Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 2: 1107 - 1116

DOI: https://doi.org/10.12418/CJAN2024.100

RESEARCH PAPER

Effects of Different Microecological Preparations on Growth Performance and Serum Biochemical, Immune and Antioxidant Indexes of Lambs

  • MA Wenbin , 1, 2 ,
  • WANG Yan 2 ,
  • CHEN Xiangyu 2 ,
  • ABUXIAHEMAN Mubalake 2 ,
  • YUAN Cen 3 ,
  • QIN Rongyan 2 ,
  • LIU Yanfeng 2 ,
  • ZHANG Zhijun 2 ,
  • LIU Limeng 1, 2 ,
  • WANG Lele 2 ,
  • WANG Wenqi , 2, *
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  • 1 College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China
  • 2 Feed Research Institute, Xinjiang Academy of Animal Science, Urumqi 830000, China
  • 3 Bachu Anxin Animal Husbandry Co., Ltd., Bachu 843800, China
* professor, E-mail:

Received date: 2023-08-22

  Online published: 2024-02-01

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Abstract

The aim of this experiment was to study the effects of different microecological preparations on growth performance and serum biochemical, immune and antioxidant indexes of lambs, to provide data reference for the improvement of growth performance and health condition of lambs. Forty-eight newborn lambs with the same age, similar weight [(3.00±0.20) kg] and good health condition were selected and randomly divided into 3 groups with 4 replicates in each group and 4 lambs in each replicate. The control group was fed a starter, the test group 1 was fed the starter supplemented with 2.2% microecological preparation Ⅰ (Clostridium butyricum:Bacillus subtilis=4:5), and the test group 2 was fed the starter supplemented with 2.2% microecological preparation Ⅱ (Clostridium butyricum:Bacillus subtilis:oligofructose=4:5:5). The experiment lasted for 40 days. The results showed as follows: 1) there were no significant differences in average daily milk intake, average daily starter intake and average daily gain of lambs during 15 to 34 days of age, 35 to 54 days of age and 15 to 54 days of age among all groups (P>0.05). 2) The serum lactate dehydrogenase (LDH) activity and β-hydroxybutyric acid(β-HB) content of test group 1 were significantly higher than those of the control group (P<0.05), and the serum alanine aminotransferase (ALT) activity of test group 2 was significantly higher than that of the control group (P<0.05). 3) The serum immunoglobulin G (IgG) content of test group 1 and test group 2 was significantly higher than that of the control group (P<0.05), the contents of immunoglobulin M (IgM) and interleukin-4 (IL-4) in serum of test group 2 were significantly higher than those of the control group (P<0.05), and the contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in serum of test group 1 and test group 2 were significantly lower than those of the control group (P<0.05). 4) The serum glutathione peroxidase (GSH-Px), catalase (CAT) activities and total antioxidant capacity (T-AOC) of test group 1 and test group 2 were significantly higher than those of the control group (P<0.05), the serum malondialdehyde (MDA) content of test group 1 and test group 2 was significantly lower than that of the control group (P<0.05), and the serum nitric oxide (NO) content of test group 2 was significantly higher than that of the control group (P<0.05). In conclusion, dietary different microecological preparations can enhance the immunity and improve the antioxidant capacity of lambs, and the starter supplemented with 2.2% microecological preparation Ⅱ (Clostridium butyricum:Bacillus subtilis:oligofructose=4:5:5) has batter effects.

