Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 10: 6335 - 6344

DOI: https://doi.org/10.12418/CJAN2024.539

RESEARCH PAPER

Effects of Melittin on Growth Performance, Antioxidant Capacity and Intestine Health of Broilers

  • HU Yue , 1 ,
  • ZHAN Yuming 2 ,
  • MENG Chunyan 3 ,
  • LI Yupeng 4, 5, 6 ,
  • QIAO Jiayun , 7, * ,
  • LI Haihua , 1, *
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  • 1 Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China
  • 2 Shandong Provincial Key Laboratory of Quality Safety Monitoring and Risk Assessment for Animal Products, Shandong Center for Quality Control of Feed and Veterinary Drug, Ji’nan 250100, China
  • 3 Zhongji Hi-Tech (Beijing) Biotechnology Co., Ltd., Beijing 102200, China
  • 4 Institute of Animal Science and Veterinary, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China
  • 5 Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology, Tianjin 300381, China
  • 6 Tianjin Engineering Research Center of Animal Healthy Farming, Tianjin 300381, China
  • 7 Tianjin Key Laboratory of Conservation and Utilization of Animal Diversity, College of Life Sciences, Tianjin Normal University, Tianjin 300387, China
*QIAO Jiayun, professor, E-mail: ;
LI Haihua, associate professor, E-mail:

Received date: 2024-04-11

  Online published: 2024-10-14

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Abstract

This experiment was conducted to investigate the effects of melittin on growth performance, antioxidant capacity and intestine health of broilers, and to determine the appropriate dosage of melittin. A total of 160 1-day-old 817 broilers were randomly divided into 4 groups with 5 replicates per group and 8 broilers per replicate. Broilers in the control group (CON group) were fed a basal diet, and those in the experimental groups were fed the basal diets supplemented with 0.05 (low-dose group, group A), 0.10 (medium-dose group, group B) and 0.15 g/kg (high-dose group, group C) melittin, respectively. The experiment lasted for 42 days. The results showed as follows: 1) compared with CON group, the average daily feed intake (ADFI) of broilers in groups B and C was significantly increased from 22 to 42 days of age and 1 to 42 days of age (P<0.05). 2) Compared with CON group, at 21 days of age, the serum malondialdehyde (MDA) content of broilers in experimental groups was significantly decreased (P<0.05); at 42 days of age, the serum superoxide dismutase (SOD) activity in group B was significantly increased (P<0.05), and the serum MDA content was significantly decreased (P<0.05). 3) Compared with CON group, at 21 days of age, the serum interleukin-6 (IL-6) content of broilers in groups B and C was significantly decreased (P<0.05); at 42 days of age, the serum contents of tumor necrosis factor-α (TNF-α) and IL-6 in groups B and C were significantly decreased (P<0.05). 4) Compared with CON group, at 21 days of age, the duodenal villus height of broilers in groups B and C were significantly increased (P<0.05), the jejunal villus height and villus height to crypt depth ratio (V/C) in group B were significantly increased (P<0.05), and the ileal crypt depth in groups B and C were significantly decreased and V/C was significantly increased (P<0.05); at 42 days of age, the jejunal villus height in group B was significantly increased (P<0.05). 5) Compared with CON group, at 21 days of age, the jejunal trypsin activity of broilers in groups B and C was significantly increased (P<0.05); at 42 days of age, the activities of jejunal lipase and trypsin in groups B and C were significantly increased (P<0.05). 6) Compared with CON group, at 21 days of age, the jejunal zonula occludens-1 (ZO-1) mRNA relative expression level of broilers in experimental groups was significantly increased (P<0.05), and the jejunal occludin (OCLN) mRNA relative expression level in groups B and C was significantly increased (P<0.05); at 42 days of age, the jejunal OCLN mRNA relative expression level in experimental groups was significantly increased (P<0.05). In conclusion, dietary melittin can improve the antioxidant capacity and anti-inflammatory capacity of broilers, promote intestinal morphological development, improve intestinal digestive enzyme activity, maintain intestinal barrier function, and thus improve their growth performance, and the feeding effect is better when the dosage of melittin is 0.10 g/kg.

Key words： melittin; broilers; growth performance; antioxidant capacity; intestinal morphology and structure

Cite this article

HU Yue , ZHAN Yuming , MENG Chunyan , LI Yupeng , QIAO Jiayun , LI Haihua . Effects of Melittin on Growth Performance, Antioxidant Capacity and Intestine Health of Broilers[J]. Chinese Journal of Animal Nutrition, 2024 , 36(10) : 6335 -6344 . DOI: 10.12418/CJAN2024.539

