Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (22): 4917-4930.doi: 10.3864/j.issn.0578-1752.2021.22.017

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles    

Effects of Glucose Oxidase on Growth Performance, Immune Functions and Intestinal Health of Ducks Challenged by Escherichia coli

LIU Jiao(),CHEN ZhiMin,ZHENG AiJuan,LIU GuoHua,CAI HuiYi,CHANG WenHuan()   

  1. Feed Research Institute Chinese Academy of Agricultural Sciences/Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Beijing 100081
  • Received:2020-06-09 Accepted:2021-09-27 Online:2021-11-16 Published:2021-11-19
  • Contact: WenHuan CHANG E-mail:853652746@qq.com;changwenhuan@caas.cn

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Abstract:

【Objective】 This experiment was conducted to study the effects of glucose oxidase (GOD) on growth performance, immune functions and intestinal health of ducks challenged by Escherichia coli (E. coli) O88 and its mechanism to find a substitute for antibiotics to prevent duck colibacillosis. 【Method】 A total of 144 one-day-old healthy male lean Peking ducklings were selected and randomly divided into three groups with six replicates and eight ducks per replicate. Ducks were fed the three diets supplemented with nothing (Control group), 30 mg·kg-1virginiamycin (Antibiotic group) and 200 U·kg-1 GOD in basal diet, respectively. On day 7, all ducks were orally taken 0.2 mL E. coli O88 (3×109 CFU/mL) twice, 8 hours apart. The experiment lasted for 28 days. 【Result】The results showed as follows: 1) Compared with the control group, adding antibiotic or GOD in the diet significantly increased the average daily gain and average daily feed intake of 1 to 14-day-old attacking ducks (P<0.01). 2) The number of serum white blood cell of 28-day-old attacking ducks was significantly reduced and the percentage of serum lymphocyte was significantly increased by adding antibiotic and GOD in diet (P<0.01), and the number of serum red blood cell of 28-day- old attacking ducks was significantly reduced by adding GOD in diet (P<0.05). 3) Serum MDA at day 14 (P<0.05) and CAT contents at day 28 (P<0.01) were significantly reduced by adding antibiotic and GOD, serum T-AOC at day 28 was significantly increased (P<0.05), and CAT at day 14 showed a tendency to decrease by adding GOD (P=0.087). 4) Supplementation of antibiotic and GOD significantly decreased endotoxin of 14 and 28-day-old attacking ducks (P<0.01). 5) Adding antibiotic or GOD significantly decreased the concentration of IL-1β and IL-6 of ducks at day 14 and 28 and TNF-α at d 28 (P<0.05), no significant differences between the GOD and the antibiotic groups（P>0.05）. 6) The concentration of serum diamine oxidase and D-lactic of 14 and 28-day-old attacking ducks were significantly reduced by adding antibiotic and GOD (P<0.05), no significant differences between the GOD and the antibiotic groups (P>0.05). 7) The addition of GOD increased the number of unique OTUs in ileum, reduced the contents of E. coli, and increased the relative abundance of probiotics such as Lactobacillus and Bifidobacterium. 【Conclusion】 In conclusion, addition of GOD in the diet can balance the structure of intestinal flora, reduce the production of bacterial endotoxin and decrease oxidative stress, and then maintain the integrity of intestinal mucosa and prevent activation of inflammatory signal pathway by endotoxin and occurrence of inflammatory reaction, so as to improve intestinal health and growth of ducks infected with E. coli. The GOD can be used as antibiotics substitute to prevent or alleviate colibacillosis of ducks.

