Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 4: 2524 - 2532

DOI: https://doi.org/10.12418/CJAN2023.236

RESEARCH PAPER

Effects of Dietary Rhodiola Extract Supplemental Level on Growth Performance, Immune Performance and Antioxidant Capacity of Meat Rabbits

  • LIU Gongyan , 1 ,
  • SUN Haitao 1 ,
  • GAO Shuxia 1 ,
  • BAI Liya 1 ,
  • HAO Lihong 1 ,
  • WANG Yanping 1 ,
  • HU Hongmei 1 ,
  • NING Liangchuan 2 ,
  • WANG Cheng , 1, *
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  • 1 Key Laboratory of Livestock and Poultry Multiomics of Ministry of Agriculture and Rural Affairs, Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Husbandry and Veterinary, Shandong Academy of Agricultural Sciences, Jinan 250100, China
  • 2 Shandong Bora Lilai Bioengineering Co., Ltd., Tai'an 271000, China
*associate professor, E-mail:

Received date: 2022-09-30

  Online published: 2023-04-12

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Abstract

The purpose of this experiment was to study the effects of dietary rhodiola extract supplemental level on growth performance, immune performance and antioxidant capacity of meat rabbits, so as to provide theoretical reference for rhodiola extract as a new green feed additive in rabbit production. A total of 400 twenty-eight-day-old weaned commercial Hyla rabbits were randomly divided into 4 groups with 10 replicates in each group and 10 rabbits in each replicate. Rabbits were fed diets supplemented with 0 (control group), 0.5%, 1.0% and 2.0% rhodiola extract, respectively. The pre-experimental period lasted for 7 days, and the experimental period lasted for 30 days. The results showed as follows: 1) the mortality rate of 1.0% and 2.0% supplement level groups was significantly lower than that of the control group (P<0.05). 2) The spleen weight and spleen index 0.5% supplement level group were significantly higher than those of the control group (P<0.05). 3) The contents of immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin A (IgA), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and interleukin-6 (IL-6) in serum of 2.0% supplement level group were significantly higher than those of the control group (P<0.05). 4) The contents of secretory immunoglobulin A (sIgA), IgG, IgM, TNF-α and IFN-γ in intestine of 0.5%, 1.0% and 2.0% supplement level groups were significantly higher than those of the control group (P<0.05), and the content of IL-6 in intestine of 0.5% supplement level group was significantly higher than that of the control group (P<0.05). 5) The total antioxidant capacity in intestine of 0.5%, 1.0% and 2.0% supplement level groups was significantly higher than that of the control group (P<0.05), the activity of glutathione peroxidase in intestine of 0.5% supplement level group was significantly higher than that of the control group (P<0.05), and the content of malondialdehyde in intestine of 1.0% and 2.0% supplement level groups was significantly higher than that of the control group (P<0.05). In conclusion, dietary rhodiola extract can improve the intestinal antioxidant capacity of meat rabbits, increase the body non-specific immune performance by increasing the contents of immunoglobulins and immune factors, reduce the mortality, and the suitable supplemental level is 0.5% to 1.0%.

Key words： rhodiola extract; rabbits; growth performance; immune performance; antioxidant capacity

Cite this article

LIU Gongyan , SUN Haitao , GAO Shuxia , BAI Liya , HAO Lihong , WANG Yanping , HU Hongmei , NING Liangchuan , WANG Cheng . Effects of Dietary Rhodiola Extract Supplemental Level on Growth Performance, Immune Performance and Antioxidant Capacity of Meat Rabbits[J]. Chinese Journal of Animal Nutrition, 2023 , 35(4) : 2524 -2532 . DOI: 10.12418/CJAN2023.236

