Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 6: 3594 - 3606

DOI: https://doi.org/10.12418/CJAN2023.335

RESEARCH PAPER

Effects of Plant Essential Oil Replacement of Aureomycin on Growth Performance, Serum Biochemical Indices and Intestinal Health of Weaned Piglets

  • LIU Xiang ,
  • ZHAO Yirun ,
  • WANG Jiayi ,
  • ZHANG Fan ,
  • LIU Nian ,
  • MA Xiaokang , *
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  • College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China
*lecturer, E-mail:

Received date: 2022-11-28

  Online published: 2023-06-08

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Abstract

This experiment was conducted to investigate the effects of dietary supplementation of plant essential oil replacement of aureomycin on growth performance, serum biochemical indices, intestinal digestive enzyme activities, mucosal inflammatory factor contents, intestinal morphology and volatile fatty acid contents of weaned piglets. A total of 24 healthy Duroc×Landrace×Yorkshire weaned piglets with similar body weight (7.17±0.25) kg at the age of 28 days were randomly divided into 3 groups: control group (basal diet), essential oil group (supplemented with 300 mg/kg plant essential oil) and aureomycin group (supplemented with 75 mg/kg aureomycin) were divided into 8 replicates per group and 1 piglet per replicate. The experiment lasted for 28 days, with 1 to 14 days as the early stage and 15 to 28 days as the late stage. The results showed as follows: at the early stage of the experiment, the average daily gain, ratio of weight gain to feed, serum immunoglobulin A content, immunoglobulin M contents, superoxide dismutase activity and total antioxidant activity in essential oil group were significantly higher than those in control group (P<0.05). The diarrhea rate, serum malondialdehyde, D-lactic acid contents and diamine oxidase activity in essential oil group were significantly lower than those in control group (P<0.05). At the later stage of the experiment, the average daily gain, the ratio of weight gain to feed and serum superoxide dismutase activity in essential oil and aureomycin groups were significantly higher than those in control group (P<0.05), but had no significant difference with aureomycin group (P>0.05). Serum malondialdehyde content and diamine oxidase activity in essential oil group were significantly lower than those in control group (P<0.05). During the whole experiment period, the average daily gain, the ratio of weight gain to feed, the apparent digestibilities of crude protein and gross energy, the activities of chymotrypsin and trypsin in jejunum, the ratio of villus height to crypt depth in jejunum, the contents of butyric acid in cecum and total volatile fatty acids in colon in essential oil group were significantly higher than those in control group (P<0.05), but had no significant differences compared with aureomycin group (P>0.05). The diarrhea rate and the contents of interleukin-1β and tumor necrosis factor-α in jejunum mucosa in essential oil group were significantly lower than those in control group (P<0.05), but had no significant difference with aureomycin group (P>0.05). In conclusion, dietary supplementation of plant essential oil can improve serum immune level and antioxidant capacity of weaned piglets, increase intestinal digestive enzyme activity and volatile fatty acid contents, improve intestinal morphology and intestinal permeability, reduce diarrhea rate, and improve nutrient digestibility and growth performance of weaned piglets. These results suggest that plant essential oil has the same effect as aureomycin and can be used as a substitute for aureomycin in weaned piglets’ diets.

Key words： plant essential oil; weaned piglets; growth performance; serum immune; digestive enzyme activity; volatile fatty acids

Cite this article

LIU Xiang , ZHAO Yirun , WANG Jiayi , ZHANG Fan , LIU Nian , MA Xiaokang . Effects of Plant Essential Oil Replacement of Aureomycin on Growth Performance, Serum Biochemical Indices and Intestinal Health of Weaned Piglets[J]. Chinese Journal of Animal Nutrition, 2023 , 35(6) : 3594 -3606 . DOI: 10.12418/CJAN2023.335