Key words: glucose oxidase, duck, Escherichia coli challenge, growth performance, immune function, intestinal health

Table 1

Experimental grouping and attack methods"

组别Groups 攻毒剂量Dosage (mL) 菌液接种量 Inoculation quantity(CFU) 攻毒次数Time
T1 LB肉汤培养基 1 - 1
T2 大肠杆菌O88 E.coli O88 0.05 0.5×108 1
T3 大肠杆菌O88 E.coli O88 0.05 0.5×108 2
T4 大肠杆菌O88 E.coli O88 0.10 1×108 1
T5 大肠杆菌O88 E.coli O88 0.10 1×108 2
T6 大肠杆菌O88 E.coli O88 0.20 3×108 1
T7 大肠杆菌O88 E.coli O88 0.20 3×108 2
T8 大肠杆菌O88 E.coli O88 0.60 6×108 1
T9 大肠杆菌O88 E.coli O88 0.60 6×108 2
T10 大肠杆菌O88 E.coli O88 1.00 1×109 1
T11 大肠杆菌O88 E.coli O88 1.00 1×109 2

Table 2

Composition and nutrient levels of basal diets (air- dry basis) %"

项目
Items		 1—14 日龄
1 to 14 days of age (%)		 15—28 日龄
15 to 28 days of age (%)
原料 Ingredients
玉米 Corn 58.90 67.98
豆粕 Soybean meal 32.42 22.78
棉籽粕 Cottonseed meal 2.00 4.00
大豆油 Soybean oil 2.28 1.10
石粉 Limestone 1.07 1.14
磷酸氢钙 CaHPO4 1.71 1.49
食盐 NaCl 0.30 0.30
L-赖氨酸 L-Lys 0.17 0.09
DL-蛋氨酸 DL-Met 0.14 0.12
维生素预混料 Vitamin premix1) 0.03 0.03
微量元素预混料 Microelement premix2) 0.10 0.10
沸石粉 Zeolite powder 0.87 0.87
合计 Total 100.00 100.00
营养水平 Nutrient level
代谢能 ME (MJ·kg-1) 12.14 12.14
粗蛋白质 CP 20.00 17.50
钙 Ca 0.90 0.85
总磷 TP 0.65 0.60
有效磷 AP 0.45 0.40
赖氨酸 Lys 1.10 0.85
蛋氨酸 Met 0.45 0.40
蛋氨酸+半胱氨酸 Met + Cys 0.80 0.71

Fig. 1

Effects of different infecting methods on growth performance of 7 to 14 days of age ducks A: Effects of different infecting methods on average daily gain of 7 to 14 days of age ducks; B: Effects of different infecting methods on feed/gain of 7 to 14 days of age ducks; C: Effects of different infecting methods on mortality of 7 to 14 days of age ducks; T: Treatment"

Table 3

16s rDNA PCR primers used to quantify Escherichia coli"

目标基因
Gene name		 引物序列（5′-3′）
Primers sequence (5′-3′) *
大肠杆菌
Escherichia coli		 F: GTTAATACCTTTGCTCATTGA
R: ACCAGGGTATCTAATCCTGTT

Table 4

Effects of glucose oxidase on growth performance of ducks infected with Escherichia coli (n=6)"

项目
Items		 对照组
Control group		 抗生素组
Antibiotic group		 葡萄糖氧化酶组
Glucose oxidase group		 标准误
SEM		 P值
P-value
1—14 日龄 1 to 14 days of age
14日龄体重BW at 14 days (kg) 0.60 0.62 0.64 0.006 0.178
平均日增重ADG (g) 38.17Bb 41.17Aa 42.27Aa 0.558 <0.001
平均日采食量ADFI (g) 50.21Bb 54.32Aa 54.93Aa 0.691 <0.001
料重比F / G 1.32 1.32 1.30 0.009 0.674
14—28日龄14 to 28 days of age
平均日增重ADG (g) 77.96 80.11 81.26 1.528 0.697
平均日采食量ADFI (g) 137.67 140.62 139.81 1.847 0.817
料重比F / G 1.77 1.76 1.73 0.016 0.614
1—28 日龄 1 to 28 days of age
28日龄体重BW at 28 days (kg) 1.70 1.74 1.77 0.025 0.525
平均日增重ADG (g) 58.94 60.32 61.43 0.889 0.549
平均日采食量ADFI (g) 92.20 94.92 94.36 1.043 0.560
料重比F / G 1.57 1.58 1.54 0.011 0.404