消化道疾病是家兔最为多发的疾病,再加上流行性兔腹胀病的发生,断奶仔兔的死亡率可高达60%,严重影响养殖场的经济效益[1-2]。在家兔生产上,抗生素曾被广泛用于控制断奶仔兔死亡率。然而,抗生素在畜禽养殖中产生的副作用引起人们的广泛关注,我国也于2020年全面禁止抗生素作为饲料添加剂在饲料中添加使用。因此,研究开发替代抗生素的新型绿色饲料添加剂成为当务之急。红景天(Rhodiola rosea L.)为景天科、红景天属多年生草本植物,主要生长于在西藏等恶劣的高山环境中。红景天作为一种传统的药食两用植物,富含蛋白质、脂肪、碳水化合物、维生素、微量元素等多种营养物质。另外,从红景天中分离的景天庚糖、红景天苷、酪醇、黄酮等多种活性成分,具有抗氧化、免疫调节、抗炎作用[3⇓-5]。Cheng[6]研究发现,在红沼泽小龙虾饲料中添加红景天多糖能够提高小龙虾增重,改善生长性能。李龙等[7]研究报道,红景天提取物能够促进肉鸡外周血白细胞免疫球蛋白和细胞因子的产生,改善肉鸡的免疫机能。李鹏飞等[8]研究发现,红景天苷能够通过影响调节性T细胞进而抑制小鼠肿瘤的生长。此外,红景天苷能够通过调控miR-217-5p/YY1相关因子2(YY1 associated factor 2)减轻重症急性胰腺炎诱导的胰腺损伤和炎症反应[9]。然而,关于红景天及其提取物在家兔生产上的相关研究报道较少。因此,本试验以商品伊拉肉兔为研究对象,探讨饲粮中红景天提取物添加水平对家兔生长性能、免疫性能和抗氧化能力的影响,为红景天提取物作为新型绿色饲料添加剂在肉兔生产上使用提供理论参考。

1 材料与方法

1.1 试验设计和饲养管理

试验选用28日龄断奶的商品伊拉肉兔400只(公母各占1/2),随机分为4组,每组10个重复,每个重复10只。各组分别饲喂红景天提取物(以乙醇作为溶剂从红景天中提取制备)添加水平为0(对照组)、0.5%、1.0%、2.0%的试验饲粮。基础饲粮参考《肉兔营养需要量》(NY/T 4049—2021)[10]饲养标准配制而成,其组成及营养水平见表1。试验兔在封闭兔舍每笼5只饲养(公母分开),预试期7 d,正试期30 d。试验期间,试验兔出现疾病问题,不采取任何治疗措施。
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) %

原料Ingredients 含量Content 营养水平Nutrient levels2) 含量Content
玉米Corn 5.0 消化能DE/(MJ/kg) 10.23
豆粕Soybean meal 8.0 干物质DM 89.82
大麦Barley 6.0 粗蛋白质CP 16.12
麸皮Wheat bran 15.0 粗脂肪EE 2.80
玉米胚芽粕Corn germ meal 16.0 粗纤维CF 17.38
玉米皮Corn husk 17.0 中性洗涤纤维NDF 38.74
苜蓿草粉Alfalfa meal 15.0 酸性洗涤纤维ADF 23.08
豆秸粉Soybean straw powder 7.0 酸性洗涤木质素ADL 6.29
稻壳粉Rice hull powder 8.0 粗灰分Ash 9.03
磷酸氢钙CaHPO4 1.5 钙Ca 0.95
氯化钠NaCl 0.5 总磷TP 0.45
预混料Premix1) 1.0 赖氨酸Lys 0.60
合计Total 100.0 蛋氨酸+半胱氨酸Met+Cys 0.65

1)预混料为每千克饲粮提供 The premix provided for the following per kilogram of the diet:VA 8 000 IU,VD3 1 500 IU,VE 45 mg,VK3 2.0 mg,VB1 1.0 mg,VB2 3.0 mg,VB6 1.5 mg,烟酸 nicotinic acid 30 mg,泛酸 pantothenic acid 50 mg,叶酸 folic acid 0.5 mg,氯化胆碱 choline chloride 100 mg,Fe 50 mg,Cu 10 mg,Zn 50 mg,Mn 10 mg,I 0.5 mg,Se 0.05 mg,赖氨酸 Lys 1.5 g,蛋氨酸 Met 0.5 g,其余为麦饭石载体补足 the rest was supplemented by medical stone carrier。

2)营养水平除消化能外均为实测值。Nutrient levels were measured values except DE.