近几十年来,我国养猪生产趋向现代化、集约化和专业化方向发展,为了有效控制疾病、提高母猪生产力、提高饲料利用率、促进仔猪的生长发育,通常对仔猪进行早期隔离断奶[1]。然而,断奶往往会造成仔猪心理、饲粮和环境等诸多应激,导致仔猪消化和免疫系统功能紊乱,使仔猪肠道菌群失调、采食量下降、生长速度缓慢、腹泻,甚至出现死亡,给养猪生产带来较大的经济损失[2-3]。在生产中,饲粮添加亚治疗剂量的金霉素能有效缓解这一困难,但抗生素在应用中也存在一系列问题,如使细菌产生耐药性、降低畜禽免疫力、引发菌群失调、引发内源性和双重感染、产生毒副作用导致在动物产品和环境中残留等问题,严重地威胁了人类的健康安全[4⇓⇓-7]。欧盟在2006年就禁止在饲粮中使用抗生素作为生长促进剂[5],美国也在2013年开始禁止饲用抗生素[8],2015年我国也明令禁止促生长抗生素在饲粮中的使用[9]。因此,在减抗、禁抗的大背景趋势下,研发绿色、安全、高效的替抗饲料添加剂成为当今研究的热点。植物精油是一种天然、易得、无毒无残留的绿色饲料添加剂[10],含有大量具有抗菌、抗氧化、增强机体免疫力等生理功能的活性物质[11],能有效改善断奶仔猪生长性能、抗氧化能力和免疫系统,调节肠道健康,降低腹泻率[12],是替抗饲料添加剂的典型代表。因此,本试验旨在探讨植物精油替代金霉素对断奶仔猪生长性能、营养物质表观消化率、肠道消化酶活性、血清生化指标和肠道健康的影响,为评价饲粮添加植物精油替代金霉素的效果提供参考,具有一定的研究意义和应用价值。

1 材料与方法

1.1 试验材料

试验中使用的植物精油主要成分为13.5%百里香酚和4.5%肉桂醛,由麦芽糊精包被。

1.2 试验动物

选用28日龄、平均体重为(7.17±0.25) kg的健康“杜×长×大”断奶仔猪(公猪)24头。

1.3 试验设计和试验饲粮

试验根据体重及健康状况等均匀分布原则,随机分为3组:对照组(基础饲粮)、植物精油组(添加300 mg/kg植物精油)、金霉素组(添加75 mg/kg的金霉素),每组8个重复,每个重复1头仔猪(公猪)。试验期28 d,1~14 d为试验前期,15~28 d为试验后期。试验所需饲粮严格参照NRC(2012)配制,饲粮营养水平均满足仔猪生长需要,基础饲粮组成及营养水平见表1。
表1 基础饲粮组成及营养水平(饲喂基础)

Table 1 Composition and nutrient levels of basal diets (as-fed basis)%

项目
Items		 前期
Prophase		 后期
Post phase
原料 Ingredients
玉米 Corn 52.49 56.13
豆粕 Soybean meal 20.00 20.00
膨化全脂大豆 Full fat soybean 9.00 8.00
鱼粉 Fish meal 5.11 4.00
乳清粉 Whey powder 7.00 7.00
豆油 Soybean oil 3.19 2.08
磷酸氢钙 CaHPO4 0.65 0.50
石粉 Limestone 0.88 0.78
食盐 NaCl 0.30 0.30
L-赖氨酸盐酸盐 L-lysine·HCl 0.37 0.30
DL-蛋氨酸 DL-methionine 0.09 0.07
L-苏氨酸 L-threonine 0.14 0.08
L-色氨酸 L-tryptophan 0.03 0.01
三氧化二铬 Cr2O3 0.25 0.25
预混料 Permix1) 0.50 0.50
合计 Total 100.00 100.00
营养水平 Nutrient levels2)
消化能 DE/(MJ/kg) 14.82 14.60
粗蛋白质 CP 20.56 18.84
钙 Ca 0.81 0.72
总磷 TP 0.59 0.53
标准回肠可消化赖氨酸 SID Lys 1.35 1.24
标准回肠可消化蛋氨酸 SID Met 0.40 0.36
标准回肠可消化苏氨酸 SID Thr 0.81 0.74
标准回肠可消化色氨酸 SID Trp 0.23 0.22

1) 预混料为每千克饲粮提供The premix provided the following per kg of diets: VA 12 000 IU,VD3 2 500 IU,VE 30 IU,VK3 30 mg,VB1 1.5 mg,VB2 4 mg,VB6 3 mg, VB12 12 μg, 氯化胆碱 choline chloride 400 mg,叶酸 folic acid 0.7 mg,烟酸 nicotinic acid 40 mg,泛酸 pantothenic acid 15 mg,生物素 biotin 0.1 mg,Fe 90 mg,Cu 8.8 mg,Mn 40 mg,Zn 100 mg,I 0.35 mg,Se 0.3 mg。

2) 粗蛋白质为实测值,钙、总磷、消化能和标准回肠可消化氨基酸为计算值。CP was a measured value, while Ca, TP, DE and SID amino acids were calculated values.

