Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 7: 4276 - 4286

DOI: https://doi.org/10.12418/CJAN2023.398

RESEARCH PAPER

Effects of Tannic Acid on Growth Performance, Intestinal Barrier Function and Antioxidant Capacity of Piglets Infected with Porcine Epidemic Diarrhea Virus

  • WANG Sitian , 1 ,
  • XU Pengtao 1 ,
  • ZHENG Liyun 1 ,
  • QI Ya 1 ,
  • LIU Zhipeng 1 ,
  • HE Lai 1 ,
  • LIU Cheng’ao 1 ,
  • HOU Yongqing 1 ,
  • GUO Shuangshuang 1 ,
  • TONG Qingfang 2 ,
  • ZHANG Zhengfan , 1, ** ,
  • DING Binying , 1, **
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  • 1 Engineering Research Center of Feed Protein Resources on Agricultural By-Products, Hubei Key Laboratory of Animal Nutrition and Feed Science, Wuhan Polytechnic University, Wuhan 430023, China
  • 2 Wuhan Jiangxia District Animal Disease Prevention and Control Center, Wuhan 430072, China
** ZHANG Zhengfan, associate professor, E-mail: ;
DING Binying, professor, E-mail:

*Contributed equally

Received date: 2023-01-16

  Online published: 2023-07-11

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Abstract

This study was conducted to investigate the effects of tannic acid (TA) on the growth performance, intestinal barrier function and antioxidant capacity of piglets infected with porcine epidemic diarrhea virus (PEDV). Twenty-four 7-day-old healthy piglets (Duroc×Landrace×Yorkshire), with an initial body weight of (2.72±0.22) kg, were divided into 4 groups with 6 replicates per group, and 1 pig per replicate. The groups were as follows: control group, TA group, PEDV group and PEDV+TA group. The experiment was divided into 2 stages as pre-infected stage (days 1 to 8) and infected stage (days 9 to 12). On days 1 to 8, piglets in the TA group and PEDV+TA group were orally administered with 190 mg/kg BW TA, and piglets in the control group and PEDV group were orally administered with liquid milk replacer at the same volume; on day 9, piglets in the PEDV group and PEDV+TA group were orally administered PEDV suspension at a dose of 106 TCID50 (50% tissue culture infection dose) per pig, piglets in the control group and TA group were orally administered with phosphate buffered solution at the same volume. The results showed that, before PEDV infection, the average daily gain (ADG) of piglets in TA group was significantly increased compared with the control group (P<0.05). After PEDV infection, the ADG of piglets was significantly decreased (P<0.05), and the feed/gain (F/G) and diarrhea index were significantly increased (P<0.05); the serum D-xylose content was significantly decreased (P<0.05), and serum diamine oxidase activity and intestinal damage scores of duodenum, jejunum, ileum and colon were significantly increased (P<0.05); the villus height (VH) of duodenum, jejunum and ileum and the villus area (VA) of duodenum and ileum were significantly decreased (P<0.05), the crypt depth (CD) of jejunum, ileum and colon was significantly increased (P<0.05), and the VH/CD of duodenum, jejunum and ileum was significantly decreased (P<0.05); the activities of glutathione peroxidase (GSH-Px), catalase (CAT) and total superoxide dismutase (T-SOD) in serum were significantly decreased (P<0.05), the activities of GSH-Px in jejunum, ileum and colon, T-SOD in ileum and colon, and total antioxidant capacity (T-AOC) in jejunum were significantly decreased (P<0.05), and the contents of malondialdehyde (MDA) in jejunum and hydrogen peroxide (H2O2) in duodenum and colon were significantly increased (P<0.05). TA administration could significantly mitigate the increase of F/G, diarrhea index, serum alanine aminotransferase (ALT) activity and CD of duodenum, jejunum, ileum and colon of piglets by PEDV infection (P<0.05), significantly mitigate the decrease of ADG, jejunal VH, VA and VH/CD of duodenum and jejunum (P<0.05), and significantly increase the activities of GSH-Px in duodenum and jejunum, colonic T-SOD and T-AOC of duodenum and colon of PEDV-infected piglets (P<0.05). Overall, administration of 190 mg/kg BW TA can improve the ADG and decrease the diarrhea rate of neonatal piglets, and mitigate the intestinal injury of piglets caused by PEDV infection by increasing antioxidant capacity and improving intestinal morphology.

Key words： tannic acid; PEDV; growth performance; intestinal barrier function; antioxidant capacity

Cite this article

WANG Sitian , XU Pengtao , ZHENG Liyun , QI Ya , LIU Zhipeng , HE Lai , LIU Cheng’ao , HOU Yongqing , GUO Shuangshuang , TONG Qingfang , ZHANG Zhengfan , DING Binying . Effects of Tannic Acid on Growth Performance, Intestinal Barrier Function and Antioxidant Capacity of Piglets Infected with Porcine Epidemic Diarrhea Virus[J]. Chinese Journal of Animal Nutrition, 2023 , 35(7) : 4276 -4286 . DOI: 10.12418/CJAN2023.398

猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)属于冠状病毒科,可引起仔猪急性腹泻,因其高传染性和高死亡率给生猪养殖业造成巨大经济损失[1-2]。禁抗限抗背景下,高锌饲粮预防断奶仔猪腹泻的弊端已日益显露,如土壤锌富集和畜产品高锌残留等问题[3]。目前,已有研究证实益生菌、益生素和有机酸等饲料添加剂可以降低断奶仔猪腹泻率,但对PEDV引起的腹泻的改善效果不稳定[4]。
植物提取物或其衍生物可能是抗生素的潜在替代品。其中,单宁酸(tannins,TA)作为多酚类化合物,因其收敛性强,具有抗腹泻、抗菌、抗氧化、调节肠道微生物稳态等作用而受到广泛关注[5]。TA已被证明可以通过与肠道黏膜黏蛋白结合形成保护膜,从而起到防治腹泻、提高生长性能的作用[6-7]。此外,TA还可以增加肠道绒毛高度,降低隐窝深度,从而改善肠道形态结构完整性,修复肠道损伤[6,8-9],但TA在PEDV感染仔猪上的研究较少。因此,本试验拟通过建立仔猪PEDV感染模型,研究TA对PEDV感染仔猪生长性能、肠道屏障功能和抗氧化能力的影响,以探讨其作用机制。

1 材料与方法

1.1 试验材料

试验仔猪购于武汉绿色巨农农牧股份有限公司。代乳料由上海高得饲料有限公司提供。PEDV(YN株)由华中农业大学微生物重点实验室提供。五倍子TA由五峰赤诚生物科技有限股份公司提供,TA含量为80%。

1.2 试验设计

试验共选用24头健康状况良好、遗传背景一致、2~3胎次、平均体重为(2.72±0.22) kg的7日龄杜×长×大仔猪,随机分为4组,分别为对照组、TA组、PEDV组和PEDV+TA组,每组6个重复,每个重复1头猪。试验期12 d,分为感染前(第1~8天)和感染后(第9~12天)2个阶段。在试验第1~8天,TA组和PEDV+TA组仔猪口腔灌服190 mg/kg BW TA(190 mg单宁酸溶于1 mL液体代乳料中配成悬浊液,每千克体重灌服1 mL悬浊液),对照组和PEDV组仔猪口腔灌服相同体积的液体代乳料;试验第9天,PEDV组和PEDV+TA组仔猪口腔灌服1 mL 106 TCID50/头PEDV悬液(TCID50为50%组织培养感染剂量),对照组和TA组仔猪口腔灌服等体积的磷酸盐缓冲溶液。试验用代乳料主要成分为全脂奶粉和大豆磷脂,其营养水平见表1。试验饲粮为代乳料+0.1‰商品维生素预混料,其中商品维生素预混料可为每千克代乳料提供维生素A 12 500 IU、维生素D3 2 800 IU、维生素E 30 IU、维生素K 5 mg、维生素B1 3 mg、维生素B2 10 mg、维生素B6 8 mg、烟酸40 mg、维生素B12 40 μg、D-泛酸 15 mg、叶酸1 mg、生物素0.08 mg。
表1 代乳料营养水平(风干基础)

Table 1 Nutrient levels of milk replacer (air-dry basis) %

项目
Item		 粗蛋白质
CP		 粗灰分
Ash		 粗纤维
CF		 水分
Moisture		 赖氨酸
Lys		 食盐
NaCl		 钙
Ca		 总磷
TP
代乳料Milk replacer ≥20.0 ≤9.0 ≤1.0 ≤10.0 ≥1.4 0.3~1.5 0.4~1.1 ≥0.3

1.3 饲养管理

本试验在湖北省武汉市农业科学院畜牧兽医研究所进行,采用2栋饲养条件完全一致的全封闭式猪舍[舍内温度为(32±2) ℃,相对湿度为60%~70%],试验仔猪均单栏饲养于0.8 m×1.0 m的漏缝式地板圈内,每天清扫猪舍1次,并用聚维酮碘消毒液喷洒消毒栏舍,严格防止2栋猪舍之间发生交叉感染。仔猪每天饲喂5次(饲喂时间为07:30、11:00、15:00、18:30、21:00),代乳料用温水(45~55 ℃)冲泡,早、中、晚各喂1次温水。

1.4 样品采集与处理

于试验第12天06:00,所有仔猪均口服10% D-木糖(1 mL/kg BW),1 h后对所有仔猪进行前腔静脉采血并保存于肝素钠抗凝管中。全群仔猪采血后肌肉注射50 mg/kg BW戊巴比妥钠,麻醉后进行屠宰取样。取十二指肠、空肠、回肠和结肠各肠道中间段(长度为10~15 cm),纵向剖开肠段,用预冷的生理盐水冲洗肠道内容物,刮取肠黏膜快速置于液氮中冷冻,后转移至-80 ℃保存。

1.5 检测指标与方法

1.5.1 生长性能

每天记录仔猪采食量,于试验第1、9和12天07:30给隔夜禁食的仔猪进行空腹称重,按照如下公式计算平均日增重(ADG)、平均日采食量(ADFI)和料重比(F/G):
ADG=总增重/试验天数;
ADFI=总采食量/试验天数;
F/G=ADFI/ADG。

1.5.2 腹泻评分

试验期间视觉观察并记录仔猪每天的粪便形态。按照粪便形态进行评分,分为4个等级:0=干燥成形,1=糊状,2=半液体状,3=液体状,计算每日平均值,得分≥2视为腹泻[10]。按如下公式计算腹泻指数:
腹泻指数(%)=(每头仔猪粪便评分之和/观察总天数)×100。