Key words： microecological preparations; lambs; growth performance; serum biochemical indexes; immune; antioxidant

Cite this article

MA Wenbin , WANG Yan , CHEN Xiangyu , ABUXIAHEMAN Mubalake , YUAN Cen , QIN Rongyan , LIU Yanfeng , ZHANG Zhijun , LIU Limeng , WANG Lele , WANG Wenqi . Effects of Different Microecological Preparations on Growth Performance and Serum Biochemical, Immune and Antioxidant Indexes of Lambs[J]. Chinese Journal of Animal Nutrition, 2024 , 36(2) : 1107 -1116 . DOI: 10.12418/CJAN2024.100

随着人们生活水平的提高,新时代的消费者对动物产品的“量”和“质”都有了更高的期待。抗生素虽然有效减少了动物的疾病,但是对大众的健康有着很大的威胁,而微生态制剂因其在使用过程中安全、无毒、不会产生耐药性且不会污染环境,所以是良好的替抗产品[1]。微生态制剂是利用正常微生物或促进微生物生长的物质制成的活性微生物制剂。也就是说,一切能促进正常微生物菌群生长繁殖的及抑制致病菌生长繁殖的微生物制剂都称为微生态制剂[2]。由于微生态制剂具有调节胃肠道之功效,可快速构建胃肠道微生态平衡,因此无论在婴儿、老人,还是在新生畜禽的应用中都可以很好地促进其胃肠道健康[3]。
《饲料添加剂品种目录(2013)》允许使用的饲料级微生物包括丁酸梭菌(Clostridium butyricum)和枯草芽孢杆菌(Bacillus subtilis)等。丁酸梭菌属于厌氧微生物,可在无氧和酸性环境中保持活性,在畜禽生产应用领域可提高机体免疫力,促进动物胃肠道健康[4];枯草芽孢杆菌具有绿色无污染、安全性好、无残留等优点,在畜牧生产中得到广泛应用[5]。肖克权等[6]研究发现,在麻鸡饲粮中添加丁酸梭菌和枯草芽孢杆菌制剂可提高麻鸡的生长性能。低聚果糖是一种典型的益生元,具有促进机体生长、降低腹泻率和死亡率、调控肠道菌群结构等作用,可促进动物健康生长[7]。饲喂枯草芽孢杆菌和粪肠球菌等可改善羔羊的肠道菌群,降低羔羊腹泻率[8]。Obianwuna等[9]研究表明,在蛋鸡饲粮中添加丁酸梭菌和低聚果糖可提高其生长性能和免疫功能等。Xu等[10]研究发现,枯草芽孢杆菌和低聚果糖的组合在牛的生产应用方面可增加活性氧的产生,从而促进细胞凋亡。羔羊存在成活率低[11]、多疾病、易腹泻、免疫力低下等问题[12],多数研究都以枯草芽孢杆菌直接加入到其饲粮中来解决这些问题。因此,本试验主要研究不同微生态制剂对羔羊生长性能及血清生化、免疫、抗氧化指标的影响,旨在为羔羊饲粮中添加适宜微生态制剂提供数据参考。

1 材料与方法

1.1 试验材料

微生态制剂由某生物科技有限公司研发,包括微生态制剂Ⅰ(丁酸梭菌:枯草芽孢杆菌=4:5)和微生态制剂Ⅱ(丁酸梭菌:枯草芽孢杆菌:低聚果糖=4:5:5),其中,丁酸梭菌有效活菌数≥5×1011 CFU/kg,枯草芽孢杆菌有效活菌数≥1013 CFU/kg,低聚果糖纯度为96.9%。

1.2 试验时间和地点

动物饲养试验于2022年7—9月在新疆喀什地区巴楚县的巴楚安欣牧业有限责任公司的第二羊场开展。

1.3 试验设计

试验选取日龄相同、体重[(3.00±0.20) kg]相近、健康状况良好的初生湖羊羔羊48只,随机分为3组,每组4个重复,每个重复4只羊。对照组饲喂开食料,试验1组在开食料中添加2.2%的微生态制剂Ⅰ,试验2组在开食料中添加2.2%的微生态制剂Ⅱ。试验期40 d。开食料参考《肉羊营养需要量》(NY/T 816—2021)并与实际应用相结合进行配制,开食料组成及营养水平见表1。
表1 开食料组成及营养水平(风干基础)

