Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 8: 4988 - 4997

DOI: https://doi.org/10.12418/CJAN2023.463

RESEARCH PAPER

Effects of Nano Zinc Oxide on Blood Biochemical, Antioxidant and Immune Indexes and Free Amino Acid Contents in Piglets

  • CUI Yiyan , 1 ,
  • TIAN Zhimei 1 ,
  • LIU Zhichang 1 ,
  • RONG Ting 1 ,
  • LI Zhenming 1 ,
  • YU Miao 1 ,
  • MA Xianyong 1, 2 ,
  • YE Xiufeng , 1, *
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  • 1 Guangdong Engineering Technology Research Center of Animal Meat Quality and Safety Control and Evaluation, Guangdong Provincial Key Laboratory of Animal Breeding and Nutrition, Key Laboratory of Animal Nutrition and Feed Science in South China, Ministry of Agriculture and Rural Affairs, State Key Laboratory of Livestock and Poultry Breeding, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
  • 2 Guangdong Laboratory for Lingnan Modern Agriculture, Maoming Branch, Maoming 525000, China
*senior livestock specialist, E-mail:

Received date: 2023-02-17

  Online published: 2023-08-10

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Abstract

This study investigated the effects of nano zinc oxide (N-ZnO) on blood biochemical, antioxidant and immune indexes and free amino acid contents in piglets. A total of 144 piglets [Duroc×Landrace×Large White, weaned at 21 days of age; initial weight (6.15±0.01) kg] were assigned to 6 groups with 6 replicates in each group and 4 pigs per replicate. The groups were: antibiotic (ANTI) group (basal diet+75 mg/kg aureomycin), zinc oxide (ZnO) group (basal diet+1 600 mg/kg ZnO), and N-ZnO groups (basal diet + 200, 300, 400 or 500 mg/kg N-ZnO, named as N200, N300, N400 and N500 groups, respectively). After 26 d, the blood was collected to determine plasma biochemical and antioxidant indexes and free amino acid contents, as well as serum immunoglobulin (Ig) contents. The results showed as follows. 1) compared with ANTI and ZnO groups, the plasma aspartate aminotransferase activity was significantly decreased in N-ZnO groups (P<0.05), and the activities of alanine aminotransferase and alkaline phosphatase were significantly higher in ZnO group than those in ANTI and N-ZnO groups (P<0.05). 2) The plasma glutathione peroxidase activity was significantly higher in ANTI, ZnO and N500 groups than that in the rest groups (P<0.05). Compared with ZnO and N500 groups, plasma total antioxidant capacity was significantly decreased in N200 to N400 groups (P<0.05). 3) Compared with ANTI group, the serum IgA content was significantly increased in N200 to N400 groups and ZnO group (P<0.05). Compared with ANTI and ZnO groups, the serum IgG content in N200 to N300 groups and the IgM content in N400 group were significantly increased (P<0.05). With the increase of N-ZnO additive amount, the contents of IgA and IgG in serum were decreased linearly and quadratically (P<0.05), and IgG content was increased linearly and quadratically (P<0.05). 4) Compared with ANTI and ZnO groups, the plasma histidine content was significantly increased in N200 to N400 groups (P<0.05), and the contents of proline, β-alanine, and hydroxyproline were significantly decreased (P<0.05). With the increase of N-ZnO additive amount, the contents of arginine and citrulline in plasma were decreased quadratically (P<0.05), while the contents of serine, glycine and hydroxyproline were increased quadratically or linearly (P<0.05). In conclusion, dietary N-ZnO improves the liver function, antioxidant capacity, immune function, and alters free amino acid profile in plasma of piglets.