1.2 样品采集与制备

试验结束当天,每个重复选取1只肉兔屠宰,心脏采血后分离血浆,同时采集回肠样品用于肠道免疫和抗氧化指标的测定。

1.3 测定指标和方法

1.3.1 生长性能

在试验正式开始和结束当日,称量每个重复家兔重量,计算初始体重、终末体重和平均日增重(ADG)。统计试验期间每个重复的采食量,计算平均日采食量(ADFI)。料重比(F/G)为ADFI/ADG。以重复为单位,计算试验期间家兔死亡率[11]。

1.3.2 免疫器官发育

试验结束前1天晚上禁食,自由饮水,试验结束当天每个重复选取1只试验兔屠宰,记录宰前活体重、胸腺重、脾脏重、圆小囊重和蚓突重,并计算相应免疫器官指数。

1.3.3 血清和肠道免疫指标

将回肠样品在磷酸盐缓冲液(PBS)中破碎匀浆,离心后分离上清液用于免疫指标的测定。采用酶联免疫吸附测定(ELISA)试剂盒测定血清和肠道中免疫球蛋白A(IgA)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)和白细胞介素-6(IL-6)含量,试剂盒购自武汉华美生物工程有限公司。使用ELISA试剂盒测定血清和肠道中免疫球蛋白G(IgG)、免疫球蛋白M(IgM)和分泌型免疫球蛋白A(sIgA)含量,试剂盒购自武汉云克隆科技股份有限公司。

1.3.4 肠道抗氧化指标

将回肠样品在PBS中破碎匀浆,离心后分离上清液用BCA试剂盒测定蛋白浓度后,使用南京建成生物工程研究所生产的相应试剂盒测定肠道中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量和总抗氧化能力(T-AOC)。

1.4 数据处理

用SAS 9.4统计软件中的GLM程序进行方差分析,用Duncan氏法进行多重比较,同时采用正交多项式分析不同添加水平的红景天提取物对各指标的线性和二次曲线效应,结果以平均值(mean)和均方根误差(RMSE)表示,P<0.05为差异显著。

2 结果

2.1 饲粮中红景天提取物添加水平对肉兔生长性能的影响

由表2可知,饲粮中红景天提取物添加水平对肉兔终末体重、平均日增重、平均日采食量和料重比无显著影响(P>0.05)。饲粮中红景天提取物添加水平对肉兔死亡率有显著影响(P<0.05),呈现显著二次曲线变化(P=0.021);其中,0.5%、1.0%和2.0%添加水平组死亡率较对照组分别降低了25%(P>0.05)、50%(P<0.05)和38%(P<0.05),以1.0%添加水平组效果最佳。
表2 饲粮中红景天提取物添加水平对肉兔生长性能的影响

Table 2 Effects of dietary rhodiola extract supplemental level on growth performance of meat rabbits

项目
Items		 红景天提取物添加水平
Rhodiola extract supplemental level/%		 均方根
误差
RMSE		 P值P-value
0(对照
Control)		 0.5 1.0 2.0 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
初始体重IBW/g 1 077.5 1 077.5 1 072.5 1 095.0 39.594 0.606 0.419 0.304
终末体重FBW/g 2 428.6 2 321.7 2 420.3 2 444.7 174.236 0.395 0.695 0.128
平均日增重ADG/(g/d) 45.03 41.47 44.93 44.99 5.606 0.412 0.825 0.215
平均日采食量ADFI/(g/d) 162.80 159.90 162.40 163.30 9.782 0.869 0.587 0.850
料重比F/G 3.64 3.94 3.72 3.67 0.582 0.650 0.857 0.207
死亡率Mortality rate/% 20.00a 15.00ab 10.00b 12.50b 12.430 0.031 0.179 0.021