1.4 饲养管理

试验仔猪自由采食和饮水,试验期间每天不定时巡视,观察记录采食、饮水和健康等情况,试验过程中按常规程序进行驱虫和免疫,试验期内定期对猪舍进行清洁消毒。采用恒温控制仪自动调节舍内环境温度并定时开窗通风。

1.5 样品采集与制备

试验开始前,每组取500 g饲粮样品,用于测定饲粮中营养成分含量。试验第26~28天,收集各组仔猪粪便,并将其混匀,65 ℃烘72 h,回潮24 h,饲粮样品和烘干的粪样分别粉碎过40目筛,装袋待检。
试验第14和28天,各组全部仔猪前腔静脉采血10 mL,室温下静置30 min,在4 ℃下3 000 r/min离心10 min制备血清,于-20 ℃保存待检。
试验第28天,每组屠宰8头仔猪,采集十二指肠、空肠和回肠肠段样品,置于10%中性福尔马林缓冲液中,用于组织形态分析;取相同位置的十二指肠、空肠和回肠黏膜样品,经液氮速冻后-80 ℃冰箱保存,用于消化道酶活性和炎性细胞因子含量测定;采集盲肠和结肠食糜,液氮速冻后-80 ℃冰箱保存,用于挥发性脂肪酸含量测定。

1.6 检测指标及测定方法

1.6.1 生长性能与腹泻率

试验第1、14和28天,分别称量经空腹处理后的仔猪个体重和每个重复的饲粮重量,计算平均日增重(ADG)、平均日采食量(ADFI)和增重耗料比(G∶F)。
ADG=(终末体重-初始体重)/试验天数;
ADFI=各组仔猪总耗料量/(试验天数×试验仔猪数量);
G∶F=ADG/ADFI。
腹泻判定参照Tan等[13]的方法,根据每天08:00和16:00每头仔猪的粪便外观进行目测评估。
腹泻率(%)=[仔猪腹泻头次总和/(仔猪头数×试验天数)]×100。

1.6.2 营养物质表观消化率

饲粮和粪便中干物质和粗蛋白质含量分别按照GB/T 6435—2006[14]和GB/T 6432—1994[15]方法进行测定,总能使用氧弹式测热仪,按照ISO 9831:1998方法进行测定。使用三氧化二铬作为外源指示剂,采用原子吸收光谱仪测定饲粮样和粪样中铬含量。根据指示剂法计算营养物质表观消化率。

1.6.3 血清生化指标

血清中免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、D-乳酸(D-LA)、丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、二胺氧化酶(DAO)活性及总抗氧化能力(T-AOC)均使用南京建成生物工程研究所试剂盒,按照说明书检测方法测定。

1.6.4 消化酶活性

制备组织匀浆时将样品解冻,取样品约0.5 g,在冰冷的生理盐水中漂洗,用滤纸滤干,称重;用移液管量取预冷的0.9%生理盐水,按照组织块重量,生理盐水质量比为1∶9的比例加入生理盐水;用组织匀浆机15 000 r/min研磨组织;将制备好的10%匀浆用低温离心机4 000 r/min离心10 min,取上清-20 ℃冻存备用,使用二喹啉甲酸法测定组织匀浆液中的蛋白含量。十二指肠、空肠和回肠消化酶活性均使用南京建成生物工程研究所试剂盒,按照说明书检测方法测定。

1.6.5 肠道黏膜炎性因子含量

组织匀浆制备同上,肠道黏膜炎性因子白细胞介素-1β(IL-1β)、白细胞介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)和γ-干扰素(IFN-γ)含量均使用南京建成生物工程研究所试剂盒,按照说明书检测方法测定。

1.6.6 肠道形态

采用酒精脱水,二甲苯透明,石蜡包埋,切成6 μm切片,苏木精-伊红(HE)染色(苏木素染核和伊红染胞质),然后在每个部位组织切片上,选取至少5个典型视野(绒毛清晰完整),用真彩图像分析软件采集图像,并量取绒毛高度和隐窝深度[16]。

1.6.7 肠道食糜挥发性脂肪酸含量

将食糜在4 ℃下解冻并混匀,称取回肠和盲肠食糜样品1.0 g左右于10 mL离心管中,加超纯水至8 mL,将食糜充分打碎后超声20 min,然后将离心管在4 ℃条件下10 000 r/min离心15 min。取上清液用超纯水稀释100倍,用注射器取1.5 mL稀释液过滤到上机瓶中,使用气相色谱仪进行挥发性脂肪酸含量的测定。

