Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 3: 1599 - 1611

DOI: https://doi.org/10.12418/CJAN2024.141

RESEARCH PAPER

Effects of Mangosteen and Pigeonpea Extracts on Growth Performance, Plasma Biochemical Indices, Intestinal Health and Meat Quality of Chickens

  • FAN Qiuli , 1 ,
  • TAO Zhengguo 2 ,
  • LI Hui 2 ,
  • GOU Zhongyong 1 ,
  • WANG Yibing 1 ,
  • LIN Xiajing 1 ,
  • YE Jinling 1 ,
  • ZHANG Sai 1 ,
  • JIANG Shouqun , 1, *
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  • 1 Guangdong Provincial Key Laboratory of Animal Breeding and Nutrition, Key Laboratory of Animal Nutrition and Feed Science in South China, Ministry of Agriculture and Rural Affairs, State Key Laboratory of Livestock and Poultry Breeding, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
  • 2 Guangzhou Leader Biotechnology Co., Ltd., Guangzhou 510663, China
*professor, E-mail:

Received date: 2023-10-17

  Online published: 2024-03-13

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Abstract

This experiment was conducted to investigate the effects of dietary mangosteen and pigeonpea extracts on growth performance, plasma biochemical indices, intestinal mucosal cytokine contents and cecal microflora and meat quality of 818 chickens, to provide reference for their application in chickens. A total of 600 one-day-old 818 male chickens were randomly divided into control group, antibiotic group, mangosteen extract group and pigeonpea extract group according to the principle of consistent body weight with 6 replicates in each group and 25 chickens in each replicate. The chickens in the 4 groups were fed a basal diet and the basal diet supplemented with 500 mg/kg enramycin, 30 mg/kg mangosteen extract and 300 mg/kg pigeonpea extract, respectively. The experimental period was 40 days. The results showed as follows, compared with the control group: 1) the final body weight and average daily gain of chickens in antibiotic group and mangosteen extract group were significantly increased (P<0.05). 2) The contents of total protein (TP), triiodothyronine (T3) and thyroxine (T4) as well as total antioxidant capacity (T-AOC) in plasma in mangosteen extract group were significantly increased (P<0.05), and the plasma T-AOC was significantly higher than that in antibiotic group (P<0.05). Meanwhile, the contents of glucose (GLU), uric acid (UA) and malondialdehyde (MDA) in plasma in mangosteen extract group were significantly decreased (P<0.05), and the plasma GLU content was significantly lower than that in antibiotic group (P<0.05). The contents of triglyceride (TG) and total cholesterol (TC) in plasma in pigeonpea extract group were significantly decreased (P<0.05), which were significantly lower than those in antibiotic group (P<0.05). 3) The contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in jejunal mucosa in mangosteen and pigeonpea extract groups were significantly decreased (P<0.05), and the IL-6 content in jejunal mucosa in mangosteen extract group was significantly lower than that in antibiotic group (P<0.05). The interleukin-10 (IL-10) content in jejunal mucosa in mangosteen extract group was significantly increased (P<0.05), which was significantly higher than that in antibiotic group (P<0.05). The Enterococcus relative abundance in cecum in pigeonpea extract group was significantly increased (P<0.05), which was higher than that in antibiotic group (P<0.05). 4) The breast muscle color brightness (L*) value after slaughter for 45 min and 24 h and drip loss in mangosteen and pigeonpea extract groups were significantly decreased (P<0.05), the breast muscle color redness (a*) value was significantly increased (P<0.05), and the a* value in breast muscle in mangosteen extract group was significantly higher than that in antibiotic group (P<0.05). In conclusion, 30 mg/kg mangosteen extract can improve growth performance, antioxidant capacity, intestinal immune function and relative abundance of beneficial microflora, improve meat color and reduce drip loss; 300 mg/kg pigeonpea extract can reduce blood lipid content, improve intestinal immune function, improve meat color and reduce drip loss; 30 mg/kg mangosteen extract can replace antibiotic added into the diet for 818 chickens from 1 to 40 days of age.[Chinese Journal of Animal Nutrition, 2024, 36(3):1599-1611]

Key words： mangosteen extract; pigeonpea extract; 818 chickens; growth performance; plasma biochemical indices; intestinal health; meat quality

Cite this article

FAN Qiuli , TAO Zhengguo , LI Hui , GOU Zhongyong , WANG Yibing , LIN Xiajing , YE Jinling , ZHANG Sai , JIANG Shouqun . Effects of Mangosteen and Pigeonpea Extracts on Growth Performance, Plasma Biochemical Indices, Intestinal Health and Meat Quality of Chickens[J]. Chinese Journal of Animal Nutrition, 2024 , 36(3) : 1599 -1611 . DOI: 10.12418/CJAN2024.141