Key words： nano zinc oxide; piglets; antioxidant capacity; free amino acid

Cite this article

CUI Yiyan , TIAN Zhimei , LIU Zhichang , RONG Ting , LI Zhenming , YU Miao , MA Xianyong , YE Xiufeng . Effects of Nano Zinc Oxide on Blood Biochemical, Antioxidant and Immune Indexes and Free Amino Acid Contents in Piglets[J]. Chinese Journal of Animal Nutrition, 2023 , 35(8) : 4988 -4997 . DOI: 10.12418/CJAN2023.463

断奶应激会导致仔猪肠道功能障碍、免疫稳态受损、炎症增加以及产生氧化应激,使仔猪腹泻增加和生长不良。仔猪断奶后前2周,使用药理剂量氧化锌(zinc oxide,ZnO)和抗生素是预防、治疗仔猪腹泻的普遍措施[1]。抗生素和高剂量ZnO能改善肠上皮组织结构和免疫系统,从而提高仔猪生长性能[2-3]。但是,用高剂量ZnO或抗生素饲喂仔猪将增加多次抗药性和重金属污染的风险[4-5]。我国农业农村部第2625号和第194号公告明确规定,ZnO在仔猪断奶后使用剂量不能超过1 600 mg/kg锌,不得在饲粮中添加促生长抗生素。然而,减少ZnO或抗生素可能会增加腹泻发生率,降低仔猪生长性能,继而减少养猪业的经济效益。因此,寻找高剂量ZnO和抗生素的替代品用于缓解断奶应激具有重要意义。
纳米氧化锌(nano zinc oxide,N-ZnO)是直径在1~100 nm的ZnO产品,具有独特的体积效应和表面效应。N-ZnO体积小,更容易被吸收。与ZnO相比,N-ZnO具有更高的生物利用度[6]。此外,N-ZnO具有抗菌、抗氧化、免疫调节以及促进生长等功效[7⇓-9]。这意味着使用比普通ZnO更少量的N-ZnO作为动物饲粮中的锌源,可以产生同等甚至更好的效果。这也有助于减少浪费和环境退化[10]。使用低剂量的N-ZnO可能解决高剂量ZnO带来的负面影响,而且本课题组同期试验表明,添加不同剂量的N-ZnO对仔猪生长性能无显著影响[11]。因此,本试验研究不同剂量的N-ZnO对仔猪血液生化、抗氧化、免疫指标以及游离氨基酸含量的影响,旨在为低剂量N-ZnO的应用提供参考。

1 材料与方法

1.1 试验设计

选取初始体重为(6.15±0.01) kg的21日龄杜×长×大断奶仔猪144头,按体重随机分为6组,每组6个重复,每个重复4头猪(公母各占1/2)。具体分组情况如下:抗生素组(ANTI组,基础饲粮+56 g/t 金霉素)、ZnO组(基础饲粮+1 600 mg/kg ZnO)、N200组(基础饲粮+200 mg/kg N-ZnO)、N300组(基础饲粮+300 mg/kg N-ZnO)、N400组(基础饲粮+400 mg/kg N-ZnO)、N500组(基础饲粮+500 mg/kg N-ZnO)。ZnO和N-ZnO由广东某动物保健品有限公司提供,N-ZnO平均粒径为70 nm。基础饲粮参照NRC(2012)标准配制,基础饲粮组成及营养水平见表1[11]。
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 35.31
膨化玉米Extruded corn 15.00
发酵豆粕Fermented soybean meal 9.00
膨化大豆Extruded soybean 10.00
鱼粉Fish meal 4.00
乳清粉Whey powder 11.00
大豆皮Soybean hulls 5.00
大豆油Soybean oil 1.20
血浆蛋白粉Plasma protein powder 3.00
白砂糖White granulated sugar 2.00
氯化胆碱Choline chloride 0.20
食盐NaCl 0.45
二水磷酸氢钙CaHPO4·2H2O 0.62
石粉Limestone 0.65
L-赖氨酸盐酸盐L-Lys·HCl 0.60
L-蛋氨酸L-Met 0.22
L-苏氨酸L-Thr 0.21
L-色氨酸L-Trp 0.04
预混料Premix1) 1.50
合计Total 100.00
营养水平Nutrient levels2)
消化能DE/(MJ/kg) 14.90
粗蛋白质CP 19.20
钙Ca 0.68
总磷TP 0.56
有效磷AP 0.39
赖氨酸Lys 1.57
蛋氨酸+半胱氨酸Met+Cys 0.89
苏氨酸Thr 0.97
色氨酸Trp 0.26

1)预混料为每千克饲粮提供 Premix provided the following per kg of the diet:VA 12 400 IU,VD3 2 800 IU,VE 30 IU,VK 5 mg,VB12 40 μg,VB1 3 mg,VB2 10 mg,烟酸 nicotinic acid 40 mg,D-泛酸 D-pantothenic acid 15 mg,叶酸 folic acid 1 mg,VB6 8 mg,生物素 biotin 0.08 mg,Fe 120 mg,Cu 16 mg,Mn 70 mg,Zn 120 mg,I 0.7 mg,Se 0.48 mg。

2)营养水平为计算值。Nutrient levels were calculated values.