Fig. 2

Effects of glucose oxidase on blood routine of 28 days of age ducks infected by Escherichia coli (n=6) A: The red blood cells content of 28 days of age ducks; B: The white blood cells content of 28 days of age ducks; C: The lymphocyte proportion of 28 days of age ducks; D: The granulocyte absolute value of 28 days of age ducks; CON: control group; ANT: antibiotic group; GOD: glucose oxidase group; RBC: red blood cells; WBC: white blood cells; LYM: lymphocyte; GRA: granulocyte. Value columns with different small letters mean significant difference (P<0.05). The same as below"

Table 5

Effects of glucose oxidase on antioxidant function of ducks infected by Escherichia coli (n=6)"

项目
Items		 对照组
Control group		 抗生素组
Antibiotic group		 葡萄糖氧化酶组
Glucose oxidase group		 标准误
SEM		 P值
P-value
14日龄 14 days of age
总抗氧化能力T-AOC (U·mL-1) 23.27 18.96 20.17 0.875 0.118
总超氧化物歧化酶T-SOD (U·mL-1) 107.02 111.92 114.58 2.199 0.394
丙二醛MDA (nmol·mL-1) 5.79a 5.12b 5.13b 0.117 0.030
过氧化氢酶CAT (U·mL-1) 4.27 4.01 2.94 0.276 0.087
28日龄 28 days of age
总抗氧化能力T-AOC (U·mL-1) 16.68b 19.76ab 22.57a 0.887 0.015
总超氧化物歧化酶T-SOD (U·mL-1) 116.58 117.80 121.43 2.615 0.790
丙二醛MDA (nmol·mL-1) 4.75 4.24 4.65 0.164 0.440
过氧化氢酶CAT (U/·mL-1) 5.14Aa 3.59Bb 2.62Bb 0.335 0.004

Table 6

Effects of different feed additives on serum endotoxin of ducks infected with Escherichia coli EU/mL (n=6)"

项目
Items		 日龄
Age		 对照组
Control group		 抗生素组
Antibiotic group		 葡萄糖氧化酶组
Glucose oxidase group		 标准误
SEM		 P值
P-value
内毒素
Endotoxin		 14 0.628Aa 0.563Bb 0.584Bb 0.009 0.003
28 0.602Aa 0.559Bb 0.553Bb 0.008 0.008

Fig. 3

Effects of glucose oxidase on jejunal cytokine concentration of ducks infected by Escherichia coli (n=6) A: The jejunal TLR4 concentration of ducks; B: The jejunal TNF-α concentration of ducks; C: The jejunal IL-1β concentration of ducks; D: The jejunal IL-6 concentration of ducks; CON: Control group; ANT: Antibiotic group; GOD: Glucose oxidase group; D: Days old"

Fig. 4

Effects of glucose oxidase on intestinal permeability of ducks infected by Escherichia coli (n=6) A: The concentration of DAO in serum of ducks; B: The concentration of D-LA in serum of ducks; CON: Control group; ANT: Antibiotic group; GOD: Glucose oxidase group; DAO: Diamine oxidase; D-LA: D-lactic acid; D: Days old"

Fig. 5

Alpha diversity of ileum microbial community (n=6) A: Simpson index of OUT level; B: Shannon index of OUT level. CON: Control group; GOD: Glucose oxidase group"

Fig. 6

Venn figure of ileum microbial community (n=6) CON: Control group; GOD: Glucose oxidase group"

Fig. 7

Similarity analysis of ileum microbial community (PCoA plot) CON: Control group; GOD: Glucose oxidase group"

Fig. 8

The relative abundance of main taxa in the ileum at different taxonomic levels (n=6) CON: Control group; GOD: Glucose oxidase group"

Fig. 9

Effects of glucose oxidase on ileum Escherichia coli concentration of 14 days of age ducks infected by Escherichia coli (n=6) CON: Control group; GOD: Glucose oxidase group"

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