在畜禽生产中合理使用抗生素对各种疾病的防治具有重要意义,但滥用抗生素会严重破坏肠道菌群,导致免疫系统发育迟缓和免疫功能障碍[1]。此外,过度使用抗生素会促进细菌的耐药性,从而对动物和人类健康产生负面影响。抗生素的残留和耐药性已成为一个日益严重的问题,我国自2020年7月1日起,根据农业农村部194号公告,饲料中全面禁止添加抗生素。因此,寻找绿色、安全的抗生素替代品已成为当下的研究热点。抗菌肽存在于生物体的组织和细胞中,具有广谱抗菌作用,是先天免疫系统的重要分子。抗菌肽通过与病原菌膜性结构吸附、融合,改变其通透性,从而导致其内容物外泄而致其死亡。由于其抗菌机制与抗生素不同,可降低细菌耐药性的风险,抗菌肽被认为是替代抗生素的理想选择[2]。近年来,抗菌肽在猪和家禽生产中被广泛用作新的抗菌剂[3]。研究表明,复合抗菌肽在提高断奶仔猪生长性能和养分表观消化率、改善粪便菌群以及降低腹泻率等方面具有明显优势,可作为抗生素生长促进剂的潜在替代品[4]。还有研究表明,饲粮添加抗菌肽A3和P5可提高肉鸡生长性能,其效果与抗生素相似[5]。此外,饮用水添加抗菌肽也可以提高肉鸡的生长性能[6]。
蜂毒素(melittin)是从蜜蜂毒素中分离出来的一种抗菌肽,拥有独特的结构,富含活性成分,具有抗菌、抗癌和抗炎等多种药学特性[7]。研究表明,在种蛋中注射蜂毒素可提高其孵化率,增强雏鸡的免疫性能[8]。还有研究表明,蜂毒素可调节家兔体内生化指标及抗氧化能力,改善繁殖性能、免疫机能和健康状况[9]。目前,尽管已有不少关于抗菌肽在畜禽饲粮中添加应用的研究报道,但是蜜蜂源抗菌肽(蜂毒素)对肉鸡生长发育的影响却知之甚少。因此,本研究通过在饲粮中添加不同剂量的蜂毒素来探究其对肉鸡生长性能、抗氧化能力和抗炎能力以及肠道形态发育、消化能力和屏障功能的影响,并确定其适宜的添加水平,为蜂毒素在肉鸡饲粮中的应用提供理论依据。

1 材料与方法

1.1 试验设计和饲养管理

选取1日龄817杂交肉鸡160只,随机分成4个组,每组5个重复,每个重复8只鸡。对照组(CON组)饲喂基础饲粮;试验组分别在饲喂基础饲粮的基础上添加0.05(低剂量组,A组)、0.10(中剂量组,B组)和0.15 g/kg(高剂量组,C组)的蜂毒素。蜂毒素为市售产品,主要成分为蜂毒肽,高效液相色谱法测定其含量≥98%);蜂毒素添加量参考Xing等[10]并有所改进。基础饲粮以豆粕和玉米为主要原料,参考《鸡饲养标准》(NY/T 33—2004)配制而成,其组成及营养水平见表1。试验期42 d。本试验通过天津师范大学伦理委员会审查批准(伦理批准编号:2024051401)。
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		 22~42日龄
22 to 42
days of age
原料 Ingredients
玉米 Corn 54.55 55.56
豆粕 Soybean meal 38.00 35.50
大豆油 Soybean oil 3.20 5.00
磷酸氢钙 CaHPO4 1.40 1.25
石粉 Limestone 1.18 1.11
氯化钠 NaCl 0.30 0.30
DL-蛋氨酸 DL-Met 0.24 0.20
L-赖氨酸 L-Lys 0.13 0.08
预混料 Premix1) 1.00 1.00
合计 Total 100.00 100.00
营养水平 Nutrient levels2)
代谢能 ME/(MJ/kg) 12.28 12.79
粗蛋白质 CP 20.62 19.64
钙 Ca 0.87 0.80
总磷 TP 0.62 0.58
有效磷 AP 0.38 0.35
赖氨酸 Lys 1.26 1.16
苏氨酸 Thr 0.81 0.76
蛋氨酸+胱氨酸 Met+Cys 0.91 0.84

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 5 000 IU,VE 30 IU,VK 3 mg,VB1 2 mg,VB2 8 mg,VB6 3 mg,VB12 10 μg,Fe 40 mg,Zn 100 mg,Mn 100 mg,Cu 15 mg,Se 0.35 mg,I 1 mg,烟酸 nicotinic acid 30 mg,泛酸 pantothenic acid 12 mg,叶酸 folic acid 0.7 mg,生物素 biotin 0.12 mg。

2)代谢能和有效磷为计算值,参考《中国饲料成分及营养价值表(2022年第33版)》计算;粗蛋白质、钙、总磷和氨基酸为实测值,分别参考国家标准GB/T 6432—2018、GB/T 6436—2018、GB/T 6437—2018和GB/T 18246—2019进行测定。ME and AP were calculated values referred to the Tables of Feed Composition and Nutritive Values in China (33rd edition, 2022); CP, Ca, TP and amino acids were measured values referred to national standards of GB/T 6432—2018, GB/T 6436—2018, GB/T 6437—2018 and GB/T 18246—2019, respectively.