1.2 饲养管理

试验在广东省农业科学院动物科学研究所仔猪舍进行。试验仔猪在漏缝高床上饲养,自由采食和饮水。各组饲养管理和环境条件一致。消毒及免疫程序按照猪场管理进行。

1.3 样品采集

试验结束当天空腹采血。每组选6头接近平均体重的猪进行前腔静脉采血,抗凝采血真空管收集血浆,不添加抗凝剂采血真空管收集血清,-80 ℃保存。

1.4 血浆生化指标的测定

血浆葡萄糖(GLU)、尿素氮(UN)、白蛋白(ALB)、总蛋白(TP)、总胆固醇(TCHO)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)含量以及谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)活性根据南京建成生物工程研究所试剂盒说明书进行检测。

1.5 血浆抗氧化指标的测定

血浆总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、铜锌超氧化物歧化酶(CuZn-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量根据南京建成生物工程研究所试剂盒说明书进行检测。

1.6 血清免疫指标的测定

血清中免疫球蛋白(Ig)A、IgG、IgM的含量根据北京方程生物科技有限公司酶联免疫吸附试验(ELISA)试剂盒说明书进行检测。

1.7 血浆游离氨基酸含量的测定

血浆与10%磺基水杨酸按照1∶3的比例混匀,然后在4 ℃下,12 000×g离心15 min,取上清过0.22 μm滤膜,通过氨基酸分析仪(L-8900,日立,日本)测定游离氨基酸含量。

1.8 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),以Tukey法进行多重比较。通过正交多项式对比评估了随着N-ZnO添加量增加的线性和二次效应。试验结果以平均值和均值标准误(SEM)表示,P<0.05表示差异显著,0.05<P≤0.10表示差异有显著的趋势。

2 结果

2.1 纳米氧化锌对仔猪血浆生化指标的影响

由表2可知,与ANTI组和ZnO组相比,N-ZnO组血浆AST活性显著降低(P<0.05)。ZnO组血浆ALT和ALP活性显著高于ANTI组、N-ZnO组(P<0.05),N-ZnO组间血浆AST、ALT、ALP活性差异不显著(P>0.05)。各组间其他血浆生化指标均无显著差异(P>0.05)。
表2 纳米氧化锌对仔猪血浆生化指标的影响

Table 2 Effects of N-ZnO on plasma biochemical indexes in piglets

项目
Items		 组别Groups 均值
标准误
SEM		 P值P-value
ANTI ZnO N200 N300 N400 N500 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
葡萄糖GLU/(mmol/L) 3.79 4.96 3.90 3.87 3.27 4.13 0.66 0.652 0.980 0.558
总蛋白TP/(g/L) 49.62 50.73 51.53 49.67 51.04 49.74 1.13 0.827 0.542 0.846
白蛋白ALB/(g/L) 32.93 36.92 39.07 36.87 36.40 37.39 1.83 0.354 0.526 0.416
尿素氮UN/(mmol/L) 13.82 15.37 14.52 13.70 13.37 13.73 1.22 0.919 0.691 0.696
甘油三酯TG/(mmol/L) 0.35 0.34 0.33 0.36 0.30 0.44 0.05 0.609 0.273 0.298
总胆固醇
TCHO/(mmol/L)		 2.13 1.79 2.15 1.93 2.02 1.80 0.15 0.432 0.242 0.976
低密度脂蛋白胆固醇
LDL-C/(mmol/L)		 1.11 0.73 1.20 0.98 0.94 1.03 0.12 0.173 0.378 0.250
高密度脂蛋白胆固醇
HDL-C/(mmol/L)		 0.82 0.89 0.99 0.90 0.77 0.88 0.07 0.512 0.293 0.297
谷草转氨酶AST/(U/L) 25.88a 23.59a 14.73b 14.62b 10.26b 12.27b 1.76 0.039 0.353 0.706
谷丙转氨酶ALT/(U/L) 17.38b 24.82a 17.40b 14.37b 15.20b 15.68b 2.26 0.038 0.673 0.375
碱性磷酸酶ALP/(U/L) 16.96b 22.63a 16.10b 17.37b 17.12b 14.92b 1.56 0.049 0.554 0.223

同行数据肩标无字母或相同字母表示差异不显著(P>0.05),不同小写字母表示差异显著(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 small letter superscripts mean significant difference (P<0.05). The same as below.

