研究论文

博落回提取物对育肥湖羊公羔生长性能及瘤胃发酵参数、酶活性和微生物数量的影响

  • 高秀杰 , 1 ,
  • 郝小燕 1 ,
  • 何甜 1 ,
  • 张暄梓 1 ,
  • 田光元 1 ,
  • 项斌伟 2 ,
  • 张建新 , 1, *
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  • 1 山西农业大学动物科学学院,太谷 030801
  • 2 右玉县农业农村和水利发展中心,右玉 037200
*张建新,教授,博士生导师,E-mail:

高秀杰(2001—),男,山西朔州人,硕士研究生,从事饲料资源开发和利用研究。E-mail:

Copy editor: 靳爽

收稿日期: 2025-04-11

  网络出版日期: 2025-11-14

基金资助

国家现代农业产业技术体系建设专项资金资助(CARS-38)

山西省现代农业产业技术体系建设专项(2024CYJSTX14-09)

山西省科技成果转化引导专项(202404021301048)

2025年度“特”“优”农业高质量发展科技支撑工程项目(TYGC25-75)

山西农业大学“产学研融合推进工程”项目(2024CXYRH-023)

Effects of Macleaya cordata Extract on Growth Performance, Rumen Fermentation Parameters, Enzyme Activities and Microbial Numbers of Fattening Male Hu Lambs

  • GAO Xiujie , 1 ,
  • HAO Xiaoyan 1 ,
  • HE Tian 1 ,
  • ZHANG Xuanzi 1 ,
  • TIAN Guangyuan 1 ,
  • XIANG Binwei 2 ,
  • ZHANG Jianxin , 1, *
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  • 1 College of Animal Science, Shanxi Agricultural University, Taigu 030801, China
  • 2 Youyu County Agriculture Rural and Water Conservancy Development Center, Youyu 037200, China
*professor, E-mail:

Received date: 2025-04-11

  Online published: 2025-11-14

摘要

本试验旨在探究博落回提取物(MCE)对育肥湖羊公羔生长性能及瘤胃发酵参数、酶活性和微生物数量的影响。试验选取48只体况良好、体重[(27.50±1.25) kg]相近的4月龄纯种湖羊公羔,随机分为4组,每组12只。对照组饲喂基础饲粮,试验组分别在基础饲粮中添加300(300 MCE组)、600(600 MCE组)和900 mg/kg(900 MCE组)的MCE。试验期75 d,其中预试期15 d,正试期60 d。结果表明:1)600 MCE组羔羊终末体重和平均日增重显著高于对照组(P<0.05),料重比显著低于对照组(P<0.05)。2)600 MCE组瘤胃液丙酸浓度显著高于对照组和300 MCE组(P<0.05),瘤胃液戊酸和氨态氮(NH3-N)浓度显著低于对照组(P<0.05);600 MCE组和900 MCE组瘤胃液乙丙比显著低于对照组(P<0.05)。3)600 MCE组瘤胃液蛋白酶活性显著低于对照组(P<0.05),瘤胃液羧甲基纤维素酶活性显著高于对照组和300 MCE组(P<0.05);300 MCE组、600 MCE组和900 MCE组瘤胃液α-淀粉酶活性均显著高于对照组(P<0.05);600 MCE组和900 MCE组瘤胃液果胶酶活性显著高于对照组(P<0.05)。4)600 MCE组瘤胃产甲烷菌数量显著低于对照组和300 MCE组(P<0.05),瘤胃黄色瘤胃球菌数量显著高于对照组和300 MCE组(P<0.05);600 MCE组和900 MCE组瘤胃嗜淀粉瘤胃杆菌和产琥珀酸丝状杆菌数量显著高于对照组(P<0.05),栖瘤胃普雷沃氏菌数量显著低于对照组(P<0.05)。综上所述,饲粮中添加MCE可以促进育肥湖羊公羔瘤胃发酵,降低瘤胃液NH3-N浓度,改变瘤胃微生物数量及酶活性,从而提高饲料利用率,促进羔羊生长。在本试验条件下,MCE的适宜添加量为600 mg/kg。

本文引用格式

高秀杰 , 郝小燕 , 何甜 , 张暄梓 , 田光元 , 项斌伟 , 张建新 . 博落回提取物对育肥湖羊公羔生长性能及瘤胃发酵参数、酶活性和微生物数量的影响[J]. 动物营养学报, 2025 , 37(11) : 7681 -7691 . DOI: 10.12418/CJAN2025.624

