Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 1: 603 - 610

DOI: https://doi.org/10.3969/j.issn.1006-267x.2023.01.057

RESEARCH PAPER

Effects of Glucose Oxidase on Growth Performance, Nutrient Apparent Digestibility and Intestinal Morphology of Growing Female Minks

  • WANG Luyi ,
  • YU Xiaojun ,
  • GUO Zhihao ,
  • LI Yalin ,
  • ZHEN Shibo ,
  • WANG Guang ,
  • WANG Lihua , *
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  • College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
* professor, E-mail:

Received date: 2022-06-01

  Online published: 2023-01-11

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Abstract

The present experiment was conducted to determine the effects of glucose oxidase (GOD) on growth performance, nutrient apparent digestibility and intestinal morphology of growing female minks. A total of 150 red eye white minks (female minks) at 75 days of age were selected and randomly divided into 5 groups with 10 replicates in each group and 3 minks in each replicate. Minks in five groups were fed basal diets supplemented with 0 (control group), 50, 100, 200 and 400 U/kg GOD, respectively. The pre-experimental period lasted for 1 week, and the experimental period lasted for 8 weeks. The experimental period was divided into early stage (1 to 4 weeks) and later stage (5 to 8 weeks). The results showed as follows: 1) compared with the control group, during early stage of the experiment, the final body weight, average daily gain and average daily feed intake of minks of 100 U/kg GOD supplementation group were significantly increased (P<0.05), and the ration of feed to gain was significantly decreased (P<0.05); during later stage of the experiment, the final body weight of minks of 50, 100 and 400 U/kg GOD supplementation groups was significantly increased (P<0.05). 2) Compared with the control group, during early stage of the experiment, the dry matter apparent digestibility of 50, 100, 200 and 400 U/kg GOD supplementation groups was significantly increased (P<0.05), and the crude protein apparent digestibility of 50 U/kg GOD supplementation group was significantly increased (P<0.05); during later stage of the experiment, the dry matter and crude protein apparent digestibility of 100 U/kg GOD supplementation group was significantly increased (P<0.05). 3) Compared with the control group, the duodenal trypsin activity of 50, 100 and 200 U/kg GOD supplementation groups was significantly increased (P<0.05). 4) Compared with the control group, the villus height and villus height/crypt depth in duodenum of 50, 100, 200 and 400 U/kg GOD supplementation groups were significantly increased (P<0.05), the crypt depth in jejunum of 100 U/kg GOD supplementation group was significantly decreased (P<0.05), and the villus height/crypt depth in jejunum of 100, 200 and 400 U/kg GOD supplementation groups was significantly increased (P<0.05). In conclusion, dietary supplemented with 100 U/kg GOD can improve the growth performance, nutrient apparent digestibility and duodenal trypsin activity, and improve the intestinal morphology.

Key words： glucose oxidase; minks; growth performance; apparent digestibility; intestinal morphology

Cite this article

WANG Luyi , YU Xiaojun , GUO Zhihao , LI Yalin , ZHEN Shibo , WANG Guang , WANG Lihua . Effects of Glucose Oxidase on Growth Performance, Nutrient Apparent Digestibility and Intestinal Morphology of Growing Female Minks[J]. Chinese Journal of Animal Nutrition, 2023 , 35(1) : 603 -610 . DOI: 10.3969/j.issn.1006-267x.2023.01.057

葡萄糖氧化酶(glucose oxidase,GOD)能专一氧化β-D-葡萄糖成葡萄糖酸和过氧化氢,同时消耗大量氧气,使肠道形成厌氧环境,促进有益菌繁殖[1]。过氧化氢能直接抑制大肠杆菌等有害菌生长。葡萄糖酸能降低胃肠道内pH,形成酸性环境促进有益菌繁殖,提高胃蛋白酶活性,促进养分消化吸收。GOD也能促进胃肠道内食糜发酵生成短链脂肪酸,为肠道的健康发育和有益菌繁殖提供能量[2-3]。有研究表明,饲粮中添加GOD可以改善畜禽肠道组织形态[4],提高消化酶活性,改善动物养分消化吸收能力[5],提高动物的生长性能[6]。水貂的育成期正值每年的6月至9月中旬,处于一年中气候最炎热的时期,也是水貂肠道疾病的多发期。肠道是机体养分消化、吸收和免疫的主要器官,肠道环境的健康不仅有利于养分的消化和吸收,更有助于增强机体抗病能力[7]。本试验通过在育成期水貂饲粮中添加不同水平的GOD,探究GOD对育成期母水貂生长性能、养分表观消化率及肠道组织形态的影响,确定GOD在育成期母水貂上的应用效果及适宜添加水平。