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

项目 Items 含量 Content
原料 Ingredients
玉米 Corn 42.20
麸皮 Wheat bran 3.50
玉米淀粉 Corn starch 4.10
豆粕 Soybean meal 27.00
预混料 Premix1) 2.50
苜蓿 Alfalfa 20.00
小苏打 NaHCO3 0.20
食盐 NaCl 0.50
合计 Total 100.00
营养水平 Nutrient levels2)
干物质 DM 95.62
粗蛋白质 CP 19.26
粗脂肪 EE 1.97
中性洗涤纤维 NDF 23.31
酸性洗涤纤维 ADF 15.79
粗灰分 Ash 6.98
钙 Ca 0.76
磷 P 0.47
总能 GE/(MJ/kg) 17.06

1)预混料为每千克开食料提供 Premix provided the following per kg of the starter:VA 30 000 IU,VD 5 000 IU,VE 40 IU,Cu (CuSO4·5H2O) 10 mg,Zn (ZnSO4·H2O) 60 mg,Co (CoCl2·6H2O) 0.15 mg,Mn (MnSO4·5H2O) 65 mg,Fe (FeSO4·7H2O) 50 mg,Se (Na2SeO3) 0.2 mg,I(KI)0.4 mg。

2)营养水平为实测值。Nutrient levels were measured values.

1.4 饲养管理

试验前改造圈舍,将哺乳母羊圈舍一分为二,实现母子分群饲养,并对圈舍进行消毒、杀菌。羔羊1~7日龄,母羊和羔羊在同一圈舍,羔羊随母哺乳;羔羊8~14日龄,母羊每天08:00—09:00、13:00—14:00、18:00—19:00进行3次合圈哺乳,其余时间母羊和羔羊分离。羔羊15~54日龄为试验期,羔羊15~34日龄,母羊每天08:00—09:00、18:00—19:00进行2次合圈哺乳,其余时间母羊和羔羊分离,为羔羊补饲开食料;羔羊35~54日龄,母羊每天08:00—09:00进行1次合圈哺乳,其余时间母羊和羔羊分离,为羔羊补饲开食料。母羊和羔羊分离期间,羔羊自由采食开食料、自由饮水。

1.5 样品采集与指标测定

1.5.1 饲粮营养水平

饲粮干物质(DM)含量采用105 ℃灼烧法测定,钙(Ca)含量采用火焰原子吸收光谱法(GB 5009.92—2016)测定,其他营养水平参考《饲料分析及饲料质量检测技术》中的方法测定。

1.5.2 生长性能

于羔羊15、34和54日龄晨饲前对羔羊进行空腹称重,计算体重和平均日增重(ADG);试验期内每天监测开食料采食量,计算平均日开食料采食量(ADSI);于羔羊34、54日龄合圈之前给羔羊穿上尿不湿并称重,合圈之后再次称重,测定羔羊的平均日羊奶摄入量(ADMI),34日龄羊奶摄入量视为15~34日龄平均日羊奶摄入量,54日龄羊奶摄入量视为35~54日龄平均日羊奶摄入量。

1.5.3 血清生化、免疫、抗氧化指标

正试期结束时,每组随机选取8只羔羊,每个重复选取2只羔羊,共选取24只。空腹从静脉采血10 mL,室温静置样品2 h,并以3 000 r/min离心15 min,取上清液,-20 ℃保存待测。
血清生化指标:肌酸激酶(CK)、乳酸脱氢(LDH)、谷草转氨酶(AST)、谷丙转氨酶(ALT)、碱性磷酸酶(ALP)活性和β-羟丁酸(β-HB)含量。
血清免疫指标:免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和白细胞介素-4(IL-4)含量。
血清抗氧化指标:谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性和丙二醛(MDA)、一氧化氮(NO)含量及总抗氧化能力(T-AOC)。
血清CK、LDH、AST、ALT、ALP活性和β-HB、IgA、IgG、IgM含量采用比色法测定,试剂盒购自中生北控股份有限公司,仪器为全自动生化仪(BS-420,深圳迈瑞医疗国际有限公司);血清TNF-α、IL-1β、IL-6、IL-4、MDA、NO含量和GSH-Px、CAT活性及T-AOC采用酶免法测定,试剂盒购自北京华英生物技术研究所,仪器为酶标分析仪(DR-200BS,无锡华卫德朗仪器有限公司)。以上指标均委托北京华英生物技术研究所检测。

