Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 4: 2278 - 2286

DOI: https://doi.org/10.12418/CJAN2025.193

RESEARCH PAPER

Effects of Zinc Oxide-Loaded Montmorillonite on Growth Performance, Diarrhea Rate and Intestinal Health of Weaned Piglets

  • MA Mingqi ,
  • LI Yang ,
  • WU Qiong ,
  • DING Xiaoling ,
  • ZHANG Cheng , *
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  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
* professor, E-mail:

Received date: 2024-09-20

  Online published: 2025-04-15

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Abstract

The aim of this experiment was to investigate the effects of zinc oxide-loaded montmorillonite (ZnO-MMT) on growth performance, diarrhea rate and intestinal health of weaned piglets. A total of 210 weaned piglets with an average body weight of (8.15±0.05) kg at 28 days of age were randomly divided into 7 groups with 6 replicates per group and 5 pigs per replicate. Piglets in the control group (CON group) were fed a basal diet, those in the zinc oxide group (ZnO group) were fed the basal diet supplemented with 1 600 mg/kg zinc oxide (ZnO, containing 1 440 mg/kg zinc), and those in the other five groups were fed the basal diet supplemented with 600 (ZnO-MMT1 group), 1 200 (ZnO-MMT2 group), 1 800 (ZnO-MMT3 group), 2 400 (ZnO-MMT4 group) and 3 000 mg/kg (ZnO-MMT5 group) ZnO-MMT (containing 20.4, 40.8, 61.2, 81.6 and 102.0 mg/kg zinc in sequence), respectively. The experiment lasted for 14 days. The results showed as follows: 1) compared with CON group, the final body weight, average daily feed intake (ADFI) and average daily gain (ADG) of weaned piglets in ZnO group, ZnO-MMT2 group and ZnO-MMT3 group were significantly increased (P<0.05), and the ratio of feed to gain (F/G) in ZnO-MMT2 group and ZnO-MMT3 group was significantly decreased (P<0.05); the diarrhea rate in ZnO group and all ZnO-MMT supplemental groups was significantly decreased (P<0.05). 2) Compared with CON group, the serum D-lactic acid (D-LA) content of weaned piglets in ZnO group and ZnO-MMT2 to ZnO-MMT5 groups was significantly decreased (P<0.05), with the lowest in ZnO-MMT3 group; meanwhile, the serum diamine oxidase (DAO) activity in ZnO group, ZnO-MMT2 group and ZnO-MMT3 group was significantly decreased (P<0.05). 3) Compared with CON group, the villus height and the ratio of villus height to crypt depth (V/C) of weaned piglets in ZnO group and ZnO-MMT3 group were significantly increased (P<0.05). 4) Compared with CON group, the mRNA relative expression levels of zonula occludens-1 (ZO-1) and interleukin-10 (IL-10) in jejunum of weaned piglets in ZnO group and ZnO-MMT3 group were significantly increased (P<0.05), and the interleukin-1β (IL-1β) mRNA relative expression level in jejunum was significantly decreased (P<0.05); the mRNA relative expression levels of occludin and superoxide dismutase (SOD) in jejunum in ZnO-MMT3 group were significantly increased (P<0.05). In conclusion, dietary ZnO and ZnO-MMT supplementation have similar effects on alleviating diarrhea and intestinal inflammatory response, improving growth performance and intestinal antioxidant capacity as well as enhancing intestinal mucosal barrier function of weaned piglets, and ZnO-MMT can effectively replace ZnO in the production of weaned piglets. The optimum supplemental level of ZnO-MMT is 1 800 mg/kg under the experimental conditions.