1 材料与方法

1.1 试验材料

试验用GOD(活性10 000 U/g)购自济南某饲料公司。基础饲粮为自配鲜饲料。水貂选自青岛市某水貂养殖基地。

1.2 试验方法

1.2.1 试验设计

选取75日龄红眼白貂(母水貂)150只,随机分为5组,每组10个重复,每个重复3只水貂。各组饲粮中GOD添加水平分别为0(对照组)、50、100、200和400 U/kg。预试期1周,正试期8周。正试期分为前期(1~4周)和后期(5~8周)。基础饲粮参照NRC(1982)和文献[8-9]配制,其组成及营养水平见表1。
表1 基础饲粮组成及营养水平

Table 1 Composition and nutrient levels of basal diets %

项目
Items		 含量Content
1~4周1 to 4 weeks 5~8周5 to 8 weeks
原料(饲喂基础) Ingredients (as feed basis)
海杂鱼Sea fish 32 29
鸡架Chicken skeleton 20 20
鳕鱼排Cod bone 15 15
鸡蛋Egg 10 12
鸡肠Chicken intestines 6 9
鸡头Chicken head 5 5
膨化玉米Extruded corn 3 3
预混料Premix1) 3 3
豆粕Soybean meal 2 1
豆油Soybean oil 2 2
鸡肝Chicken livers 1 1
猪血球蛋白粉Pig blood powder globulin 1
合计Total 100 100
营养水平(干物质基础) Nutrient levels (DM basis)2)
干物质DM 30.19 31.34
代谢能ME/(MJ/kg) 16.22 16.74
粗脂肪EE 17.64 19.56
粗蛋白质CP 36.93 34.07
粗灰分Ash 13.52 12.21
钙Ca 3.63 3.09
总磷TP 1.86 1.65

1)预混料组成同宇晓军等[10]。Premix composition was the same as YU et al[10].

2)代谢能为计算值,参照《水貂配合饲料》(LS/T 3403—1992);其他营养水平为实测值。ME was a calculated value, which reference to Compound Feed of Mink (LS/T 3403—1992); while the other nutrient levels were measured values.

1.2.2 饲养管理

试验于2021年7—9月在青岛市某水貂养殖基地进行,试验水貂已完成犬瘟热和细小病毒疫苗的接种。水貂采用双列棚舍饲养,为自然光照,每天05:00、14:00和20:00各饲喂1次,饲粮为黏稠状,放置于笼顶供水貂自由采食,笼内设有水盒,保证水貂自由饮水。

1.2.3 测定指标及方法

试验正式开始时、试验中期和结束时对水貂逐只进行空腹称重,试验期间每天准确记录给料量和剩料量,计算平均日增重(ADG)、平均日采食量(ADFI)和料重比(F/G),公式如下:
平均日增重(g/d)=(终末体重-初始体重)/(试验天数×水貂数量);
平均日采食量(g/d)=(初始料量-剩余料量)/(试验天数×水貂数量);
料重比=平均日采食量/平均日增重。
分别在正式试验后第3周和第7周采用内源指示剂法进行消化试验,每组6个重复,参照李丹丹等[11]的方法处理粪便、饲粮样品,参照相应标准测定粪样和饲粮样品中盐酸不溶灰分(GB/T 23742—2009)、水分(GB/T 6435—2014)、粗蛋白质(GB/T 6432—2018)、粗脂肪(GB/T 6433—2006)和粗灰分(GB/T 6438—2007)含量,并计算养分表观消化率,公式如下:
饲粮中某养分表观消化率(%)=100-(A1/A2)×(B2/B1)。
式中:A1为饲粮中盐酸不溶灰分含量;A2为粪样中盐酸不溶灰分含量;B1为饲粮中该养分含量;B2为粪样中该养分含量。
试验结束后,每组选6只水貂(来自不同重复)进行屠宰,取十二指肠内容物用于测定胰蛋白酶、脂肪酶和α-淀粉酶活性,具体方法参照王群等[12]的方法。取十二指肠和空肠制作组织切片,参照宇晓军等[13]的方法,在50倍显微镜下观察,应用ZEN 2011(Blue version)软件测量十二指肠和空肠绒毛高度和隐窝深度,计算绒毛高度/隐窝深度(V/C)。