本试验在山东某肉鸡场进行,采用3层重叠笼养方式饲养。试验开始第1周温度控制在(33±1) ℃,随后逐步降温至(25±1) ℃;试验期间24 h光照,试验鸡常规免疫,自由采食和饮水。

1.2 样品采集

分别于试验第21和42天,每个重复随机抽取1只鸡,颈静脉采血并分离血清,置于-80 ℃保存,用于后续相关指标的检测。试验鸡采血后屠宰,采集肠道样品,用生理盐水浸洗后,一部分固定在4%多聚甲醛固定液中用于组织形态结构观察;另一部分液氮速冻后置于-80 ℃中保存,用于消化酶活性测定及实时荧光定量PCR的检测。

1.3 测定指标及方法

1.3.1 生长性能测定

分别于试验第1、21和42天早上空腹(前1天晚上断粮)称量鸡的体重和剩余饲粮重,计算平均日增重(ADG)、平均日采食量(ADFI)和料重比(F/G)。

1.3.2 血清抗氧化和炎性指标测定

采用试剂盒(南京建成生物工程研究所)测定血清超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性以及丙二醛(MDA)含量;并采用酶联免疫吸附测定(ELISA)试剂盒(美国BD公司)测定血清肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)含量。

1.3.3 肠道形态结构观察

将固定好的肠道组织样品进行脱水、石腊包埋处理,并以5 μm进行切片,苏木素-伊红染色后,采用中性树胶封固,使用Revolve正倒置一体显微镜观察切片,测量肠道绒毛高度和隐窝深度,并计算绒隐比(绒毛高度/隐窝深度)。

1.3.4 空肠消化酶活性测定

按照南京建成生物工程研究所试剂盒说明书,将冻存的肠道样品进行研磨,制备匀浆液,离心分离上清液后测定淀粉酶、胰蛋白酶和脂肪酶活性。

1.3.5 空肠内紧密连接蛋白mRNA相对表达量测定

根据NCBI数据库中肉鸡闭锁小带蛋白-1(ZO-1)、闭合蛋白(occludin,OCLN)和甘油醛-3-磷酸脱氢酶(GAPDH)的mRNA序列设计引物,引物序列信息见表2。参照刘雪姣等[11]的方法,按照试剂盒说明书对组织进行RNA提取、RNA反转录、反应体系的制备和实时荧光定量PCR。以GAPDH为内参基因,通过2-△△Ct方法计算ZO-1和OCLN的mRNA相对表达量。
表2 引物序列信息

Table 2 Primer sequences information

基因
Genes		 引物序列
Primer sequences (5'—3')		 产物大小
Product
size/bp		 退火温度
Annealing
temperature/℃		 GenBank
序列号
GenBank No.
闭锁小带蛋白-1
ZO-1		 F:TAAAGCCATTCCTGTAAGCC
R:GTTTCACCTTTCTCTTTGTCC		 243 62 XM_040706827
闭合蛋白
OCLN		 F:TCATCGCCTCCATCGTCTAC
R:TCTTACTGCGCGTCTTCTGG		 240 62 NM_205128
甘油醛-3-磷酸脱氢酶
GAPDH		 F:CCCCCATGTTTGTGATGGGT
R:TGATGGCATGGACAGTGGTC		 162 60 NM_204305

F为上游引物,R为下游引物。

F was forward primer, and R was reverse primer.

1.4 数据统计与分析

试验数据采用Excel进行整理,然后利用SPSS 20.0软件进行单因素方差分析(one-way ANOVA)和LSD多重比较,结果数据以平均值和均值标准误(SEM)表示,统计显著水平为P<0.05。

2 结果

2.1 蜂毒素对肉鸡生长性能的影响

由表3可知,22~42日龄和1~42日龄,与CON组相比,B组和C组肉鸡ADFI显著提高(P<0.05),A组ADFI无显著差异(P>0.05);与A组相比,B组ADFI显著提高(P<0.05)。
表3 蜂毒素对肉鸡生长性能的影响

Table 3 Effects of melittin on growth performance of broilers

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
CON A B C



1~21日龄
1 to 21 days of age		 末重 FBW/g 572.82 575.80 593.17 589.34 3.53 0.085
平均日增重 ADG/g 24.97 25.26 27.76 26.32 0.42 0.054
平均日采食量 ADFI/g 37.64 38.31 39.60 39.12 0.29 0.053
料重比 F/G 1.42 1.43 1.48 1.43 0.02 0.677



22~42日龄
22 to 42 days of age		 末重 FBW/g 1 803.07 1 808.14 1 815.63 1 811.72 1.89 0.087
平均日增重 ADG/g 59.14 59.48 60.93 60.04 0.29 0.148
平均日采食量 ADFI/g 104.77c 105.23bc 106.85a 106.54ab 0.32 0.027
料重比 F/G 1.72 1.74 1.76 1.74 0.01 0.364



