Chinese Journal of Animal Nutrition >

2026 , Vol. 38 >Issue 6: 4339 - 4351

DOI: https://doi.org/10.12418/CJAN2026.348

RESEARCH PAPER

Effects of Combined Supplementation of Clostridium butyricum Culture and Aspergillus oryzae Culture in Late Gestation Diet on Growth Performance and Serum Biochemical Indicators of Ewes and Growth and Development of Their Offspring Lambs

  • LIU Yanfeng , 1 ,
  • ZHAO Linbo 1 ,
  • LI Jiguo 2 ,
  • Nazakaitiayi Taxibolati 1 ,
  • DONG Zehua 1 ,
  • WANG Lele 1 ,
  • WANG Wenqi , 1, **
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  • 1 Xinjiang Key Laboratory of Herbivorous Livestock Feed Biotechnology, Feed Research Institute, Xinjiang Uygur Autonomous Region Academy of Animal Science, Urumqi 830011, China
  • 2 Management Committee of Modern Agricultural Industrial Park in Fuhai County, Fuhai 836400, China
** professor, E-mail:

*Contributed equally

Received date: 2025-11-14

  Online published: 2026-06-13

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Abstract

This study aimed to investigate the effects of combined supplementation of Clostridium butyricum culture (CBC) and Aspergillus oryzae culture (AOC) in late gestation diet on growth performance and serum biochemical indicators of ewes, as well as on the growth and development of their offspring lambs. Forty-five healthy Altay ewes with singleton pregnancies at day 121 of gestation, weighing (71.64±2.83) kg, were randomly divided into 3 groups with 3 replicates per group and 5 ewes per replicate. Ewes in the three groups were fed a basal diet (CK group), or experimental diets supplemented with 0.21% CBC (CBC group) and 0.21% CBC+0.50% AOC (CBC+AOC group), respectively, for 30 days (from day 121 to 150 of gestation). The results showed as follows: 1) compared with the CK group, the body weight of ewes in the CBC+AOC group on day 150 of gestation and day 30 postpartum was significantly increased (P<0.05). From day 121 to 150 of gestation, the average daily gain (ADG) in the CBC+AOC group and CBC groups was extremely significantly increased (P<0.01). 2) Compared with the CK group, the serum contents of total protein (TP), β-hydroxybutyric acid (β-HB), glucose (GLU) and triglyceride (TG) in the CBC+AOC group on day 150 of gestation were extremely significantly increased (P<0.01), while the serum urea content was significantly decreased (P<0.05); moreover, the serum β-HB and GLU contents in the CBC+AOC group on day 30 postpartum were extremely significantly increased (P<0.01). The serum GLU content in the CBC group on day 150 of gestation and day 30 postpartum was extremely significantly increased (P<0.01). 3) Compared with the CK group, the total antioxidant capacity (T-AOC), and the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) in serum of ewes in the CBC+AOC group on day 150 of gestation were significantly increased (P<0.05), the serum malondialdehyde (MDA) content was extremely significantly decreased (P<0.01), and the serum diamine oxidase (DAO) activity and D-lactic acid (D-LA) content were significantly decreased (P<0.05); on day 30 postpartum, serum CAT and SOD activities of ewes in the CBC+AOC group were significantly increased (P<0.05), serum GSH-Px activity was extremely significantly increased (P<0.01), and serum D-LA content was significantly decreased (P<0.05); in the CBC group, serum SOD activity on day 150 of gestation was significantly increased (P<0.05), serum MDA content was extremely significantly decreased (P<0.01), and serum GSH-Px activity on day 30 postpartum was extremely significantly increased (P<0.01). 4) The body oblique length and withers height of lambs in the CBC+AOC group on days 1 and 30 after birth were extremely significantly higher than those in the CK group and CBC group (P<0.01), and the withers height on day 60 after birth was significantly higher than that in the CK group and CBC group (P<0.05). The chest girth of lambs in the CBC+AOC group on days 1, 30 and 60 after birth was significantly increased (P<0.05). 5) Compared with the CK group, the serum TP content of lambs in the CBC+AOC group on day 30 after birth was extremely significantly increased (P<0.01), while the serum contents of urea and total cholesterol (TC) were significantly decreased (P<0.05), and the serum GLU content was significantly increased (P<0.05); on day 60 after birth, the serum urea content of lambs in the CBC+AOC group was extremely significantly decreased (P<0.01), and the serum TP and albumin (ALB) contents were significantly increased (P<0.05); the serum T-AOC and the activities of GSH-Px and CAT of lambs in the CBC+AOC group on day 30 after birth were significantly increased (P<0.05), and the serum T-AOC and the activities of GSH-Px, SOD and CAT of lambs on day 60 after birth were extremely significantly increased (P<0.01); the serum MDA content of lambs in the CBC+AOC group and CBC group on day 30 after birth was extremely significantly decreased (P<0.01). In conclusion, combined supplementation of CBC and AOC in late gestation diet can effectively increase the ADG of ewes, enhance their antioxidant capacity, immune function and intestinal barrier function, promote protein metabolism, improve energy metabolism, and benefit the early growth and development of offspring lambs. Moreover, the combined supplementation of CBC and AOC shows better effects than supplementation with CBC alone.