Key words： zinc oxide-loaded montmorillonite; weaned piglets; growth performance; diarrhea; intestinal health

Cite this article

MA Mingqi , LI Yang , WU Qiong , DING Xiaoling , ZHANG Cheng . Effects of Zinc Oxide-Loaded Montmorillonite on Growth Performance, Diarrhea Rate and Intestinal Health of Weaned Piglets[J]. Chinese Journal of Animal Nutrition, 2025 , 37(4) : 2278 -2286 . DOI: 10.12418/CJAN2025.193

断奶应激会导致仔猪肠道功能受损、腹泻率升高以及生长性能降低等问题[1]。长期以来,将抗生素添加到动物饲粮中可抑制肠道有害菌繁殖,从而提高其生长性能。然而,“禁抗”的实施使得寻求可缓解仔猪断奶应激、降低腹泻率的抗生素替代品成为当务之急[2]。饲粮中添加氧化锌(ZnO)作为一种防治断奶仔猪腹泻的传统方法,在生产中应用广泛[3]。药理剂量的ZnO可有效缓解仔猪应激性腹泻,提高其生长性能,并增强肠道屏障功能和抗氧化能力[4]。然而,高剂量ZnO的添加可能带来金属毒性及环境污染风险等问题[5]。为此,研发应对仔猪断奶应激且更为安全有效的饲料添加剂成为当前研究的热点。
蒙脱石(montmorillonite,MMT)因其良好的稳定性及较强的离子交换能力,常被用作控制药物分子释放速度的理想载体[6]。据报道,通过离子交换法和共沉淀法制备的载氧化锌蒙脱石(ZnO-MMT)不仅孔体积增大,还展现出优异的抑菌性能,对金黄色葡萄球菌、沙门氏菌和大肠杆菌均有较好的抗菌活性[7]。研究表明,饲粮中添加ZnO-MMT可改善肉鸡肠道屏障功能,并提高其生长性能[8]。Hu等[9]报道,将ZnO-MMT添加到饲粮中可使断奶仔猪腹泻率显著降低,提高其生长性能并维持健康的肠道功能。尽管如此,将ZnO-MMT应用在断奶仔猪上的研究仍十分有限,还需开展相关研究。因此,本试验旨在研究饲粮中添加ZnO-MMT对断奶仔猪生长性能、腹泻率和肠道健康的影响,以期为ZnO-MMT在生猪养殖中的实际应用提供依据。

1 材料与方法

1.1 试验材料

ZnO-MMT由本实验室制备,方法如下:取MMT添加到硫酸锌(ZnSO4)与氢氧化钠(NaOH)的混合溶液中(MMT、ZnSO4和NaOH的摩尔比为1.0∶1.5∶1.8),在60 ℃下磁力搅拌6 h,离心后去上清液,加入去离子水清洗3次,烘干,即获得ZnO-MMT。ZnO-MMT中的ZnO负载量为4.2%。

1.2 试验设计

本研究动物试验经安徽农业大学实验动物福利伦理委员会批准(批准号:AHAUXMSQ2023678)。试验选取210头平均体重为(8.15±0.05) kg的28日龄断奶仔猪,随机分为7组,每组6个重复,每个重复5头猪。对照组(CON组)饲喂基础饲粮,氧化锌组(ZnO组)饲喂在基础饲粮的基础上添加1 600 mg/kg ZnO(含锌1 440 mg/kg)的饲粮,其余5组分别饲喂在基础饲粮的基础上添加600(ZnO-MMT1组)、1 200(ZnO-MMT2组)、1 800(ZnO-MMT3组)、2 400(ZnO-MMT4组)和3 000 mg/kg(ZnO-MMT5组)ZnO-MMT(分别含锌20.4、40.8、61.2、81.6和102.0 mg/kg)的饲粮。试验期14 d。基础饲粮参照NRC(2012)仔猪生长所需营养要求配制,其组成及营养水平见表1。
表1 基础饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of the basal diet (air-dry basis) %

项目Items 含量Content
原料Ingredients
玉米Corn 31.70
膨化玉米Extruded corn 32.60
膨化大豆Extruded soybean 15.00
大豆浓缩蛋白Soybean protein concentrate 11.00
鱼粉Fish meal 3.00
蔗糖Sucrose 2.50
葡萄糖Glucose 1.00
石粉Limestone 0.90
磷酸氢钙CaHPO4 0.60
大豆油Soybean oil 0.50
L-赖氨酸盐酸盐L-Lys·HCl 0.30
氯化钠NaCl 0.30
DL-蛋氨酸DL-Met 0.05
氯化胆碱Choline chloride(50%) 0.10
预混料Premix1) 0.45
合计Total 100.00
营养水平Nutrient levels2)
消化能DE/(MJ/kg) 14.78
粗蛋白质CP 19.49
钙Ca 0.71
标准全肠道可消化磷STTD P 0.33
赖氨酸Lys 1.21
苏氨酸Thr 0.66
蛋氨酸+半胱氨酸Met+Cys 0.61