1~42日龄
1 to 42 days of age		 平均日增重 ADG/g 42.76 43.28 44.01 43.82 0.20 0.091
平均日采食量 ADFI/g 71.96c 72.81bc 73.78a 73.52ab 0.24 0.015
料重比 F/G 1.64 1.65 1.66 1.65 0.01 0.956

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。

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

2.2 蜂毒素对肉鸡血清抗氧化指标的影响

由表4可知,21日龄时,与CON组相比,各试验组肉鸡血清MDA含量均显著降低(P<0.05),且各试验组之间无显著差异(P>0.05)。42日龄时,与CON组相比,B组肉鸡血清SOD活性显著提高(P<0.05),血清MDA含量显著降低(P<0.05);A组和C组血清SOD活性和MDA含量均无显著差异(P>0.05)。
表4 蜂毒素对肉鸡血清抗氧化指标的影响

Table 4 Effects of melittin on serum antioxidant indices of broilers

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
CON A B C
21日龄 21 days of age
超氧化物歧化酶 SOD/(U/mL) 72.64 75.14 79.99 81.07 1.47 0.133
谷胱甘肽过氧化物酶 GSH-Px/(U/mL) 788.04 803.20 835.72 830.97 7.70 0.074
丙二醛 MDA/(nmol/mL) 3.13a 2.91b 2.81b 2.88b 0.04 0.033
42日龄 42 days of age
超氧化物歧化酶 SOD/(U/mL) 96.19b 100.20ab 107.01a 104.24ab 1.51 0.046
谷胱甘肽过氧化物酶 GSH-Px/(U/mL) 879.66 921.93 938.27 930.09 10.09 0.169
丙二醛 MDA/(nmol/mL) 3.73a 3.43ab 3.23b 3.41ab 0.06 0.029

2.3 蜂毒素对肉鸡血清炎性指标的影响

由表5可知,21日龄时,与CON组相比,B组和C组肉鸡血清IL-6含量显著降低(P<0.05),A组血清IL-6含量无显著差异(P>0.05)。42日龄时,与CON组相比,B组和C组肉鸡血清TNF-α和IL-6含量均显著降低(P<0.05),A组血清TNF-α和IL-6含量无显著差异(P>0.05)。
表5 蜂毒素对肉鸡血清炎性指标的影响

Table 5 Effects of melittin on serum inflammatory indices of broilerspg/mL

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
CON A B C
21日龄 21 days of age
肿瘤坏死因子-α TNF-α 71.96 70.51 64.57 65.73 1.43 0.194
白细胞介素-6 IL-6 44.51a 39.53ab 31.31c 33.01bc 1.61 0.023
42日龄 42 days of age
肿瘤坏死因子-α TNF-α 72.76a 68.91ab 64.57b 63.60b 1.32 0.037
白细胞介素-6 IL-6 42.08a 37.12ab 30.83c 32.25bc 1.29 0.013

2.4 蜂毒素对肉鸡肠道形态结构的影响

由表6可知,21日龄时,与CON组相比,B组和C组肉鸡十二指肠绒毛高度显著提高(P<0.05),B组空肠绒毛高度和V/C值显著提高(P<0.05),B组和C组回肠隐窝深度显著降低而V/C值显著提高(P<0.05)。42日龄时,与CON组相比,B组肉鸡空肠绒毛高度显著提高(P<0.05),A组和C组空肠绒毛高度无显著差异(P>0.05)。
表6 蜂毒素对肉鸡肠道形态结构的影响

Table 6 Effects of melittin on intestinal morphology and structure of broilers

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
CON A B C
21日龄 21 days of age
十二指肠 Duodenum
绒毛高度 Villus height/μm 1 282.22c 1 323.30bc 1 410.43ab 1 495.21a 23.71 0.001
隐窝深度 Crypt depth/μm 134.80 134.00 125.77 128.85 6.26 0.958
绒隐比 V/C 10.10 10.51 11.55 11.96 0.58 0.678
空肠 Jejunum
绒毛高度 Villus height/μm 879.23b 912.21b 1 063.90a 982.28ab 25.59 0.037
隐窝深度 Crypt depth/μm 86.20 79.04 67.68 73.43 3.30 0.239
绒隐比 V/C 10.49b 11.65b 15.79a 14.25ab 0.76 0.040
回肠 Ileum
绒毛高度 Villus height/μm 572.31 612.81 653.60 625.41 13.69 0.210
隐窝深度 Crypt depth/μm 62.68a 57.16ab 51.03b 53.68b 1.57 0.037
绒隐比 V/C 9.16b 10.75ab 13.15a 11.76a 0.49 0.020
42日龄 42 days of age
十二指肠 Duodenum
绒毛高度 Villus height/μm 1 353.32 1 403.28 1 477.24 1 503.46 23.78 0.087
隐窝深度 Crypt depth/μm 147.09 140.65 136.76 135.02 4.04 0.758
绒隐比 V/C 9.38 10.11 10.94 11.29 0.37 0.272
空肠 Jejunum
绒毛高度 Villus height/μm 957.38b 1 031.61ab 1 228.33a 1 126.73ab 37.54 0.045
隐窝深度 Crypt depth/μm 93.52 90.85 87.42 89.28 2.19 0.819
绒隐比 V/C 10.46 11.56 13.43 12.09 0.48 0.175
回肠 Ileum
绒毛高度 Villus height/μm 715.06 752.65 786.14 767.22 12.77 0.253
隐窝深度 Crypt depth/μm 66.83 64.09 55.83 56.84 2.02 0.144
绒隐比 V/C 10.90 11.79 14.28 13.89 0.53 0.061