1.7 统计方法

试验数据采用Excel 2010进行初步处理,采用SPSS 21.0软件进行单因素方差分析,Duncan氏法进行多重比较,结果以平均值和均值标准误表示,以P<0.05为差异显著判断标准。

2 结果与分析

2.1 植物精油对断奶仔猪生长性能和腹泻率的影响

由表2可见,在试验前期,精油组ADG显著高于对照组(P<0.05),而与金霉素组无显著差异(P>0.05);精油组G∶F显著高于其他2组(P<0.05);在试验前期(1~14 d)、全期(1~28 d),精油组和金霉素组腹泻率显著低于对照组(P<0.05);在试验后期(15~28 d)、全期,精油组和金霉素组ADG和G∶F显著高于对照组(P<0.05)。
表2 植物精油对断奶仔猪生长性能和腹泻率的影响

Table 2 Effects of plant essential oil on growth performance and diarrhea rate of weaned piglets (n=8)

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
初始体重 IBW/kg 7.17 7.17 7.18 0.08 0.99
第14天体重 BW at day 14/kg 11.04 11.57 11.33 0.16 0.09
第28天体重 BW at day 28/kg 16.31b 17.40a 17.37a 0.17 <0.01
第1~14天 Days 1 to 14
平均日增重 ADG/g 276.70b 314.38a 296.34ab 7.82 0.01
平均日采食量 ADFI/g 460.18 469.55 470.71 13.36 0.83
增重耗料比 G∶F 0.60b 0.67a 0.63b 0.01 <0.01
腹泻率 Diarrhea rate/% 6.36a 2.12b 2.34b 0.64 <0.01
第15~28天 Days 15 to 28
平均日增重 ADG/g 375.98b 416.25a 431.43a 9.59 <0.01
平均日采食量 ADFI/g 674.02 683.13 693.13 20.08 0.80
增重耗料比 G∶F 0.56b 0.61a 0.62a 0.01 <0.01
腹泻率 Diarrhea rate/% 2.34 1.00 1.56 0.56 0.27
第1~28天 Days 1 to 28
平均日增重 ADG/g 326.34b 365.31a 363.88a 5.24 <0.01
平均日采食量 ADFI/g 567.10 576.34 581.92 8.82 0.50
增重耗料比 G∶F 0.58b 0.63a 0.63a 0.01 <0.01
腹泻率 Diarrhea rate/% 4.35a 1.56b 1.95b 0.47 <0.01

同行数据肩标不同小写字母表示差异显著(P<0.05)。下表同。

Values in the same row with different small letter superscripts mean significant difference (P<0.05). The same as below.

2.2 植物精油对断奶仔猪全肠道营养物质表观消化率的影响

由表3可见,与对照组相比,精油组和金霉素组粗蛋白质和总能表观消化率显著提高(P<0.05)。
表3 植物精油对断奶仔猪营养物质表观消化率的影响

Table 3 Effects of plant essential oils on nutrient apparent digestibility of weaned piglets (n=8)%

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
干物质 DM 81.23 81.58 81.50 0.39 0.80
粗蛋白质 CP 78.12b 80.42a 80.02a 0.54 0.02
总能 GE 79.80b 81.20a 81.28a 0.37 0.02

2.3 植物精油对断奶仔猪血清生化指标的影响

由表4可见,试验第14天,与对照组相比,精油组仔猪血清IgA和IgM含量显著提高(P<0.05),而与金霉素组之间无显著差异(P>0.05);精油组和金霉素组仔猪血清SOD活性和T-AOC显著提高(P<0.05),而MDA含量显著降低(P<0.05);精油组和金霉素组仔猪血清D-LA含量显著降低(P<0.05),精油组仔猪血清DAO活性显著低于对照组和金霉素组(P<0.05)。
表4 植物精油对断奶仔猪血清生化指标的影响

Table 4 Effects of plant essential oils on serum biochemical indices of weaned piglets (n=8)