1)预混料为每千克饲粮提供 The premix provided the following per kg of the diet:VA 16 000 IU,VD3 3 000 IU,VE 38.5 mg,VK 3.5 mg,VB1 3.5 mg,VB2 10.0 mg,VB6 5.5 mg,VB12 0.10 mg,叶酸 folic acid 3.0 mg,烟酰胺 niacinamide 35 mg,生物素 biotin 0.25 mg,泛酸 pantothenic acid 30 mg,Cu (CuSO4·5H2O) 6 mg,Fe (FeSO4·H2O) 100 mg,Mn (MnSO4·5H2O) 40 mg,Zn (ZnSO4· H2O) 100 mg,I 0.14 mg,Se (Na2SeO3) 0.35 mg。

2)消化能和标准全肠道可消化磷为计算值,其余为实测值。DE and STTD P were calculated values, while the others were measured values.

1.3 指标测定及方法

1.3.1 饲粮营养水平

饲粮消化能根据《猪营养需要量》(GB/T 39235—2020)中各原料的消化能计算,标准全肠道可消化磷根据《中国饲料成分及营养价值表(2020年第31版)》中各原料的磷含量及其标准全肠道消化率计算;饲粮中粗蛋白质(GB/T 6432—2018)、钙(GB/T 6436—2018)和氨基酸(GB/T 18246—2019)含量参照相应国标方法测定。

1.3.2 生长性能和腹泻率测定

以重复为单位记录断奶仔猪每日采食量,汇总后结合体重数据计算平均日增重(ADG)、平均日采食量(ADFI)和料重比(F/G)。
仔猪粪便以目测评估方式进行评分记录,粪便评分标准如下[10]:0分=正常粪便(条状或颗粒状),1分=软粪便(基本成形但湿润松软),2分=半液体粪便(水分较高但无粪水分离),3分=液体状粪便(水分极高且粪水分离)。当粪便评分在2分及以上时,仔猪被认为是腹泻。
腹泻率(%)=100×(腹泻仔猪数×腹泻天数)/(仔猪总数×试验天数)。

1.3.3 血清肠道通透性指标测定

饲养试验结束后,禁食12 h,每个重复选取接近平均体重的1头仔猪进行前腔静脉采血6 mL,于4 ℃、3 000×g条件下离心10 min,取血清于-20 ℃保存;采用南京建成生物工程研究所生产的试剂盒测定血清D-乳酸(D-LA)含量和二胺氧化酶(DAO)活性。

1.3.4 肠道形态测定

仔猪采血后屠宰,分离空肠,剪取约2.0 cm肠段固定于4%多聚甲醛溶液以制备肠道切片,采用光学显微镜测量空肠绒毛高度(VH)和隐窝深度(CD),并计算绒隐比(V/C)。

1.3.5 空肠黏膜紧密连接、抗氧化和炎症相关基因表达测定

刮取空肠黏膜样品并提取总RNA(TRIzol试剂,Yeasen,中国),检测合格后将RNA反转录为cDNA(Hifair® Ⅱ第1链试剂盒,Yeasen,中国)。使用Hieff®荧光定量试剂盒(Yeasen,中国)和荧光定量仪进行PCR扩增。引物序列参照本课题组前期发表的文献[11],由生工生物工程(上海)股份有限公司合成,以甘油醛-3-磷酸脱氢酶(GAPDH)作为内参基因,测定闭合蛋白(occludin)、封闭蛋白(claudin)-1、claudin-2、闭锁小带蛋白(ZO)-1、ZO-2、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、白细胞介素(IL)-1β、IL-6、IL-8、IL-10和肿瘤坏死因子-α(TNF-α)的mRNA相对表达量。

1.4 数据处理及统计分析

采用SPSS 19.0软件进行单因素方差分析(one-way ANOVA)和Duncan氏法多重比较,采用正交多项式对比来评估ZnO-MMT添加水平的线性和二次效应;结果数据以平均值和均值标准误(SEM)表示,P<0.05表示差异显著。