Abstract

This experiment was conducted to investigate the effects of Macleaya cordata extract (MCE) on growth performance, rumen fermentation parameters, enzyme activities and microbial numbers of fattening male Hu lambs. A total of 48 healthy 4-month-old purebred male Hu lambs with similar body weight [(27.50±1.25) kg] were randomly divided into 4 groups, with 12 lambs per group. The control group was fed a basal diet, while the experimental groups were fed the basal diets supplemented with 300 (300 MCE group), 600 (600 MCE group) and 900 mg/kg (900 MCE group) of MCE, respectively. The experiment lasted for 75 days, including a 15-day preliminary feeding period and a 60-day formal feeding period. The results showed as follows: 1) the final body weight and average daily gain of lambs in the 600 MCE group were significantly higher than those in the control group (P<0.05), while the feed-to-gain ratio was significantly lower than that in the control group (P<0.05). 2) The rumen propionate concentration in the 600 MCE group was significantly higher than that in the control group and 300 MCE group (P<0.05), while the rumen valerate and ammonia nitrogen (NH3-N) concentrations were significantly lower than those in the control group (P<0.05); the rumen acetate-to-propionate ratio in the 600 MCE group and 900 MCE group was significantly lower than that in the control group (P<0.05). 3) The rumen protease activity in the 600 MCE group was significantly lower than that in the control group (P<0.05), and the rumen carboxymethyl cellulase activity was significantly higher than that in the control group and 300 MCE group (P<0.05); the rumen α-amylase activity in the 300 MCE group, 600 MCE group and 900 MCE group was significantly higher than that in the control group (P<0.05); the rumen pectinase activity in the 600 MCE group and 900 MCE group was significantly higher than that in the control group (P<0.05). 4) The rumen methanogen number in the 600 MCE group was significantly lower than that in the control group and 300 MCE group (P<0.05), and the rumen Ruminococcus flavefaciens number was significantly higher than that in the control group and 300 MCE group (P<0.05); the rumen Ruminobacter amylophilus and Fibrobacter succinogenes numbers in the 600 MCE group and 900 MCE group were significantly higher than those in the control group (P<0.05), while the rumen Prevotella ruminicola number was significantly lower than that in the control group (P<0.05). In conclusion, diet supplemented with MCE can promote rumen fermentation, reduce rumen NH3-N concentration, alter rumen microbial numbers and enzyme activities of fattening male Hu lambs, thereby improving feed utilization efficiency and promoting lamb growth. Under the conditions of this experiment, the optimal supplementation level of MCE is 600 mg/kg.

随着羊肉销售量的持续增长,从断奶羔羊开始进行集约化育肥已成为普遍的生产方式。在过去,为了提升动物的生产性能与经济效益,养殖业普遍采用在饲粮中添加抗生素的方式提高饲料利用率和促进生长[1]。然而,抗生素过度使用导致的畜禽产品中药物残留问题日益突出,已成为制约畜牧业可持续发展的关键挑战之一[2]。在这一背景下,畜牧业迫切需要寻找既能有效替代抗生素、又具备更高安全性的新型饲料添加剂。博落回提取物(Macleaya cordata extract,MCE)是从博落回植物中提取的一种主要成分为血根碱和白屈菜红碱的异喹啉类生物碱,具有抗菌、抗炎、抗氧化、抗病毒和杀虫等多种生物学功能[3]。研究表明,MCE能够减轻小鼠的氧化应激并增强先天免疫反应[4]。Li等[5]研究发现,在妊娠后期母猪及宫内发育迟缓仔猪饲粮中添加MCE,可通过调节免疫应答机制有效缓解热应激诱导的氧化损伤。罗鑫宇等[6]研究发现,饲粮中添加40 mg/kg博落回散制剂可显著提高番鸭生长性能,促进免疫器官发育,并增强机体免疫功能及抗氧化能力。Hussein等[7]研究发现,饲粮中添加异喹啉类生物碱添加剂能够有效预防肉鸡坏死性肠炎的发生,并可作为抗生素替代物提升肉鸡生长性能及改善肉品质。此外,血根碱可通过提高奶山羊和犊牛血清中免疫球蛋白G(IgG)、免疫球蛋白A(IgA)及白细胞介素-2(IL-2)的含量,进而增强其免疫功能[8-9]。在早期断奶山羊中,MCE的摄入还会影响Toll样受体及下消化道信号通路相关基因的表达[10]。以上研究表明,MCE在增强动物免疫力、促进动物生长、预防畜禽疾病、改善畜禽产品品质方面具有广阔应用前景。然而,不同物种及同一物种不同生产阶段对MCE的消化、代谢、残留情况乃至性能反应存在差异,仍需进一步研究。尤其是在公羔育肥阶段,关于MCE对瘤胃发酵特性、酶活性及微生物群落结构的影响尚不明确。因此,本研究以湖羊公羔为试验对象,探究MCE对其生长性能及瘤胃发酵参数、酶活性和微生物数量的影响,以期为MCE在反刍动物生产中的合理应用提供理论依据。