Key words： Clostridium butyricum culture; Aspergillus oryzae culture; late gestation; ewes; lambs; growth performance; serum biochemical indicators

Cite this article

LIU Yanfeng , ZHAO Linbo , LI Jiguo , Nazakaitiayi Taxibolati , DONG Zehua , WANG Lele , WANG Wenqi . Effects of Combined Supplementation of Clostridium butyricum Culture and Aspergillus oryzae Culture in Late Gestation Diet on Growth Performance and Serum Biochemical Indicators of Ewes and Growth and Development of Their Offspring Lambs[J]. Chinese Journal of Animal Nutrition, 2026 , 38(6) : 4339 -4351 . DOI: 10.12418/CJAN2026.348

母羊妊娠后期的营养调控对保障母羊健康与羔羊生长发育具有关键作用,此阶段羔羊在母体内快速发育,而母羊因生理变化导致干物质采食量和营养摄入下降,进而影响羔羊的初生重与成活率[1-2]。因此,优化母羊妊娠后期营养调控策略尤为重要。微生物制剂是由单一或多种活微生物组成的培养物,作为一种绿色、安全的饲料添加剂,其可通过增强动物抗氧化能力、改善免疫功能和调节胃肠道微生态平衡等途径促进动物健康[3-4]。其中,丁酸梭菌(Clostridium butyricum)和米曲霉(Aspergillus oryzae)作为具有改善生长性能的益生菌,在肉牛、猪和鸡的饲养中已表现出良好效果[5]。丁酸梭菌为革兰氏阳性厌氧杆菌,可发酵多种碳水化合物如葡萄糖、蔗糖、果糖及淀粉,主要代谢产物包括丁酸、乙酸和乳酸等短链脂肪酸[6]。丁酸梭菌培养物(Clostridium butyricum culture,CBC)是丁酸梭菌发酵产物的干燥混合物,包含灭活菌体、发酵底物以及丁酸等多种代谢产物[7],因此其潜在功效可能超出单一丁酸或菌体的效果。作为一种多功能发酵制品,CBC在反刍动物中的潜在应用价值尚未得到充分探索。米曲霉属于好气性真菌,其菌丝为多细胞结构,可分泌多种酶类,包括蛋白酶、纤维素酶、淀粉酶和果胶酶等[8]。米曲霉培养物(Aspergillus oryzae culture,AOC)是以米曲霉为菌种通过特定发酵工艺制备的真菌制剂,富含多种消化酶、氨基酸、矿物质、维生素及未知免疫活性因子[9]。Guo等[10]在成年湖羊饲粮中添加不同水平(10、20和40 g/d)的AOC,发现20和40 g/d的AOC可提高饲粮干物质消化率,并增加瘤胃总挥发性脂肪酸(TVFA)浓度。然而,关于在妊娠后期母羊饲粮中联合使用CBC与AOC能否协同改善母羊生长性能、促进子代羔羊生长发育,目前仍缺乏系统研究。为此,本研究通过在母羊妊娠后期饲粮中单独添加CBC或联合添加CBC与AOC,系统评价其对母羊生长性能、血清生化指标及其子代羔羊生长发育和血清生化指标的影响,旨在为这2种微生物制剂在妊娠母羊饲粮中的合理应用提供科学依据。

1 材料与方法

1.1 伦理声明

本研究所有动物程序均经新疆维吾尔自治区畜牧科学院饲料研究所审查委员会批准,批准编号为No.720250304。

1.2 试验时间和材料

饲养试验于2025年2月15日至2025年5月15日在新疆福海种羊场阿勒泰羊繁育中心进行,试验羊和饲料原料均由该中心提供。试验所用CBC和AOC由合作企业提供,CBC和AOC的活性成分及营养成分见表1。
表1 CBC和AOC的活性成分及营养成分(风干基础)

Table 1 Active components and nutritional components of CBC and AOC (air-dry basis)

项目 Items 含量 Content
丁酸梭菌培养物 CBC
有效孢子 Viable spores/(CFU/g) 1.6×109
甘露聚糖 Mannan/% 0.30
粗灰分 Ash/% 17.26
干物质 DM/% 94.00
粗蛋白质 CP/% 8.03
酸溶蛋白 Acid-soluble protein/% 4.11
米曲霉培养物 AOC
有效孢子 Viable spores/(CFU/g) ≥8.5×108
中性蛋白酶 Neutral protease/(U/g) 3 540
糖化酶 Amylase/(U/g) 1 560
木聚糖酶 Xylanase/(U/g) 597
纤维素酶 Cellulase/(U/g) 401

1.3 试验设计

选取45只体重[(71.64±2.83) kg]相近、怀有单羔的妊娠121 d的健康阿勒泰母羊,随机分为3组,即CK组、CBC组、CBC+AOC组,每组3个重复,每个重复5只母羊。CK组饲喂基础饲粮,CBC组饲喂添加0.21% CBC的试验饲粮,CBC+AOC组饲喂同时添加0.21% CBC和0.50% AOC(CBC、AOC的添加量分别参考崔锦等[11]和张对红[12]并结合合作企业的产品推荐量确定)的试验饲粮,持续饲喂30 d(妊娠第121~150天),之后所有母羊统一饲喂不含有CBC和AOC的常规饲粮。

1.4 饲粮配制与饲养管理

参照《肉羊营养需要量》(NY/T 816—2021)中母羊营养需要量并结合实际生产配制母羊妊娠后期基础饲粮,精粗比为3∶7,在基础饲粮配方基础上分别添加0.21% CBC和0.21%CBC+0.50% AOC并适当调整其他原料的用量,配制与基础饲粮等氮等能的试验饲粮。各组饲粮组成及营养水平见表2。试验期间,母羊每日分别于09:00和19:00采用全混合日粮(TMR)形式进行饲喂,保证剩料量不低于总饲喂量的5%,并保证自由饮水,定期清理圈舍。
表2 各组饲粮组成及营养水平(风干基础)