1.5.3 血清生化指标

使用全自动生化分析仪(日立HITEC-7100,上海曼普生物科技有限公司)测定血清生化指标,试剂盒购于富士胶片和光纯耀化学有限公司。

1.5.4 肠道屏障功能指标

参照Vilaseca等[11]描述的方法进行肠道损伤评分,分为4个评分等级:无肉眼可见出血点记0分;偶有损伤或有零星出血点记1分;有肉眼可见明显损伤即肠道出现溃疡或肠壁变薄变透记2分;肠道出现溃疡、糜烂,肠壁严重变薄变透并充血记3分。使用光学显微镜(Olympus,日本)对肠道组织切片进行观察,选取10根不同走向且完整的绒毛,测量绒毛高度(villous height,VH)、隐窝深度(crypt depth,CD)以及绒毛表面积(villous area,VA),计算绒毛高度/隐窝深度(VH/CD)。
采用南京建成生物工程研究所生产的试剂盒测定血清D-木糖含量和二胺氧化酶(DAO)活性。

1.5.5 血清和肠道抗氧化指标

采用南京建成生物工程研究所生产的试剂盒使用多功能酶标仪(Spectra Max M5,Molecular Devices)测定血清和肠道组织总抗氧化能力(T-AOC)与谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)和总超氧化物歧化酶(T-SOD)活性以及丙二醛(MDA)和过氧化氢(H2O2)含量。

1.6 数据统计分析

试验数据先采用Excel 2010软件进行预处理,再使用SPSS 23.0软件对仔猪感染前生长性能数据进行单因素方差分析,对感染后的试验数据进行双因素方差分析(two-way ANOVA),组间多重比较用Duncan氏法进行。试验结果以平均值±标准差体现,P<0.05表示显著差异,0.05≤P<0.10表示有趋势。

2 结果

2.1 TA对仔猪生长性能的影响

TA对仔猪生长性能的影响见表2和表3。感染PEDV前,灌服TA仔猪的ADG显著提高(P<0.05),ADFI和FCR无显著变化(P>0.05)。感染PEDV后,仔猪ADG显著下降(P<0.05),F/G显著升高(P<0.05);灌服TA显著提高了PEDV感染仔猪的ADG(P<0.05),显著降低了F/G(P<0.05),且TA与PEDV在F/G上存在交互效应(P<0.05)。
表2 TA对仔猪生长性能(第1~8天)的影响

Table 2 Effects of TA on growth performance (days 1 to 8) of piglets (n=6)

项目Items 对照组Control group TA组TA group P值P-value
平均日增重ADG/g 81.54±3.92b 94.10±6.94a <0.001
平均日采食量ADFI/g 90.51±9.70 90.34±10.95 0.342
料重比F/G 1.07±0.09 0.96±0.17 0.146

同行数据肩标不同字母表示差异显著(P<0.05)。下表同。

In the same row, values with different letter superscripts mean significant difference (P<0.05). The same as below.

表3 TA对PEDV感染仔猪生长性能(第9~12天)的影响

Table 3 Effects of TA on growth performance (days 9 to 12) of piglets infected with PEDV

项目
Items		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
平均日增重ADG/g 122.29±11.69 134.00±29.46 35.43±9.91 55.23±13.44 0.029 <0.001 0.556
平均日采食量ADFI/g 93.60±14.26 93.80±7.91 82.41±35.68 85.05±16.07 0.613 0.747 0.881
料重比F/G 0.74±0.15c 0.69±0.10c 2.58±0.21a 1.54±0.11b <0.001 <0.001 <0.001

-PEDV:未感染猪流行性腹泻病毒;+PEDV:感染猪流行性腹泻病毒;-TA:未灌服单宁酸;-TA:灌服单宁酸;PEDV:猪流行性腹泻病毒效应;TA:单宁酸效应;PEDV×TA:猪流行性腹泻病毒与单宁酸的交互效应。下表同。

-PEDV: uninfected with PEDV; +PEDV: infected with PEDV; -TA: non-administered with TA; -TA: administered with TA; PEDV: PEDV effect; TA: TA effect; PEDV×TA: interaction effect of PEDV and TA. The same as below.

2.2 TA对PEDV感染仔猪腹泻指数的影响

TA对PEDV感染仔猪腹泻指数的影响见表4。感染PEDV后,仔猪的腹泻指数显著升高(P<0.05);灌服TA可显著降低PEDV感染仔猪的腹泻指数(P<0.05)。
表4 TA对PEDV感染仔猪腹泻指数(第9~12天)的影响

Table 4 Effects of TA on diarrhea index (days 9 to 12) of piglets infected with PEDV

项目
Item		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
腹泻指数
Diarrhea index		 0±0b 0±0b 0.85±0.42a 0.23±0.20b <0.001 0.004 0.004

2.3 TA对PEDV感染仔猪血清生化指标的影响

由表5可知,感染PEDV后,仔猪血清总胆固醇(TC)、磷(P)、低密度脂蛋白(LDL)和高密度脂蛋白(HDL)含量显著下降(P<0.05),血清甘油三酯(TG)和葡萄糖(GLU)含量显著升高(P<0.05);灌服TA能显著降低感染PEDV仔猪血清谷草转氨酶(ALT)活性(P<0.05),且TA与PEDV在血清ALT活性上存在交互效应(P<0.05)。仔猪血清总胆红素(TB)、总蛋白(TP)、白蛋白(ALB)、钙(Ca)、肌酐(CREA)和直接胆红素(DB)含量及碱性磷酸酶(ALP)、γ-谷氨酰转移酶(GGT)和乳酸脱氢酶(LDH)活性各组之间无显著差异(P>0.05)。
表5 TA对PEDV感染仔猪血清生化指标的影响