1 材料与方法

1.1 试验材料

MCE购自湖南某生物资源股份有限公司,其活性成分为血根碱、白屈菜红碱等异喹啉类生物碱(生物碱含量≥0.375%,其余为载体)。

1.2 试验设计及饲粮

动物试验程序经山西农业大学实验动物伦理委员会批准(批准号:SXAU-EAW-2024G.0S.004017286)。采用单因素完全随机设计,选取48只体况良好、体重[(27.50±1.25) kg]相近的4月龄纯种湖羊公羔,随机分为4组,每组12只。对照组饲喂基础饲粮,试验组分别在基础饲粮中添加300(300 MCE组)、600(600 MCE组)和900 mg/kg(900 MCE组)的MCE。试验期75 d,其中预试期15 d,正试期60 d。基础饲粮参照NY/T 816—2021中体重30 kg、日增重300 g的生长育肥羊营养需求配制,其组成及营养水平见表1
表1 基础饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of the basal diet (air-dry basis)%

项目Items 含量Content
原料Ingredients
玉米Corn 31.5
玉米胚芽粕Corn germ meal 9.0
喷浆玉米皮Sprayed corn bran 5.0
豆粕Soybean meal 9.5
葵花籽饼Sunflower seed meal 4.0
小米糠Millet bran 1.0
次粉Wheat middling 5.0
大豆皮Soybean hull 15.0
稻壳粉Rice hull powder 15.0
预混料Premix1) 5.0
合计Total 100.0
营养水平Nutrient levels2)
代谢能ME/(MJ/kg) 9.19
干物质DM 96.10
粗灰分Ash 6.15
粗脂肪EE 3.30
粗蛋白质CP 14.09
中性洗涤纤维NDF 40.22
酸性洗涤纤维ADF 25.67

1)预混料为每千克饲粮提供 The premix provided the following per kg of the diet:VA 20 000 IU,VD 4 000 IU,VE 40 IU,Fe 55 mg,Cu 15 mg,Mn 50 mg,Zn 25 mg,Se 0.3 mg,I 0.5 mg,Co 0.2 mg。

2)代谢能为计算值[11],其余为实测值。ME was a calculated value[11], while the others were measured values.

1.3 饲养管理

本试验于2024年6—10月在山西省朔州市右玉县宏宇牧业科技小院试验基地开展。试验开始前对羊舍进行清洗和消毒,对试验羊进行分组、编号、驱虫和免疫。待羊群生理指标恢复正常后,进行空腹称重并做好个体标记,随后进行随机分组。疫苗接种结束1周后,将试验羊转入独立圈舍进行单栏(规格为2.2 m×1.0 m)饲养,预试期自此正式开始。预试期内通过3次空腹称重动态调整组间均重,确保正式试验开始时各组试验羊体重无显著差异(P>0.05)。
在正式试验期间,每日使用分析天平精确称量每只试验羊饲粮中的MCE添加量。为确保MCE的有效摄入,先将其与60 g粉状基础饲粮在不锈钢容器中充分混匀后置于空料槽,待羊只舔食完毕后再饲喂剩余颗粒饲粮。每日于07:00和17:00各饲喂1次,羊只自由采食和饮水。

1.4 样品采集

试验结束当天,按常规流程完成饲喂,待羊只采食结束3 h后,采用口腔探管法采集瘤胃液。为避免唾液污染的影响,弃去前5~10 mL瘤胃液,对后续收集的80 mL瘤胃液立即进行pH检测。随后将样品分装至离心管中,一部分经液氮速冻后转至-80 ℃保存,用于瘤胃微生物群落定量分析及酶活性检测;另一部分置于-20 ℃保存,用于瘤胃发酵参数的测定。

