.17 13.21±0.19
7D 13.11±0.15 13.22±0.01 13.06±0.31 12.94±0.07
能量代谢Energy metabolism -14D 5.76±0.06 5.78±0.04 5.65±0.09 5.70±0.14 0.73 0.75 0.86
-7D 5.64±0.19 5.78±0.04 5.89±0.05 5.80±0.06
0D 5.66±0.05 5.68±0.00 5.75±0.05 5.61±0.05
3D 5.71±0.12 5.82±0.13 5.65±0.15 5.65±0.23
7D 5.86±0.06 5.70±0.16 5.77±0.11 5.62±0.07
脂质代谢Lipid metabolism -14D 4.31±0.26AB 4.11±0.27 5.13±0.48 4.62±0.20 0.08 0.60 0.41
-7D 3.79±0.16B 4.16±0.14 4.25±0.23 4.34±0.17
0D 4.99±0.33A 4.75±0.11 4.31±0.18 4.46±0.19
3D 4.70±0.10AB 4.58±0.04 4.76±0.26 4.67±0.22
7D 4.46±0.46AB 4.60±0.39 4.95±0.58 4.21±0.36

3 讨论

3.1 复合益生菌对羔羊营养物质表观消化率的影响

断奶应激会导致羔羊营养物质吸收利用率降低[14]。益生菌可有效降解饲粮中难以消化的大分子物质及抗营养因子,有助于提升养分消化率[15]。益生菌作为代谢调节剂被应用于畜禽养殖有显著效果,其中使用低剂量丁酸梭菌可以显著提高羔羊对淀粉、蛋白质和脂肪的消化率[16]。本试验结果显示,羔羊饲粮中添加复合益生菌可有效提高羔羊对饲粮中大部分营养物质的表观消化率。研究发现,益生菌随饲粮进入瘤胃定殖后具有一定固氮能力,能够促进氮利用微生物的增殖,增加含氮物质的吸收和利用,减少粪中氮的排出,从而提高蛋白质的消化利用率[17]。研究表明,脂肪和能量的消化率与蛋白质的消化率呈正相关,益生菌对DM和GE表观消化率的提高作用主要归因于对NDF和氮消化的增强[18-19]。本研究发现,饲粮添加1.0%复合益生菌显著提高羔羊DM、CP和EE表观消化率,且该组NDF和ADF表观消化率分别比对照组提高了17.04%和5.71%;而饲粮添加1.5%复合益生菌对营养物质表观消化率无显著影响,这与Dobrowolski等[20]针对不同剂量益生菌对火鸡肠道影响的研究结果类似,说明适量添加益生菌有利于改善饲粮营养物质消化率,但并不是添加越多越好。研究表明,细菌主要定殖在肠道皱褶表面,由于表面有限,菌株需要依靠其触须附着在肠道上皮细胞表面,部分学者认为当单次补充菌株剂量适宜时,有利于补充的菌株定殖;剂量过大,则会刺激动物肠道表面固有的细菌大量增殖,使得补充的菌株就很难在肠道表面定殖,其益生效果也就不显著;还有学者认为单次补充菌株剂量太大,固有菌和补充菌相互间竞争营养,最终导致补充的菌株营养不足,定殖能力变差[21],但具体原因还需进一步明确。