Table 5 Effects of TA on serum biochemical indices of piglets infected with PEDV

项目
Items		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
总胆红素TB/(mg/dL) 0.36±0.22 0.28±0.14 0.30±0.22 0.29±0.18 0.743 0.555 0.673
总蛋白TP/(g/L) 5.72±0.68 6.45±0.58 5.88±0.38 6.10±0.84 0.695 0.054 0.278
白蛋白ALB/(g/L) 2.81±0.34 2.82±0.21 2.56±0.29 2.61±0.46 0.086 0.833 0.884
谷丙转氨酶
AST/(U/L)		 36.89±9.04 42.00±5.83 30.69±6.64 28.29±6.47 0.001 0.615 0.170
谷草转氨酶
ALT/(U/L)		 52.09±4.35b 52.17±7.70b 62.13±6.75a 48.00±5.03b 0.222 0.006 0.006
碱性磷酸酶
ALP/(U/L)		 741.2±213.1 685.0±211.9 594.4±225.5 740.9±209.8 0.585 0.588 0.229
总胆固醇
TC/(mg/dL)		 191.5±37.3 206.1±37.5 101.9±26.2 141.5±33.0 <0.001 0.039 0.322
甘油三酯
TG/(mg/dL)		 38.5±28.6 30.6±15.9 57.8±20.6 48.3±14.4 0.025 0.272 0.916
葡萄糖
GLU/(mg/dL)		 69.78±16.04 85.08±13.13 85.53±16.13 98.14±9.22 0.013 0.015 0.804
钙Ca/(mg/dL) 10.09±0.16 10.53±0.21 10.66±1.29 10.36±0.24 0.501 0.825 0.228
磷P/(mg/dL) 9.12±0.50 9.45±1.11 7.50±0.44 8.38±0.66 <0.001 0.028 0.293
肌酐CREA/(mg/dL) 0.92±0.79 0.67±0.27 0.60±0.17 0.69±0.08 0.310 0.580 0.274
高密度脂蛋白
HDL/(mg/dL)		 94.9±15.5 112.7±17.7 46.1±12.9 66.8±17.9 <0.001 0.004 0.814
低密度脂蛋白
LDL/(mg/dL)		 148.8±41.3 147.5±40.3 64.2±23.7 95.9±26.5 <0.001 0.225 0.188
γ-谷氨酰转移酶
GGT/(U/L)		 37.6±6.03 38.8±13.9 29.1±14.3 41.9±14.9 0.585 0.171 0.257
直接胆红素
DB/(mg/dL)		 0.18±0.08 0.16±0.06 0.19±0.13 0.17±0.07 0.696 0.620 0.973
乳酸脱氢酶
LDH/(U/L)		 790.4±117.9 772.5±101.7 691.6±67.1 766.7±100.6 0.156 0.433 0.206

2.4 TA对PEDV感染仔猪肠道损伤评分和肠道吸收功能指标的影响

由表6可知,感染PEDV后,仔猪小肠(十二指肠、空肠、回肠)和结肠肠道损伤评分均显著升高(P<0.05),血清D-木糖含量显著降低(P<0.05),血清DAO活性显著上升(P<0.05);灌服TA显著降低感染PEDV仔猪空肠肠道损伤评分(P<0.05),有降低十二指肠(P=0.080)和空肠(P=0.056)肠道损伤评分的趋势。
表6 TA对PEDV感染仔猪肠道损伤评分和肠道吸收功能指标的影响

Table 6 Effects of TA on intestinal damage score and intestinal absorption function indexes of piglets infected with PEDV

项目
Items		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
肠道损伤评分Intestinal damage score/分
十二指肠Duodenum 0±0 0±0 1.33±0.26 1.05±0.27 <0.001 0.080 0.080
空肠Jejunum 0.17±0.41 0±0 1.80±0.49 1.08±0.18 <0.001 0.004 0.056
回肠Ileum 0±0 0±0 1.17±0.41 1.12±0.45 <0.001 0.842 0.842
结肠Colon 0±0 0±0 1.50±0.55 1.08±0.34 <0.001 0.128 0.128
肠道吸收功能指标Intestinal absorption function indexes
二胺氧化酶
DAO/(U/L)		 18.86±0.43 18.33±2.90 22.58±1.73 20.27±1.80 0.002 0.091 0.277
D-木糖
D-xylose/(mmol/L)		 3.23±0.12 3.38±0.33 1.83±0.08 2.02±0.25 <0.001 0.052 0.793

2.5 TA对PEDV感染仔猪肠道形态的影响

由表7可知,感染PEDV后,仔猪小肠各肠段VH以及十二指肠和回肠VA显著降低(P<0.05),空肠、回肠和结肠CD显著增加(P<0.05),小肠各肠段VH/CD均显著下降(P<0.05);灌服TA可显著增加感染PEDV仔猪十二指肠和空肠VH(P<0.05),显著降低小肠各肠段和结肠CD(P<0.05),显著增加十二指肠VA(P<0.05),显著增加小肠各肠段VH/CD(P<0.05);TA与PEDV在空肠VH,小肠各肠段和结肠CD,十二指肠、空肠和回肠VH/CD以及十二指肠和空肠VA上存在交互效应(P<0.05)。
表7 TA对PEDV感染仔猪肠道形态的影响