1.5 指标测定

1.5.1 饲粮营养物质含量

饲粮干物质(DM)、粗灰分(Ash)和粗脂肪(EE)含量分别参照GB/T 6435—2014、GB/T 6438—2007和GB/T 6433—2006的方法测定;粗蛋白质(CP)含量参照GB/T 6432—2018,采用凯氏定氮法测定;中性洗涤纤维(NDF)含量参照GB/T 20806—2022,采用中性洗涤剂法测定;酸性洗涤纤维(ADF)含量参照NY/T 1459—2022,采用酸性洗涤剂法测定。

1.5.2 生长性能

正试期间,每天精确记录每只羊的投料量与剩料量,计算平均日采食量(ADFI);在正式试验的第1天和第60天晨饲前,对全部试验羊进行空腹称重,分别记为初始体重(IBW)和终末体重(FBW),计算平均日增重(ADG),并根据平均日采食量和平均日增重计算料重比(F/G)。

1.5.3 瘤胃发酵参数

瘤胃液挥发性脂肪酸(VFA)浓度参照Wang等[12]的气相色谱法,使用Agilent 7980B气相色谱仪(安捷伦科技有限公司,美国)测定;氨态氮(NH3-N)浓度参照金亚倩等[13]的比色法进行测定。

1.5.4 瘤胃酶活性

瘤胃液中纤维二糖酶、羧甲基纤维素酶、α-淀粉酶、木聚糖酶、蛋白酶及果胶酶活性参照Agarwal等[14]的方法进行测定。

1.5.5 瘤胃微生物数量

采用实时荧光定量PCR技术对目标微生物进行定量分析。瘤胃微生物总DNA通过锆珠研磨法提取,使用赛默飞世尔科技公司生产的超微量核酸检测仪测定DNA浓度与纯度,并用TE缓冲液将所有样本DNA浓度统一稀释至100 ng/mL。目标微生物的特异性引物由深圳华大基因股份有限公司合成,引物序列见表2。标准品由山西农业大学动物营养研究室提供,参照Li等[15]描述的方法,经10倍系列稀释构建标准曲线模板。荧光定量PCR反应使用TaKaRa公司生产的SYBR Premix Ex Taq Ⅱ试剂盒(含Tli RNaseH Plus酶),在ABI StepOne Plus实时荧光定量PCR系统上进行。反应体系总体积为20.0 μL,包含2.0 μL DNA模板、10.0 μL SYBR® Premix Ex TaqTM、0.8 μL上游引物(10 μmol/L)、0.8 μL下游引物(10 μmol/L)、0.4 μL ROX Reference Dye Ⅱ以及6.0 μL灭菌蒸馏水。
表2 引物序列

Table 2 Primer sequences

项目
Items
引物序列
Primer sequences (5'—3')
产物大小
Product size/bp
参考文献
References
产甲烷菌
Methanogens
F:TTCGGTGGATCDCARAGRGC
R:GBARGTCGWAWCCGTAGAATCC
140 Zhang等[16]
嗜淀粉瘤胃杆菌
Ruminobacter amylophilus
F:CTGGGGAGCTGCCTGAATG
R:GCATCTGAATGCGACTGGTTG
100 Jami等[17]
栖瘤胃普雷沃氏菌
Prevotella ruminicola
F:CATCCTATAGCGGTAAACCTTTGG
R:GAAAGTCGGATTAATGCTCTATGTTG
74 Jami等[17]
溶纤维丁酸弧菌
Butyrivibrio fibrisolvens
F:TAACATGAGAGTTTGATCCTGGCTC
R:CGTTACTCACCCGTCCGC
136 Forster等[18]
产琥珀酸丝状杆菌
Fibrobacter succinogenes
F:GGCGGGATTGAATGTACCTTGAGA
R:TCCGCCTGCCCCTGAACTATC
204 Denman等[19]
白色瘤胃球菌
Ruminococcus albus
F:CCCTAAAAGCAGTCTTAGTTCG
R:CCTCCTTGCGGTTAGAACA
176 Koike等[20]
黄色瘤胃球菌
Ruminococcus flavefaciens
F:ATTGTCCCAGTTCAGATTGC
R:GGCGTCCTCATTGCTGTTAG
173 Denman等[19]
总菌
Total bacteria
F:CGGTGAATACGTTCYCGG
R:CGWTACCTTGTTACGACTT
123 Denman等[19]