Table 2 Composition and nutrient levels of diets for each group (air-dry basis) %

项目
Items		 组别 Groups
CK CBC CBC+AOC
原料 Ingredients
玉米 Corn 18.30 18.26 18.17
棉籽粕 Cottonseed meal 6.30 6.29 6.25
小麦麸 Wheat bran 3.30 3.29 3.28
豆粕 Soybean meal 1.20 1.20 1.19
苜蓿干草 Alfalfa hay 41.00 40.91 40.71
小麦秸秆 Wheat straw 28.70 28.64 28.49
磷酸氢钙 CaHPO4 0.40 0.40 0.40
食盐 NaCl 0.40 0.40 0.40
预混料 Premix1) 0.40 0.40 0.40
丁酸梭菌培养物 CBC 0.21 0.21
米曲霉培养物 AOC 0.50
合计 Total 100.00 100.00 100.00
营养水平 Nutrient levels2)
代谢能 ME/(MJ/kg) 8.17 8.15 8.11
干物质 DM 90.54 90.36 89.89
粗蛋白质 CP 13.07 13.05 12.98
中性洗涤纤维 NDF 52.11 52.00 51.74
酸性洗涤纤维 ADF 37.53 37.46 37.27
钙 Ca 1.03 1.03 1.03
磷 P 0.56 0.56 0.56

1)预混料为每千克饲粮提供The premix provided the following per kg of diets:VA 10 000 IU,VD3 6 000 IU,VE 55 IU,Fe 50 mg,Mn 15 mg,Zn 15 mg,Cu 4.5 mg,I 0.22 mg,Se 0.09 mg。
2)代谢能参考NY/T 816—2021计算,其余均为实测值。ME was calculated according to NY/T 816—2021, while the others were measured values.

1.5 指标测定

1.5.1 饲粮营养成分的测定

分别参照GB/T 6534—2014、GB/T 6432—2018、GB/T 6436—2018、GB/T 6437—2018、GB/T 20806—2022和NY/T 1459—2022测定饲粮中干物质(DM)、粗蛋白质(CP)、钙(Ca)、磷(P)、中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)的含量。

1.5.2 母羊生长性能的测定

试验期间每天记录给料量和晨饲前剩料量,以计算平均干物质采食量(ADMI);分别于母羊妊娠第121、150天和产后第30、60天清晨空腹称量体重,计算母羊体增重(BWG)、平均日增重(ADG)、产后失重、失重率和料重比(F/G)。
ADMI(g/d)=(给料量×饲粮DM含量-剩料量×剩料DM含量)/饲养天数;
BWG(kg)=终末体重-初始体重;
ADG(g/d)=(终末体重-初始体重)/饲养天数;
产后失重(kg)=产后第30天体重-妊娠第150天体重;
失重率(%)=(产后失重/母羊妊娠第150天体重)×100;
F/G=ADMI/ADG。

1.5.3 母羊和羔羊血清生化指标的测定

于母羊妊娠第150天、产后第30天及羔羊出生后第30天、第60天晨饲前,每个重复随机选择2只母羊或羔羊(每组6只),使用普通真空采血管静脉采血5 mL后,快速使用离心机1 788.56×g离心10 min,用一次性胶头滴管将上层血清吸出,-20 ℃保存待测。
使用迈瑞BS-420全自动生化分析仪测定血清蛋白质和能量代谢相关指标,包括总蛋白(TP)、白蛋白(ALB)、尿素(UREA)、β-羟丁酸(β-HB)、葡萄糖(GLU)、总胆固醇(TC)和甘油三酯(TG)含量,上述指标测定所用试剂盒均购于南京建成生物工程研究所。
使用华卫德朗DR-200BS酶标分析仪,采用比色法测定血清抗氧化相关指标[总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)与谷胱甘肽过氧化物酶(GSH-Px)活性、丙二醛(MDA)含量],采用酶联免疫吸附测定法测定免疫[肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-4(IL-4)、白细胞介素-6(IL-6)]和肠道健康相关指标[二胺氧化酶(DAO)活性和D-乳酸(D-LA)含量]。上述指标检测所用试剂盒均为北京华英生物技术研究所生产。

1.5.4 羔羊生长性能和体尺指标的测定

于羔羊出生后第1、30和60天晨饲前空腹称量体重,计算羔羊的ADG;同时,使用卷尺测定羔羊体斜长、体高、胸围和管围。

1.6 数据统计与分析

试验数据采用SPSS 26.0软件进行单因素方差分析(one-way ANOVA),并采用Duncan氏法进行组间多重比较。P<0.01表示差异极显著,P<0.05表示差异显著,P>0.05表示差异不显著。试验数据以平均值和均值标准误(SEM)形式给出。

2 结果

2.1 妊娠后期饲粮中联合添加CBC和AOC对母羊生长性能的影响

由表3可知,与CK组相比,CBC+AOC组母羊妊娠第150天和产后第30天的体重均显著升高(P<0.05)。妊娠第121~150天的体增重和ADG极显著升高(P<0.01),妊娠第121~150天的DMI、F/G极显著降低(P<0.01);CBC组妊娠第121~150天的ADG和体增重均极显著提升(P<0.01),妊娠第121~150天的F/G极显著降低(P<0.01)。此外,CBC+AOC组母羊妊娠第121~150天的体增重和ADG极显著高于CBC组(P<0.01),同时DMI、F/G极显著低于CBC组(P<0.01)。
表3 妊娠后期饲粮中联合添加CBC和AOC对母羊生长性能的影响