2.5 蜂毒素对肉鸡空肠消化酶活性的影响

由表7可知,21日龄时,与CON组相比,B组和C组肉鸡空肠胰蛋白酶活性显著提高(P<0.05),A组空肠胰蛋白酶活性无显著差异(P>0.05)。42日龄时,与CON组相比,B组和C组肉鸡空肠脂肪酶和胰蛋白酶活性均显著提高(P<0.05),A组空肠脂肪酶和胰蛋白酶活性无显著差异(P>0.05);此外,B组和C组之间空肠脂肪酶和胰蛋白酶活性无显著差异(P>0.05)。
表7 蜂毒素对肉鸡空肠消化酶活性的影响

Table 7 Effects of melittin on digestive enzyme activities in jejunum of broilers

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
CON A B C
21日龄 21 days of age
淀粉酶 AMS/(U/mg prot) 0.97 1.12 1.23 1.20 0.40 0.077
脂肪酶 LPS/(U/g prot) 22.94 24.86 28.05 27.32 0.76 0.054
胰蛋白酶 Trypsin/(U/mg prot) 1 867.23c 1 925.06bc 2 118.39ab 2 178.80a 42.94 0.013
42日龄 42 days of age
淀粉酶 AMS/(U/mg prot) 1.22 1.32 1.43 1.40 0.03 0.071
脂肪酶 LPS/(U/g prot) 83.12b 94.23ab 110.09a 107.47a 3.45 0.017
胰蛋白酶 Trypsin/(U/mg prot) 3 016.62b 3 865.61ab 4 046.19a 4 499.10a 213.24 0.032

2.6 蜂毒素对肉鸡空肠紧密连接蛋白mRNA相对表达量的影响

由表8可知,21日龄时,与CON组相比,各试验组肉鸡空肠ZO-1 mRNA相对表达量均显著提高(P<0.05),且各试验组之间无显著差异(P>0.05);B组和C组空肠OCLN mRNA相对表达量显著提高(P<0.05)。42日龄时,与CON组相比,各试验组肉鸡空肠OCLN mRNA相对表达量均显著提高(P<0.05),且各试验组之间无显著差异(P>0.05)。
表8 蜂毒素对肉鸡空肠紧密连接蛋白mRNA相对表达量的影响

Table 8 Effects of melittin on mRNA relative expression levels of tight junction proteins in jejunum of broilers

项目
Items		 组别 Groups 均值标准误
SEM		 P值
P-value
CON A B C
21日龄 21 days of age
闭锁小带蛋白-1 ZO-1 1.05b 1.26a 1.39a 1.36a 0.04 0.012
闭合蛋白 OCLN 1.07b 1.23ab 1.34a 1.28a 0.03 0.017
42日龄 42 days of age
闭锁小带蛋白-1 ZO-1 1.08 1.30 1.41 1.38 0.05 0.071
闭合蛋白 OCLN 1.08b 1.26a 1.37a 1.31a 0.03 0.010

3 讨论

3.1 蜂毒素对肉鸡生长性能的影响

抗菌肽在先天免疫中发挥着重要作用,并由于其独特的抗菌作用机制,能够降低耐药性产生的倾向[12],已被用作预防疾病的候选抗菌药物。研究表明,给断奶仔猪饲喂抗菌肽,可提高其ADG,并降低仔猪腹泻率[13]。还有研究表明,饲喂蜂毒素可以提高鹌鹑的采食量和产蛋率[14]。本研究发现,饲粮添加蜂毒素能在一定程度上提高肉鸡末重和采食量,促进肉鸡的生长,且中剂量组(B组)饲粮添加0.10 g/kg蜂毒素提高肉鸡生长性能的效果较为明显。