天然植物提取物因绿色、安全和环保等特点已被广泛应用于畜禽无抗养殖。倒捻子素又称山竹黄酮,是一种从山竹果壳中提取的双苯吡酮类化合物,主要分为α、β、γ-倒捻子素,不仅具有植物激素样作用,同时可抑制环氧化酶和神经鞘磷脂酶、诱导细胞凋亡,在炎症、肿瘤和激素相关疾病治疗方面具有一定的效果[1⇓-3]。木豆素是从木豆叶中提取的一种芪类化合物,具有抑菌、抗炎、抗氧化、抗肿瘤和降血糖等药理作用[4-5]。研究发现,0.4~1.6 μg/mL α-倒捻子素可显著抑制金黄色葡萄球菌诱发的奶牛乳腺上皮细胞炎症反应[6];5 μmol/L α-倒捻子素可通过调节Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路炎症因子的表达抑制脂多糖(LPS)诱导的IEC-6细胞炎症[7];100 mg/(kg·d) α-倒捻子素可提高免疫抑制小鼠免疫器官指数和脾脏淋巴细胞增殖率以及自然杀伤(NK)细胞活性[8];3.75~15.00 μmol/L α-倒捻子素可通过磷脂酰肌醇-3-羟激酶(PI3K)/蛋白激酶B(AKT)/过氧化物酶体增殖物受体γ共激活因子-1α(PGC-1α)信号通路介导沉默信息调节因子2相关酶3(SIRT3)失活,从而减轻碘酸钠诱导的细胞毒性和氧化损伤[9];1~8 μmol/L木豆素可通过抑制氧化和内质网应激介导的凋亡来保护皮质酮诱导的PC12细胞损伤[10];0.05~0.50 μmol/L木豆素可通过调节核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路相关基因和蛋白表达水平提高HepG2细胞的抗氧化作用[11]。基于α-倒捻子素和木豆素具有以上抗炎、抗氧化和提高免疫功能等作用,且其在肉鸡上的应用研究目前尚无报道。因此,本试验根据前期预试验结果,以1~40日龄818肉鸡为试验对象,研究30 mg/kg山竹提取物(α-倒捻子素)和300 mg/kg木豆提取物(木豆素)对其生长性能、脂质代谢、抗氧化能力、免疫功能、肠道健康和肉品质的影响,为山竹和木豆提取物在肉鸡健康、高效养殖中的应用提供科学依据。

1 材料与方法

1.1 试验材料

山竹提取物有效成分为α-倒捻子素,含量50%;木豆提取物有效成分为木豆素,含量2%。

1.2 试验设计

选用600只初始体重(38.16±0.03) g且状态良好的1日龄818肉鸡公雏,按照体重一致原则随机分为4个组,分别为对照组(饲喂基础饲粮)、抗生素组(饲喂基础饲粮+500 mg/kg恩拉霉素)、山竹提取物组(饲喂基础饲粮+30 mg/kg山竹提取物)、木豆提取物组(饲喂基础饲粮+300 mg/kg木豆提取物),每组6个重复,每个重复25只。试验期40 d,分为1~21日龄和22~40日龄2个阶段。

1.3 试验饲粮

试验采用玉米-豆粕型基础饲粮,不同饲养阶段饲粮营养水平参考《鸡饲养标准》(NY/T 33—2004)配制,且不同组相同饲养阶段营养水平保持一致。抗生素、山竹和木豆提取物按照试验设计的添加水平等量替代预混料中的统糠。基础饲粮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 basal diets (air-dry basis) %

项目
Items		 1~21日龄
1 to 21 days of age		 22~40日龄
22 to 40 days of age
原料 Ingredients
玉米 Corn (8.7% CP) 59.00 62.00
豆粕 Soybean meal (46.0% CP) 17.90 14.40
花生粕 Peanut meal (47.8% CP) 12.00 10.50
次粉 Wheat shorts (14.9% CP) 2.80 4.00
玉米蛋白粉 Corn gluten meal (63.5% CP) 2.00 2.00
大豆油 Soybean oil 1.20 2.50
石粉 Limestone 1.36 1.17
磷酸氢钙 CaHPO4 1.80 1.65
L-赖氨酸盐酸盐 L-lysine·HCl (98.5%) 0.32 0.27
DL-蛋氨酸 DL-methionine (99.0%) 0.32 0.21
氯化钠 NaCl 0.30 0.30
预混料 Premix1) 1.00 1.00
合计 Total 100.00 100.00
营养水平 Nutrient levels2)
表观代谢能 AME/(MJ/kg) 12.13 12.55
粗蛋白质 CP 21.31 19.07
赖氨酸 Lys 1.15 1.00
蛋氨酸+半胱氨酸 Met+Cys 0.91 0.76
钙 Ca 0.93 0.84
总磷 TP 0.63 0.57
非植酸磷 NPP 0.45 0.40