Table 3 Effects of combined supplementation of CBC and AOC in late gestation diet on growth performance of ewes

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC







体重
BW/kg		 妊娠第121天
Day 121 of gestation		 71.87 71.38 71.67 0.666 0.962
妊娠第150天
Day 150 of gestation		 75.00b 76.65ab 78.87a 0.750 0.049
产后第30天
Day 30 postpartum		 63.65b 66.03ab 69.05a 0.804 0.012
产后第60天
Day 60 postpartum		 68.23 70.93 74.18 1.038 0.054



体增重
BWG/kg		 妊娠第121~150天
Days 121 to 150 of gestation		 3.13Bc 5.10Bb 7.20Aa 0.475 <0.001
产后第30~60天
Days 30 to 60 postpartum		 4.58 4.90 5.13 0.455 0.568



平均日增重
ADG/(g/d)		 妊娠第121~150天
Days 121 to 150 of gestation		 158.07C 242.86B 342.86A 21.869 <0.001
产后第30~60天
Days 30 to 60 postpartum		 152.72 163.33 171.11 15.158 0.432
产后失重
Postpartum weight
loss/kg		 妊娠第150天至产后第30天
Day 150 of gestation to day 30 postpartum		 -11.35 -10.62 -9.82 0.580 0.488
失重率
Weight loss rate/%		 妊娠第150天至产后第30天
Day 150 of gestation to day 30 postpartum		 15.17 13.70 12.45 0.740 0.343
平均干物质采食量
ADMI/(kg/d)		 妊娠第121~150天
Days 121 to 150 of gestation		 1.86Aa 1.83Aa 1.74Bb 0.100 <0.001
料重比
F/G		 妊娠第121~150天
Days 121 to 150 of gestation		 11.75A 7.55B 5.08C 0.281 <0.001

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),不同大写字母表示差异极显著(P<0.01)。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), and with different capital letter superscripts mean extremely significant difference (P<0.01). The same as below.

2.2 妊娠后期饲粮中联合添加CBC和AOC对母羊血清生化指标的影响

2.2.1 妊娠后期饲粮中联合添加CBC和AOC对母羊血清蛋白质和能量代谢相关指标的影响

由表4可知,与CK组相比,CBC+AOC组母羊妊娠第150天血清TP含量极显著提升(P<0.01),同时血清UREA含量显著降低(P<0.05);CBC+AOC组和CBC组母羊妊娠第150天和产后第30天血清β-HB含量极显著降低(P<0.01),同时血清GLU含量极显著升高(P<0.01);CBC+AOC组和CBC组母羊妊娠第150天血清TG含量极显著升高(P<0.01)。此外,CBC+AOC组母羊妊娠第150天血清TP含量显著高于CBC组(P<0.05)。
表4 妊娠后期饲粮中联合添加CBC和AOC对母羊血清蛋白质和能量代谢相关指标的影响

Table 4 Effects of combined supplementation of CBC and AOC in late gestation diet on serum protein and energy metabolism related indicators of ewes

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC
蛋白质代谢相关指标 Protein metabolism related indicators

总蛋白
TP/(g/L)		 妊娠第150天 Day 150 of gestation 64.14Bb 66.89ABb 69.56Aa 0.842 0.005
产后第30天 Day 30 postpartum 63.34 65.35 67.11 1.013 0.346

白蛋白
ALB/(g/L)		 妊娠第150天 Day 150 of gestation 26.27 28.25 29.31 0.871 0.325
产后第30天 Day 30 postpartum 24.22 26.05 28.83 0.967 0.145

尿素
UREA/(mmol/L)		 妊娠第150天 Day 150 of gestation 6.42a 5.57ab 4.18b 0.421 0.047
产后第30天 Day 30 postpartum 5.61 4.52 3.37 0.465 0.143
能量代谢相关指标 Energy metabolism related indicators

β-羟丁酸
β-HB/(mmol/L)		 妊娠第150天 Day 150 of gestation 0.61Aa 0.50Bb 0.46Bb 0.023 0.002
产后第30天 Day 30 postpartum 0.67Aa 0.53Bb 0.46Bb 0.031 0.004

葡萄糖
GLU/(mmol/L)		 妊娠第150天 Day 150 of gestation 3.03Bb 3.72Aa 3.97Aa 0.155 0.010
产后第30天 Day 30 postpartum 2.80Bb 3.68Aa 3.92Aa 0.171 0.003

甘油三酯
TG/(mmol/L)		 妊娠第150天 Day 150 of gestation 0.74Bb 0.83Aa 0.85Aa 0.017 0.003
产后第30天 Day 30 postpartum 0.67 0.72 0.75 0.026 0.321

总胆固醇
TC/(mmol/L)		 妊娠第150天 Day 150 of gestation 1.84 1.79 1.75 0.024 0.290
产后第30天 Day 30 postpartum 1.88 1.66 1.57 0.064 0.128