3.2 复合益生菌对羔羊瘤胃发酵参数的影响

瘤胃参与营养物质的消化吸收是其内部强大的微生物区系主导的发酵过程,研究认为羔羊瘤胃消化能力的强弱是由其发酵功能的发育情况决定的[22]。植物性饲料经过微生物作用产生VFA,瘤胃微生物对含氮物质的利用使NH3-N浓度升高,瘤胃内容物的变化直接影响其pH[23]。pH、NH3-N和VFA是衡量瘤胃内环境和发育程度的重要指标。pH作为瘤胃发酵的直观指标,受多种瘤胃内容物影响,能够反映反刍动物瘤胃发酵能力和饲粮营养物质消化情况[24]。研究发现,补充益生菌和益生元可以稳定瘤胃pH,降低动物发生亚急性瘤胃酸中毒的风险[25]。本试验中,各组间瘤胃pH无显著差异,但随着试验时间的延长呈上升趋势,说明羔羊断奶使瘤胃pH升高[26]。这可能与断奶前后羔羊采食方式的转变有关,采食植物性饲料会引起更多的唾液分泌,唾液中含有大量碳酸氢钠,对瘤胃液具有缓冲作用[27]。瘤胃微生物对蛋白氮、非蛋白氮及内源氮的利用和饲粮中碳水化合物的降解和吸收情况决定了瘤胃NH3-N和TVFA浓度[28]。本试验结果发现,各复合益生菌添加组间羔羊瘤胃发酵参数无显著差异,且瘤胃NH3-N和TVFA浓度高于对照组,说明益生菌可加强瘤胃发酵功能,提高瘤胃对营养物质的消化利用。卢晨阳[29]研究发现,羔羊饲喂复合益生菌制剂能够提高细胞代谢相关代谢通路水平,瘤胃中氨基酸代谢、碳水化合物代谢、脂质代谢等相关的菌群丰度显著升高。本试验发现,瘤胃NH3-N和TVFA浓度在断奶前第7天、断奶当天和断奶后第3天间无显著变化,部分研究者也观察到羔羊断奶前后瘤胃NH3-N浓度不会出现显著变化,断奶前后羔羊对饲粮中氮的转化能力和瘤胃微生物利用氮合成微生物蛋白之间相互调节并影响VFA浓度[30]。本试验结果表明,瘤胃NH3-N和TVFA浓度在断奶前后几天不会出现显著变化,可能是羔羊断奶后采食量增加,瘤胃发酵气体受食糜通过速率和瘤胃内容物缓冲能力影响的结果[23]。不过,断奶后第7天瘤胃NH3-N和TVFA浓度显著高于断奶前第14天,说明断奶后一段时间,随着断奶应激的缓解,羔羊对饲粮氮和植物性纤维的消化能力提高。当瘤胃中分解纤维素的菌群丰度较高时,瘤胃降解纤维素的能力较强,此时瘤胃乙酸浓度较高;当分解淀粉的菌群成为优势菌群时,瘤胃优先分解淀粉,此时瘤胃丙酸浓度上升。本试验中,各组间瘤胃乙酸和丙酸浓度无显著差异,说明两类分解碳水化合物菌群的相对丰度在不同组间无显著差异[31]。此外,瘤胃乙酸、丙酸和丁酸浓度随着试验的进行先降低再升高,并在断奶当天处于较低水平,说明断奶会降低瘤胃中部分短链脂肪酸浓度,这可能是羔羊断奶应激影响了瘤胃微生物代谢,导致与分解碳水化合物的菌群在断奶前后相对丰度出现差异有关[30,32]。

3.3 复合益生菌对羔羊瘤胃微生物区系的影响

瘤胃微生物对羔羊的健康和生长性能至关重要。研究表明,瘤胃微生物区系容易受到多种因素的影响,大部分饲料添加剂是通过调节瘤胃微生物来提高动物健康状态和生长性能[33]。本试验通过对瘤胃微生物进行16S rRNA检测分析发现,复合益生菌处理和采样时间对Chao1指数有显著交互作用。1.0%复合益生菌添加组瘤胃微生物Chao1指数和Observed_species指数在断奶当天以及断奶后第3天和第7天均显著高于对照组,说明1.0%复合益生菌提高了羔羊断奶后瘤胃微生物的丰富度和菌种数目。对照组Chao1指数和Shannon指数较低,说明断奶使瘤胃菌群多样性和丰富度降低。进一步分析羔羊断奶前后瘤胃微生物在门水平上的组成发现,厚壁菌门和拟杆菌门共占总菌的80%以上,是优势菌。厚壁菌门和拟杆菌门主要参与碳水化合物发酵,反刍动物在开始采食饲粮后,这两类菌群的相对丰度会快速升高[34]。本试验发现,羔羊断奶后第3天瘤胃厚壁菌门和拟杆菌门相对丰度较断奶前有所下降,结合瘤胃VFA浓度的结果,说明羔羊断奶后瘤胃消化碳水化合物菌群相对丰度的下降,是造成羔羊断奶对饲粮消化能力下降的原因之一。
此外,厚壁菌门和拟杆菌门直接影响饲料利用率,其相对丰度与反刍动物的日增重呈正相关[35],且有助于乙酸的产生[36]。厚壁菌门有助于消化碳水化合物并产生有机酸,其可为机体提供多种维生素,提高免疫水平,缓解炎症,增强胃肠道屏障功能并产生生物拮抗作用,有效改善机体蛋白质代谢[37]。本试验中,羔羊断奶后第3天瘤胃拟杆菌门和厚壁菌门相对丰度降低,直接导致瘤胃对营养物质的消化利用率下降,而添加复合益生菌可以提高瘤胃菌群丰富度和菌种数目,使得发酵碳水化合物、脂肪和蛋白质的菌群相对丰度提高,从而使得羔羊对营养物质的消化利用率提高。对瘤胃微生物进行功能预测发现,1.0%复合益生菌添加组断奶后第3天碳水化合物代谢功能相关基因丰度显著高于对照组,这与本试验中营养物质表观消化率和瘤胃发酵参数的结果基本吻合,表明羔羊饲粮添加1.0%复合益生菌有利于调节瘤胃菌群结构,从而缓解断奶应激对羔羊的影响,提高羔羊对营养物质的消化利用。

