Effects of Oregano Carvacrol and Bacillus subtilis on Growth Performance, Nutrient Digestion and Metabolism, and Antioxidant Ability of Growing Wusuli Raccoon Dogs

PAN Chenxi; PAN Chenchong; LIU Jie; LI Xin; LI Wei; DENG Lufang; ZONG Wenli; WANG Hongna; REN Erjun

doi:10.12418/CJAN2024.336

2024 , Vol. 36 >Issue 6: 3931 - 3941

DOI: https://doi.org/10.12418/CJAN2024.336

RESEARCH PAPER

Effects of Oregano Carvacrol and Bacillus subtilis on Growth Performance, Nutrient Digestion and Metabolism, and Antioxidant Ability of Growing Wusuli Raccoon Dogs

PAN Chenxi ^,¹ ,
PAN Chenchong ¹ ,
LIU Jie ²^,³ ,
LI Xin ²^,³ ,
LI Wei ²^,³ ,
DENG Lufang ⁴ ,
ZONG Wenli ⁴ ,
WANG Hongna ¹ ,
REN Erjun ^,²^,³^,^*

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¹ College of Life Science and Food Engineering, Hebei University of Engineering, Handan 056038, China
² Shijiazhuang Academy of Agricultural and Forestry Sciences, Shijiazhuang 050041, China
³ Hebei Fur Animal Breeding Technology Innovation Center, Shijiazhuang 050041, China
⁴ Feed Branch of Beijing Sanyuan Seed Industry Technology Co., Ltd., Beijing 101105, China

^*professor, E-mail: renerjun2012@163.com

Received date: 2023-10-23

Online published: 2024-06-07

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Abstract

This experiment was conducted to study the effects of oregano eugenol and Bacillus subtilis on growth performance, nutrient digestion and metabolism, and antioxidant ability of growing Wusuli raccoon dogs. A 3×3 two-factor design was used in the experiment. A total of 160 healthy (70±5)-day-old raccoon dogs were selected and randomly divided into 10 groups, and there were 4 replicates in each group and 4 raccoon dogs in each replicate. The ratio of males to females in each replicate was 1∶1. The raccoon dogs in control group were fed a basal diet supplemented with antibiotics (100 mg/kg quinocetone+500 mg/kg chlortetracycline); those in test groups Ⅰ, Ⅱ and Ⅲ were fed the basal diet supplemented with 100 mg/kg oregano carvacrol, and 150, 300 and 450 mg/kg Bacillus subtilis, respectively; those in test groups Ⅳ, Ⅴ and Ⅵ were fed the basal diet supplemented with 200 mg/kg oregano carvacrol, and 150, 300 and 450 mg/kg Bacillus subtilis, respectively; those in test groups Ⅶ, Ⅷ and Ⅸ were fed the basal diet supplemented with 300 mg/kg oregano carvacrol, and 150, 300 and 450 mg/kg Bacillus subtilis, respectively. The pre-trial period was 7 days and the trial period was 82 days. The results showed as follows: 1) there were no significant differences in the initial weight, final weight, average daily gain, average daily feed intake and feed to gain ratio between the test groups and the control group (P>0.05), and the final weight and feed to gain ratio of the test group Ⅲ were the best. 2) The apparent digestibility of dry matter of test groups Ⅰ and Ⅲ was significantly higher than that of the control group (P<0.05). The apparent digestibility of crude protein of test group Ⅳ, Ⅷ and Ⅸ was significantly higher than that of the control group (P<0.05). There was no significant difference in the apparent digestibility of ether extract among groups (P>0.05). 3) The nitrogen deposition, net protein utilization rate and biological value of protein of test group IV were significantly lower than those of the control group (P<0.05), but there were no significant differences between the other test groups and the control group (P>0.05). 4) Serum glutathione peroxidase (GSH-Px) activity in test group Ⅷ was the highest, which was not significantly different compared with control group and test groups Ⅳ, Ⅸ, but significantly higher than that of other groups (P<0.05). Serum malondialdehyde (MDA) content in test groups Ⅱ and Ⅶ was significantly higher than that of test groups Ⅳ, Ⅵ and Ⅸ (P<0.05), but there were no significant differences between the test groups and the control group (P>0.05). In summary, the effects of appropriate oregano carvacrol and Bacillus subtilis to the diet on the growth performance, nutrient digestion and metabolism, and antioxidant ability of growing Wusuli raccoon dogs are similar to antibiotics. Under the conditions of this experiment, the optimal supplemental levels of oregano carvacrol and Bacillus subtilis in diets of growing Wusuli raccoon dogs are 100 and 450 mg/kg, respectively.