2.2.2 妊娠后期饲粮中联合添加CBC和AOC对母羊血清抗氧化、免疫和肠道健康相关指标的影响

由表5可知,与CK组相比,CBC+AOC组母羊妊娠第150天血清T-AOC、GSH-Px和CAT活性均显著提高(P<0.05);CBC+AOC组和CBC组母羊妊娠第150天血清SOD活性均显著提升(P<0.05),且CBC+AOC组在产后第30天仍保持显著提升(P<0.05);CBC+AOC组和CBC组母羊产后第30天血清GSH-Px活性均极显著提升(P<0.01),妊娠第150天血清MDA含量均极显著降低(P<0.01);CBC+AOC组产后第30天血清CAT活性显著提高(P<0.05)。此外,CBC+AOC组母羊产后第30天血清GSH-Px、CAT活性显著高于CBC组(P<0.05)。与CK组相比,CBC+AOC组母羊妊娠第150天血清TNF-α含量显著降低(P<0.05),且在产后第30天仍保持极显著降低(P<0.01);CBC+AOC组母羊妊娠第150天血清IL-1β含量极显著降低(P<0.01),且在产后第30天仍保持显著降低(P<0.05);CBC+AOC组母羊产后第30天血清IL-6含量极显著降低(P<0.01)。此外,CBC+AOC组母羊妊娠第150天血清IL-1β含量极显著低于CBC组(P<0.01),在产后第30天仍保持显著低于CBC组(P<0.05),同时产后第30天血清IL-6含量极显著低于CBC组(P<0.01)。与CK组相比,CBC+AOC组妊娠第150天血清DAO活性以及妊娠第150天和产后第30天血清D-LA含量均显著降低(P<0.05)。
表5 妊娠后期饲粮中联合添加CBC和AOC对母羊血清抗氧化、免疫和肠道健康相关指标的影响

Table 5 Effects of combined supplementation of CBC and AOC in late gestation diet on serum antioxidant, immune and intestinal health related indicators of ewes

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC
抗氧化相关指标 Antioxidant related indicators

总抗氧化能力
T-AOC/(U/mL)		 妊娠第150天 Day 150 of gestation 8.53b 9.40ab 10.10a 0.284 0.036
产后第30天 Day 30 postpartum 8.43 8.74 9.18 0.182 0.257

超氧化物歧化酶
SOD/(U/mL)		 妊娠第150天 Day 150 of gestation 81.39b 85.33a 86.05a 0.868 0.027
产后第30天 Day 30 postpartum 72.63b 74.81ab 78.90a 1.048 0.032

谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 妊娠第150天 Day 150 of gestation 157.07b 166.86ab 174.64a 3.020 0.020
产后第30天 Day 30 postpartum 111.04Bc 127.14Ab 140.94Aa 4.381 0.004

丙二醛
MDA/(nmol/mL)		 妊娠第150天 Day 150 of gestation 5.05Aa 3.93Bb 3.15Bc 0.280 0.002
产后第30天 Day 30 postpartum 4.50 4.36 3.69 0.181 0.145

过氧化氢酶
CAT/(U/mL)		 妊娠第150天 Day 150 of gestation 41.62b 45.55ab 46.86a 0.979 0.039
产后第30天 Day 30 postpartum 32.01b 32.55b 39.05a 1.400 0.045
免疫相关指标 Immune related indicators/(pg/mL)

肿瘤坏死因子-α
TNF-α		 妊娠第150天 Day 150 of gestation 58.80a 55.72ab 51.39b 1.224 0.011
产后第30天 Day 30 postpartum 60.06A 54.00B 47.95C 1.762 0.004

白细胞介素-1β
IL-1β		 妊娠第150天 Day 150 of gestation 22.79Aa 21.36Aa 17.61Bb 0.750 0.006
产后第30天 Day 30 postpartum 26.12a 24.44a 20.47b 0.922 0.016

白细胞介素-4
IL-4		 妊娠第150天 Day 150 of gestation 5.31 5.24 6.23 0.269 0.259
产后第30天 Day 30 postpartum 4.52 4.77 5.85 0.261 0.072

白细胞介素-6
IL-6		 妊娠第150天 Day 150 of gestation 126.19 111.66 111.04 3.321 0.191
产后第30天 Day 30 postpartum 130.07Aa 124.44Aa 118.29Bb 1.748 0.005
肠道健康相关指标 Intestinal health related indicators

二胺氧化酶
DAO/(U/L)		 妊娠第150天 Day 150 of gestation 2.98a 2.71ab 2.36b 0.113 0.033
产后第30天 Day 30 postpartum 2.71 2.46 2.35 0.067 0.129

D-乳酸
D-LA/(mg/dL)		 妊娠第150天 Day 150 of gestation 2.98a 2.71ab 2.35b 0.113 0.033
产后第30天 Day 30 postpartum 3.18a 2.91ab 2.40b 0.131 0.029

2.3 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊生长发育的影响

2.3.1 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊生长性能的影响

由表6可知,与CK组相比,CBC+AOC组羔羊出生后第1天、第30天和第60天体重极显著增加(P<0.01),出生后第1~30天ADG极显著升高(P<0.01)。
表6 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊生长性能的影响

Table 6 Effects of combined supplementation of CBC and AOC in late gestation diet of ewes on growth performance of lambs

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC


体重
BW/kg		 出生后第1天 Day 1 after birth 5.68Bb 5.92Bb 6.48Aa 0.117 0.005
出生后第30天 Day 30 after birth 12.99Bb 13.51Bb 14.89Aa 0.234 <0.001
出生后第60天 Day 60 after birth 20.92Bb 21.50Bb 24.16Aa 0.446 0.004