1)预混料为1~21日龄每千克饲粮提供 The premix provided the following per kg of the diet for 1 to 21 days of age:VA 8 000 IU,VB1 2 mg,VB2 8 mg,烟酸 nicotinic acid 35 mg,VB6 3.5 mg,VB12 0.01 mg,VD 1 000 IU,VE 20 IU,VK 0.5 mg,生物素 biotin 0.18 mg,泛酸 pantothenic acid 10 mg,叶酸 folic acid 0.55 mg,氯化胆碱 choline chloride 1 300 mg,Fe 100 mg,Cu 8 mg,Mn 120 mg,Zn 100 mg,I 0.7 mg,Se 0.3 mg。预混料为22~40日龄每千克饲粮提供 The premix provided the following per kg of the diet for 22 to 40 days of age:VA 6 000 IU,VB1 2 mg,VB2 5 mg,烟酸 nicotinic acid 30 mg,VB6 3 mg,VB12 0.01 mg,VD 750 IU,VE 10 IU,VK 0.5 mg,生物素 biotin 0.15 mg,泛酸 pantothenic acid 10 mg,叶酸 folic acid 0.55 mg,氯化胆碱 choline chloride 1 000 mg,Fe 80 mg,Cu 8 mg,Mn 100 mg,Zn 80 mg,I 0.7 mg,Se 0.3 mg。

2)表观代谢能、氨基酸和非植酸磷为计算值,粗蛋白质、钙和总磷为实测值。AME, amino acid and NPP were calculated values, while CP, Ca and TP were measured values.

1.4 饲养管理

试验鸡于地面平养,地面铺放谷壳,自由采食颗粒料和饮水。各组饲养管理和环境条件一致,每天自然光照和人工光照相结合,白天自然光照,夜间人工光照,保持鸡舍清洁卫生。按照常规操作程序和免疫流程进行饲养和免疫,每天观察鸡只健康状况、采食量和死亡鸡情况。

1.5 样品采集与制备

试验第40天20:00断料供水,次日08:00每重复挑选接近平均体重的2只鸡,翅静脉采集全血5 mL于抗凝管中,3 500 r/min离心10 min分离血浆,于-80 ℃保存备测血浆生化指标。采血后的试鸡颈部放血致死,剖取空肠,剪取中间肠段用生理盐水清洗干净内容物,滤纸吸干表面水分,载玻片刮取黏膜装于2 mL灭菌管中于-20 ℃保存备测细胞因子含量。采集盲肠内容物于灭菌的5 mL离心管中,于-20 ℃保存备测微生物区系。剖取2侧胸肌,去除表面筋膜和脂肪,左侧用于测定屠宰后45 min和24 h肉色以及pH,右侧用于测定滴水损失和剪切力。

1.6 测定指标及方法

1.6.1 饲粮营养水平

饲粮表观代谢能、氨基酸和非植酸磷含量依据《中国饲料成分及营养价值表(2019年第30版)》各原料数值进行计算;粗蛋白质、钙和总磷含量分别参照GB/T 6432—2018[12]、GB/T 6436—2018[13]和GB/T 6437—2018[14]方法进行测定。

1.6.2 生长性能

试验第40天20:00断料供水,次日08:00以重复为单位称重,统计耗料量,计算平均日采食量(ADFI)、平均日增重(ADG)、料重比(F/G)和死亡率。
死亡率(%)=(死亡鸡数量/总鸡数量)×100。

1.6.3 血浆生化指标

血浆总蛋白(TP)、尿酸(UA)、葡萄糖(GLU)、甘油三酯(TG)、总胆固醇(TC)、丙二醛(MDA)含量以及总超氧化物歧化酶(T-SOD)活性和总抗氧化能力(T-AOC)测定试剂盒均购买于南京建成生物工程研究所,各指标测定步骤和结果计算严格按照说明书进行。血浆三碘甲腺原氨酸(T3)和甲状腺素(T4)酶联免疫吸附测定(ELISA)试剂盒购买于广州步前生物科技有限公司,具体测定方法和结果计算严格按照说明书进行。以上所有血浆生化指标均在多功能酶标仪(SYNERGY H1,BioTek,美国)上读数。

1.6.4 空肠黏膜细胞因子含量

空肠黏膜样品10%匀浆液的制备以及总蛋白质含量测定方法参考范秋丽等[15]的方法操作,肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)和白细胞介素-22(IL-22)ELISA试剂盒均购买于广州步前生物科技有限公司,具体测定方法和结果计算严格按照说明书进行。