4 结论

饲粮添加复合益生菌可以提高羔羊营养物质表观消化率,并通过调节瘤胃菌群促进瘤胃发酵产生VFA,从而缓解断奶应激对羔羊的影响。本试验条件下,饲粮添加1.0%复合益生菌效果较佳。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
王世琴. 断奶应激及饲喂白藜芦醇和地衣芽孢杆菌对羔羊胃肠道发育及其微生物区系的影响[D]. 博士学位论文. 兰州: 兰州大学, 2020.

WANG S Q. Effects of weaning stress and supplementation of resveratrol and Bacillus licheniformis on gastrointestinal development and microbiota in lambs[D]. Ph.D. Thesis. Lanzhou: Lanzhou University, 2020. (in Chinese)

[2]
李冲. 断奶应激对羔羊消化道结构与功能发育的影响及其机理研究[D]. 博士学位论文. 兰州: 兰州大学, 2019.

LI C. Effects of weaning stress on the structural and functional development of digestive tract of lamb and its mechanism[D]. Ph.D.Thesis. Lanzhou: Lanzhou University, 2019. (in Chinese)

[3]
HILL C, GUARNER F, REID G, et al. The international scientific association for probiotics and prebiotics consensus statement on the scope and appropriate use of the term probiotic[J]. Nature Reviews: Gastroenterology & Hepatology, 2014, 11(8):506-514.

[4]
HAMDON H A, KASSAB A Y, VARGAS-BELLO-PÉREZ E, et al. Using probiotics to improve the utilization of chopped dried date palm leaves as a feed in diets of growing Farafra lambs[J]. Frontiers in Veterinary Science, 2022,9:1048409.

[5]
KHOLIF A E, ANELE A, ANELE U Y. Microbial feed additives in ruminant feeding[J]. AIMS Microbiology, 2024, 10(3):542-571.

DOI PMID

[6]
王兴岗, 郝小燕, 张暄梓, 等. 不同精粗比饲粮中添加凝结芽孢杆菌对绵羊羔羊瘤胃发酵和免疫功能的影响[J]. 动物营养学报, 2024, 36(3):1769-1781.

DOI

WANG X G, HAO X Y, ZHANG X Z, et al. Effects of adding Bacillus coagulans into diets with different concentrate to forage ratios on rumen fermentation and immune function of lambs[J]. Chinese Journal of Animal Nutrition, 2024, 36(3):1769-1781. (in Chinese)

[7]
MEALE S J, LI S C, AZEVEDO P, et al. Weaning age influences the severity of gastrointestinal microbiome shifts in dairy calves[J]. Scientific Reports, 2017, 7(1):198.

DOI PMID

[8]
LIAO R R, XIE X H, LV Y H, et al. Ages of weaning influence the gut microbiota diversity and function in Chongming white goats[J]. Applied Microbiology and Biotechnology, 2021, 105(9):3649-3658.

[9]
LI Y Y, LI X, NIE C X, et al. Effects of two strains of Lactobacillus isolated from the feces of calves after fecal microbiota transplantation on growth performance,immune capacity,and intestinal barrier function of weaned calves[J]. Frontiers in Microbiology, 2023,14:1249628.

[10]
CHEN H, GUO B B, YANG M R, et al. Response of growth performance,blood biochemistry indices,and rumen bacterial diversity in lambs to diets containing supplemental probiotics and Chinese medicine polysaccharides[J]. Frontiers in Veterinary Science, 2021,8:681389.

[11]
GART E V, LAWHON S D, SUCHODOLSKI J S, et al. 157 the relationship of weaning stress, sex and temperament on fecal microbiota and metabolites in Brahman calves[J]. Journal of Animal Science, 2019, 97(Suppl.1):47.