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 饲粮中红景天提取物添加水平对肉兔免疫器官发育的影响

由表3可知,饲粮中红景天提取物添加水平对肉兔脾脏重和脾脏指数影响显著(P<0.05),呈现显著二次曲线变化(P=0.013和P=0.011);其中,0.5%添加水平组脾脏重和脾脏指数最高,显著高于对照组(P<0.05)。饲粮中红景天提取物添加水平对肉兔胸腺重、圆小囊重、蚓突重及胸腺指数、圆小囊指数、蚓突指数无显著影响(P>0.05)。
表3 饲粮中红景天提取物添加水平对肉兔免疫器官发育的影响

Table 3 Effects of dietary rhodiola extract supplemental level on immune organ development of meat rabbits

项目
Items		 红景天提取物添加水平
Rhodiola extract supplemental level/%		 均方根
误差
RMSE		 P值P-value
0(对照
Control)		 0.5 1.0 2.0 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
胸腺重Thymus weight/g 4.00 4.30 3.90 4.70 1.015 0.305 0.138 0.124
脾脏重Spleen weight/g 1.60b 1.87a 1.79ab 1.78ab 0.207 0.042 0.082 0.013
圆小囊重
Sacculus rotundus weight/g		 3.40 3.40 3.60 3.40 0.843 0.935 0.668 0.905
蚓突重
Vermicular process weight/g		 10.50 10.00 10.70 9.40 2.977 0.768 0.680 0.875
胸腺指数Thymus index/(g/kg) 1.91 2.08 1.86 2.22 0.473 0.311 0.192 0.295
脾脏指数Spleen index/(g/kg) 0.76b 0.90a 0.86ab 0.85ab 0.128 0.001 0.176 0.011
圆小囊指数
Sacculus rotundus index/(g/kg)		 1.61 1.64 1.71 1.62 0.378 0.933 0.734 0.885
蚓突指数
Vermicular process index/(g/kg)		 5.00 4.76 4.95 4.48 1.134 0.737 0.469 0.755

2.3 饲粮中红景天提取物添加水平对肉兔血清免疫指标的影响

由表4可知,饲粮中红景天提取物添加水平对肉兔血清中免疫球蛋白(IgG、IgM和IgA)和免疫因子(TNF-α、IFN-γ和IL-6)含量影响显著(P<0.05),呈现显著线性和二次曲线变化(P<0.05);其中,2.0%添加水平组血清IgG、IgM、IgA、TNF-α、IFN-γ和IL-6含量显著高于对照组(P<0.05)。
表4 饲粮中红景天提取物添加水平对肉兔血清免疫指标的影响

Table 4 Effects of dietary rhodiola extract supplemental level on serum immune indexes of meat rabbits

项目
Items		 红景天提取物添加水平
Rhodiola extract supplemental level/%		 均方根
误差
RMSE		 P值P-value
0(对照
Control)		 0.5 1.0 2.0 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
免疫球蛋白G IgG/(μg/mL) 7.38b 7.87a 8.28a 8.26a 0.467 <0.001 <0.001 <0.001
免疫球蛋白M IgM/(ng/mL) 639.01b 666.57ab 686.52a 699.77a 41.677 0.015 0.002 0.005
免疫球蛋白A IgA/(ng/mL) 112.28c 120.09b 127.21a 130.41a 6.671 <0.001 <0.001 <0.001
肿瘤坏死因子-α
TNF-α/(ng/mL)		 2.69c 2.98bc 3.20ab 3.34a 0.352 0.002 <0.001 <0.001
干扰素-γ IFN-γ/(pg/mL) 293.99c 349.68ab 324.12bc 378.29a 49.616 0.004 0.002 0.010
白细胞介素-6 IL-6/(pg/mL) 241.95c 285.94b 287.94b 330.25a 29.505 <0.001 <0.001 <0.001

2.4 饲粮中红景天提取物添加水平对肉兔肠道免疫指标的影响

由表5可知,饲粮中红景天提取物添加水平对肉兔肠道中sIgA、IgG、IgM和免疫因子(TNF-α、IFN-γ和IL-6)含量影响显著(P<0.05),除IL-6含量外,均呈现显著线性和二次曲线变化(P<0.05);其中,0.5%、1.0%和2.0%添加水平组肠道中sIgA、IgG、IgM、TNF-α和IFN-γ含量显著高于对照组(P<0.05)。肠道中IL-6含量呈现先升高后降低二次曲线变化(P=0.025),其中,0.5%添加水平组肠道中IL-6含量最高,显著高于对照组和1.0%添加水平组(P<0.05)。
表5 饲粮中红景天提取物添加水平对肉兔肠道免疫指标的影响