2.2 纳米氧化锌对仔猪血浆抗氧化指标的影响

由表3可知,与ANTI、ZnO、N500组相比,N200、N300、N400组血浆GSH-Px活性显著降低(P<0.05),N200、N300、N400组间差异不显著(P>0.05)。与ZnO组和N500组相比,N200、N300、N400组血浆T-AOC降低(P<0.05),ANTI组血浆T-AOC与各组间差异均不显著(P>0.05)。
表3 纳米氧化锌对仔猪血浆抗氧化指标的影响

Table 3 Effects of N-ZnO on plasma antioxidant indexes in piglets

项目
Items		 组别Groups 均值
标准误
SEM		 P值P-value
ANTI ZnO N200 N300 N400 N500 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
丙二醛MDA/(nmol/L) 2.07 2.55 2.00 1.83 2.29 1.78 0.38 0.774 0.908 0.666
过氧化氢酶CAT/(U/mL) 6.03 6.27 4.87 5.32 4.50 4.80 1.26 0.920 0.859 0.963
谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 254.39a 258.45a 206.51b 191.37b 220.59b 252.09a 10.24 0.001 0.199 0.647
总抗氧化能力
T-AOC/(U/mL)		 2.08ab 2.54a 1.62b 1.43b 1.41b 2.45a 0.23 0.044 0.278 0.243
超氧化物歧化酶
SOD/(U/mL)		 179.42 172.67 187.22 178.26 173.29 180.78 4.40 0.257 0.327 0.114
铜锌超氧化物歧化酶
CuZn-SOD/(U/mL)		 79.14 85.91 84.13 88.10 84.27 81.02 5.80 0.925 0.627 0.586

2.3 纳米氧化锌对仔猪血清免疫指标的影响

由表4可知,与ANTI组相比,N200、N300、N400、ZnO组血清IgA含量显著提高(P<0.05),ZnO组与N200、N300组血清IgA含量差异不显著(P>0.05)。与ANTI组和ZnO组相比,N200组和N300组血清IgG含量显著提高(P<0.05)。与ANTI组和ZnO组相比,N400组血清IgM含量显著提高(P<0.05)。随着N-ZnO添加量的增加,血清中IgA、IgG含量呈线性降低和二次曲线变化(P<0.05),血清中IgM含量呈线性增加和二次曲线变化(P<0.05)。
表4 纳米氧化锌对仔猪血清免疫指标的影响

Table 4 Effects of N-ZnO on serum immune indexes in piglets μg/mL

项目
Items		 组别Group 均值
标准误
SEM		 P值P-value
ANTI ZnO N200 N300 N400 N500 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
免疫球蛋白A IgA 89.76c 112.16a 111.44ab 107.99ab 105.88b 90.48c 1.93 <0.001 <0.001 0.007
免疫球蛋白G IgG 485.33d 575.75c 690.03a 656.63b 600.56c 506.80d 10.43 <0.001 <0.001 <0.001
免疫球蛋白M IgM 96.68d 111.30b 90.43e 105.13c 117.46a 100.91cd 1.97 <0.001 <0.001 <0.001

2.4 纳米氧化锌对仔猪血浆游离氨基酸含量的影响

由表5可知,与ANTI组和ZnO组相比,N300组血浆精氨酸含量和N200、N300、N400组的组氨酸含量显著提高(P<0.05),N200组丝氨酸含量显著降低(P<0.05)。N200、N300、N400组血浆脯氨酸、β-丙氨酸、羟脯氨酸含量显著低于ANTI组和ZnO组(P<0.05),N500组与ANTI组、ZnO组差异不显著(P>0.05)。随着N-ZnO添加量的提高,仔猪血浆组氨酸(0.05<P≤0.10)、精氨酸(P<0.05)、瓜氨酸(P<0.05)、丝氨酸(P<0.05)含量呈二次曲线变化,色氨酸(0.05<P≤0.10)、甘氨酸(P<0.05)、羟脯氨酸(P<0.05)、总非必需氨基酸(0.05<P≤0.10)含量呈线性增加。
表5 纳米氧化锌对仔猪血浆游离氨基酸含量的影响