1.6 数据统计分析

采用Excel 2021处理试验数据,并采用SPSS 26.0软件进行单因素方差分析,再通过Duncan氏法进行多重比较,结果用“平均值±标准差”表示,P<0.05表差异显著,P>0.05表差异不显著。

2 结果

2.1 不同微生态制剂对羔羊生长性能的影响

由表2可知,各组之间15、34和54日龄体重及15~34日龄、35~54日龄、15~54日龄ADMI、ADSI、ADG均无显著差异(P>0.05)。
表2 不同微生态制剂对羔羊生长性能的影响

Table 2 Effects of different microecological preparations on growth performance of lambs

项目
Items		 日龄
Days of age		 组别 Groups
对照 Control 1 2


体重
BW/kg		 15 4.89±0.62 4.36±0.41 4.62±0.51
34 7.91±0.30 7.20±0.76 7.15±0.81
54 12.34±0.60 11.24±1.44 11.22±0.76


平均日羊奶摄入量
ADMI/(mL/d)		 15~34 410.00±31.62 420.00±18.26 422.50±33.37
35~54 420.83±29.46 418.13±27.01 418.75±24.79
15~54 415.42±24.35 419.06±23.29 420.63±28.93


平均日开食料采食量
ADSI/(g/d)		 15~34 61.33±5.27 57.87±4.85 52.86±5.55
35~54 278.86±12.73 258.43±18.03 272.55±20.93
15~54 170.00±10.94 158.15±15.56 162.70±16.89


平均日增重
ADG/(g/d)		 15~34 155.94±12.00 140.31±13.65 127.66±10.80
35~54 188.60±16.79 202.10±19.88 203.31±20.89
15~54 184.61±15.00 171.85±16.24 165.48±8.58

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

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

2.2 不同微生态制剂对羔羊血清生化、免疫和抗氧化指标的影响

2.2.1 不同微生态制剂对羔羊血清生化指标的影响

由表3可知,各组之间血清CK、AST和ALP活性均无显著差异(P>0.05)。试验1组血清LDH活性显著高于对照组(P<0.05),试验2组血清ALT含量显著高于对照组(P<0.05),试验1组血清β-HB含量显著高于对照组(P<0.05)。
表3 不同微生态制剂对羔羊血清生化指标的影响

Table 3 Effects of different microecological preparations on serum biochemical indexes of lambs

项目
Items		 组别 Groups
对照 Control 1 2
肌酸激酶 CK/(U/L) 92.24±24.21 110.43±31.52 108.13±29.35
乳酸脱氢酶 LDH/(U/L) 298.17±75.25b 402.46±56.59a 364.35±32.19ab
谷草转氨酶 AST/(U/L) 48.57±15.12 44.30±13.07 56.58±9.14
谷丙转氨酶 ALT/(U/L) 7.10±1.60b 8.51±1.35ab 9.07±0.58a
碱性磷酸酶 ALP/(U/L) 403.16±117.39 402.85±73.18 384.39±40.69
β-羟丁酸 β-HB/(mmol/L) 0.17±0.06b 0.30±0.09a 0.18±0.05b

2.2.2 不同微生态制剂对羔羊血清免疫指标的影响

由表4可知,各组之间血清IgA含量无显著差异(P>0.05)。试验1组、试验2组血清IgG含量显著高于对照组(P<0.05),试验2组血清IgM含量显著高于对照组(P<0.05),试验1组、试验2组血清TNF-α含量显著低于对照组(P<0.05),试验1组、试验2组血清IL-1β和IL-6含量显著低于对照组(P<0.05),试验2组血清IL-4含量显著高于对照组(P<0.05)。
表4 不同微生态制剂对羔羊免疫指标的影响