Table 7 Effects of TA on intestinal morphology of piglets infected with PEDV

项目
Items		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
绒毛高度VH/μm
十二指肠Duodenum 205.36±18.12 237.18±19.40 141.53±13.65 192.75±35.03 <0.001 0.002 0.392
空肠Jejunum 198.16±22.75b 238.23±29.78a 75.85±9.44c 77.93±3.94c <0.001 0.020 0.033
回肠Ileum 183.57±16.89 186.66±16.37 93.82±14.69 100.19±22.09 <0.001 0.521 0.823
隐窝深度CD/μm
十二指肠Duodenum 80.56±9.98c 117.00±7.06b 147.42±13.75a 105.96±20.98b 0.665 <0.001 <0.001
空肠Jejunum 98.39±11.18c 119.60±14.53b 135.68±18.41a 41.72±3.00d <0.001 0.001 <0.001
回肠Ileum 91.00±9.59b 93.16±8.49b 155.52±7.43a 54.09±12.92c <0.001 0.005 <0.001
结肠Colon 180.38±9.24b 180.80±18.65b 244.23±24.61a 168.07±6.82b 0.001 <0.001 <0.001
绒毛面积VA/μm2
十二指肠Duodenum 37 635.71
±9 025.89a		 41 885.70
±8 945.03a		 17 689.61
±1 412.14b		 38 062.24
±2 354.16a		 0.001 0.001 0.019
空肠Jejunum 23 158.10
±3 634.04b		 34 418.41
±7 872.79a		 20 353.70
±1 621.11c		 10 000.89
±424.16c		 0.804 <0.001 <0.001
回肠Ileum 25 161.34
±3 881.32		 29 307.30
±2 313.82		 12 297.98
±2 843.91		 14 625.96
±5 546.19		 <0.001 0.053 0.569
绒毛高度/隐窝深度VH/CD
十二指肠Duodenum 2.55±0.35a 2.00±0.17b 0.96±0.22c 2.03±0.20b 0.012 <0.001 <0.001
空肠Jejunum 2.01±0.05a 2.01±0.06a 0.56±0.34b 1.86±0.16a <0.001 <0.001 <0.001
回肠Ileum 2.02±0.04a 2.00±0.03a 0.60±0.11b 1.88±0.20a <0.001 <0.001 <0.001

2.6 TA对PEDV感染仔猪血清和肠道抗氧化指标的影响

由表8可知,感染PEDV后,仔猪血清CAT和T-SOD活性显著下降(P<0.05);灌服TA能显著提高感染PEDV仔猪血清T-AOC(P<0.05),但对血清GSH-Px、CAT、T-SOD活性和MDA含量无显著影响(P>0.05)。
表8 TA对PEDV感染仔猪血清抗氧化指标的影响

Table 8 Effects of TA on serum antioxidant indexes of piglets infected with PEDV

项目
Items		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 345.7±32.5 352.6±25.3 312.7±16.8 341.1±27.8 0.050 0.114 0.328
过氧化氢酶
CAT/(U/mL)		 3.50±0.01 4.11±0.97 2.94±0.28 3.15±0.15 0.002 0.061 0.347
总超氧化物歧化酶
T-SOD/(U/mL)		 82.01±2.05 84.05±1.45 76.71±5.02 78.61±4.86 0.002 0.208 0.963
丙二醛
MDA/(nmol/mL)		 2.92±0.58 2.86±0.44 3.15±0.53 3.05±0.38 0.307 0.700 0.922
总抗氧化能力
T-AOC/(mmol/L)		 0.20±0.02 0.28±0.06 0.20±0.03 0.25±0.03 0.224 <0.001 0.317
由表9可知,感染PEDV后,仔猪十二指肠、空肠、结肠GSH-Px活性与回肠、结肠T-SOD活性以及空肠T-AOC显著下降(P<0.05),空肠MDA含量与十二指肠和结肠H2O2含量显著增加(P<0.05);灌服TA能显著提高PEDV感染仔猪十二指肠、空肠和结肠GSH-Px活性与空肠和结肠T-SOD活性以及十二指肠T-AOC(P<0.05),显著降低空肠MDA含量(P<0.05);TA与PEDV在十二指肠、空肠和结肠GSH-Px活性与空肠和结肠T-SOD活性以及十二指肠和结肠T-AOC上存在交互效应(P<0.05)。
表9 TA对PEDV感染仔猪肠道抗氧化指标的影响

Table 9 Effects of TA on intestinal antioxidant indexes of piglets infected with PEDV