2 结果与分析

2.1 ZnO-MMT对断奶仔猪生长性能和腹泻率的影响

由表2可知,与CON组相比,ZnO组、ZnO-MMT2组和ZnO-MMT3组断奶仔猪末重、ADFI和ADG显著提高(P<0.05),且ZnO-MMT2组和ZnO-MMT3组F/G显著降低(P<0.05);ZnO组和各ZnO-MMT添加组腹泻率均显著降低(P<0.05)。ZnO-MMT添加水平与断奶仔猪末重、ADFI、ADG、F/G和腹泻率均呈二次相关(P<0.05),且ZnO-MMT3组具有最高的ADG和末重以及最低的F/G。
表2 ZnO-MMT对断奶仔猪生长性能和腹泻率的影响

Table 2 Effects of ZnO-MMT on growth performance and diarrhea rate of weaned piglets

项目
Items		 组别Groups 均值
标准误
SEM		 P值P-value
CON ZnO ZnO-
MMT1		 ZnO-
MMT2		 ZnO-
MMT3		 ZnO-
MMT4		 ZnO-
MMT5		 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
初重
Initial BW/kg		 8.14 8.13 8.14 8.14 8.15 8.14 8.15 0.010 0.972 0.454 0.877
末重
Final BW/kg		 11.47c 12.12ab 11.63c 12.27a 12.32a 11.83bc 11.67c 0.060 0.001 0.121 0.001
平均日采食量
ADFI/g		 438.93c 499.45a 454.03bc 498.42a 495.65a 464.25bc 473.10ab 4.680 0.001 0.014 0.001
平均日增重
ADG/g		 237.90d 285.07ab 249.63cd 295.10a 297.71a 263.30bc 250.90cd 4.480 0.001 0.135 0.001
料重比
F/G		 1.86ab 1.75bcd 1.82abc 1.69cd 1.67d 1.77abcd 1.90a 0.020 0.004 0.948 0.001
腹泻率
Diarrhea rate/%		 32.06a 13.71c 23.12b 10.37c 9.31c 7.55c 8.17c 1.560 0.001 0.001 0.001

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

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

2.2 ZnO-MMT对断奶仔猪血清肠道通透性指标的影响

由表3可知,与CON组相比,ZnO组、ZnO-MMT2组、ZnO-MMT3组、ZnO-MMT4组和ZnO-MMT5组断奶仔猪血清D-LA含量显著降低(P<0.05),且以ZnO-MMT3组最低;同时,ZnO组、ZnO-MMT2组和ZnO-MMT3组血清DAO活性显著降低(P<0.05)。与ZnO组相比,ZnO-MMT2组、ZnO-MMT3组、ZnO-MMT4组和ZnO-MMT5组血清D-LA含量和DAO活性差异均不显著(P>0.05)。ZnO-MMT添加水平与血清D-LA含量呈线性和二次相关(P<0.05),与血清DAO活性呈二次相关(P<0.05)。综上所述,饲粮中添加1 800 mg/kg ZnO-MMT效果较佳,并以此添加水平进行后续试验。
表3 ZnO-MMT对断奶仔猪血清肠道通透性指标的影响

Table 3 Effects of ZnO-MMT on serum intestinal permeability indices of weaned piglets

项目
Items		 组别Groups 均值
标准误
SEM		 P值P-value
CON ZnO ZnO-
MMT1		 ZnO-
MMT2		 ZnO-
MMT3		 ZnO-
MMT4		 ZnO-
MMT5		 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
D-乳酸
D-LA/(μg/mL)		 25.16a 9.07c 19.39ab 11.60c 8.93c 10.60c 14.26bc 1.180 <0.001 <0.001 <0.001
二胺氧化酶
DAO/(U/L)		 11.29a 6.11c 9.66ab 6.74bc 7.66bc 8.17abc 8.95abc 0.450 0.026 0.100 0.014