1.3 数据统计分析

试验数据采用SPSS 25.0软件进行单因素方差分析(one-way ANOVA),并采用Duncan氏法进行多重比较,结果均以平均值±标准差表示,P<0.05表示差异显著。

2 结果

2.1 GOD对育成期母水貂生长性能的影响

由表2可知,GOD显著影响育成期母水貂试验前期的末重、平均日增重、平均日采食量和料重比(P<0.05),显著影响试验后期的末重(P<0.05)。试验前期,与对照组相比,饲粮中添加100 U/kg GOD可显著提高末重和平均日采食量(P<0.05),饲粮中添加100、200和400 U/kg GOD可显著提高平均日增重(P<0.05),饲粮中添加100和200 U/kg GOD可显著降低料重比(P<0.05)。试验后期,与对照组相比,饲粮中添加50、100和400 U/kg GOD可显著提高末重(P<0.05)。
表2 GOD对育成期母水貂生长性能的影响

Table 2 Effects of GOD on growth performance of growing female minks

项目
Items		 GOD添加水平GOD supplemental levels/(U/kg) P值
P-value
0 50 100 200 400
前期Early stage
初重IBW/g 952.79±66.83 976.32±110.71 975.54±96.61 947.93±101.16 962.48±127.56 0.80
末重FBW/g 1 164.54±111.12b 1 221.32±141.58ab 1 278.15±89.25a 1 209.63±114.50ab 1 235.30±178.22ab 0.04
平均日增重ADG/g 7.56±2.80c 8.75±2.23bc 10.81±3.12a 9.35±2.60ab 9.74±3.33ab <0.01
平均日采食量
ADFI/g		 234.60±15.21bc 231.87±18.96b 248.12±23.02a 242.83±18.05ab 234.77±20.78bc <0.01
料重比F/G 33.05±7.98a 27.50±4.54ab 23.43±3.22b 26.60±4.48b 27.56±9.50ab 0.03
后期Later stage
末重FBW/g 1 289.71±172.18b 1 399.28±198.39a 1 469.85±141.25a 1 379.30±163.64ab 1 397.48±193.71a 0.01
平均日增重ADG/g 3.68±2.46 5.23±3.30 5.64±2.56 5.00±3.72 4.77±2.13 0.19
平均日采食量
ADFI/g		 208.00±27.50 226.32±43.83 224.44±30.02 210.94±35.15 217.16±31.86 0.07
料重比F/G 56.10±15.99 43.42±12.96 45.58±19.37 49.16±17.12 49.22±13.75 0.47

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 GOD对育成期母水貂养分表观消化率的影响

由表3可知,GOD显著影响育成期母水貂试验前期和后期的干物质和粗蛋白质表观消化率(P<0.05),对粗脂肪、粗灰分表观消化率无显著影响(P>0.05)。试验前期,与对照组相比,饲粮中添加50、100、200和400 U/kg GOD可显著提高干物质表观消化率(P<0.05),饲粮中添加50 U/kg GOD可显著提高粗蛋白质表观消化率(P<0.05)。试验后期,与对照组相比,饲粮中添加50、100、200 U/kg GOD可显著提高干物质表观消化率(P<0.05),饲粮中添加100 U/kg GOD可显著提高粗蛋白质表观消化率(P<0.05)。
表3 GOD对育成期母水貂养分表观消化率的影响

Table 3 Effects of GOD on nutrient apparent digestibility of growing female minks%

项目
Items		 GOD添加水平GOD supplemental levels/(U/kg) P值
P-value
0 50 100 200 400
前期Early stage
干物质DM 71.12±0.11e 73.24±0.10d 72.57±0.08c 72.31±0.14b 71.93±0.16a <0.01
粗蛋白质CP 82.89±0.56b 84.23±0.82a 83.39±0.82b 83.10±0.53b 82.91±0.48b 0.01
粗脂肪EE 95.22±0.89 95.51±0.73 94.66±1.17 94.00±1.59 93.91±1.50 0.12
粗灰分Ash 23.07±2.27 26.28±1.89 24.58±1.97 24.20±2.62 24.76±3.89 0.35
后期Later stage
干物质DM 73.12±0.10c 73.82±0.08b 73.90±0.09ab 73.95±0.14a 73.09±0.07c <0.01
粗蛋白质CP 82.00±0.95b 82.64±0.48ab 83.42±0.47a 82.86±0.89ab 82.47±0.84ab 0.04
粗脂肪EE 96.08±0.47 95.99±0.68 96.15±0.69 96.11±0.59 96.61±0.69 0.48
粗灰分Ash 30.40±2.40 28.96±1.75 30.40±3.72 31.25±1.48 31.65±2.29 0.39