Table 4 Effects of different microecological preparations on serum immune indexes of lambs

项目
Items		 组别 Groups
对照 Control 1 2
免疫球蛋白A IgA/(g/L) 1.67±0.13 1.78±0.06 1.82±0.14
免疫球蛋白G IgG/(g/L) 17.97±0.82b 23.35±1.52a 24.83±1.83a
免疫球蛋白M IgM/(g/L) 1.43±0.19b 1.60±0.14b 1.74±0.12a
肿瘤坏死因子-α TNF-α/(pg/mL) 54.56±5.29a 44.28±7.99b 36.19±7.80b
白细胞介素-1β IL-1β/(pg/mL) 30.96±4.13a 24.79±5.94b 18.67±2.57c
白细胞介素-6 IL-6/(pg/mL) 127.56±11.22a 117.62±7.02b 94.26±2.88c
白细胞介素-4 IL-4/(pg/mL) 8.14±1.50b 9.30±0.48b 11.93±2.51a

2.2.3 不同微生态制剂对羔羊血清抗氧化指标的影响

由表4可知,试验1组、试验2组血清GSH-Px和CAT活性显著高于对照组(P<0.05),试验1组、试验2组血清T-AOC显著高于对照组(P<0.05),试验1组、试验2组血清MDA含量显著低于对照组(P<0.05),试验2组血清NO含量显著高于对照组(P<0.05)。
表5 不同微生态制剂对羔羊血清抗氧化指标的影响

Table 5 Effects of different microecological preparations on serum antioxidant indexes of lambs

项目
Items		 组别 Groups
对照 Control 1 2
谷胱甘肽过氧化物酶 GSH-Px/(U/mL) 269.87±37.67b 342.53±17.94a 370.29±32.02a
总抗氧化能力 T-AOC/(U/mL) 5.45±0.74b 6.96±0.96a 7.65±1.36a
过氧化氢酶 CAT/(U/mL) 30.16±5.11c 40.95±2.74b 47.59±2.64a
丙二醛 MDA/(nmol/mL) 3.76±0.13a 3.02±0.30b 2.50±0.35c
一氧化氮 NO/(μmol/L) 38.03±7.18b 47.13±12.01b 60.15±6.70a

3 讨论

3.1 不同微生态制剂对羔羊生长性能的影响

生长性能是最直接反映不同微生态制剂影响羔羊生长的客观指标[13]。微生态制剂及其代谢产物进入羔羊消化道后[14],可改善胃肠道内环境[15],促进羔羊消化和吸收营养物质[16]。此外,微生态制剂在羔羊生长过程中还可以改善其胃肠道微生态平衡[17],减少腹泻的发生[18]。张磊等[19]研究发现,草本复合微生态制剂、猪源性乳酸菌微生态制剂和芽孢杆菌微生态制剂都促进了动物的生长性能。张学虎等[20]研究表明,全混合日粮(TMR)中添加枯草芽孢杆菌、纤维素酶和果寡糖等可促进小尾寒羊生长,促进机体对多种营养物质的吸收。姚逸安等[21]研究发现,补饲地衣芽孢杆菌、枯草芽孢杆菌和屎肠球菌可提高羔羊的生长性能。
在本试验中,不同微生态制剂对羔羊的体重、ADSI、ADMI、ADG影响不显著,但是随着羔羊生长阶段的推移,微生态制剂增强了羔羊的采食量,但是没有明显的增长趋势,可能与某一菌群的变化有关,具体原因需进一步研究,再加上本试验在夏天进行,高温干燥,因此羔羊增重较慢。