Table 5 Effects of N-ZnO on plasma free amino acid contents in piglets ng/μL

项目
Items		 组别Groups 均值
标准误
SEM		 P值P-value
ANTI ZnO N200 N300 N400 N500 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
缬氨酸Valine 15.85 15.19 13.55 17.88 15.53 13.07 4.00 0.959 0.830 0.368
异亮氨酸Isoleucine 7.99 12.72 7.21 8.74 9.02 9.43 1.29 0.113 0.267 0.656
亮氨酸Leucine 21.29 19.44 17.24 22.16 18.09 18.53 2.94 0.794 0.902 0.399
苯丙氨酸Phenylalanine 13.67 12.24 12.07 15.34 13.17 13.93 1.43 0.599 0.663 0.280
色氨酸Tryptophan 2.50 3.19 3.04 3.42 3.33 4.17 0.63 0.827 0.053 0.576
赖氨酸Lysine 45.37 53.95 38.37 46.43 39.80 47.74 6.65 0.700 0.410 0.905
组氨酸Histidine 3.09b 2.46b 5.53a 7.03a 5.72a 5.15a 0.51 0.048 0.481 0.078
精氨酸Arginine 7.07c 7.49c 11.42b 18.05a 13.70ab 11.70b 1.32 0.012 0.224 0.022
丝氨酸Serine 25.93a 26.15a 14.50b 19.88ab 22.73ab 19.15ab 2.26 0.032 0.064 0.017
甘氨酸Glycine 88.05 105.03 69.18 76.04 78.25 95.04 9.08 0.110 0.038 0.577
丙氨酸Alanine 54.95 60.82 54.78 48.39 52.15 58.13 4.80 0.497 0.501 0.257
谷氨酸Glutamate 22.64 23.40 27.04 19.19 22.02 20.94 3.19 0.680 0.430 0.332
谷氨酰胺Glutamine 43.42 48.86 39.85 39.97 40.09 45.74 6.83 0.917 0.593 0.734
天冬氨酸Aspartic acid 0.81 1.59 1.30 1.87 1.98 2.59 0.65 0.544 0.551 0.975
天冬酰胺Asparagine 2.12 2.68 1.90 1.76 1.74 2.43 0.29 0.154 0.195 0.174
酪氨酸Tyrosine 10.62 10.06 10.90 12.27 10.40 11.69 1.39 0.905 0.965 0.914
脯氨酸Proline 27.29a 30.77a 23.04b 23.82b 20.99b 25.04ab 0.90 0.040 0.617 0.375
鸟氨酸Ornithine 9.71 12.21 9.84 12.53 11.01 11.50 1.88 0.892 0.693 0.434
牛磺酸Taurine 26.23 31.20 23.90 23.29 17.72 23.51 3.79 0.359 0.748 0.472
瓜氨酸Citrulline 5.18 5.80 3.29 6.07 4.74 4.15 0.81 0.269 0.927 0.043
β-丙氨酸β-alanine 6.63a 5.65a 2.06b 1.67b 1.67b 3.91ab 0.73 0.001 0.093 0.061
羟脯氨酸Hydroxyproline 20.22ab 23.26a 8.05c 12.05bc 13.19bc 18.08ab 1.79 <0.001 0.004 0.882
α-氨基已二酸
α-aminoadipic acid		 6.64 4.64 5.68 5.43 6.23 5.14 1.30 0.909 0.884 0.799
α-氨基正丁酸
α-amino-n-butyric acid		 1.54 1.48 1.67 2.17 1.75 2.07 0.49 0.972 0.863 0.835
胱硫醚Cystathionine 20.47 19.98 14.31 20.15 18.41 18.21 2.92 0.740 0.536 0.286
总必需氨基酸TEAA 131.14 134.24 121.28 153.16 131.09 135.10 16.37 0.825 0.883 0.336
总非必需氨基酸TNEAA 366.50 400.76 311.78 326.34 325.13 364.40 20.70 0.055 0.093 0.582
总氨基酸TAA 497.64 535.00 433.06 479.50 456.22 499.49 31.88 0.358 0.227 0.888