1.6.5 盲肠微生物区系

参考Tao等[16]的方法进行盲肠微生物DNA提取、16S rDNA V3~V4可变区域扩增、文库构建、测序和物种注释,分析计算基于操作分类单元(OTU)的盲肠微生物门、属水平相对丰度、α多样性和β多样性。

1.6.6 肉品质

左侧胸肌分别使用色差仪(CR-410型,美能达公司,日本)和便携式pH计(Testo-205,深圳市吉格机电设备有限公司)测定屠宰后45 min和24 h肉色及pH。右侧胸肌在固定位置取10 g左右称重记录,悬挂于吹气后的封口袋中,4 ℃冷库保存24 h后取出称重记录,计算滴水损失;剩余右侧胸肌4 ℃保存24 h后置于90 ℃水浴锅中,待样品中心温度升至70 ℃取出,流水冷却后取样,满足宽度(1.0 cm)×厚度(1.0 cm)×长度(1.5 cm),采用质构仪(Instron 4411型,英斯特朗公司,美国)测定剪切力。
滴水损失(%)=100×(滴水前重量-滴水后重量)/滴水前重量。

1.7 数据统计分析

试验数据采用SPSS 21.0软件中的one-way ANOVA过程进行单因素方差分析,当处理效应差异显著时进行Duncan氏法多重比较,P<0.05为差异显著,试验结果数据均用平均值和均值标准误(SEM)表示。

2 结果与分析

2.1 山竹和木豆提取物对1~40日龄肉鸡生长性能的影响

由表2可知,与对照组相比,抗生素组和山竹提取物组肉鸡终末体重和平均日增重均显著升高(P<0.05)。不同组间肉鸡平均日采食量、料重比和死亡率均无显著差异(P>0.05)。
表2 山竹和木豆提取物对1~40日龄肉鸡生长性能的影响

Table 2 Effects of mangosteen and pigeonpea extracts on growth performance of chickens from 1 to 40 days of age

项目
Items		 对照组
Control
group		 抗生素组
Antibiotic
group		 山竹提取物组
Mangosteen extract
group		 木豆提取物组
Pigeonpea extract
group		 均值
标准误
SEM		 P值
P-value
初始体重 Initial BW/g 38.16 38.14 38.19 38.15 0.01 0.885
终末体重Final BW/g 1 204.50b 1 259.72a 1 267.60a 1 235.67ab 13.45 0.047
平均日增重 ADG/g 29.16b 30.54a 30.74a 29.94ab 0.34 0.048
平均日采食量 ADFI/g 56.07 58.59 58.34 57.35 0.50 0.302
料重比 F/G 1.92 1.90 1.90 1.92 0.01 0.515
死亡率 Mortality/% 1.60 2.40 0.00 0.00 0.37 0.283

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

Different letter superscripts in peer data indicate significant difference (P<0.05), and the same letter or no letter superscripts indicate not significant difference (P>0.05). The same as below.

2.2 山竹和木豆提取物对肉鸡血浆生化指标的影响

由表3可知,与对照组相比,抗生素组和山竹提取物组肉鸡血浆TP含量显著升高(P<0.05);山竹提取物组血浆UA、GLU和MDA含量显著降低(P<0.05),且血浆GLU含量显著低于抗生素组(P<0.05);木豆提取物组血浆TG和TC含量显著降低(P<0.05),且显著低于抗生素组(P<0.05);抗生素组、山竹提取物组和木豆提取物组血浆T3含量显著升高(P<0.05);山竹提取物组血浆T4含量和T-AOC均显著升高(P<0.05),且血浆T-AOC显著高于抗生素组(P<0.05)。不同组间血浆T-SOD活性无显著差异(P>0.05)。
表3 山竹和木豆提取物对肉鸡血浆生化指标的影响

Table 3 Effects of mangosteen and pigeonpea extracts on plasma biochemical indices of chickens

项目
Items		 对照组
Control
group		 抗生素组
Antibiotic
group		 山竹提取物组
Mangosteen extract
group		 木豆提取物组
Pigeonpea extract
group		 均值
标准误
SEM		 P值
P-value
总蛋白 TP/(g/L) 14.73b 17.82a 18.46a 17.06ab 0.46 0.015
尿酸 UA/(μmol/L) 237.32a 172.47ab 160.17b 202.27ab 11.45 0.038
葡萄糖 GLU/(mmol/L) 15.69a 14.41a 11.80b 13.29ab 0.46 0.017
甘油三酯 TG/(mmol/L) 0.66a 0.61a 0.57a 0.34b 0.06 0.003
总胆固醇 TC/(mmol/L) 3.09a 3.12a 2.69ab 2.37b 0.08 0.002
三碘甲腺原氨酸
T3/(pmol/L)		 148.37b 188.17a 177.83a 176.47a 4.75 0.045
甲状腺素 T4/(pmol/L) 742.40b 825.78ab 884.68a 750.39b 16.40 0.008
丙二醛 MDA/(nmol/mL) 3.59a 3.03ab 2.52b 2.92ab 0.19 0.035
总超氧化物歧化酶
T-SOD/(U/mL)		 303.58 316.12 316.12 320.57 6.35 0.725
总抗氧化能力
T-AOC/(U/mL)		 8.72b 8.27b 12.00a 9.76ab 0.46 0.016