平均日增重
ADG/(g/d)		 出生后第1~30天
Days 1 to 30 after birth		 243.67Bb 253.22Bb 280.11Aa 5.328 0.006
出生后第31~60天
Days 31 to 60 after birth		 264.39 266.39 309.00 9.835 0.107

2.3.2 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊体尺指标的影响

由表7可知,CBC+AOC组羔羊出生后第1和30天的体斜长和体高极显著高于CK组和CBC组(P<0.01),且出生后第60天体高显著高于CK组和CBC组(P<0.05);CBC+AOC组羔羊出生后第1、30和60天胸围均较CK组显著提升(P<0.05)。
表7 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊体尺指标的影响

Table 7 Effects of combined supplementation of CBC and AOC in late gestation diet of ewes on body size indicators of lambscm

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC


体斜长
Body oblique length		 出生后第1天 Day 1 after birth 50.50Bb 52.67Bb 55.50Aa 0.656 0.002
出生后第30天 Day 30 after birth 62.83Bb 63.83Bb 66.67Aa 0.525 0.002
出生后第60天 Day 60 after birth 87.50 88.67 92.00 0.957 0.136


体高
Withers height		 出生后第1天 Day 1 after birth 39.33C 44.33B 46.33A 0.788 <0.001
出生后第30天 Day 30 after birth 49.50Bb 51.17Bb 54.50Aa 0.587 <0.001
出生后第60天 Day 60 after birth 59.00b 60.50b 63.00a 0.595 0.011


胸围
Chest girth		 出生后第1天 Day 1 after birth 44.00b 46.67ab 48.00a 0.667 0.034
出生后第30天 Day 30 after birth 58.83b 59.67b 62.33a 0.553 0.015
出生后第60天 Day 60 after birth 70.00b 72.50ab 77.83a 0.991 0.022


管围
Circumference of
cannon bone		 出生后第1天 Day 1 after birth 6.67 6.92 7.33 0.124 0.079
出生后第30天 Day 30 after birth 7.67 7.75 7.92 0.092 0.560
出生后第60天 Day 60 after birth 8.33 8.42 8.75 0.090 0.136

2.4 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊血清生化指标的影响

2.4.1 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊血清蛋白质和能量代谢相关指标的影响

由表8可知,与CK组相比,CBC+AOC组羔羊出生后第30天血清TP含量极显著提升(P<0.01),出生后第60天血清TP和ALB含量均显著提升(P<0.05)。与CK组相比,CBC+AOC组羔羊出生第30天血清UREA和TC含量显著降低(P<0.05),出生第30天血清GLU含量显著增加(P<0.05),出生第60天血清UREA含量极显著降低(P<0.01)。
表8 母羊妊娠后期添加CBC和AOC对羔羊血清蛋白质和能量代谢相关指标的影响

Table 8 Effects of combined supplementation of CBC and AOC in late gestation diet of ewes on serum protein and energy metabolism related indicators of lambs

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC
蛋白质代谢相关指标 Protein metabolism related indicators

总蛋白 TP/(g/L)		 出生后第30天 Day 30 after birth 56.55Bb 57.18Bb 59.45Aa 0.403 0.002
出生后第60天 Day 60 after birth 58.15b 60.89ab 63.37a 0.875 0.039

白蛋白ALB/(g/L)		 出生后第30天 Day 30 after birth 31.86 31.96 34.61 0.782 0.282
出生后第60天 Day 60 after birth 31.42b 33.47ab 35.34a 0.664 0.043

尿素UREA/(mmol/L)		 出生后第30天 Day 30 after birth 9.70a 7.59ab 5.99b 0.563 0.015
出生后第60天 Day 60 after birth 10.98Aa 8.39Bb 6.54Bc 0.608 0.004
能量代谢相关指标 Energy metabolism related indicators/(mmol/L)

葡萄糖GLU		 出生后第30天 Day 30 after birth 4.86b 5.06ab 5.78a 0.163 0.042
出生后第60天 Day 60 after birth 5.19 5.97 6.12 0.407 0.633

甘油三酯TG		 出生后第30天 Day 30 after birth 0.43 0.45 0.64 0.075 0.486
出生后第60天 Day 60 after birth 0.54 0.62 0.80 0.047 0.058

总胆固醇TC		 出生后第30天 Day 30 after birth 2.46b 2.44b 1.52a 0.178 0.035
出生后第60天 Day 60 after birth 1.50 1.89 1.94 0.163 0.510

2.4.2 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊血清抗氧化、免疫和肠道健康相关指标的影响

由表9可知,与CK组相比,CBC+AOC组羔羊出生后第30天血清T-AOC及GSH-Px、CAT活性均显著提升(P<0.05),且均在出生后第60天达到极显著提升(P<0.01);CBC+AOC组羔羊出生后第30和60天血清SOD活性均极显著提升(P<0.01);CBC+AOC组和CBC组羔羊出生后第30天血清MDA活性极显著降低(P<0.01)。此外,CBC+AOC组羔羊第30和60天血清SOD和CAT活性显著(P<0.05)或极显著(P<0.01)高于CBC组。与CK组相比,CBC+AOC组羔羊出生后第30和60天血清TNF-α含量显著降低(P<0.05),同时血清IL-6含量极显著降低(P<0.01),出生后第30天血清IL-1β含量极显著降低(P<0.01),出生后第60天血清IL-4含量极显著增加(P<0.01);CBC组羔羊出生后第30天血清IL-1β、IL-6含量极显著降低(P<0.01)。此外,CBC+AOC组羔羊出生后第30天血清IL-1β、IL-6含量极显著低于CBC组(P<0.01),出生后第60天血清IL-4含量极显著高于CBC组(P<0.01)。与CK组相比,CBC+AOC组羔羊出生后第30天血清DAO活性极显著降低(P<0.01),出生后第60天血清DAO活性以及出生后第30和60天血清D-LA含量显著降低(P<0.05);CBC组羔羊出生后第30天血清DAO活性极显著降低(P<0.01)。此外,CBC+AOC组羔羊出生后第30天血清DAO活性极显著低于CBC组(P<0.01)。
表9 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊血清抗氧化、免疫和肠道健康相关指标的影响