3.2 蜂毒素对肉鸡抗氧化能力和抗炎能力的影响

活性氧(ROS)自由基是生物体内正常细胞代谢的副产物[15]。正常情况下,机体的抗氧化系统可以消除ROS,但当无法消除多余的ROS时,ROS就会氧化体内的核酸、蛋白质和类脂体,出现氧化应激导致的细胞损伤[16]。机体内的抗氧化酶包括SOD和GSH-Px等可以清除ROS,因此,SOD和GSH-Px常用于评价机体抗氧化能力的强弱[16]。MDA是脂质氧化的终产物,可影响线粒体呼吸链复合物及线粒体内关键酶活性,加剧膜损伤。因此,检测MDA的含量可反映机体脂质过氧化的程度,并间接地反映细胞损伤的程度[17]。Liu等[18]研究发现,饲喂抗菌肽可提高断奶仔猪血清SOD和GSH-Px活性,改善仔猪的生长性能、抗氧化能力和免疫功能。本研究与前人研究结果相似,中剂量组肉鸡血清SOD活性显著提高,血清MDA含量显著降低,表明饲喂蜂毒素可促进抗氧化酶的产生,降低机体氧化应激反应,增强肉鸡的抗氧化能力。
细胞因子不仅在免疫和非免疫细胞的增殖和分化中发挥作用,还在调节肠道炎症中发挥重要作用[19]。若促炎细胞因子(如IL-6和TNF-α)在畜禽体内表达异常,会增加肠上皮的通透性,引起肠道病理损伤和免疫功能下降,并导致多种炎症级联反应[20]。因此,免疫能力的增强表现为促炎细胞因子IL-6和TNF-α含量的降低[21]。研究表明,蜂毒素可降低葡聚糖硫酸钠(DSS)诱导的结肠炎模型小鼠体内炎性因子的含量,从而缓解溃疡性结肠炎的症状[22]。本试验结果表明,饲粮添加0.10 g/kg蜂毒素可降低肉鸡血清IL-6和TNF-α含量,表明蜂毒素可提高肉鸡的抗炎能力,从而保护肉鸡肠道健康免受炎症损伤。

3.3 蜂毒素对肉鸡肠道健康的影响

肠道是动物最大的消化吸收器官,也是最重要的免疫器官。肉鸡的生长性能与肠道健康密切相关。肠道形态结构完整性、消化酶活性和紧密连接蛋白水平等常用于评价肠道健康情况[23]。据报道,肠道绒毛高度、隐窝深度和V/C值是衡量小肠消化和吸收功能的重要指标[24]。肠道绒毛高度提高会促进肠道对营养物质的吸收;隐窝深度代表肠道上皮细胞的生成率,隐窝越浅表明细胞成熟度越好,肠上皮组织分泌消化液的功能就越好,反之,隐窝越深其能力就越弱;V/C值在一定程度上可以反映肠道吸收功能的强弱,其值越高表示肠道消化吸收能力越强[25]。Zhang等[26]研究发现,饲粮添加抗菌肽可以改善肉鸡生长性能和免疫功能,并提高肠绒毛高度,改善肠道健康。本试验中,饲粮添加0.10 g/kg蜂毒素能够提高肉鸡小肠绒毛高度和V/C值,并降低隐窝深度,表明蜂毒素在一定程度上可以促进肠道形态发育,提高肉鸡对营养物质的消化吸收能力。
消化酶活性的高低对营养物质消化和动物生长具有重要意义[27]。消化酶将大分子营养物质水解成更容易消化的小分子物质,有助于肠道更有效地吸收利用营养成分,从而提高饲料利用率,改善动物的生长性能[28]。淀粉酶是将淀粉和多糖水解为二糖和低聚糖的关键酶[29];蛋白质和肽链可被胰蛋白酶水解成易于吸收的多肽和氨基酸[30];甘油单酯和游离脂肪酸可以被生物体有效吸收,并通过脂肪酶从脂质中分解[31]。研究发现,饲粮添加抗菌肽可提高肉鸡肠道糜蛋白酶、胃蛋白酶和脂肪酶活性,有利于肉鸡生长性能的提高[32]。本研究中,饲粮添加0.10 g/kg蜂毒素可显著提高肉鸡空肠胰蛋白酶和脂肪酶活性,有助于增加肠道的代谢吸收能力,促进肉鸡生长发育。
肠道屏障的完整性主要由上皮细胞紧密连接支持,是维护肠道健康和稳态的基础。紧密连接蛋白包括细胞内支架蛋白和跨膜蛋白,如密封蛋白(claudin)、OCLN和ZO-1,若紧密连接蛋白破坏会导致细胞间通透性增加,进而导致疾病的发生[33]。Xie等[34]研究发现,抗菌肽与植物精油联合添加到肉鸡饲粮中,可显著提高肉鸡生长性能,并显著提高肠道ZO-1的mRNA相对表达量。Daneshmand等[35]研究发现,饲粮添加重组抗菌肽可改善肉鸡肠道损伤,降低死亡率,并提高空肠紧密连接相关蛋白claudin-1和OCLN的mRNA相对表达量。本研究发现,饲粮添加0.10 g/kg蜂毒素能够上调肉鸡空肠中OCLN和ZO-1的mRNA相对表达量,结合本研究中蜂毒素对肠道形态发育的积极作用,表明中剂量蜂毒素在改善肉鸡肠道屏障功能和完整性方面具有较佳效果。

4 结论

饲粮添加蜂毒素可以提高肉鸡抗氧化能力和抗炎能力,促进肠道形态发育,提高肠道消化酶活性,维持肠道屏障功能,从而提高其生长性能,且当蜂毒素添加量为0.10 g/kg时饲喂效果较佳。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
NOVOA RAMA E, BAILEY M, KUMAR S, et al. Characterizing the gut microbiome of broilers raised under conventional and no antibiotics ever practices[J]. Poultry Science, 2023, 102(8):102832.