Table 5 Effects of dietary rhodiola extract supplemental level on intestinal immune indexes of meat rabbits ng/g

项目
Items		 红景天提取物添加水平
Rhodiola extract supplemental level/%		 均方根
误差
RMSE		 P值P-value
0(对照
Control)		 0.5 1.0 2.0 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
分泌型免疫球蛋白A sIgA 229.47b 314.70a 300.92a 294.70a 38.219 <0.001 0.013 0.001
免疫球蛋白G IgG 14.47b 23.85a 22.80a 24.53a 5.923 0.002 0.002 0.002
免疫球蛋白M IgM 21.96c 33.45b 43.74a 49.97a 7.236 <0.001 <0.001 <0.001
肿瘤坏死因子-α TNF-α 15.27b 22.52a 22.77a 22.88a 3.816 <0.001 <0.001 <0.001
干扰素-γ IFN-γ 57.62b 73.53a 69.70a 67.58a 6.992 <0.001 0.034 0.001
白细胞介素-6 IL-6 1.27b 1.68a 1.36b 1.45ab 0.301 0.031 0.485 0.025

2.5 饲粮中红景天提取物添加水平对肉兔肠道抗氧化指标的影响

由表6可知,饲粮中红景天提取物添加水平对肉兔肠道中总抗氧化能力、谷胱甘肽过氧化物酶活性和丙二醛含量影响显著(P<0.05),均呈现显著线性和二次曲线变化(P<0.05);其中,0.5%、1.0%和2.0%添加水平组肠道中总抗氧化能力显著高于对照组(P<0.05),0.5%添加水平组肠道中谷胱甘肽过氧化物酶活性显著高于对照组(P<0.05),1.0%和2.0%添加水平组肠道中丙二醛含量显著低于对照组和0.5%添加水平组(P<0.05)。
表6 饲粮中红景天提取物添加水平对肉兔肠道抗氧化指标的影响

Table 6 Effects of dietary rhodiola extract supplemental level on intestinal antioxidant indexes of meat rabbits

项目
Items		 红景天提取物添加水平
Rhodiola extract supplemental level/%		 均方根
误差
RMSE		 P值P-value
0(对照
Control)		 0.5 1.0 2.0 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
总抗氧化能力
T-AOC/(mmol/g prot)		 0.89b 2.93a 3.00a 2.96a 1.535 0.008 0.004 0.001
谷胱甘肽过氧化物酶
GSH-Px/(U/mg prot)		 11.16b 13.98a 12.20ab 10.73b 2.443 0.025 0.012 0.023
超氧化物歧化酶
SOD/(U/mg prot)		 279.06 303.14 310.72 306.60 52.943 0.547 0.193 0.216
丙二醛MDA/(nmol/mg prot) 0.74a 0.73a 0.50b 0.46b 0.166 <0.001 <0.001 0.002