1.6 数据统计分析

试验数据经Excel 2019初步整理后,采用SPSS 26.0软件进行单因素方差分析(one-way ANOVA),差异显著的指标采用Duncan氏法进行多重比较,并进行线性和二次回归分析。结果以平均值和均值标准误(SEM)表示,P<0.05表示差异显著,0.05≤P<0.10表示差异有显著趋势。

2 结果

2.1 饲粮中添加MCE对育肥湖羊公羔生长性能的影响

表3可知,饲粮中添加MCE对羔羊平均日采食量没有显著影响(P>0.05)。600 MCE组羔羊终末体重和平均日增重显著高于对照组(P<0.05),料重比显著低于对照组(P<0.05)。随着MCE添加量的增加,羔羊终末体重和平均日增重均呈线性升高(P<0.05),料重比呈线性降低(P<0.05)。
表3 饲粮中添加MCE对育肥湖羊公羔生长性能的影响

Table 3 Effects of diet supplemented with MCE on growth performance of fattening male Hu lambs

项目
Items
组别Groups SEM PP-value
对照
Control
300 MCE 600 MCE 900 MCE 处理
Treatment
线性
Linear
二次
Quadratic
初始体重IBW/kg 27.56 27.19 27.96 27.50 0.235 0.678 0.958 0.748
终末体重FBW/kg 39.99b 40.19b 43.69a 41.94ab 0.503 0.021 0.025 0.279
平均日采食量ADFI/(kg/d) 1.55 1.51 1.64 1.63 0.039 0.584 0.304 0.874
平均日增重ADG/(g/d) 207.1b 216.6b 262.1a 244.5ab 7.15 0.015 0.008 0.281
料重比F/G 7.56a 7.04a 6.25b 6.72ab 0.172 0.043 0.024 0.127

同行数据肩标无字母或相同小写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 small 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.

2.2 饲粮中添加MCE对育肥湖羊公羔瘤胃发酵参数的影响

表4可知,饲粮中添加MCE对瘤胃液pH、总挥发性脂肪酸(TVFA)、乙酸、丁酸、异丁酸、异戊酸浓度均无显著影响(P>0.05)。随着MCE添加量的增加,瘤胃液丙酸浓度呈二次曲线变化的趋势(P=0.076);600 MCE组瘤胃液丙酸浓度显著高于对照组和300 MCE组(P<0.05),与900 MCE组无显著差异(P>0.05)。随着MCE添加量的增加,瘤胃液戊酸浓度呈线性降低(P<0.05);600 MCE组瘤胃液戊酸浓度显著低于对照组(P<0.05)。600 MCE组和900 MCE组瘤胃液乙丙比显著低于对照组(P<0.05)。随着MCE添加量的增加,瘤胃液NH3-N浓度呈线性和二次曲线变化(P<0.05);600 MCE组瘤胃液NH3-N浓度显著低于对照组(P<0.05)。
表4 饲粮中添加MCE对育肥湖羊公羔瘤胃发酵参数的影响

Table 4 Effects of diet supplemented with MCE on rumen fermentation parameters of fattening male Hu lambs

项目
Items
组别Groups SEM PP-value
对照
Control
300 MCE 600 MCE 900 MCE 处理
Treatment
线性
Linear
二次
Quadratic
pH 5.64 5.68 5.77 5.78 0.044 0.661 0.236 0.829
总挥发性脂肪酸TVFA/(mmol/L) 96.89 98.65 104.89 101.49 1.552 0.296 0.156 0.410
乙酸Acetate/(mmol/L) 51.86 52.54 54.64 51.48 0.967 0.690 0.918 0.350
丙酸Propionate/(mmol/L) 25.85c 29.02bc 34.44a 31.18ab 1.033 0.015 0.011 0.076
丁酸Butyrate/(mmol/L) 15.23 13.27 13.00 15.73 0.674 0.393 0.835 0.096
戊酸Valerate/(mmol/L) 3.35a 3.04ab 2.22b 2.59ab 0.153 0.036 0.018 0.217
异丁酸Isobutyate/(mmol/L) 0.23 0.32 0.23 0.21 0.023 0.354 0.441 0.228
异戊酸Isovalerate/(mmol/L) 0.37 0.46 0.36 0.30 0.025 0.182 0.123 0.199
乙丙比A∶P 2.02a 1.82ab 1.61b 1.68b 0.056 0.033 0.011 0.167
氨态氮NH3-N/(mg/dL) 18.94a 16.47ab 13.41b 15.95ab 0.690 0.027 0.031 0.043