2.3 山竹和木豆提取物对肉鸡空肠黏膜细胞因子含量的影响

由表4可知,与对照组相比,山竹提取物组和木豆提取物组肉鸡空肠黏膜TNF-α和IL-6含量均显著降低(P<0.05),且山竹提取物组空肠黏膜IL-6含量显著低于抗生素组(P<0.05);山竹提取物组空肠黏膜IL-10含量显著升高(P<0.05),且显著高于抗生素组(P<0.05)。不同组间空肠黏膜IL-22含量无显著差异(P>0.05)。
表4 山竹和木豆提取物对肉鸡空肠黏膜细胞因子含量的影响

Table 4 Effects of mangosteen and pigeonpea extracts on cytokine contents in jejunal mucosa of chickensng/g

项目
Items		 对照组
Control
group		 抗生素组
Antibiotic
group		 山竹提取物组
Mangosteen extract
group		 木豆提取物组
Pigeonpea extract
group		 均值
标准误
SEM		 P值
P-value
肿瘤坏死因子-α TNF-α 12.67a 10.19ab 7.64b 7.77b 0.59 0.001
白细胞介素-6 IL-6 3.20a 3.10ab 2.25c 2.54bc 0.12 0.007
白细胞介素-10 IL-10 5.49b 6.08b 7.71a 6.35b 0.24 0.004
白细胞介素-22 IL-22 3.67 3.74 4.43 4.73 0.20 0.181

2.4 山竹和木豆提取物对肉鸡盲肠微生物区系的影响

2.4.1 α多样性分析

本试验24个盲肠内容物样品共获得1 882 616条高质量序列,平均长度为412.22 nt。OTU聚类结果显示,24个样品共获得14 914个OTU,聚类为252个门、431个纲、857个目、1 237个科以及2 221个属。由表5可知,不同组间肉鸡盲肠微生物的Shannon指数、Simpson指数、Chao1指数、Ace指数和Coverage指数均无显著差异(P>0.05)。
表5 山竹和木豆提取物对肉鸡盲肠微生物α多样性的影响

Table 5 Effects of mangosteen and pigeonpea extracts on microbial α diversity in cecum of chickens

项目
Items		 对照组
Control
group		 抗生素组
Antibiotic
group		 山竹提取物组
Mangosteen extract
group		 木豆提取物组
Pigeonpea extract
group		 均值
标准误
SEM		 P值
P-value
Shannon指数 Shannon index 4.83 4.72 4.96 5.08 1.27 0.799
Simpson指数 Simpson index 0.86 0.86 0.88 0.91 0.01 0.417
Chao1指数 Chaol index 648.95 598.25 673.52 639.03 31.77 0.906
Ace指数 Ace index 644.62 591.85 669.82 671.84 37.09 0.908
Coverage指数 Coverage index 0.995 0.997 0.997 0.995 0.00 0.818

2.4.2 β多样性分析

如图1所示,基于加权UniFrac(图1-A)和非加权UniFrac(图1-B)距离主坐标分析(PCoA)结果表明,主成分(PC)1和PC2对样品差异的贡献值分别为41.55%、23.96%和13.11%、11.48%。山竹提取物组和木豆提取物组样品和抗生素组样品有明显的分开,重叠部分不多。加权UniFrac综合主要物种进化关系、种类和其相对丰度分析,结果显示4个组样品菌群组成有相似部分,又有不同部分;非加权UniFrac主要对物种的存在和进化关系分析,结果显示抗生素组、山竹提取物和木豆提取物组的物种比对照组的更为丰富。综上说明,抗生素、山竹提取物和木豆提取物的添加改变了盲肠微生物中菌群组成,且菌群物种更为丰富。
图1 基于加权UniFrac(A)和非加权UniFrac(B)距离主坐标分析

C:对照组样品 samples in control group;A:抗生素组样品 samples in antibiotic group;M:山竹提取物组样品 samples in mangosteen group;P:木豆提取物组样品 samples in pigeonpea group。

Fig.1 Principal coordinates analysis based on weighted UniFrac (A) and unweighted UniFrac (B) distances