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
第14天 Day 14
免疫球蛋白 G IgG/(g/L) 6.66 7.47 6.81 0.27 0.12
免疫球蛋白 A IgA/(g/L) 0.87b 1.07a 0.97ab 0.04 <0.01
免疫球蛋白 M IgM/(g/L) 0.42b 0.55a 0.57a 0.02 <0.01
超氧化物歧化酶 SOD/(U/mL) 119.57b 139.95a 136.67a 4.73 0.02
丙二醛 MDA/(nmol/mL) 4.73a 2.58c 3.28b 0.22 <0.01
谷胱甘肽过氧化物酶 GSH-Px/(U/mL) 748.29 768.99 764.96 24.23 0.82
总抗氧化能力 T-AOC/(U/mL) 8.52b 10.90a 10.44a 0.55 0.02
D-乳酸 D-LA/(mg/L) 7.45a 5.98b 6.36b 0.24 <0.01
二胺氧化酶 DAO/(U/L) 10.57a 4.88c 7.12b 0.31 <0.01
第28天 Day 28
免疫球蛋白 G IgG/(g/L) 9.93 10.53 10.10 0.35 0.48
免疫球蛋白 A IgA/(g/L) 1.19 1.20 1.20 0.02 0.98
免疫球蛋白 M IgM/(g/L) 0.79b 0.88ab 0.95a 0.04 0.04
超氧化物歧化酶 SOD/(U/mL) 134.49b 159.88a 157.08a 4.93 <0.01
丙二醛 MDA/(nmol/mL) 4.66a 3.08b 3.30b 0.31 <0.01
谷胱甘肽过氧化物酶 GSH-Px/(U/mL) 719.73 763.27 751.49 25.85 0.48
总抗氧化能力 T-AOC/(U/mL) 6.71 7.52 7.28 0.62 0.64
D-乳酸 D-LA/(mg/L) 9.40 8.68 8.63 0.40 0.34
二胺氧化酶 DAO/(U/L) 14.49a 7.94c 10.15b 0.34 <0.01
试验第28天,与对照组相比,金霉素组仔猪血清IgM含量显著提高(P<0.05),但与精油组之间无显著差异(P>0.05);精油组和金霉素组仔猪血清SOD活性显著提高(P<0.05),而MDA含量显著降低(P<0.05);精油组仔猪血清DAO活性显著低于对照组和金霉素组(P<0.05)。

2.4 植物精油对断奶仔猪肠道消化酶活性的影响

由表5可见,与对照组相比,精油组和金霉素组仔猪空肠黏膜的糜蛋白酶和胰蛋白酶活性显著提高(P<0.05)。
表5 植物精油对断奶仔猪肠道消化酶活性的影响

Table 5 Effects of plant essential oils on intestinal digestive enzyme activities of weaned piglets (n=8)U/mg

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
十二指肠 Duodenum
糜蛋白酶 Chymotrypsin 5.89 7.06 7.33 0.45 0.10
胰蛋白酶 Trypsin 1 762.90 2 002.60 2 045.40 91.10 0.11
淀粉酶 Amylase 0.16 0.17 0.19 0.01 0.12
空肠 Jejunum
糜蛋白酶 Chymotrypsin 7.93b 10.82a 11.22a 0.43 <0.01
胰蛋白酶 Trypsin 1 239.09b 1 598.43a 1 749.66a 57.51 <0.01
淀粉酶 Amylase 0.20 0.22 0.23 0.01 0.32
回肠 Ileum
糜蛋白酶 Chymotrypsin 6.03 6.85 7.06 0.33 0.12
胰蛋白酶 Trypsin 1 569.60 1 827.80 1 854.20 87.60 0.08
淀粉酶 Amylase 0.12 0.13 0.14 0.02 0.80

2.5 植物精油对断奶仔猪肠道黏膜炎性因子含量的影响

由表6可见,与对照组相比,精油组和金霉素组的仔猪空肠黏膜中IL-1β和TNF-α的含量显著降低(P<0.05)。
表6 植物精油对断奶仔猪肠道黏膜炎性因子含量的影响

Table 6 Effects of plant essential oil on intestinal mucosal inflammatory factor contents of weaned piglets (n=8)pg/mg

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
十二指肠 Duodenum
白细胞介素-1β IL-1β 14.00 12.61 11.61 1.32 0.46
白细胞介素-2 IL-2 3.82 3.79 3.03 0.31 0.17
肿瘤坏死因子-α TNF-α 24.96 23.77 19.13 1.68 0.06
γ-干扰素 IFN-γ 25.06 23.21 22.63 1.11 0.30
空肠Jejunum
白细胞介素-1β IL-1β 15.15a 12.12b 11.40b 0.75 <0.01
白细胞介素-2 IL-2 2.40 1.99 1.91 0.17 0.12
肿瘤坏死因子-α TNF-α 12.68a 10.56b 9.49b 0.65 0.01
γ-干扰素 IFN-γ 25.84 23.33 22.42 1.79 0.40
回肠Ileum
白细胞介素-1β IL-1β 13.15 12.89 12.87 0.54 0.92
白细胞介素-2 IL-2 1.68 1.63 1.62 0.08 0.83
肿瘤坏死因子-α TNF-α 19.76 19.00 18.02 1.13 0.57
γ-干扰素 IFN-γ 29.61 27.67 25.80 1.13 0.09