2.3 饲粮中添加MCE对育肥湖羊公羔瘤胃酶活性的影响

表5可知,饲粮中添加MCE对瘤胃液纤维二糖酶和木聚糖酶活性无显著影响(P>0.05)。随着MCE添加量的增加,瘤胃液蛋白酶活性呈先降低后升高的二次曲线变化(P<0.05);600 MCE组瘤胃液蛋白酶活性显著低于对照组(P<0.05)。300 MCE组、600 MCE组和900 MCE组瘤胃液α-淀粉酶活性均显著高于对照组(P<0.05)。600 MCE组瘤胃液羧甲基纤维素酶活性显著高于对照组和300 MCE组(P<0.05)。600 MCE组和900 MCE组瘤胃液果胶酶活性显著高于对照组(P<0.05)。
表5 饲粮中添加MCE对育肥湖羊公羔瘤胃酶活性的影响

Table 5 Effects of diet supplemented with MCE on rumen enzyme activities of fattening male Hu lambsU/mL

项目
Items
组别Groups SEM PP-value
对照
Control
300 MCE 600 MCE 900 MCE 处理
Treatment
线性
Linear
二次
Quadratic
纤维二糖酶Cellobiase 0.68 0.69 0.75 0.73 0.015 0.250 0.134 0.492
羧甲基纤维素酶
Carboxymethyl cellulase
0.14bc 0.13c 0.22a 0.19ab 0.012 0.010 0.009 0.638
木聚糖酶Xylanase 0.61 0.66 0.76 0.70 0.026 0.189 0.113 0.234
果胶酶Pectinase 1.88b 2.02ab 2.49a 2.56a 0.108 0.047 0.008 0.867
α-淀粉酶α-amylase 1.01b 1.36a 1.49a 1.41a 0.066 0.036 0.017 0.071
蛋白酶Protease 1.36a 1.23a 1.02b 1.21ab 0.041 0.017 0.066 0.016

2.4 饲粮中添加MCE对育肥湖羊公羔瘤胃微生物数量的影响

表6可知,饲粮中添加MCE对瘤胃溶纤维丁酸弧菌、白色瘤胃球菌和总菌数量均无显著影响(P>0.05)。600 MCE组瘤胃产甲烷菌数量显著低于对照组和300 MCE组(P<0.05)。600 MCE组和900 MCE组瘤胃嗜淀粉瘤胃杆菌和产琥珀酸丝状杆菌数量显著高于对照组(P<0.05),栖瘤胃普雷沃氏菌数量显著低于对照组(P<0.05)。600 MCE组瘤胃黄色瘤胃球菌数量显著高于对照组和300 MCE组(P<0.05)。
表6 饲粮中添加MCE对育肥湖羊公羔瘤胃微生物数量的影响

Table 6 Effects of diet supplemented with MCE on rumen microbial numbers of fattening male Hu lambs

项目
Items
组别Groups SEM PP-value
对照
Control
300 MCE 600 MCE 900 MCE 处理
Treatment
线性
Linear
二次
Quadratic
产甲烷菌
Methanogens/(×108拷贝数/mL)
1.21a 1.12a 0.81b 1.01ab 0.051 0.021 0.023 0.116
嗜淀粉瘤胃杆菌
Ruminobacter amylophilus/
(×108拷贝数/mL)
2.94b 3.64ab 4.06a 3.90a 0.152 0.030 0.010 0.109
栖瘤胃普雷沃氏菌
Prevotella ruminicola/
(×1010拷贝数/mL)
3.87a 2.91ab 1.85b 2.38b 0.265 0.030 0.012 0.111
溶纤维丁酸弧菌
Butyrivibrio fibrisolvens/
(×1010拷贝数/mL)
1.35 2.17 3.04 2.17 0.290 0.249 0.204 0.151
产琥珀酸丝状杆菌
Fibrobacter succinogenes/
(×1010拷贝数/mL)
2.62b 2.77ab 3.11a 3.00a 0.070 0.041 0.013 0.296
白色瘤胃球菌
Ruminococcus albus/
(×108拷贝数/mL)
0.87 0.93 1.12 1.02 0.049 0.332 0.159 0.441
黄色瘤胃球菌
Ruminococcus flavefaciens/
(×109拷贝数/mL)
1.71c 1.88bc 2.19a 2.05ab 0.060 0.015 0.007 0.121
总菌
Total bacteria/(×1011拷贝数/mL)
2.67 2.93 3.08 3.00 0.090 0.429 0.173 0.367