3 讨论

3.1 纳米氧化锌对仔猪血浆生化指标的影响

血液生化指标是评估动物健康状况的重要指标。给大鼠灌胃N-ZnO(87.5、175、350 mg/kg 体重)未改变大鼠血液生化指标[12]。本研究与上述研究结果相似,各组仔猪血浆TCHO、TG、GLU等生化指标均无显著差异。同期研究表明,N-ZnO、ZnO、ANTI组仔猪生长性能、腹泻率、皮肤、毛色等健康指标差异不显著[11]。这与血浆生化指标的变化结果一致,说明各组间仔猪健康状况相似。毛俊舟等[13]发现,凹土N-ZnO降低仔猪血液TCHO和TG含量,提高HDL-C含量。由此可见,不同来源或结构(70 nm N-ZnO与凹土N-ZnO[13])的N-ZnO对血液生化指标的影响不一致。
AST和ALT活性常用作肝脏毒性的标准量度。饲喂1 600 mg/kg ZnO比饲喂N-ZnO的仔猪具有更高的血浆AST、ALT、ALP活性,与Bai等[14]和Cho等[15]研究结果一致。高剂量ZnO引起肝细胞损伤,导致血浆转氨酶活性增加。研究报道,添加400、800、1 200 mg/kg N-ZnO对仔猪血清AST、ALT、乳酸脱氢酶活性无显著影响[16-17]。本试验与上述研究结果相似,N-ZnO组间血浆转氨酶活性差异不显著,且转氨酶活性低于ZnO组,可见200~500 mg/kg N-ZnO对仔猪肝脏功能无损害作用。Hosseini-Vardanjani等[18]和Cho等[15]报道增加锌的吸收可以提高血清中ALP活性。ZnO组添加了1 600 mg/kg ZnO,其中锌含量高于ANTI组和N-ZnO组(200~500 mg/kg),这可能是ZnO组血浆ALP活性提高的原因。

3.2 纳米氧化锌对仔猪血浆抗氧化指标的影响

锌是许多抗氧化酶的基本元素,影响酶的活性与稳定性[19]。断奶应激通常与氧化应激有关,表现为MDA含量升高以及抗氧化酶活性降低。SOD、CAT、GSH-Px是重要的抗氧化酶,MDA是脂质过氧化、氧化损伤的重要指标[20]。研究认为,高剂量ZnO和低剂量N-ZnO能改善动物抗氧化能力[21⇓-23]。Long等[4]发现,3 000 mg/kg ZnO和500 mg/kg N-ZnO均能显著提高仔猪血清GSH-Px活性,降低MDA含量。N-ZnO能增加肉鸡血浆GSH-Px活性[24]和鹌鹑血清SOD、GSH-Px活性[25]。本试验也发现,ZnO组和N500组仔猪血浆GSH-Px活性、T-AOC显著提高,可见ZnO和500 mg/kg N-ZnO能改善仔猪抗氧化能力。血浆抗氧化结果与小肠抗氧化能力[11]变化一致,500 mg/kg N-ZnO的添加量少于1 600 mg/kg ZnO,但能起到相似的抗氧化效果。这是因为N-ZnO比ZnO具有更高的生物利用度[26],在相同剂量下N-ZnO的抗氧化能力优于ZnO[8]。本试验结果与Sun等[27]研究结果相似,即600 mg/kg N-ZnO与2 000 mg/kg ZnO具有相似的抗氧化作用。此外,N-ZnO添加量低于500 mg/kg未能改善仔猪的抗氧化能力,可能与锌对参与抗氧化防御系统的酶活性的必要性有关[28],需要足够量的锌才能起到相应的抗氧化效果。