2.3 GOD对育成期母水貂十二指肠消化酶活性的影响

由表4可知,GOD显著影响育成期母水貂十二指肠胰蛋白酶活性(P<0.05),对十二指肠脂肪酶和α-淀粉酶活性无显著影响(P>0.05)。与对照组相比,饲粮中添加50、100和200 U/kg GOD可显著提高十二指肠胰蛋白酶活性(P<0.05)。
表4 GOD对育成期母水貂十二指肠消化酶活性的影响

Table 4 Effects of GOD on duodenum digestive enzyme activities of growing female minks

项目
Items		 GOD添加水平GOD supplemental levels/(U/kg) P值
P-value
0 50 100 200 400
胰蛋白酶
Trypsin/(U/mg prot)		 791.46±87.23c 952.51±34.07b 1 260.15±89.85a 1 015.66±143.69b 806.12±74.91c <0.01
脂肪酶
Lipase/(U/g prot)		 19.87±3.64 21.08±1.96 25.24±5.05 22.15±5.42 22.10±2.33 0.22
α-淀粉酶
α-amylase/(U/mg prot)		 0.29±0.01 0.29±0.07 0.31±0.01 0.29±0.29 0.30±0.02a 0.42

2.4 GOD对育成期母水貂肠道组织形态的影响

由表5可知,GOD显著影响育成期母水貂十二指肠绒毛高度、V/C以及空肠绒毛高度、隐窝深度、V/C(P<0.05)。与对照组相比,饲粮中添加50、100、200和400 U/kg GOD可显著提高十二指肠绒毛高度和V/C(P<0.05);饲粮中添加200 U/kg GOD可显著提高空肠绒毛高度(P<0.05),饲粮中添加100 U/kg GOD可显著降低空肠隐窝深度(P<0.05),饲粮中添加100、200和400 U/kg GOD可显著提高空肠V/C(P<0.05)。
表5 GOD对育成期母水貂肠道组织形态的影响

Table 5 Effects of GOD on intestinal morphology of growing female minks

项目
Items		 GOD添加水平GOD supplemental levels/(U/kg) P值
P-value
0 50 100 200 400
十二指肠Duodenum
绒毛高度Villus height/μm 715.04±84.75c 890.68±64.28a 894.55±89.71a 913.59±49.21a 797.33±68.81b <0.01
隐窝深度Crypt depth/μm 582.28±64.82 554.69±87.13 511.62±55.11 545.14±70.74 519.82±25.13 0.06
绒毛高度/隐窝深度V/C 1.24±0.16c 1.65±0.33ab 1.76±0.18a 1.70±0.23ab 1.54±0.28b <0.01
空肠Jejunum
绒毛高度Villus height/μm 824.88±77.09b 817.51±59.17b 851.84±37.71b 1 010.17±108.87a 890.39±100.88b <0.01
隐窝深度Crypt depth/μm 534.95±53.79ab 528.15±64.43abc 475.32±48.97c 549.77±68.76a 489.75±58.19bc 0.02
绒毛高度/隐窝深度V/C 1.55±0.11b 1.57±0.19b 1.81±0.26a 1.86±0.29a 1.85±0.37a 0.01