3 讨论

3.1 饲粮中添加MCE对育肥湖羊公羔生长性能的影响

MCE具有抗炎、抗菌、抗氧化及增强免疫力等多种生物学功能。其主要活性成分之一血根碱,作为一种高效的饲用抗生素替代品,长期应用于畜禽饲粮中可有效提高畜禽生长性能和免疫力[21-22]。Jiao等[23]研究表明,饲粮中添加500 mg/d MCE可通过改善瘤胃免疫功能和抗氧化能力,显著提高羔羊生长性能;Estrada-Angulo等[24]也发现,在母羊饲粮中添加MCE能有效缓解热应激,进而改善其生长性能。上述研究结果与本试验结果一致。本试验中,随着MCE添加量的增加,羔羊的终末体重与平均日增重均呈线性升高,料重比呈线性降低,其中600 MCE组料重比最低。这表明在育肥湖羊公羔饲粮中添加MCE能够明显提高饲料利用率,在一定程度上降低养殖成本。有研究报道,MCE中的血根碱能够直接与细胞内烟酰胺腺嘌呤二核苷酸(NADPH)发生反应,使相关酶失去活性,从而增强机体抗氧化能力,减少动物应激反应[25]。动物摄入MCE还可增强肠道屏障功能[26],减轻肠道炎症反应[10],改善肠道形态[27],进而促进营养物质的消化与利用。因此,本试验中MCE对羔羊生长性能的改善作用,可能与其促进小肠营养物质吸收有关,后续将进一步探究MCE对育肥湖羊公羔肠道发育的具体影响机制。然而,Toprak[28]和Küçükoflaz等[29]研究发现,饲粮中添加MCE对西门塔尔犊牛和荷斯坦犊牛的采食量和饲料转换率无显著影响;Chen等[30]的研究也发现,对早期断奶山羊补饲300 mg/d MCE,其干物质采食量和日增重均未发生显著变化。综合来看,MCE对动物生长性能的影响存在差异,这可能与MCE的补充剂量、试验动物品种及所处生产阶段等因素有关。

3.2 饲粮中添加MCE对育肥湖羊公羔瘤胃发酵参数的影响

反刍动物瘤胃中的VFA由微生物分解碳水化合物产生,其中乙酸、丙酸和丁酸等短链脂肪酸经瘤胃壁吸收后,可满足动物机体70%~80%的能量需求[31]。瘤胃功能及其内环境稳定性通常可通过瘤胃pH、VFA和NH3-N浓度等发酵参数进行评估。本试验中,饲粮中添加MEC对羔羊瘤胃液pH及TVFA浓度均未产生显著影响。虽然瘤胃液中TVFA、乙酸和丁酸浓度变化不显著,但在MCE添加量为600 mg/kg时,瘤胃液丙酸浓度显著升高,同时乙丙比显著下降,表明适量添加MCE可使羔羊瘤胃发酵模式转向丙酸型发酵。丙酸型发酵有利于淀粉的分解与利用,提高能量利用效率,改善动物生长性能[32]。这一结果也与本试验中瘤胃嗜淀粉瘤胃杆菌数量增加和α-淀粉酶活性升高的现象一致。湖羊体外瘤胃发酵试验表明,当MCE浓度为5 mg/dL、发酵24 h时,发酵液中丙酸浓度显著提高[33]。然而,Jiao等[23]和Ling等[34]研究发现,饲粮中添加MCE对断奶羔羊及哺乳期奶山羊瘤胃液丙酸浓度无显著影响,但会显著提高丁酸浓度,与本试验结果存在差异。这可能是由于试验对象及各试验所用饲粮组成不同所致。同时,本试验中饲粮添加MCE降低了羔羊瘤胃液戊酸浓度,与Aguilar-Hernández等[35]的研究结果一致。戊酸浓度可反映瘤胃内多肽及氨基酸的降解程度[36],而本试验中瘤胃液戊酸浓度随MCE添加量增加呈线性降低,提示MCE可能减少了饲粮蛋白质在瘤胃内的降解,使过瘤胃蛋白含量增加;这一推测也与本试验中添加MCE组瘤胃液NH3-N浓度降低的结果相吻合。从机制上看,栖瘤胃普雷沃氏菌是瘤胃内主要的蛋白质降解菌,本试验中该菌数量减少直接导致瘤胃液蛋白酶活性下降,进而降低饲粮蛋白质的瘤胃降解效率;而瘤胃液NH3-N主要来源于蛋白质的瘤胃降解过程[37],因此蛋白质降解减少使得NH3-N生成量降低。已有报道指出,从博落回中提取的血根碱可通过减少饲粮蛋白质在瘤胃内的降解,增加非氨态氮向小肠的流量[35];Zhang等[38]也发现,饲粮中添加血根碱可降低犊牛瘤胃液NH3-N浓度。综上所述,MCE可能通过抑制瘤胃微生物对饲粮蛋白质的降解,提高过瘤胃蛋白比例,增加小肠中可代谢蛋白的供应量,进而促进羔羊生长。