3.2 不同微生态制剂对羔羊血清生化、免疫和抗氧化指标的影响

血清生化指标可反映羔羊的生理机能和免疫能力。彭涛等[22]研究发现,在基础饲粮中添加复合益生菌制剂,可提高山羊血清总蛋白和球蛋白含量。曾晓等[23]试验表明,在基础饲粮中添加复合益生菌可改善奶牛血清生化指标。AST、ALT和ALP在机体中主要负责代谢蛋白质,进而为机体提供营养支持[24]。AST和ALT参与机体蛋白质代谢,正常情况下,血清中它们的活性较低[25]。而本试验中试验2组由于加入了低聚果糖,血清ALT活性显著升高,可能是试验2组微生态制剂的组合效应所致。ALP主要影响消化代谢,可反映机体生长性能,其活性升高是肝功能下降的体现[26]。本试验结果显示,与对照组相比,试验1组、试验2组血清ALP活性有所降低,表明微生态制剂可促进羔羊机体肝脏的发育。β-HB是胴体的一种成分,其含量升高代表脂肪代谢出现了异常[27]。而本试验中试验1组血清β-HB含量较对照组显著升高,说明不利于羔羊机体的脂肪代谢;试验2组血清β-HB含量与对照组相近,说明试验2组效果更好。
羔羊的免疫系统本来就弱,加上试验期正值夏日,在夏季高温环境下,其免疫能力更低。免疫球蛋白是机体免疫的抗体之一,它能特异性识别并消灭细菌及病毒[28]。血液中最多的免疫球蛋白是IgG,它可最大限度地降低致病菌对宿主细胞的影响[29]。本试验结果表明,试验1组、试验2组血清IgA、IgG和IgM含量均有明显升高,其中试验1组血清IgG含量显著高于对照组,试验2组血清IgG和IgM含量显著高于对照组。TNF-α和白细胞介素都是衡量机体早期免疫的重要指标[30]。TNF-α是一种促炎细胞因子,参与正常的炎症反应和免疫反应,机体发生应激等行为时,其含量升高[31]。本试验结果表明,试验1组、试验2组血清TNF-α含量显著低于对照组,说明微生态制剂可有效降低羔羊炎症发生概率,减小应激。郭爽等[32]研究显示,大鼠应激后,血清白细胞介素-2(IL-2)、TNF-α和IL-4等含量均会上升。免疫球蛋白分布于肠道黏膜上,可阻止病原菌的入侵[33],促进吞噬细胞发挥作用[34],并对抗原发挥免疫作用[35]。白细胞介素和机体免疫功能呈正相关[36],复合菌培养物和β-葡聚糖可显著提高肉羊血清TNF-α和IL-1β含量[37]。本试验结果表明,试验1组、试验2组血清IL-1β、IL-6含量均显著低于对照组,试验2组血清IL-4含量显著高于对照组,证实微生态制剂可促进羔羊的免疫功能。
抗氧化指标是检验机体抗氧化能力的一种重要指标。它可以反映机体内环境的稳定性,对动物的抗氧化功能具有重要作用[38]。GSH-Px、T-AOC、CAT、MDA和NO是检验羔羊抗氧化能力的重要指标。SOD和GSH-Px是清除体内自由基最重要的酶之一[39],CAT主要催化分解过氧化氢[40],T-AOC反映羔羊抗氧化的总体水平[41],MDA可反映体内氧化反应对羔羊的损耗情况[42]。钱明珠等[43]研究证实,在饲粮中添加复合微生态制剂可显著降低犊牛血清MDA含量。范晶等[44]研究表明,在饲粮中添加0.15%单宁可提高断奶羔羊抗氧化能力。陈浅等[45]试验表明,丁酸梭菌具有提高机体抗氧化能力的作用,与本试验相符。在本试验中,试验1组、试验2组血清GSH-Px、CAT活性和T-AOC显著高于对照组,血清MDA含量显著低于对照组,说明微生态制剂提高了羔羊抗氧化能力,达到了试验效果。

4 结论

饲粮中添加不同微生态制剂对羔羊生长性能无显著影响,但可增强羔羊机体免疫力,提高抗氧化能力。本试验条件下,开食料中添加2.2%的微生态制剂Ⅱ(丁酸梭菌:枯草芽孢杆菌:低聚果糖=4:5:5)的效果更好。

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