3.3 纳米氧化锌对仔猪血清免疫指标的影响

Ig是体液免疫的重要指标。锌在体液免疫反应与先天免疫功能中起重要作用[29]。N-ZnO能改善动物免疫功能[30],如提高绵羊血清IgG含量[8]和鹌鹑血清IgG、IgM含量[25]。研究发现,ZnO和N-ZnO(400~600 mg/kg)改善仔猪血清IgM、IgG含量[27]。本试验中,ZnO和N-ZnO(200~400 mg/kg)显著提高血清IgG、IgA含量,与上述研究结果相似。可见低剂量N-ZnO与高剂量ZnO均可改善仔猪的免疫功能。另外,随着N-ZnO添加量的增加,仔猪血清IgM含量线性增加,但IgA、IgG含量线性减少。本试验结果与Sun等[27]研究结果有相似之处,仔猪血清IgG、IgM含量随着N-ZnO添加量的增加而线性增加。Ig含量的不同变化可能与仔猪试验时间和N-ZnO来源的不同有关。ANTI组与N500组血清Ig含量均低于其他组,与小肠中Ig含量的变化结果[11]一致。锌有助于免疫功能的调节,影响多种信号通路[31]。还需要更多的研究来解释N-ZnO对仔猪机体免疫功能的影响。

3.4 纳米氧化锌对仔猪血浆游离氨基酸含量的影响

血浆游离氨基酸含量可反映机体各种生理、病理状况[32]。组氨酸是一种抗炎和抗氧化因子,血浆低组氨酸含量与炎症有关[33]。与ANTI组和ZnO组相比,添加N-ZnO提高了仔猪血浆组氨酸含量,N-ZnO可能缓解了仔猪的炎症反应,改善仔猪机体免疫功能(N200、N300、N400组的血清IgA、IgG含量显著高于ANTI组)。ANTI组和ZnO组中血浆组氨酸含量较低,可能引起了仔猪炎症,这需要进行更多的研究来证实。
甘氨酸、脯氨酸、羟脯氨酸在营养和新陈代谢中起着重要作用,对动物的健康、生长至关重要[34]。甘氨酸是一种专门的生糖氨基酸,分解代谢产生丙酮酸,用于合成GLU[32,35]。脯氨酸是三羧酸循环能量代谢的来源,在蛋白质合成、抗氧化、免疫反应中发挥多种作用[32]。羟脯氨酸是甘氨酸的主要前体和抗氧化分子,是组织生长发育的重要底物[34,36]。甘氨酸和丝氨酸可以快速相互转化[37]。β-丙氨酸用于增强能量代谢,是骨骼肌中肌肽形成的限制因素[38]。N500组的血浆羟脯氨酸、脯氨酸、β-丙氨酸含量与ANTI组、ZnO组相似,意味着补充500 mg/kg N-ZnO能达到与ANTI和ZnO相似效果,促进仔猪生长发育。N200、N300、N400组的血浆甘氨酸、丝氨酸、羟脯氨酸含量低于ANTI组、ZnO组。但是,随着N-ZnO添加量的增加,仔猪血浆中这些氨基酸含量显著增加,即仔猪生长发育随着补充N-ZnO的剂量增加而提高。有研究报道,与25、50 mg/kg N-ZnO相比,100 mg/kg N-ZnO提高了母鸡血浆中甘氨酸的含量[39]。不同添加量的N-ZnO对血浆氨基酸含量的影响不同,这可能与机体吸收的锌浓度有关[40],需要进一步的研究来检验这些假设。
胃肠道功能受损会导致全身瓜氨酸和精氨酸含量减少[31]。随着N-ZnO添加量的增加,仔猪血浆精氨酸、瓜氨酸含量呈二次曲线变化。但是,与ANTI组和ZnO组相比,N-ZnO组血浆的精氨酸含量显著提高,即N-ZnO的添加一定程度上能改变肠道功能。同期研究也表明,饲喂N-ZnO的猪小肠SOD活性、IgA含量、绒毛高度以及绒毛高度与隐窝深度的比值大于抗生素组[11]。另外,精氨酸在尿素循环、免疫功能调节、物质合成(多胺、肌酸、一氧化氮)中起着关键作用[41]。N-ZnO上调血浆精氨酸含量反映了其可以改善仔猪机体代谢,有益于仔猪健康。

4 结论

饲粮添加N-ZnO对仔猪肝脏功能、抗氧化能力、免疫功能产生了一定益处,还改变了血浆游离氨基酸谱。

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