项目
Items		 -PEDV +PEDV P值P-value
-TA +TA -TA +TA PEDV TA PEDV×TA
谷胱甘肽过氧化物酶GSH-Px/(U/mg prot)
十二指肠Duodenum 31.98±2.16a 31.31±2.88a 19.93±3.84b 30.52±6.43a 0.001 0.008 0.003
空肠Jejunum 29.09±4.55b 24.51±1.87c 22.96±1.96c 53.58±2.91a <0.001 <0.001 <0.001
回肠Ileum 31.66±2.88 29.73±17.66 23.91±3.86 31.04±6.44 0.426 0.518 0.266
结肠Colon 4.98±0.95b 8.37±1.73a 4.74±0.48b 4.88±0.75b <0.001 0.001 0.002
过氧化氢酶CAT/(U/mg prot)
十二指肠Duodenum 7.20±2.61 6.79±1.85 4.96±0.87 5.81±0.93 0.034 0.757 0.383
空肠Jejunum 20.19±3.93 19.14±5.04 15.68±3.08 19.77±7.29 0.361 0.470 0.230
回肠Ileum 1.87±0.22 1.73±0.10 1.88±0.18 1.79±0.15 0.628 0.101 0.753
结肠Colon 3.89±0.98 3.32±0.06 3.55±0.48 3.88±0.80 0.687 0.680 0.122
总超氧化物歧化酶T-SOD/(U/mg prot)
十二指肠Duodenum 222.7±36.9 211.7±80.2 218.4±13.6 253.6±51.7 0.384 0.571 0.286
空肠Jejunum 177.1±22.0b 392.5±174.4a 209.6±45.8b 219.7±40.6b 0.079 0.008 0.014
回肠Ileum 161.1±5.3 173.6±16.3 174.7±10.7 182.2±13.3 0.036 0.055 0.626
结肠Colon 216.9±18.2b 196.9±34.0b 222.0±19.2b 300.3±30.1a <0.001 0.013 <0.001
丙二醛MDA/(nmol/mg prot)
十二指肠Duodenum 3.45±0.64 3.56±0.36 3.84±0.88 3.92±1.06 0.252 0.765 0.959
空肠Jejunum 0.54±0.12 0.39±0.12 0.61±0.02 0.50±0.07 0.026 0.002 0.616
回肠Ileum 0.65±0.08 0.68±0.09 0.64±0.07 0.64±0.12 0.537 0.752 0.696
结肠Colon 1.43±0.14 1.42±0.17 1.44±0.16 1.38±0.23 0.891 0.640 0.742
过氧化氢H2O2/(mmol/g prot)
十二指肠Duodenum 0.53±0.15 0.42±0.13 0.66±0.14 0.59±0.16 0.023 0.137 0.716
空肠Jejunum 13.03±2.93 13.80±2.12 13.62±1.78 12.82±1.97 0.834 0.988 0.401
回肠Ileum 0.86±0.47 0.86±0.30 1.02±0.16 0.86±0.47 0.477 0.629 0.620
结肠Colon 1.14±0.22 1.16±0.09 1.57±0.17 1.44±0.36 0.001 0.550 0.432
总抗氧化能力T-AOC/(mmol/g prot)
十二指肠Duodenum 52.63±9.28b 40.59±7.87b 45.14±9.67b 85.39±12.88a <0.001 0.003 <0.001
空肠Jejunum 65.11±14.32 70.20±15.74 47.33±7.65 49.24±8.95 0.001 0.488 0.752
回肠Ileum 197.8±4.3 212.9±17.7 230.1±16.2 229.3±21.8 0.002 0.295 0.248
结肠Colon 405.2±72.2b 357.1±33.6b 402.0±33.2b 541.9±149.4a 0.018 0.207 0.015

3 讨论

本试验中仔猪感染PEDV后出现急性腹泻和呕吐,解剖后小肠和结肠肠壁变薄变透明,与PEDV感染症状[12]相符,说明PEDV模型建立成功。研究表明,饲粮中添加250~1 500 mg/kg TA能显著提高仔猪的日增重,降低料重比,但添加高剂量(2 000~4 500 mg/kg)TA可能降低仔猪的采食量和日增重[7]。与前人研究结果一致,本研究发现,口腔灌服190 mg/kg BW (410 mg/kg饲粮)TA能显著改善试验仔猪的生长性能,说明TA可以有效降低PEDV感染导致的仔猪腹泻率上升和生长受阻。
动物机体血清生化指标受饲粮营养水平的影响,可以间接反映机体自身营养水平和健康状况[13]。ALT和谷草转氨酶(AST)主要存在肝细胞胞浆和线粒体中,其在血清中活性高低直接与肝细胞或线粒体的损伤程度相关[14-15]。本研究发现,感染PEDV显著提高了仔猪血清ALT活性,灌服TA后感染PEDV仔猪血清ALT活性显著下降,说明灌服TA可缓解PEDV造成的肝功能异常。血清TG和TC含量高低反映了动物对脂类的吸收和代谢能力,HDL和LDL是运输胆固醇的载体,对维持机体脂类代谢稳定起重要作用[16-17]。研究结果表明,PEDV感染导致仔猪血清TC、P、HDL和LDL含量显著下降,血清GLU含量显著升高,说明PEDV感染可能阻碍了营养物质转运,并且使仔猪糖代谢能力下降,导致生长性能的下降。
血清DAO活性和D-木糖含量能够直观反映肠道机械屏障的完整性和受损伤程度[18-19]。当肠黏膜上皮细胞受到损伤时,DAO会释放出来进入血液,导致血液中DAO活性增高,同时使D-木糖吸收能力下降,血液中D-木糖含量也会相应降低[20-21]。与上述结果一致,本研究发现仔猪感染PEDV后,血清中DAO活性显著升高,D-木糖含量显著下降;同时,十二指肠、空肠、回肠和结肠的肠道损伤评分显著升高,说明PEDV感染导致仔猪肠道屏障功能损伤,降低了肠道对营养物质的消化率,而灌服TA能在一定程度上缓解PEDV感染引起的仔猪肠道吸收功能下降。
仔猪的消化吸收功能与肠道黏膜形态完整性密切相关。小肠黏膜上皮绒毛变短、隐窝加深代表肠道黏膜上皮绒毛萎缩,吸收能力下降;VH/CD增大,代表黏膜比面积增大,消化吸收能力增强[22]。有研究表明,TA可显著提高断奶仔猪的肠道VH,降低CD,提高VH/CD[3]。本试验中,感染PEDV后,仔猪十二指肠、空肠和回肠VH以及十二指肠和回肠VA显著下降,空肠、回肠和结肠CD显著增加,十二指肠VH/CD显著下降,而灌服TA后,感染PEDV仔猪空肠VH显著上升,说明TA能有效缓解感染PEDV引起的肠黏膜萎缩,改善了肠道结构,提高了营养物质消化利用率,促进了PEDV感染后仔猪的生长。
T-SOD、GSH-Px和CAT属于机体主要的抗氧化酶系,其活性的高低直接反映了机体的抗氧化性能,并间接反映了机体清除自由基的能力[23-24]。MDA为脂质过氧化产物,其含量反映机体脂质过氧化物的生成速率和强度,MDA含量低代表动物抗氧化能力强[25]。T-AOC是代表机体酶性和非酶性抗氧化物的总体水平,直接反映机体抗氧化酶的活性及抗氧化系统的功能状态[26]。本试验结果发现,感染PEDV后,仔猪血清和肠道主要抗氧化酶活性下降,MDA含量显著上升,说明PEDV感染可引发机体氧化应激反应,造成氧化损伤;灌服TA可增强感染PEDV仔猪的机体抗氧化能力,从而改善PEDV造成的仔猪生长性能下降。