2.6 植物精油对断奶仔猪肠道形态的影响

由表7可见,与对照组相比,精油组和金霉素组的十二指肠绒毛高度以及十二指肠和空肠绒毛高度与绒毛高度/隐窝深度显著提高(P<0.05)。
表7 植物精油对断奶仔猪肠道形态的影响

Table 7 Effects of plant essential oil on intestinal morphology of weaned piglets (n=8)

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
十二指肠 Duodenum
绒毛高度 Villus height/μm 472.54b 486.54a 494.49a 4.55 0.01
隐窝深度 Crypt depth/μm 352.66 347.52 351.80 5.04 0.75
绒毛高度/隐窝深度 Villus height/crypt depth 1.34b 1.40a 1.41a 0.01 <0.01
空肠 Jejunum
绒毛高度 Villus height/μm 459.57 470.81 479.50 9.88 0.39
隐窝深度 Crypt depth/μm 355.94 340.68 342.31 5.87 0.17
绒毛高度/隐窝深度 Villus height/crypt depth 1.29b 1.38a 1.40a 0.02 0.01
回肠 Ileum
绒毛高度 Villus height/μm 373.31 383.86 376.27 6.20 0.48
隐窝深度 Crypt depth/μm 295.53 292.43 289.88 4.40 0.67
绒毛高度/隐窝深度 Villus height/crypt depth 1.26 1.31 1.30 0.02 0.24

2.7 植物精油对断奶仔猪盲肠和结肠挥发性脂肪酸含量的影响

由表8可见,与对照组相比,精油组的盲肠和结肠食糜中丙酸和丁酸含量显著提高(P<0.05),而与金霉素组之间无显著差异(P>0.05);精油组和金霉素组的结肠食糜中乙酸和总挥发性脂肪酸含量显著提高(P<0.05)。
表8 植物精油对断奶仔猪盲肠和结肠挥发性脂肪酸含量的影响

Table 8 Effects of plant essential oils on volatile fatty acid contents in caecum and colon of weaned piglets (n=8)mg/g

项目
Items		 对照组
Control
group		 精油组
Essential oil
group		 金霉素组
Chlortetracyc1ine
group		 均值标准误
SEM		 P值
P-value
盲肠 Cecum
乙酸 Acetate 2.06 2.58 2.53 0.19 0.15
丙酸 Propionate 1.26b 1.66a 1.49ab 0.09 0.03
丁酸 Butyrate 0.61b 0.75a 0.74a 0.04 0.03
戊酸 Valerate 0.19 0.19 0.18 0.01 0.72
总挥发性脂肪酸 TVFA 4.26 5.18 4.93 0.26 0.07
结肠 Colon
乙酸 Acetate 2.49b 3.04a 3.02a 0.10 <0.01
丙酸 Propionate 1.68b 2.21a 1.99ab 0.12 0.03
丁酸 Butyrate 1.02b 1.43a 1.43a 0.10 0.02
戊酸 Valerate 0.46 0.44 0.43 0.03 0.72
总挥发性脂肪酸TVFA 5.96b 7.03a 6.74a 0.22 0.01