Table 9 Effects of combined supplementation of CBC and AOC in late gestation diet of ewes on serum antioxidant, immune and intestinal health related indicators of lambs

项目
Items		 时间
Time		 组别 Groups 均值标准误
SEM		 P值
P-value
CK CBC CBC+AOC
抗氧化相关指标 Antioxidant related indicators

总抗氧化能力
T-AOC/(U/mL)		 出生后第30天 Day 30 after birth 7.33b 7.92ab 8.56a 0.191 0.020
出生后第60天 Day 60 after birth 7.64Bb 8.08ABab 8.41Aa 0.113 0.009

超氧化物歧化酶
SOD/(U/mL)		 出生后第30天 Day 30 after birth 63.25Bb 69.38Bb 78.41Aa 2.074 0.003
出生后第60天 Day 60 after birth 64.60Bc 70.46Bb 86.66Aa 2.702 <0.001

谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 出生后第30天 Day 30 after birth 95.96b 102.98ab 133.98a 6.909 0.048
出生后第60天 Day 60 after birth 114.70Bc 132.78ABb 150.86Aa 4.990 0.004

丙二醛
MDA/(nmol/mL)		 出生后第30天 Day 30 after birth 4.68Aa 3.59Bb 3.13Bb 0.184 0.001
出生后第60天 Day 60 after birth 4.91 4.68 4.03 0.276 0.421

过氧化氢酶
CAT/(U/mL)		 出生后第30天 Day 30 after birth 29.97b 30.10b 35.16a 1.011 0.046
出生后第60天 Day 60 after birth 32.74Bb 33.07Bb 46.33Aa 2.138 0.004
免疫相关指标 Immune related indicators/(pg/mL)

肿瘤坏死因子-α
TNF-α		 出生后第30天 Day 30 after birth 84.01a 73.13ab 59.12b 3.810 0.017
出生后第60天 Day 60 after birth 57.35a 50.69ab 43.82b 2.005 0.012

白细胞介素-1β
IL-1β		 出生后第30天 Day 30 after birth 35.38A 27.17B 19.52C 1.966 <0.001
出生后第60天 Day 60 after birth 27.92a 22.55ab 19.35b 1.482 0.047

白细胞介素-4
IL-4		 出生后第30天 Day 30 after birth 5.71 6.15 6.82 0.295 0.316
出生后第60天 Day 60 after birth 6.56Bb 6.82Bb 9.33Aa 0.391 0.001

白细胞介素-6
IL-6		 出生后第30天 Day 30 after birth 182.55A 155.35B 112.88C 7.742 <0.001
出生后第60天 Day 60 after birth 149.43Aa 138.06Aa 102.96Bb 6.241 0.001
肠道健康相关指标 Intestinal health related indicators

二胺氧化酶
DAO/(U/L)		 出生后第30天 Day 30 after birth 4.04A 3.22B 2.51C 0.197 0.001
出生后第60天 Day 60 after birth 2.72a 2.53ab 2.10b 0.105 0.038

D-乳酸
D-LA/(mg/dL)		 出生后第30天 Day 30 after birth 2.80a 2.28ab 1.89b 0.144 0.022
出生后第60天 Day 60 after birth 2.59a 2.06ab 1.54b 0.158 0.016

3 讨论

3.1 妊娠后期饲粮中联合添加CBC和AOC对母羊生长性能的影响

母羊在妊娠后期需同时满足胎儿生长发育和自身泌乳的营养需求,往往动员体内储存的营养物质来维持代谢平衡,加剧机体负担,容易导致体重下降、代谢疾病风险升高[13]。本研究中,CBC组和CBC+AOC组母羊在妊娠第121~150天ADG极显著增加,且产后失重降低,表明在妊娠后期补饲CBC或同时补饲CBC和AOC均有利于母羊产后体重的恢复,改善营养代谢效率;此外,CBC和AOC联合使用在提升母羊生长性能方面表现出协同增效作用,其效果优于单一使用CBC。其原因可能是丁酸梭菌产生的短链脂肪酸有助于改善肠道健康和能量代谢,而米曲霉分泌的复合酶系和未知的功能性免疫因子则提升了饲粮中养分的消化利用率[10,14]。CBC和AOC的协调作用,不仅缓解了母羊在妊娠后期的营养负平衡状态,还为其产后恢复提供了更为充分的营养保障,从而实现对母羊健康与生长性能的双重提升。