4 结论

灌服190 mg/kg BW TA能缓解PEDV感染造成的仔猪腹泻,改善肠道损伤,提高机体抗氧化能力,从而改善生长性能。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
张艳, 刘雪松, 徐婷婷, 等. 猪流行性腹泻研究进展[J]. 现代畜牧科技, 2021(6):21-24,35.

ZHANG Y, LIU X S, XU T T, et al. Research progress of porcine epidemic diarrhea[J]. Modern Animal Husbandry Science & Technology, 2021(6):21-24,35. (in Chinese)

[2]
李鹏, 吴梦郡, 余魁, 等. 幼龄仔猪PEDV感染肠道损伤模型的建立[J]. 中国畜牧兽医, 2018, 45(6):1715-1721.

LI P, WU M J, YU K, et al. The establishment of the intestinal injury model of PEDV infection in young piglets[J]. China Animal Husbandry & Veterinary Medicine, 2018, 45(6):1715-1721. (in Chinese)

[3]
邓文, 张世昌, 蔡荣斌, 等. 栗树单宁对断奶仔猪生产性能、肠道形态和通透性及抗氧化性能的影响[J]. 中国饲料, 2018(1):44-48.

DENG W, ZHANG S C, CAI R B, et al. Effects of chestnut tannins on the performance,intestinal morphology and permeability,and antioxidant status in weaned piglets[J]. China Feed, 2018(1):44-48. (in Chinese)

[4]
于文栋. 断奶仔猪腹泻的原因与后抗生素时代的综合防控措施[J]. 养殖与饲料, 2022, 21(5):87-89.

YU W D. Causes of diarrhea of weaned piglets and comprehensive prevention and control measures in the post-antibiotic era[J]. Animals Breeding and Feed, 2022, 21(5):87-89. (in Chinese)

[5]
REDONDO L M, CHACANA P A, DOMINGUEZ J E, et al. Perspectives in the use of tannins as alternative to antimicrobial growth promoter factors in poultry[J]. Frontiers in Microbiology, 2014, 5:118.

DOI PMID

[6]
YU J, SONG Y Y, YU B, et al. Tannic acid prevents post-weaning diarrhea by improving intestinal barrier integrity and function in weaned piglets[J]. Journal of Animal Science and Biotechnology, 2020, 11:87.

DOI PMID

[7]
李建光, 高炳辉, 刘倩, 等. 五倍子单宁酸对断奶仔猪腹泻和生长性能的影响[J]. 畜禽业, 2021, 32(10):15,17.

LI J G, GAO B H, LIU Q, et al. Effects of gallnut tannic acid on diarrhea and growth performance of weaned piglets[J]. Livestock and Poultry Industry, 2021, 32(10):15,17. (in Chinese)

[8]
BILIĆ-ŠOBOT D, KUBALE V, ŠKRLEP M, et al. Effect of hydrolysable tannins on intestinal morphology,proliferation and apoptosis in entire male pigs[J]. Archives of Animal Nutrition,2016,70(5):378-388.

[9]
BIAGIA G, CIPOLLINI I, PAULICKS B R, et al. Effect of tannins on growth performance and intestinal ecosystem in weaned piglets[J]. Archives of Animal Nutrition, 2010, 64(2):121-135.

DOI PMID

[10]
ZHANG Q, WU T, LI S Y, et al. Protective effect of zinc oxide and its association with neutrophil degranulation in piglets infected with porcine epidemic diarrhea virus[J]. Oxidative Medicine and Cellular Longevity, 2021, 2021:3055810.

[11]
VILASECA J, SALAS A, GUARNER F, et al. Dietary fish oil reduces progression of chronic inflammatory lesions in a rat model of granulomatous colitis[J]. Gut, 1990, 31(5):539-544.

DOI PMID

[12]
KRISHNA V D, KIM Y, YANG M, et al. Immune responses to porcine epidemic diarrhea virus (PEDV) in swine and protection against subsequent infection[J]. PLoS One, 2020, 15(4):e0231723.