3 讨论

近年来,随着人们对食品安全的关注,越来越多的中草药及植物提取物类饲料添加剂被广泛用于畜牧生产中。中草药及植物提取物类饲料添加剂成分复杂,主要的化学成分有多糖、黄酮、糖苷、有机酸和精油等[12]。红景天作为一种传统药材,具有消除疲劳、缓解抑郁症和缓解高原反应等功能[13]。最新研究发现,红景天含有红景天苷、罗塞维和酪醇等100多种活性物质,能够调节免疫球蛋白和细胞因子含量及基因表达,具有免疫调节作用[14⇓⇓-17]。李文凤等[18]研究报道,饲粮中添加10 mg/kg红景天苷能够显著降低腹水肉鸡肺动脉收缩压,增加红细胞数量,降低肉鸡死亡率,改善肉鸡生长性能。本研究发现,饲粮中添加红景天提取物在不改变肉兔生长性能前提下降低肉兔死亡率,这与红景天中红景天苷等活性物质能够提高机体非特异性免疫性能密切相关。
免疫器官对动物机体免疫性能发挥重要作用,免疫器官相对重量的变化会影响动物的免疫性能。本研究发现,饲粮中添加0.5%红景天提取物组的肉兔脾脏重量及脾脏指数显著高于对照组,这表明饲粮中添加红景天有益于肉兔免疫器官的发育。免疫器官可以分泌免疫球蛋白和细胞因子,在动物机体免疫系统中发挥重要作用,也是免疫调节的潜在靶点[19]。目前研究表明,一些中草药和植物提取物能够通过调节免疫球蛋白和细胞因子的合成发挥机体免疫调节作用[20]。张健等[21]研究发现,红景天对感染柔嫩艾美尔球虫的雏鸡有一定的预防作用,可以显著降低感染鸡球虫后雏鸡血清中白细胞介素-2(IL-2)、TNF-α含量,提高血清中IgG含量。然而,Cai等[22]通过体外和体内试验发现,红景天多糖能够提高携带肿瘤的小鼠外周血T淋巴细胞的CD4+/CD8+比值,促进血清中IL-2、TNF-α和IFN-γ的产生。本研究发现,饲粮中添加红景天提取物能够提高肉兔血清中免疫球蛋白(IgG、IgM和IgA)和免疫因子(TNF-α、IFN-γ和IL-6)含量,与前人研究报道基本一致。
大量研究表明,红景天能够参与调节应激反应系列的活动,提高动物机体抗氧化能力[23-24]。Xu等[25]研究发现,红景天多糖可以通过清除羟基和超氧阴离子自由基,使机体自由基水平保持稳定。此外,红景天苷也可以通过抑制氧化应激和炎症反应对细菌脂多糖诱导的小鼠重症急性胰腺炎起到保护作用[26]。Cao等[27]研究表明,红景天多糖在公牛精子冻融过程中发挥较强的保护作用,能够改善精液冷冻保存后的SOD、乳酸脱氢酶和谷氨酸草酰乙酸转氨酶活性。本研究发现,饲粮中添加红景天提取物能够提高肉兔肠道中谷胱甘肽过氧化物酶活性和总抗氧化能力,降低丙二醛含量,这可能与红景天中存在的红景天多糖、红景天苷等天然抗氧化活性物质有关。
肠黏膜屏障主要由上皮细胞、相邻肠细胞之间的紧密连接以及黏膜免疫系统的关键成分组成[28]。肠黏膜屏障作为一种选择性屏障,允许身体匀浆状态所需物质的吸收和排泄,并防止各种病原体或其产物通过。正常情况下,上皮细胞只允许少量完整抗原进入黏膜,在黏膜中与黏膜免疫系统相互作用,降低炎症反应。同时,肠道病原体可以激活免疫细胞,启动消除感染所需的炎症反应[29]。在炎症性肠病的情况下,抗原通过上皮层的过度渗透可能导致不适当的免疫刺激和慢性胃肠炎症[30]。此外,上皮细胞通过向管腔内分泌液体和黏液及sIgA来结合有害物质,从而发挥重要的生理防御作用[31]。sIgA是一种主要的黏膜免疫效应分子,它提供了对抗病原体的重要的第1道防线[32]。本研究发现,饲粮中红景天提取物能够影响肉兔肠道中sIgA、IgG和IgM和免疫因子(TNF-α、IFN-γ和IL-6)含量,说明红景天提却物中存在的天然抗氧化活性物质提高了肠道抗氧化能力,对改善肠道黏膜免疫性能发挥重要作用。

4 结论

红景天提取物作为新型绿色饲料添加剂能够提高肉兔抗氧化能力,通过增加肉兔血浆及肠道免疫球蛋白和免疫因子含量提高机体非特异免疫性能,在不改变生长性能的前提下降低肉兔死亡率,添加水平以0.5%~1.0%为宜。

致谢

感谢山东正宇兔业有限公司王鹏总经理及全场职工在肉兔饲养期间对试验的大力支持和帮助!

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