[2]
WANG J J, DOU X J, SONG J, et al. Antimicrobial peptides:promising alternatives in the post feeding antibiotic era[J]. Medicinal Research Reviews, 2019, 39(3):831-859.

[3]
RODRIGUES G, SOUZA SANTOS L, FRANCO O L. Antimicrobial peptides controlling resistant bacteria in animal production[J]. Frontiers in Microbiology, 2022,13:874153.

[4]
SHI J K, ZHANG P, XU M M, et al. Effects of composite antimicrobial peptide on growth performance and health in weaned piglets[J]. Animal Science Journal, 2018, 89(2):397-403.

DOI PMID

[5]
YOON J H, INGALE S L, KIM J S, et al. Effects of dietary supplementation of synthetic antimicrobial peptide-A3 and P5 on growth performance,apparent total tract digestibility of nutrients,fecal and intestinal microflora and intestinal morphology in weanling pigs[J]. Livestock Science, 2014,159:53-60.

[6]
BAO H, SHE R, LIU T, et al. Effects of pig antibacterial peptides on growth performance and intestine mucosal immune of broiler chickens[J]. Poultry Science, 2009, 88(2):291-297.

DOI PMID

[7]
WEHBE R, FRANGIEH J, RIMA M, et al. Bee venom:overview of main compounds and bioactivities for therapeutic interests[J]. Molecules, 2019, 24(16):2997.

[8]
KHALIL M H, HASSAN S S, SOLIMAN F N K, et al. In-ovo injection of melittin into Alexandria chicken eggs:a way for early immune acceleration[J]. Animal Biotechnology, 2023, 34(8):4060-4068.

[9]
ELKOMY A, EL-HANOUN A, ABDELLA M, et al. Improving the reproductive,immunity and health status of rabbit does using honey bee venom[J]. Journal of Animal Physiology and Animal Nutrition, 2021, 105(5):975-983.

[10]
XING X, ZHANG X J, FAN J Y, et al. Neuroprotective effects of melittin against cerebral ischemia and inflammatory injury via upregulation of MCPIP1 to suppress NF-κB activation in vivo and in vitro[J]. Neurochemical Research, 2024, 49(2):348-362.

[11]
刘雪姣, 李海花, 王怡梦, 等. 沙门菌感染小鼠氧化应激模型的建立及其分子机制研究[J]. 中国畜牧兽医, 2021, 48(10):3834-3844.

DOI

LIU X J, LI H H, WANG Y M, et al. Establishment of oxidative stress model in mice infected with Salmonella and its molecular mechanism[J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(10):3834-3844. (in Chinese)

[12]
MAGANA M, PUSHPANATHAN M, SANTOS A L, et al. The value of antimicrobial peptides in the age of resistance[J]. The Lancet Infectious Diseases, 2020, 20(9):e216-e230.

[13]
XIONG X, YANG H S, LI L, et al. Effects of antimicrobial peptides in nursery diets on growth performance of pigs reared on five different farms[J]. Livestock Science, 2014,167:206-210.

[14]
LI Z L, LIU R X, WANG X H, et al. Effects of melittin on laying performance and intestinal barrier function of quails[J]. Poultry Science, 2023, 102(2):102355.

[15]
JOMOVA K, RAPTOVA R, ALOMAR S Y, et al. Reactive oxygen species,toxicity,oxidative stress,and antioxidants:chronic diseases and aging[J]. Archives of Toxicology, 2023, 97(10):2499-2574.

[16]
MAS-BARGUES C, ESCRIVÁ C, DROMANT M, et al. Lipid peroxidation as measured by chromatographic determination of malondialdehyde.Human plasma reference values in health and disease[J]. Archives of Biochemistry and Biophysics, 2021,709:108941.

[17]
ZHAO Z L, XU X G, CHANG Y C, et al. Protective effect of mussel polysaccharide on cyclophosphamide-induced intestinal oxidative stress injury via Nrf2-Keap1 signaling pathway[J]. Food Science & Nutrition, 2023, 11(7):4233-4245.

[18]
LIU N, MA X K, JIANG X R. Effects of immobilized antimicrobial peptides on growth performance,serum biochemical index,inflammatory factors,intestinal morphology,and microbial community in weaning pigs[J]. Frontiers in Immunology, 2022,13:872990.

[19]
LIU H W, MENG H L, DU M M, et al. Chlorogenic acid ameliorates intestinal inflammation by inhibiting NF-κB and endoplasmic reticulum stress in lipopolysaccharide-challenged broilers[J]. Poultry Science, 2024, 103(5):103586.