3.3 饲粮中添加MCE对育肥湖羊公羔瘤胃酶活性和微生物数量的影响

瘤胃酶活性与瘤胃微生物群落结构存在紧密关联。栖瘤胃普雷沃氏菌作为瘤胃内的优势菌群,在蛋白质代谢过程中发挥关键作用,可分泌蛋白质降解酶以推动饲粮蛋白质的分解;嗜淀粉瘤胃杆菌则是瘤胃内重要的淀粉降解微生物,能通过分泌淀粉酶实现对碳水化合物的高效分解与利用。此外,由白色瘤胃球菌、黄色瘤胃球菌、产琥珀酸丝状杆菌及溶纤维丁酸弧菌组成的微生物类群,在瘤胃生态系统中展现出突出的纤维素降解活性,可通过分泌纤维素酶参与饲粮纤维的降解过程。本试验结果表明,饲粮中添加MCE显著提高了瘤胃黄色瘤胃球菌与产琥珀酸丝状杆菌数量,同时提升了瘤胃液中果胶酶和羧甲基纤维素酶活性。这一变化有助于增强羔羊对饲粮中纤维类物质的降解利用能力,为瘤胃微生物生长繁殖及羔羊机体发育提供更充足的能量[39]。Ling等[34]研究发现,饲粮中添加400 mg/kg MCE可通过改善瘤胃微生物区系,提高奶山羊对纤维的表观消化率,与本试验结果相符。与此同时,本试验中栖瘤胃普雷沃氏菌数量显著降低,导致瘤胃液蛋白酶活性下降,这可能与MCE中血根碱和白屈菜红碱的作用有关。已有研究表明,血根碱可通过干扰细菌分裂过程中收缩环(Z环)内丝状温度敏感蛋白Z(FtsZ)的组装动态来抑制细菌分裂[40];白屈菜红碱则能破坏细菌细胞膜上的通道结构,导致细胞内蛋白质外渗[41],二者共同作用抑制细菌蛋白质合成。甲烷是一种温室气体,反刍动物在瘤胃发酵过程中会产生大量甲烷,导致2%~15%的饲粮能量以甲烷形式损失,因此减少甲烷产生对提高饲粮能量利用效率有重要意义。本试验中,饲粮中添加MCE会使瘤胃液丙酸浓度显著升高,瘤胃微生物合成丙酸过程中会消耗氢,而产甲烷菌需利用氢生成甲烷,因此氢的消耗间接抑制了甲烷的生成[42]。Jia等[43]研究发现,饲粮中添加MCE可降低瘤胃中7个古菌属和9个古菌物种的相对丰度,从而减少甲烷产量,这与本试验中产甲烷菌数量显著降低的结果相吻合。综上可知,MCE对减少育肥湖羊公羔瘤胃甲烷生成及提升能量利用效率具有积极作用。

4 结论

饲粮中添加MCE可以促进育肥湖羊公羔瘤胃发酵,降低瘤胃液NH3-N浓度,改变瘤胃微生物数量及酶活性,从而提高饲料利用效率,促进羔羊生长。在本试验条件下,MCE的适宜添加量为600 mg/kg。
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