[12]
中华人民共和国农业农村部.肉羊营养需要量:NY/T 816—2021[S]. 北京: 中国农业出版社, 2021.

Ministry of Agriculture and Rural Affairs of the People's Republic of China.Nutrient requirements of meat-type sheep and goats:NY/T 816—2021[S]. Beijing: China Agriculture Press, 2021. (in Chinese)

[13]
李洪涛, 杨在宾, 杨维仁, 等. 不同日粮类型对杜泊羊瘤胃pH、氨态氮和挥发性脂肪酸浓度的影响[J]. 中国畜牧杂志, 2017, 53(3):67-72.

LI H T, YANG Z B, YANG W R, et al. Effects of different diet types on ruminal pH,NH3-N and VFA concentration of dorper sheep[J]. Chinese Journal of Animal Science, 2017, 53(3):67-72. (in Chinese)

[14]
WANG S Q, MA T, ZHAO G H, et al. Effect of age and weaning on growth performance,rumen fermentation,and serum parameters in lambs fed starter with limited ewe-lamb interaction[J]. Animals, 2019, 9(10):825.

[15]
WANG C, WEI S Y, XU B C, et al. Bacillus subtilis and Enterococcus faecium co-fermented feed regulates lactating sow's performance,immune status and gut microbiota[J]. Microbial Biotechnology, 2021, 14(2):614-627.

[16]
李稳稳. 益生菌对断奶羔羊生长性能、消化吸收能力、免疫功能和肠道微生态的影响[D]. 硕士学位论文. 合肥: 安徽农业大学, 2020.

LI W W. Effects of probiotics on growth performance,digestion and absorption ability,immune function and intestinal microecology of weaned lambs[D]. Master's Thesis. Hefei: Anhui Agricultural University, 2020. (in Chinese)

[17]
PELEGRIN-VALLS J, SERRANO-PÉREZ B, VILLALBA D, et al. Minimum effects of sampling time on the apparent digestibility of nutrients and blood protein catabolites in light lambs[J]. Master's Thesis.Animals, 2021, 11(8):2244.

[18]
GUO L, ZHANG D H, DU R F, et al. Supplementation of Aspergillus oryzae culture improved the feed dry matter digestibility and the energy supply of total volatile fatty acid concentrations in the rumen of Hu sheep[J]. Frontiers in Nutrition, 2022,9:847156.

[19]
ZAPATA O, CERVANTES A, BARRERAS A, et al. Effects of single or combined supplementation of probiotics and prebiotics on ruminal fermentation,ruminal bacteria and total tract digestion in lambs[J]. Small Ruminant Research, 2021,204:106538.

[20]
DOBROWOLSKI P, TOMASZEWSKA E, KLEBANIUK R, et al. Structural changes in the small intestine of female turkeys receiving a probiotic preparation are dose and region dependent[J]. Animal, 2019, 13(12):2773-2781.

DOI PMID

[21]
VAN NIEL C W, FEUDTNER C, GARRISON M M, et al. Lactobacillus therapy for acute infectious diarrhea in children:a Meta-analysis[J]. Pediatrics, 2002, 109(4):678-684.

[22]
WANG Y, LI Z, JIN W, et al. Isolation and characterization of ruminal yeast strain with probiotic potential and its effects on growth performance,nutrients digestibility,rumen fermentation and microbiota of Hu sheep[J]. Journal of Fungi, 2022, 8(12):1260.

[23]
张思源, 杨慧, 陈天宇, 等. 早期补饲粗料对犊牛生长性能、瘤胃发酵性能和反刍时长的影响[J]. 饲料研究, 2021, 44(23):1-5.

ZHANG S Y, YANG H, CHEN T Y, et al. Effect of providing forage to calves at early age on growth performance,rumen fermentation performance and ruminating duration[J]. Feed Research, 2021, 44(23):1-5. (in Chinese)

[24]
GODÍNEZ-OLMEDO J T, CORONA L, CASTREJÓN-PINEDA F A, et al. Optimal level of physically effective neutral detergent fibre in corn stover cracked-corn-based finishing diets on the growth performance,dietary energetics,carcase characteristics, and nutrient digestion in fattening lambs[J]. Italian Journal of Animal Science, 2022, 21(1):397-407.

[25]
张智安. 亚急性瘤胃酸中毒风险对湖羊瘤胃发酵及肌肉脂肪酸组成的影响[D]. 硕士学位论文. 兰州: 兰州大学, 2020.