2.4.3 门和属水平下的菌群结构

如图2所示,各组肉鸡盲肠微生物在门水平上主要由厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidota)组成,两者共占门水平的80%以上(图2-A)。各组肉鸡盲肠微生物在属水平上组成主要为拟杆菌属(Bacteroides)、另枝菌属(Alistipes)、巴恩斯氏菌属(Barnesiella)、魏斯氏菌属(Weissella)、双歧杆菌属(Bifidobacterium)、肠球菌属(Enterococcus)、布劳特氏菌属(Blautia)、埃希氏菌-志贺氏菌属(Escherichia-Shigella)和粪杆菌属(Faecalibacterium),分别占属水平的11.56%~35.57%、8.28%~16.07%、1.52%~7.31%、0.12%~4.20%、0.10%~3.90%、0.45%~10.74%、0.71%~5.10%、0.15%~3.62%和2.33%~7.59%(图2-B)。
图2 山竹和木豆提取物对肉鸡盲肠菌群在门(A)和属(B)水平上相对丰度的影响

C:对照组 control group;A:抗生素组 antibiotic group;M:山竹提取物组 mangosteen group;P:木豆提取物组 pigeonpea group。下图同 the same as below。

Fig.2 Effects of mangosteen and pigeonpea extracts on relative abundance of cecal microflora of chickens at phylum (A) and genus (B) levels

肉鸡盲肠微生物相对丰度大于1%的菌属见表6。与对照组相比,抗生素组肉鸡盲肠中拟杆菌属相对丰度升高,山竹提取物组盲肠拟杆菌属相对丰度降低,但差异均不显著(P>0.05);木豆提取物组盲肠肠球菌属相对丰度显著升高(P<0.05)。不同组间盲肠另枝菌属、巴恩斯氏菌属、粪杆菌属、魏斯氏菌属、双歧杆菌属、布劳特氏菌属、埃希氏菌-志贺氏菌属和粪杆菌属相对丰度均无显著差异(P>0.05)。
表6 肉鸡盲肠微生物在属水平下的优势菌群分布

Table 6 Distribution of dominant microflora in cecal microflora of chickens at genus level %

项目
Items		 对照组
Control
group		 抗生素组
Antibiotic
group		 山竹提取物组
Mangosteen extract
group		 木豆提取物组
Pigeonpea extract
group		 均值
标准误
SEM		 P值
P-value
拟杆菌属 Bacteroides 20.86ab 35.57a 11.56b 23.79ab 3.10 0.041
另枝菌属 Alistipes 13.25 11.24 16.07 8.28 2.26 0.692
巴恩斯氏菌属 Barnesiella 4.80 4.86 7.31 1.52 0.25 0.739
魏斯氏菌属 Weissella 0.12 4.20 0.47 0.25 0.10 0.429
双歧杆菌属 Bifidobacterium 3.90 1.07 0.10 0.09 0.03 0.445
肠球菌属 Enterococcus 0.45b 1.43b 1.18b 10.74a 1.27 0.004
布劳特氏菌属 Blautia 5.10 2.23 2.02 0.71 0.28 0.330
埃希氏菌-志贺氏菌属
Escherichia-Shigella		 0.15 2.17 0.24 3.62 0.04 0.535
粪杆菌属 Faecalibacterium 5.55 2.33 7.59 2.60 1.04 0.229

2.4.4 物种聚类分析

肉鸡盲肠微生物物种丰富度聚类分析如图3所示,不同组间的物种丰度聚类明显分开。饲粮添加抗生素后提高了瘤胃球菌属扭矩群(Ruminococcus_torques_group)、拟杆菌属、Mucispirillum和魏斯氏菌属相对丰度,添加山竹提取物后提高了粪杆菌属、巴恩斯氏菌属、另枝菌属、普雷沃氏菌科_UCG-001(Prevotellaceae_UCG-001)和假单胞菌属(Pseudomonas)相对丰度,添加木豆提取物后提高了埃希氏菌-志贺氏菌属、假单胞菌属和肠球菌属相对丰度。
图3 物种丰富度聚类图

Faecalibacterium:粪杆菌属;Oscillospiracea_UCG-005:颤螺菌科_UCG-005;Barnesiella:巴恩斯氏菌属;Alistipes:另枝菌属;Prevotellaceae_UCG-001:普雷沃氏菌科_UCG-001;Pediococcus:片球菌属;Escherichia-Shigella:埃希氏菌-志贺氏菌属;Pseudomonas:假单胞菌属;Enterococcus:肠球菌属;Blautia:布劳特氏菌属;Bifidobacterium:双歧杆菌属;Ruminococcus_torques_group:瘤胃球菌属扭矩群;Bacteroides:拟杆菌属;Weissella:魏斯氏菌属。