3 讨论

3.1 生长性能及腹泻率

植物精油含有芳香族挥发性物质,可以刺激仔猪嗅觉感受器、促进肠道内消化酶分泌和增强消化酶活性,从而提高仔猪采食量和饲料利用率,促进仔猪生长。目前诸多研究已经表明,植物精油能改善断奶仔猪生长性能[5,17-18]。Ariza-Nieto[19]研究发现,在断奶仔猪饲粮中添加植物精油能显著提高断奶仔猪ADFI和ADG,其效果与抗生素相似。Huang等[20]研究发现,在饲粮中添加混合精油可以替代断奶仔猪饲粮中的抗生素,提高断奶仔猪ADG,而不会对生长性能产生负面影响。Su等[21]在断奶仔猪饲粮中添加200 ppm植物精油,结果显示显著增加了仔猪ADG和营养物质消化率。本试验结果也表明,植物精油组提高了断奶仔猪ADG和G∶F,结果与上述研究报道结果基本一致。
仔猪在断奶时消化器官和免疫系统发育不全,又面临着断奶应激,往往会出现腹泻。许多研究发现在饲粮中适当添加植物精油能够降低断奶仔猪的腹泻率[5,22-23]。本试验研究结果表明,在断奶仔猪饲粮中添加300 mg/kg植物精油和75 mg/kg金霉素均显著降低了断奶仔猪的腹泻率。植物精油能降低腹泻率很大可能是因为植物精油杀菌、抑菌的特性,在消化道内能抑制病原微生物的增殖,另外植物精油还可以刺激肠道有益微生物的生长,进而改善消化道菌群结构,优化肠道中微生态环境,减少腹泻[24]。血清D-LA含量和DAO活性可作为反映肠道通透性和小肠黏膜完整性的指标,当肠黏膜屏障功能损伤时,血液中D-LA含量和DAO活性增加[25]。本试验中,饲粮中添加300 mg/kg精油显著降低了断奶仔猪血清中D-LA含量和DAO活性,这表明植物精油改善了仔猪肠道通透性,这一结果与本试验中植物精油降低腹泻率相吻合。

3.2 营养物质表观消化率

断奶仔猪生长性能的改善与营养物质消化率的提高紧密相关。Zeng等[26]研究发现,仔猪饲粮中添加精油能显著提高仔猪的干物质、粗蛋白质和能量表观消化率;Li等[27]研究发现,精油组和抗生素组断奶仔猪的干物质和粗蛋白质消化率均显著高于对照组。本试验结果也表明,精油组断奶仔猪的粗蛋白质和总能表观消化率显著高于对照组,而与金霉素组无显著差异,这与Tian等[5]研究报道的结果相似。植物精油提高营养物质表观消化率分析其原因可能是由于植物精油改善了小肠形态、增加了消化酶的分泌[28]。

3.3 血清生化指标

动物机体通过自身的体液免疫和细胞免疫来维持健康[23]。免疫球蛋白是体液免疫应答中最主要的免疫分子,血清中维持较高水平的免疫球蛋白是保证仔猪健康和平稳断奶的重要基础[24,29]。IgG是主动免疫后机体产生的主要抗体,具有抗菌、抗病毒和抗毒素等免疫活性,是动物机体抗感染的主力;IgM是机体初次接触抗原物质最早产生的抗体,也具有抗菌、抗病毒和抗毒素的功能,在机体的早期防护中占有重要地位;IgA对机体呼吸道、消化道等局部黏膜免疫起着相当重要的作用[24,29⇓-31]。在仔猪生长阶段,血清IgG、IgM和IgA的含量提高,在一定程度上反映仔猪免疫力增强[29]。本试验结果表明,精油组和金霉素组均能提高仔猪血清IgM和IgA含量,说明植物精油能在一定程度上提高仔猪机体的免疫水平。
正常情况下,动物机体维持正常的氧化还原状态。仔猪在早期断奶过程中,体内原有的氧化平衡被打破,机体抗氧化能力下降,从而产生氧化应激。植物精油中含量丰富的酚类成分通过抑制或阻止动物机体自由基链式反应的发生,使自由基活性降低并将其清除,从而提高动物机体抗氧化能力[32]。Zeng等[26]在低能量饲粮中添加植物精油显著提高了仔猪血清T-AOC,且SOD和GSH-Px活性均有提高的趋势。Xu等[33]研究表明,在饲粮中添加30 mg/kg精油,显著提高了断奶仔猪血清SOD活性,同时血清MDA含量呈下降趋势。王仁杰等[34]研究表明,在饲粮中添加450和600 mg/kg复合植物精油显著提高了断奶仔猪血清T-AOC。本研究结果表明,与对照组相比,试验第14天精油组和金霉素组仔猪血清SOD活性和T-AOC显著提高,试验第28天仔猪血清SOD活性显著提高,试验第14天和第28天仔猪血清MDA含量均显著降低,这与Zeng等[26]、Xu等[33]和王仁杰等[34]研究报道的结果相似,说明植物精油能提高仔猪机体的抗氧化能力,缓解氧化应激。