2.3 ZnO-MMT对断奶仔猪肠道形态的影响

由表4可知,与CON组相比,ZnO组和ZnO-MMT3组断奶仔猪空肠VH和V/C显著提高(P<0.05)。ZnO组和ZnO-MMT3组间空肠VH和V/C无显著差异(P>0.05)。各组间空肠CD无显著差异(P>0.05)。
表4 ZnO-MMT对断奶仔猪肠道形态的影响

Table 4 Effects of ZnO-MMT on intestinal morphology of weaned piglets

项目
Items		 组别Groups 均值标准误
SEM		 P值
P-value
CON ZnO ZnO-MMT3
绒毛高度VH/μm 275.75b 326.38a 315.10a 8.580 0.029
隐窝深度CD/μm 161.29 149.83 154.24 3.270 0.374
绒隐比V/C 1.17b 2.18a 2.05a 0.070 0.004

2.4 ZnO-MMT对断奶仔猪空肠紧密连接相关基因表达的影响

由表5可知,与CON组相比,ZnO组和ZnO-MMT3组断奶仔猪空肠ZO-1的mRNA相对表达量均显著提高(P<0.05);ZnO组空肠occludin的mRNA相对表达量无显著差异(P>0.05),但ZnO-MMT3组空肠occludin的mRNA相对表达量显著提高(P<0.05)。各组间断奶仔猪空肠ZO-2、claudin-1和claudin-2的mRNA相对表达量均无显著差异(P>0.05)。
表5 ZnO-MMT对断奶仔猪空肠紧密连接相关基因表达的影响

Table 5 Effects of ZnO-MMT on tight junction related gene expression in jejunum of weaned piglets

项目
Items		 组别Groups 均值标准误
SEM		 P值
P-value
CON ZnO ZnO-MMT3
闭锁小带蛋白-1 ZO-1 1.00b 2.06a 1.81a 0.180 0.034
闭锁小带蛋白-2 ZO-2 1.00 1.03 1.26 0.080 0.357
闭合蛋白Occludin 1.00b 1.24ab 1.62a 0.100 0.039
封闭蛋白-1 Claudin-1 1.00 1.28 1.11 0.100 0.552
封闭蛋白-2 Claudin-2 1.00 0.93 1.08 0.060 0.658

2.5 ZnO-MMT对断奶仔猪空肠抗氧化相关基因表达的影响

由表6可知,与CON组相比,ZnO-MMT3组断奶仔猪空肠SOD的mRNA相对表达量显著提高(P<0.05),且效果优于ZnO组。各组间断奶仔猪空肠GSH-Px和CAT的mRNA相对表达量无显著差异(P>0.05)。
表6 ZnO-MMT对断奶仔猪空肠抗氧化相关基因表达的影响

Table 6 Effects of ZnO-MMT on antioxidant related gene expression in jejunum of weaned piglets

项目
Items		 组别Groups 均值标准误
SEM		 P值
P-value
CON ZnO ZnO-MMT3
超氧化物歧化酶SOD 1.00b 1.40ab 1.63a 0.100 0.030
谷胱甘肽过氧化物酶GSH-Px 1.00 0.95 1.17 0.080 0.544
过氧化氢酶CAT 1.00 1.22 0.98 0.080 0.457

2.6 ZnO-MMT对断奶仔猪空肠炎症相关基因表达的影响

由表7可知,与CON组相比,ZnO组和ZnO-MMT3组断奶仔猪空肠IL-1β的mRNA相对表达量显著降低(P<0.05),空肠IL-10的mRNA相对表达量显著提高(P<0.05)。同时,ZnO组和ZnO-MMT3组间空肠IL-1β和IL-10的mRNA相对表达量无显著差异(P>0.05)。各组间空肠IL-6、IL-8和TNF-α的mRNA相对表达量均无显著差异(P>0.05)。
表7 ZnO-MMT对断奶仔猪空肠炎症相关基因表达的影响

Table 7 Effects of ZnO-MMT on inflammation related gene expression in jejunum of weaned piglets