ZHANG Z A. Effects of subacute rumen acidosis risk on rumen fermentation and composition of meat fatty acid in Hu sheep[D]. Master's Thesis. Lanzhou: Lanzhou University, 2020. (in Chinese)

[26]
刘依莎, 张巧娥, 许迟, 等. 发酵玉米芯对肉牛生长性能、瘤胃发酵参数及菌群的影响[J]. 动物营养学报, 2024, 36(2):1039-1049.

DOI

LIU Y S, ZHANG Q E, XU C, et al. Effects of fermented corn cob on growth performance,rumen fermentation parameters and microflora of beef cattle[J]. Chinese Journal of Animal Nutrition, 2024, 36(2):1039-1049. (in Chinese)

[27]
RICCI S, RIVERA-CHACON R, PETRI R M, et al. Supplementation with phytogenic compounds modulates salivation and salivary physico-chemical composition in cattle fed a high-concentrate diet[J]. Frontiers in Physiology, 2021,12:645529.

[28]
史晨迪, 赵晓雅, 田沛知, 等. 断奶期内不同饲粮对羔羊生长、营养物质表观消化率和瘤胃发酵参数的影响[J]. 动物营养学报, 2022, 34(8):5219-5233.

DOI

SHI C D, ZHAO X Y, TIAN P Z, et al. Effects of different diets on growth,nutrient apparent digestibility and rumen fermentation parameters of lambs during weaning period[J]. Chinese Journal of Animal Nutrition, 2022, 34(8):5219-5233. (in Chinese)

[29]
卢晨阳. 复合益生菌制剂对羔羊球虫感染、肠道菌群及免疫的影响[D]. 硕士学位论文. 郑州: 河南农业大学, 2022.

LU C Y. Effects of compound probiotics on coccidial infection,intestinal microbiota and immunity in lambs[D]. Master's Thesis. Zhengzhou: Henan Agricultural University, 2022. (in Chinese)

[30]
刘鹏, 李勇, 张成新, 等. 湖羊羔羊早期断奶前后瘤胃发酵与微生物区系的变化[J]. 中国畜牧兽医, 2021, 48(1):144-153.

DOI

LIU P, LI Y, ZHANG C X, et al. Variations of rumen fermentation and microflora of Hu lambs before and after early weaning[J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(1):144-153. (in Chinese)

[31]
曹文新, 李艳琴, 纪守坤, 等. 灭活酵母培养物对育肥羔羊瘤胃发酵和细菌群落及免疫指标的影响[J]. 中国畜牧杂志, 2021, 57(5):207-214.

CAO W X, LI Y Q, JI S K, et al. Effects of inactivated yeast culture on rumen fermentation,rumen bacterial diversity and immune indices of lambs[J]. Chinese Journal of Animal Science, 2021, 57(5):207-214. (in Chinese)

[32]
田林涛. 添加益生菌制剂对湖羊羔羊舍饲育肥效果及瘤胃微生物区系的影响[D]. 硕士学位论文. 杨凌: 西北农林科技大学, 2022.

TIAN L T. Effects of adding probiotics on fattening effect and rumen microflora of Hu lambs[D]. Master's Thesis. Yangling: Northwest A&F University, 2022. (in Chinese)

[33]
PALANGI V, LACKNER M. Management of enteric methane emissions in ruminants using feed additives:a review[J]. Animals, 2022, 12(24):3452.

[34]
杨蕾, 王柏辉, 靳烨. 反刍动物瘤胃菌群与机体营养沉积研究进展[J]. 黑龙江畜牧兽医, 2017(15):85-87,90.

YANG L, WANG B H, JIN Y. Rumen flora and nutrient deposition in ruminants[J]. Heilongjiang Animal Science and Veterinary Medicine, 2017(15):85-87, 90. (in Chinese)

[35]
YAXING Z, ERDENE K, ZHIBI B, et al. Effects of Allium mongolicum regel essential oil supplementation on growth performance,nutrient digestibility,rumen fermentation,and bacterial communities in sheep[J]. Frontiers in Veterinary Science, 2022,9:926721.

[36]
TIAN D D, XU X Q, PENG Q, et al. Effects of banana powder (Musa acuminata Colla) on the composition of human fecal microbiota and metabolic output using in vitro fermentation[J]. Journal of Food Science, 2020, 85(8):2554-2564.

[37]
HUANG Y L, WANG G X, LI C, et al. Periodical changes of feces microbiota and its relationship with nutrient digestibility in early lambs[J]. Animals, 2022, 12(14):1770.

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