3.2 妊娠后期饲粮中联合添加CBC和AOC对母羊血清生化指标的影响

妊娠母羊血清TP、ALB和UREA含量正常范围分别为60~79 g/L、24~30 g/L和2.83~7.11 mmol/L,本研究中各组母羊上述指标均处于正常范围内。UREA是氮代谢的终产物,血清中其含量变化可反映反刍动物机体氮循环和瘤胃蛋白质降解代谢的强度[15]。然而,CBC+AOC组母羊妊娠第150天血清TP含量极显著升高,而UREA含量显著降低,这提示CBC与AOC联合使用可促进瘤胃微生物对氨氮的利用与微生物蛋白合成,进而提高蛋白质利用率,增加氮沉积。血清β-HB是评估围产期能量负平衡和酮病风险的关键指标。酮病主要特征为血清中酮体(如β-HB)含量增高,血清GLU含量降低。本研究中,CBC组和CBC+AOC组妊娠第150天和产后30天血清β-HB含量均极显著降低,同时血清GLU含量极显著升高,其机制可能是:CBC产生的丁酸等短链脂肪酸可促进瘤胃上皮发育和挥发性脂肪酸吸收;AOC分泌的多种酶类(如淀粉酶、蛋白酶)可提高养分消化率,增加葡萄糖前体物质(如丙酸)的生成,从而改善机体能量代谢。
机体氧化应激与抗氧化状态的动态平衡是维持动物健康的关键[16]。GSH-Px、SOD和CAT作为体内主要抗氧化酶,能够有效清除氧自由基,减轻机体氧化损伤[17]。T-AOC可综合反映机体整体抗氧化水平[18];MDA是脂质过氧化的终产物,其含量降低表明机体抗脂质过氧化能力增强[19]。本研究中,CBC+AOC组母羊妊娠第150天血清MDA含量极显著降低,T-AOC显著升高,说明CBC与AOC联合使用有助于增强机体整体抗氧化能力,缓解氧化应激损伤,该作用可能与AOC含有的抗氧化活性物质有关[20-21]。Li等[22]研究发现,米曲霉发酵可显著改变苹果皮成分,提高游离酚含量,增强其抗氧化活性与益生元潜力。在许多病理状态下,TNF-α产生增多,导致机体炎症反应的加剧,最终造成组织损伤,降低机体免疫功能[23]。在本研究中,CBC+AOC组母羊产后第30天血清细胞因子IL-1β、IL-6和TNF-α含量显著或极显著降低,表明CBC与AOC联合使用可以有效抑制促炎介质的释放,从而减轻炎症反应。结合抗氧化指标结果提示,CBC与AOC联合使用可通过缓解氧化应激、增强抗氧化能力提升机体免疫功能。
D-LA和DAO是评价肠道物理屏障完整性和损伤程度的敏感指标,肠道屏障受损时二者会大量释放入血。Zhang等[24]研究发现,丁酸梭菌能够增强肠道屏障功能。本研究中,CBC+AOC组母羊妊娠第150天血清DAO活性和D-LA含量均显著降低,表明CBC与AOC联合使用可有效维持肠道黏膜结构的完整性,降低肠道通透性。完整的肠道屏障可保障营养物质充分吸收,减少内毒素等有害物质的易位,这也是CBC与AOC抑制全身炎症反应、改善生长性能的重要机制。

3.3 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊生长发育的影响

ADG是反映羔羊生长速度的常用指标,其数值越大说明羔羊生长速度越快。本试验结果发现,CBC+AOC组羔羊的初生重(出生后第1天体重)以及出生后第30、60天体重和ADG均显著提高,且血清TP、GLU含量显著增加,推测母羊妊娠后期补饲CBC和AOC可提高营养物质代谢与转运效率,使羔羊能够通过胎盘和母乳获取更充足的营养物质,进而提高初生重,并在哺乳期保持更快的生长速度。
体尺指标是评价羔羊体格结构、躯体发育和整体生长状况的重要依据。本研究发现,随日龄的增加,CBC+AOC组羔羊体斜长、体高和胸围等关键体尺指标均呈持续上升的趋势,且各阶段数值均高于CK组,尽管围指标无显著差异。这一结果进一步证实了在母羊妊娠后期补饲CBC与AOC可促进羔羊生长发育,不仅提高体重增长速度,还可促进躯体结构全面发育。

3.4 母羊妊娠后期饲粮中联合添加CBC和AOC对羔羊血清生化指标的影响

本试验结果表明,母羊妊娠后期饲粮中联合添加CBC和AOC可显著提高羔羊血清T-AOC以及SOD、GSH-Px活性,显著降低血清MDA含量。这表明,给母体补饲CBC和AOC可提高哺乳羔羊的抗氧化能力,缓解氧化应激。TNF-α、IL-6均是重要的促炎性因子。IL-4是关键的抗炎细胞因子,可调节促炎、抗炎和免疫细胞分化、激活作用,其含量升高表示机体免疫应答水平提高[25]。本试验中,母羊妊娠后期饲粮中联合添加CBC和AOC使羔羊血清IL-4含量显著增加,IL-6和TNF-α含量显著降低。这表明,CBC和AOC可通过乳汁传递,促进羔羊抗炎因子生成,抑制炎性反应,从而增强其机体抗炎与抗病能力。其机制可能在于CBC和AOC能促进肠道有益菌群的增殖,改善肠道健康并通过乳汁传递活性物质,激活抗氧化和免疫系统,最终实现促进动物生长、增强免疫力和提高抗氧化功能的效果。

4 结论

综合以上结果可以看出,妊娠后期饲粮中联合添加CBC与AOC可改善母羊的蛋白质代谢、能量平衡、抗氧化能力、免疫功能及肠道屏障功能;同时,还可通过母体-子代间的营养传递效应为羔羊的早期生长发育创造更有利的条件,这种跨代营养效应具体表现为羔羊生长性能的提升和体尺发育的改善。

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