3.4 肠道消化酶活性

消化酶活性与消化道的消化功能呈正相关[27]。消化酶在营养物质的消化中起着至关重要的作用,其活性直接反映动物的消化能力[35]。与Xu等[33]研究结果一致,本试验在断奶仔猪饲粮中添加300 mg/kg植物精油能显著提高仔猪空肠黏膜糜蛋白酶和胰蛋白酶活性。这说明植物精油能提高断奶仔猪消化酶活性,增强断奶仔猪消化能力,进而提高营养物质的消化率。植物精油可能通过改善小肠形态,进而促进内源酶的分泌,最终使小肠消化酶的活性得到提高,这也与本试验发现的植物精油能提高空肠绒毛高度/隐窝深度和营养物质表观消化率的结果相吻合,但改善酶活性的确切机制需要进一步研究。

3.5 肠道黏膜炎性因子含量

肠道炎症和肠道屏障功能损伤是断奶仔猪易发生腹泻的主要内在原因[36],细胞因子在免疫和炎症反应中发挥重要作用[37],是调节免疫和炎症反应的重要介质[38]。IL-1β和TNF-α是仔猪免疫细胞释放的典型促炎性细胞因子[39],通过参与炎症反应的过程,促进和调控急性炎症的发生发展[40];IL-2在免疫稳态过程中发挥重要作用,具有双向调节免疫的能力,在免疫缺损状态下可增进免疫功能,抑制免疫炎症反应[41⇓-43];IFN-γ主要由活化的T细胞和自然杀伤(NK)细胞产生,能够激活巨噬细胞分泌抗炎症因子[44]。本研究中,与对照组相比,精油组和金霉素组显著降低了仔猪空肠黏膜中IL-1β和TNF-α的含量,说明植物精油能通过降低肠道促炎性细胞因子的含量调节断奶仔猪肠道黏膜炎症反应,作用效果与金霉素相当,这可能也是植物精油能促进断奶仔猪生长性能和降低腹泻率的原因之一。

3.6 肠道形态

肠道是动物消化吸收营养物质的场所,健康的肠道环境是促进仔猪生长的关键。绒毛高度、隐窝深度及绒毛高度/隐窝深度是衡量肠道形态结构及完整性的重要指标[23],而肠道形态结构的完整性对于维持正常的肠道功能至关重要[5]。已经证明,断奶应激会导致仔猪肠道屏障功能和肠道形态结构损伤[45]。Zeng等[26]研究发现,在断奶仔猪饲粮中添加植物精油能显著提高仔猪空肠绒毛高度。黄金昌等[23]研究报道,与对照组相比,饲粮中添加复合植物精油能够显著提高十二指肠绒毛高度以及十二指肠和空肠绒毛高度/隐窝深度。与Zeng等[26]、扶雄锋等[46]和黄金昌等[23]研究结果一致,本研究结果表明,与对照组相比,精油组和金霉素组十二指肠的绒毛高度以及十二指肠和空肠的绒毛高度/隐窝深度均显著提高,表明植物精油可能通过改善小肠形态,增强小肠的吸收能力,进而提高仔猪的生长性能。

3.7 肠道挥发性脂肪酸含量

挥发性脂肪酸是仔猪肠道中微生物发酵碳水化合物的主要产物,可作为能源利用,促进肠上皮细胞的增殖及黏膜的生长,对维持仔猪肠道内稳态,促进仔猪肠道健康起着重要作用[12,47-48]。有研究报道,肠道中丁酸含量的增加能改善肠道免疫功能和肠道环境[49-50]。与Manzanwilla等[51]研究报道结果一致,本试验在饲粮中添加植物精油显著提高仔猪盲肠和结肠食糜中丁酸的含量。仔猪肠道中挥发性脂肪酸含量的提高,有利于肠道有益菌的生长,能改善肠道营养状况和抗炎能力[52-53]。本试验还发现,在饲粮中添加300 mg/kg植物精油能显著提高仔猪结肠食糜中乙酸和总挥发性脂肪酸含量。本试验结果表明,植物精油可以提高仔猪肠道食糜挥发性脂肪酸的含量,而盲肠和结肠食糜中挥发性脂肪酸含量变化不同可能与肠道中微生物有关。

4 结论

① 饲粮中添加植物精油可以提高断奶仔猪血清免疫水平和抗氧化能力,促进肠道健康,降低仔猪腹泻率,进而提高仔猪生长性能。
② 植物精油与金霉素作用效果相当,可以作为金霉素在断奶仔猪饲粮中的替代选择。

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