3 讨论

3.1 GOD对育成期母水貂生长性能的影响

水貂育成期正值炎热的夏季,鲜饲料在高温环境下容易滋生细菌,水貂也容易出现高温热应激,从而影响水貂的采食。有研究表明,GOD能减少动物因高温而产生的拒食、厌食、消化不良、腹泻等问题,缓解高温热应激[14-15]。本试验表明,饲粮中添加100 U/kg GOD对育成期母水貂的末重、平均日增重、平均日采食量和料重比均有显著影响。Meng等[16]和Wu等[17]研究表明,饲粮中添加GOD可以改善肠道健康,提高肉仔鸡的平均日增重。侯振平等[18]和张宏宇等[19]研究表明,饲粮中添加GOD可以提高断奶仔猪的末重和平均日增重。本试验中,饲粮中添加GOD能提高母水貂的采食量,可能是因为GOD能够氧化β-D-葡萄糖生成δ-D-葡萄糖酸内酯,δ-D-葡萄糖酸内酯具有甜味[20],而水貂喜欢甜味,由此可以推断GOD能够改善饲粮的适口性。GOD通过催化作用产生的短链脂肪酸能快速被肠道吸收并为其供能[15],促进肠道发育,保证肠道健康。这说明GOD对于水貂的促生长作用可能是通过改善肠道健康、提高消化酶活性来提高对养分的消化吸收,并通过改善饲粮适口性提高采食量来实现的。

3.2 GOD对育成期母水貂养分表观消化率和消化酶活性的影响

养分表观消化率能够反映出动物对饲粮中养分的消化吸收能力,育成期水貂生长迅速,水貂对饲粮中养分的利用程度能影响其生长发育。本试验表明,饲粮中添加100 U/kg GOD可显著提高育成期母水貂试验前期干物质表观消化率及试验后期干物质和粗蛋白质表观消化率。这与崔细鹏等[21]在白羽肉鸡上的研究结果一致。杨久仙等[4]研究表明,断奶仔猪饲粮中添加GOD能提高干物质和粗蛋白质表观消化率。养分的消化吸收能力与肠道内消化酶活性密切相关,胰蛋白酶能水解蛋白质生成小分子氨基酸,胰蛋白酶活性的提高有利于粗蛋白质的消化吸收[22]。本试验表明,饲粮中添加50、100、200 U/kg GOD可显著提高育成期母貂十二指肠胰蛋白酶活性,其中100 U/kg GOD添加组十二指肠胰蛋白酶活性显著高于50和200 U/kg GOD添加组。宋海彬等[23]研究发现,在肉鸡饲粮中添加GOD能显著提高胰蛋白酶和淀粉酶活性,优化肠壁结构,促进肠道发育。杨海涛[24]研究表明,饲粮中添加GOD能显著提高肉仔鸡十二指肠内的胰蛋白酶、脂肪酶和淀粉酶活性,维持肠道内菌群平衡,提高有益菌的数量。因此,本试验中饲粮中添加GOD提高粗蛋白质表观消化率,其原因一方面可能是GOD催化作用生成的葡萄糖酸改变肠道pH,在动物胃肠道内发挥酸化剂的功效,形成的弱酸性环境有利于消化酶活性基因的解离状态[25],适宜与底物结合,酶与底物结合力提高,反应速率提高,加快饲粮中养分的消化[26];另一方面可能是GOD能改善肠黏膜结构,促进肠道内有益菌生长,改善肠道形态结构[5],激活胃肠道内消化酶活性,进而提高养分消化率。

3.3 GOD对育成期母水貂肠道组织形态的影响

小肠绒毛高度和隐窝深度与肠道消化吸收能力有着密切联系,小肠绒毛高度提高和隐窝变浅可表明消化吸收能力增强,V/C是衡量肠道内黏膜面积的指标,V/C越大说明肠上皮表面积越大,肠道消化吸收能力越好[27-28]。本试验中,饲粮中添加GOD可提高育成期母水貂十二指肠绒毛高度和V/C,其中100 U/kg GOD添加组十二指肠V/C显著升高,空肠隐窝深度显著降低。这说明在饲粮中添加适量的GOD能够扩大肠黏膜面积,有利于肠黏膜保持完整性,进而提高肠道对养分的消化吸收。杨久仙等[4]研究表明,饲粮中添加GOD能显著提高断奶仔猪十二指肠及空肠的肠绒毛高度和V/C。GOD催化葡萄糖产生的葡萄糖酸经发酵后产生短链脂肪酸,短链脂肪酸不仅能够为肠细胞的正常生理功能提供能量,还可以通过抑制组蛋白脱乙酰酶与肠上皮细胞的相互作用或者激活游离脂肪酸受体,提高肠黏膜免疫和屏障功能[29],增加肠黏膜厚度,提高肠细胞的分泌功能,改善肠道对养分的吸收能力,促进肠道健康发育[30-31]。

4 结论

在本试验条件下,饲粮中添加100 U/kg GOD可提高育成期母水貂生长性能、养分表观消化率、十二指肠胰蛋白酶活性,改善肠道组织形态。

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