Fig.3 Species richness cluster map

2.5 山竹和木豆提取物对肉鸡肉品质的影响

由表7可知,与对照组相比,抗生素组、山竹提取物组和木豆提取物组肉鸡屠宰后45 min和24 h胸肌肉色亮度(L*)值以及滴水损失均显著降低(P<0.05);山竹提取物组和木豆提取物组屠宰后45 min和24 h胸肌肉色红度(a*)值显著升高(P<0.05),且山竹提取物组显著高于抗生素组(P<0.05)。不同组间屠宰后45 min和24 h胸肌肉色黄度(b*)值和pH以及剪切力均无显著差异(P>0.05)。
表7 山竹和木豆提取物对肉鸡肉品质的影响

Table 7 Effects of mangosteen and pigeonpea extracts on meat quality of chickens

项目
Items		 对照组
Control
group		 抗生素组
Antibiotic
group		 山竹提取物组
Mangosteen extract
group		 木豆提取物组
Pigeonpea extract
group		 均值
标准误
SEM		 P值
P-value





肉色
Meat
color

45 min		 亮度 L* 63.76a 60.18b 57.62b 59.40b 0.73 0.012
红度 a* 10.75c 11.68bc 14.71a 13.33ab 0.47 0.006
黄度 b* 19.70 16.87 17.50 17.07 0.51 0.184


24 h		 亮度 L* 68.61a 64.27b 61.71b 62.61b 0.80 0.004
红度 a* 8.44c 10.23bc 12.86a 11.88ab 0.47 0.001
黄度 b* 20.99 17.82 19.15 20.46 0.58 0.215

pH		 45 min 6.05 6.22 6.01 5.94 0.05 0.172
24 h 5.82 5.92 5.75 5.66 0.05 0.356
滴水损失 Drip loss/% 2.21a 1.91b 1.88b 1.82b 0.05 0.009
剪切力 Shear force/N 27.34 26.76 25.47 26.46 1.07 0.948

3 讨论

3.1 山竹和木豆提取物对肉鸡生长性能的影响

抗生素因具有防治动物疾病和促生长的作用,使其在养殖业中的应用飞速发展,但同时由于滥用抗生素导致的公共安全问题也日益突出,制约畜牧业的发展,因此寻找绿色、安全的抗生素替代品成为了现阶段动物营养研究领域的热点[17]。双苯吡酮类又称氧杂蒽酮类,是山竹果壳中最主要也是研究最多的活性物质,从本质上来说是黄酮类的一种特殊结构,其中已分离的化合物中α-倒捻子素含量最高[18]。木豆素属于二苯乙烯衍生物,是典型的芪类化合物,同时天然芪类化合物又是具有丰富结构和生物活性的多酚类化合物[19]。目前,α-倒捻子素和木豆素在畜禽养殖中的应用研究暂无报道,但黄酮类和多酚类已被证明可提高畜禽生长性能[20-21]。本研究结果表明,30 mg/kg山竹提取物在不影响肉鸡平均日采食量和料重比的情况下可显著提高终末体重和平均日增重,且可达到抗生素添加效果;300 mg/kg木豆提取物也提高了肉鸡终末体重和平均日增重,但效果不显著,说明α-倒捻子素和木豆素的添加对肉鸡的生长均有促进作用。

3.2 山竹和木豆提取物对肉鸡血浆生化指标的影响

血浆生化指标可反映动物生理代谢状态,其中TP和UA含量主要反映蛋白质的吸收、合成和分解,TP含量升高,UA含量降低,表明动物体对蛋白质吸收增强;GLU含量在一定程度反映了动物体能量代谢水平,其含量降低,表明机体对GLU的利用升高;TG和TC含量反映动物体脂质代谢水平,TG和TC含量降低有利于高血脂症的预防[22-23]。本研究结果表明,30 mg/kg山竹提取物显著提高了肉鸡血浆TP含量,同时降低了血浆UA和GLU含量;300 mg/kg木豆提取物显著降低了血浆TG和TC含量,说明α-倒捻子素的添加可提高机体对蛋白质和葡萄糖的利用,木豆素的添加可调节脂代谢,有利于心血管疾病的预防,此结果也与前人报道的天然芪类成分具有调节高脂血症的结果[24]相似。血液中T3和T4含量的升高使得胞内部分RNA聚合酶活性升高,影响蛋白质的合成,提高体内氮沉积,进而促进机体生长[25]。本研究结果表明,山竹和木豆提取物的添加均可升高肉鸡血浆T3含量,且山竹提取物的添加升高了血浆T4含量,此研究结果与前人报道相似,同时也与山竹提取物的添加提高了氮沉积和生长性能的结果一致。MDA含量以及T-SOD活性和T-AOC常用来作为评价机体抗氧化能力强弱的指标,MDA含量降低,T-SOD活性和T-AOC升高,表明机体抗氧化能力增强[26]。本研究结果表明,30 mg/kg山竹提取物可显著降低肉鸡血浆MDA含量,提高T-AOC,此结果与Fu等[27]应用α-倒捻子素降低LPS/D-半乳糖胺诱导的急性肝衰竭小鼠肝脏MDA含量和Tatiya-Aphiradee等[28]应用α-倒捻子素降低溃疡性结肠炎小鼠结肠MDA含量结果相似,说明山竹提取物的添加可通过降低脂质过氧化物含量和提高抗氧化酶活性提高肉鸡抗氧化能力。