项目
Items		 组别Groups 均值标准误
SEM		 P值
P-value
CON ZnO ZnO-MMT3
白细胞介素-1β IL-1β 1.00a 0.57b 0.57b 0.070 0.013
肿瘤坏死因子-α TNF-α 1.00 0.71 0.64 0.070 0.076
白细胞介素-6 IL-6 1.00 1.07 0.97 0.070 0.843
白细胞介素-8 IL-8 1.00 1.24 1.04 0.080 0.416
白细胞介素-10 IL-10 1.00b 1.83a 1.64a 0.130 0.012

3 讨论

3.1 ZnO-MMT对断奶仔猪生长性能和腹泻率的影响

断奶仔猪肠道生长发育不完善,合群、饲粮改变等多种因素均会导致肠道功能受损、菌群紊乱以及腹泻等问题[12]。药理剂量的ZnO可缓解断奶仔猪腹泻并显著提高其生长性能[13]。Peng等[14]报道,将ZnO添加到断奶仔猪饲粮中可有效维持其肠道菌群结构平衡并提高采食量。Hu等[9]研究发现,将ZnO-MMT添加到饲粮中可使断奶仔猪的ADG和ADFI显著升高。本试验结果显示,断奶仔猪饲粮中添加ZnO和ZnO-MMT均可显著提高其生长性能并缓解断奶应激导致的腹泻,提示ZnO-MMT能有效替代ZnO。此外,ZnO-MMT3组比ZnO组和其他ZnO-MMT组具有更高的ADG、末重和更低的F/G,提示饲粮中ZnO-MMT的添加水平为1 800 mg/kg时断奶仔猪的生长性能较佳。

3.2 ZnO-MMT对断奶仔猪肠道形态的影响

通常情况下,断奶应激会导致仔猪的小肠形态发生变化,包括VH降低和CD增加。小肠绒毛是肠道吸收营养物质的重要结构,VH越高,小肠上皮细胞越多,小肠吸收营养物质的能力就越强[9]。肠隐窝是由小肠上皮下陷形成的管状腺,存在于绒毛根部之间,肠道干细胞的活性和功能由VH反映[15]。因此,VH、CD及V/C是评价小肠健康和功能的3个关键形态学指标。研究表明,饲粮中添加ZnO可提高断奶仔猪小肠VH[16]。本研究结果发现,ZnO组断奶仔猪空肠VH和V/C显著高于CON组。Hu等[9]研究发现,与CON组和ZnO组相比,ZnO-MMT组仔猪具有更高的VH和V/C。与此一致,本研究结果亦表明,饲粮中添加ZnO-MMT可显著提高断奶仔猪空肠VH和V/C,且可达到高剂量ZnO的作用效果。原因可能是ZnO-MMT对肠道中的病菌及毒素具有较强的吸附能力,且将ZnO负载入MMT可使锌离子(Zn2+)在肠道内持续释放,从而维持肠道形态结构的完整性[17]。

3.3 ZnO-MMT对断奶仔猪肠道通透性的影响

肠道黏膜屏障是阻止毒素、细菌及其代谢物进入动物机体的主要屏障,其完整性对于动物健康具有十分重要的作用。DAO是一种细胞内酶,存在于哺乳动物肠绒毛细胞中;D-LA是肠道内细菌的代谢产物。肠道通透性通常由血清中D-LA含量和DAO活性来反映。前人研究揭示了断奶应激会导致仔猪肠道黏膜屏障受损,血清中DAO活性和D-LA含量升高,肠道的通透性显著增强,更易导致腹泻[18-19]。Huang等[20]研究发现,将ZnO添加到断奶仔猪饲粮中可使其肠黏膜和血清中DAO的活性显著降低,缓解腹泻症状。前人研究表明,饲粮中添加ZnO-MMT可显著降低断奶仔猪血清D-LA含量和DAO活性以及腹泻率[9]。与上述研究一致,本试验结果发现,饲粮中添加ZnO和ZnO-MMT均可显著降低断奶仔猪血清D-LA含量和DAO活性,且ZnO-MMT能达到高剂量ZnO的效果,综合而言以1 800 mg/kg的添加水平为宜。