DOI

[13]
包学太, 黎育颖, 田军权, 等. 小檗碱齐墩果酸盐对仔猪生长性能、血清生化指标和肠道健康的影响[J]. 动物营养学报, 2022, 34(3):1495-1507.

DOI

BAO X T, LI Y Y, TIAN J Q, et al. Effects of berberine-oleanolate on growth performance,serum biochemical indices and intestinal health of piglets[J]. Chinese Journal of Animal Nutrition, 2022, 34(3):1495-1507. (in Chinese)

[14]
LV Y F, TANG C H, WANG X Q, et al. Effects of dietary supplementation with palygorskite on nutrient utilization in weaned piglets[J]. Livestock Science, 2015, 174:82-86.

DOI

[15]
LIU Y Y, KONG X F, JIANG G L, et al. Effects of dietary protein/energy ratio on growth performance,carcass trait,meat quality,and plasma metabolites in pigs of different genotypes[J]. Journal of Animal Science and Biotechnology, 2015, 6(1):36.

DOI

[16]
SHARMAN M J, FERNANDEZ M L, ZERN T L, et al. Replacing dietary carbohydrate with protein and fat decreases the concentrations of small LDL and the inflammatory response induced by atherogenic diets in the Guinea pig[J]. Journal of Nutritional Biochemistry, 2008, 19(11):732-738.

DOI PMID

[17]
CHEN Y Y, GONG X X, LI G D, et al. Effects of dietary alfalfa flavonoids extraction on growth performance,organ development and blood biochemical indexes of Yangzhou geese aged from 28 to 70 days[J]. Animal Nutrition, 2016, 2(4):318-322.

DOI

[18]
WU T, LV Y, LI X N, et al. Establishment of a recombinant Escherichia coli-induced piglet diarrhea model[J]. Frontiers in Bioscience, 2018, 23(8):1517-1534.

DOI

[19]
王蕾, 易丹, 吴涛, 等. 几种仔猪肠道损伤模型的建立及肠道损伤生物标志物的研究进展[J]. 中国畜牧杂志, 2019, 55(8):6-12.

WANG L, YI D, WU T, et al. Establishment of several porcine models of intestinal injury and identified biomarkers of intestinal injury with these models[J]. Chinese Journal of Animal Science, 2019, 55(8):6-12. (in Chinese)

[20]
王蕾, 刘坚, 侯永清, 等. α-酮戊二酸对LPS慢性应激仔猪小肠黏膜形态与功能的影响[J]. 畜牧兽医学报, 2010, 41(1):46-52.

WANG L, LIU J, HOU Y Q, et al. The effects of α-ketoglutarate on intestinal mucosal morphology and function in piglets chronically challenged with lipopolysaccharide[J]. Acta Veterinaria et Zootechnica Sinica, 2010, 41(1):46-52. (in Chinese)

[21]
郭雪峰, 边连全, 付亮亮, 等. 酸化剂对早期断奶仔猪胃肠道pH和肠黏膜形态结构的影响[J]. 养猪, 2006(5):4-6.

GUO X F, BIAN L Q, FU L L, et al. The effect of acidifiers on intestinal tract pH value and morphology in piglets[J]. Swine Production, 2006(5):4-6. (in Chinese)

[22]
PUPPEL K, KAPUSTA A, KUCZYŃSKA B. The etiology of oxidative stress in the various species of animals,a review[J]. Journal of the Science of Food and Agriculture, 2015, 95(11):2179-2184.

DOI

[23]
段卫平, 李同新. 亚麻籽对断奶仔猪生长性能、抗氧化功能和血清促炎因子含量的影响[J]. 中国饲料, 2021(24):41-45.

DUAN W P, LI T X. Effect of flaxseed on growth performance,antioxidant capacity and serum pro-inflammatory cytokines of weaned piglets[J]. China Feed, 2021(24):41-45. (in Chinese)

[24]
农斯伟, 沈水宝, 伍校军, 等. 不同工艺处理的栗木水解物替代氧化锌对仔猪生长性能、血清生化和抗氧化指标的影响[J]. 饲料研究, 2020, 43(12):30-33.

NONG S W, SHEN S B, WU X J, et al. Effect of chestnut hydrolysate with different processing replaced by zinc oxide on growth performance,serum biochemical and antioxidant index of piglet[J]. Feed Research, 2020, 43(12):30-33. (in Chinese)

[25]
王福香, 李文立, 任慧英, 等. 纳米硒对肉鸡肝脏硒含量和抗氧化能力的影响[J]. 中国畜牧杂志, 2009, 45(3):27-30.

WANG F X, LI W L, REN H Y, et al. Effects of nano-selenium on the selenium concentration and antioxidant abilities in liver of broiler chickens[J]. Chinese Journal of Animal Science, 2009, 45(3):27-30. (in Chinese)

[26]
陈言言. 苜蓿皂苷对H2O2诱导IEC-6和IPEC-J2细胞氧化损伤的保护作用及其机制[D]. 硕士学位论文. 郑州: 河南农业大学, 2017.

CHEN Y Y. Protective effect of alfalfa saponins against H2O2 induced oxidative stress injury in IEC-6 and IPEC-J2 cells and its mechanism[D]. Master’s Thesis. Zhengzhou: Henan Agricultural University, 2017. (in Chinese)

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