3.3 山竹和木豆提取物对肉鸡肠道健康的影响

肠道细胞因子含量是评价机体免疫功能和肠道健康的重要指标,促炎细胞因子TNF-α和IL-6含量降低、抑炎细胞因子IL-10含量升高,表明机体免疫功能增强[29-30]。研究报道,α-倒捻子素不仅可分别降低对乙酰氨基酚引起的急性肝衰竭小鼠血清TNF-α含量和佐剂性关节炎大鼠血清中TNF-α含量[31-32],还可降低LPS刺激的人牙龈成纤维细胞中IL-6的表达水平[33]。本研究结果表明,山竹和木豆提取物的添加均可降低肉鸡空肠黏膜TNF-α和IL-6含量,且山竹提取物的添加可提高空肠黏膜IL-10含量,此结果与Huang等[34]应用木豆素降低LPS刺激小鼠巨噬细胞中TNF-α和IL-6含量的结果相似,说明山竹和木豆提取物的添加均可通过降低空肠促炎细胞因子含量提高肉鸡免疫功能。
肠道菌群复杂多样,其结构稳定是维持屏障功能的重要因素,本研究结果表明,虽然山竹和木豆提取物的添加对盲肠微生物菌群α多样性指数无显著影响,但增加了物种组成和丰富度。拟杆菌属可降解植物多糖和碳水化合物,产生乙酸盐和丙酸盐,提高肠道代谢功能;另枝菌属和粪杆菌属可产生丁酸盐,通过上调抗菌肽的表达降低机体炎症反应[35-36];巴恩斯氏菌属丰度越高,脾脏和肝脏中的边缘带B细胞和不变自然杀伤T细胞就越多[37];瘤胃球菌属能够参与食物消化,产生丙酸和丁酸维持肠道屏障功能[38];魏斯氏菌属具有类乳酸菌的作用,可被用来作为食品发酵的益生菌产生风味物质[39];肠球菌虽然可感染动物,但在多数情况下是不致病的。本研究结果表明,抗生素的添加提高了肉鸡盲肠拟杆菌属、瘤胃球菌属和魏斯氏菌属相对丰度,木豆提取物的添加提高了盲肠肠球菌属相对丰度,同时山竹提取物的添加可在一定程度上提高盲肠另枝菌属、粪杆菌属、巴恩斯氏菌属和普雷沃氏菌科_UCG-001相对丰度,此结果与Sultan等[40]报道的α-倒捻子素可作为抗菌药物降低有害病原菌的生长结果相似,同时也与柴晓莹等[41]应用葡萄糖氧化酶提高蛋鸡盲肠普雷沃氏菌科_UCG-001相对丰度降低炎症发生的结果相似,说明山竹提取物的添加可通过提高有益菌的生长改善肉鸡肠道健康。

3.4 山竹和木豆提取物对肉鸡肉品质的影响

肉色、pH 和剪切力是评价肌肉感官品质和货架期的重要指标,肌红蛋白含量是决定肌肉肉色的关键因素,其与氧结合后呈鲜红色,未与氧结合呈紫红色,肌肉的抗氧化能力越强,肉色越稳定[42]。本研究结果表明,山竹和木豆提取物的添加显著降低了肉鸡屠宰后45 min和24 h胸肌肉色L*值,升高a*值,此结果与本试验中两者的添加提高了肉鸡抗氧化能力的结果相吻合。滴水损失与肉在运输和冷冻储藏过程的水分流失相关,滴水损失减小,肉在冷冻和运输过程中的水分流失也相应减少[43]。本研究结果表明,山竹和木豆提取物的添加均显著降低了肉鸡胸肌滴水损失,以上说明两者的添加均可通过改善肉鸡胸肌肉色和滴水损失提高肉品质。

4 结论

① 30 mg/kg山竹提取物可提高肉鸡生长性能、抗氧化能力、肠道免疫功能和有益菌群相对丰度,改善肉色、降低滴水损失。
② 300 mg/kg木豆提取物可降低肉鸡血脂含量,提高肠道免疫功能,改善肉色、降低滴水损失。
③ 30 mg/kg山竹提取物可代替抗生素添加于1~40日龄818肉鸡饲粮中。

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