3.4 ZnO-MMT对断奶仔猪空肠紧密连接相关基因表达的影响

肠道紧密连接复合体位于上皮细胞的顶端,由多种蛋白质构成,occludin和claudin是其中重要的跨膜蛋白,可与胞浆蛋白ZO-1结合,在调节肠道稳态、维持上皮细胞屏障方面共同发挥关键作用,其表达异常会影响肠道的结构和功能,更易导致腹泻[21]。因此,ZO-1、occludin和claudin通常被视为判断紧密连接屏障功能的关键分子。研究表明,饲粮中添加ZnO可使断奶仔猪空肠组织中ZO-1和occludin的mRNA相对表达量显著升高,降低肠上皮细胞的通透性与肠黏膜损伤,从而改善断奶引起的腹泻[14,22]。但目前关于ZnO-MMT对断奶仔猪肠道屏障功能影响的研究十分有限。本试验结果显示,饲粮中添加ZnO和ZnO-MMT均显著提高了断奶仔猪空肠黏膜中ZO-1的mRNA相对表达量;此外,ZnO-MMT组空肠occludin的mRNA相对表达量也显著提高,表明将ZnO-MMT添加到断奶仔猪饲粮中可增强其肠道黏膜屏障完整性,降低腹泻率,并有效替代ZnO的使用。

3.5 ZnO-MMT对断奶仔猪空肠抗氧化相关基因表达的影响

断奶导致的腹泻通常与氧化应激密切相关,断奶应激导致仔猪体内抗氧化酶活性下降,肠道抗氧化功能失衡[23]。仔猪处于氧化应激时肠道中抗氧化酶GSH-Px和SOD的基因表达水平会下降,导致机体清除自由基的能力降低,进而影响其生长性能[24]。锌参与多种抗氧化酶的构成,可直接影响抗氧化酶的活性,对机体氧化还原平衡起着重要作用[25]。高剂量ZnO可通过降低断奶仔猪细胞内谷胱甘肽的氧化还原比值改善小肠氧化状态,缓解肠道损伤[26]。然而,关于ZnO-MMT对断奶仔猪肠道抗氧化能力影响的研究未见报道。本试验结果表明,尽管ZnO组和ZnO-MMT组断奶仔猪空肠GSH-Px和CAT的mRNA相对表达量与CON组相比差异不显著,但ZnO-MMT组空肠SOD的mRNA相对表达量与与CON组相比显著提高,提示饲粮中添加ZnO-MMT可增强断奶仔猪肠道抗氧化能力,这可能是其缓解仔猪应激腹泻、增强肠道屏障功能的原因。

3.6 ZnO-MMT对断奶仔猪空肠炎症相关基因表达的影响

细胞因子具有广泛的生物活性,对免疫和炎症反应具有关键调控作用,包括IL、趋化因子和肿瘤坏死因子等[27]。其中,IL-1β是促炎细胞因子,对免疫细胞的分化与增殖具有调控作用;IL-8作为趋化因子,主要介导免疫细胞的募集与激活[28];IL-6和TNF-α可强化其他促炎细胞因子,并参与炎症反应的启动与维持[29];抗炎因子IL-10可抑制过度的T细胞免疫反应,缓解炎症损伤,维持肠道稳态[30]。研究发现,断奶使仔猪肠道屏障受损,IL-6和TNF-α的表达上调[31]。本试验结果表明,饲粮中添加ZnO和ZnO-MMT可使断奶仔猪空肠黏膜中IL-1β的mRNA相对表达量显著降低,并使空肠黏膜中IL-10的mRNA相对表达量显著提高,表明饲粮中添加ZnO和ZnO-MMT均能缓解断奶仔猪肠道炎症反应。上述结果亦表明,ZnO-MMT可作为高剂量ZnO的替代产品,使断奶仔猪肠道中促炎细胞因子的表达降低,同时提高抗炎因子的表达,抑制断奶应激引发的肠道炎症反应,从而有效保护肠道形态结构及屏障功能,降低腹泻,并提高生长性能。

4 结论

饲粮中添加ZnO和ZnO-MMT在缓解断奶仔猪腹泻和肠道炎症反应,提高生长性能、肠道抗氧化能力以及改善肠道黏膜屏障功能方面具有相似的效果。因此,ZnO-MMT可有效替代ZnO应用于断奶仔猪生产。在本试验条件下,ZnO-MMT的适宜添加水平为1 800 mg/kg。

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