[20]
PARK I, OH S, NAM H, et al. Antimicrobial activity of sophorolipids against Eimeria maxima and Clostridium perfringens,and their effect on growth performance and gut health in necrotic enteritis[J]. Poultry Science, 2022, 101(4):101731.

[21]
CHANG S Y, LEE J H, OH H J, et al. Effect of different ratios of phytogenic feed additives on growth performance,nutrient digestibility,intestinal barrier integrity,and immune response in weaned pigs challenged with a pathogenic Escherichia coli[J]. Journal of Animal Science, 2023,101:skad148.

[22]
YAGHOUBI A, AMEL JAMEHDAR S, REZA AKBARI EIDGAHI M, et al. Evaluation of the therapeutic effect of melittin peptide on the ulcerative colitis mouse model[J]. International Immunopharmacology, 2022,108:108810.

[23]
STAMILLA A, MESSINA A, SALLEMI S, et al. Effects of microencapsulated blends of organics acids (OA) and essential oils (EO) as a feed additive for broiler chicken.A focus on growth performance,gut morphology and microbiology[J]. Animals, 2020, 10(3):442.

[24]
GUO S C, LIU L L, LEI J X, et al. Modulation of intestinal morphology and microbiota by dietary Macleaya cordata extract supplementation in Xuefeng black-boned chicken[J]. Animal, 2021, 15(12):100399.

[25]
李文菁, 魏凯敏, 张少秋, 等. 核桃青皮及其提取物对黄羽肉仔鸡肠道形态、黏膜抗氧化性能及微生物多样性的影响[J]. 中国畜牧兽医, 2021, 48(6):2056-2065.

DOI

LI W J, WEI K M, ZHANG S Q, et al. Effects of walnut green husk and its extract on intestinal morphology,mucosal antioxidant activity and microbial diversity of yellow-feather broilers[J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(6):2056-2065. (in Chinese)

[26]
ZHANG X H, ZHAO Q Q, WEN L J, et al. The effect of the antimicrobial peptide plectasin on the growth performance,intestinal health,and immune function of yellow-feathered chickens[J]. Frontiers in Veterinary Science, 2021,8:688611.

[27]
蔡元丽, 张维铭, 宋志刚. 溶血卵磷脂对肉鸡生长性能、肠道消化酶活性和抗氧化能力的影响[J]. 动物营养学报, 2021, 33(6):3210-3217.

DOI

CAI Y L, ZHANG W M, SONG Z G. Effects of lysophosphatidylcholine on growth performance,intestinal digestive enzyme activity and antioxidant ability of broilers[J]. Chinese Journal of Animal Nutrition, 2021, 33(6):3210-3217. (in Chinese)

[28]
OMAR A E, AL-KHALAIFAH H S, ISMAIL T A, et al. Performance,serum biochemical and immunological parameters,and digestive enzyme and intestinal barrier-related gene expression of broiler chickens fed fermented fava bean by-products as a substitute for conventional feed[J]. Frontiers in Veterinary Science, 2021,8:696841.

[29]
GALLEGO-LOBILLO P, FERREIRA-LAZARTE A, HERNÁNDEZ-HERNÁNDEZ O, et al. In vitro digestion of polysaccharides:InfoGest protocol and use of small intestinal extract from rat[J]. Food Research International, 2021,140:110054.

[30]
PATIL U, SAETANG J, ZHANG B, et al. Use of tuna visceral pepsin in combination with trypsin as digestion aid: enhanced protein hydrolysis and bioavailability[J]. Foods, 2022, 12(1):125.

[31]
KOZAN D W, DERRICK J T, LUDINGTON W B, et al. From worms to humans:understanding intestinal lipid metabolism via model organisms[J]. Biochimica et Biophysica Acta:Molecular and Cell Biology of Lipids, 2023, 1868(4):159290.

[32]
WANG F, CHEN J S, YIN Y X, et al. The effects of dietary ellagic acid supplementation on growth performance,immune response,antioxidant activity,digestive enzyme activities,and intestinal functions in yellow-feathered broilers[J]. Journal of Animal Science, 2022, 100(12):skac301.

[33]
CHU T J, YU R Y, GU Y P, et al. Kaempferol protects gut-vascular barrier from high glucose-induced disorder via NF-κB pathway[J]. The Journal of Nutritional Biochem, 2023,123:109496.

[34]
XIE Z, ZHAO Q Q, WANG H, et al. Effects of antibacterial peptide combinations on growth performance,intestinal health,and immune function of broiler chickens[J]. Poultry Science, 2020, 99(12):6481-6492.

[35]
DANESHMAND A, KERMANSHAHI H, SEKHAVATI M H, et al. Effects of cLFchimera peptide on intestinal morphology,integrity,microbiota,and immune cells in broiler chickens challenged with necrotic enteritis[J]. Scientific Reports, 2020, 10(1):17704.

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