Key words： oregano carvacrol; Bacillus subtilis; Wusuli raccoon dogs; growth performance; digestion and metabolism of nutrients; antioxidant ability

Cite this article

PAN Chenxi , PAN Chenchong , LIU Jie , LI Xin , LI Wei , DENG Lufang , ZONG Wenli , WANG Hongna , REN Erjun . Effects of Oregano Carvacrol and Bacillus subtilis on Growth Performance, Nutrient Digestion and Metabolism, and Antioxidant Ability of Growing Wusuli Raccoon Dogs[J]. Chinese Journal of Animal Nutrition, 2024 , 36(6) : 3931 -3941 . DOI: 10.12418/CJAN2024.336

抗生素自20世纪40年代被发现以来,由于可降低动物的发病率和死亡率并能提高动物的生产力被广泛应用到饲料中^[1]。但多年来的抗生素广泛使用导致动物产品中出现耐药菌和药物残留的现象^[2⇓-4],为了消除抗生素使用带来的种种危害,我国于2020年7月1日起在饲料行业全面禁止预防性抗生素的添加^[5]。“禁抗”后养殖行业不仅生产效益降低,而且养殖管理成本普遍提高。因此,寻找和开发安全、绿色、高效的替抗产品成为我国养殖业的迫切需求。目前,在动物生产中,已有研究表明益生菌、益生元、合生菌、植物精油等可作为抗生素的替代品^[6]。

牛至香酚是从植物牛至中提取的精油,其主要成分为香芹酚和百里香酚,两者均具有较强的抗菌效果^[7]。香芹酚和百里香酚能够破坏细胞膜的完整性,增加细胞膜的通透性,引起质子和钾的泄漏,最终导致膜电位的丧失^[8-9]。枯草芽孢杆菌是一种好氧菌,在肠道内消耗大量的游离氧,改善肠道内有益菌的生长环境,抑制有害菌的生长^[10-11]。因此,牛至香酚和枯草芽孢杆菌被认为是很好的抗生素替代物,但单一添加的效果有限。近年来,有研究表明益生菌和植物精油在动物饲粮中的联合添加在某些方面会达到更好的效果^[12]。Tan等^[13]的研究表明,仔猪饲粮中单独添加精油能够提高其日增重,降低腹泻率,联合添加益生菌可进一步增强精油对仔猪生长和腹泻的积极作用;饲粮中添加300 g/t精油+1 000 g/t益生菌组仔猪血清中免疫球蛋白G(IgG)含量显著提高,且仔猪粪氨排放低于抗生素组。乌苏里貉是一种原产于我国的具有较高经济价值的毛皮动物^[14]。目前针对牛至香酚和枯草芽孢杆菌作为代替抗生素的饲料添加剂在畜牧生产中的研究主要集中在猪、禽、反刍动物等,且应用效果较好。关于牛至香酚和枯草芽孢杆菌在貉生产上的联合应用还未见报道。因此,本试验通过在饲粮中添加牛至香酚和枯草芽孢杆菌,研究其对乌苏里貉生长性能、营养物质消化代谢和抗氧化能力的影响,探索其替代抗生素添加于乌苏里貉饲粮中的应用效果,为乌苏里貉无抗饲粮的研发提供理论依据。

1 材料与方法

1.1 试验材料

1.1.1 试验药品

本试验所使用的牛至香酚产品中牛至香酚含量为10%,枯草芽孢杆菌制剂中枯草芽孢杆菌活菌数≥2.0×10¹⁰ CFU/g,金霉素中有效成分含量为10%,喹烯酮中有效成分含量为50%。

1.1.2 试验动物

选取(70±5)日龄、体重相近的160只健康乌苏里貉作为试验动物,饲养于河北平山大吾乡近掌村乌苏里貉育种基地。

1.1.3 基础饲粮

基础饲粮原料由北京三元种业科技股份有限公司饲料分公司提供,基础饲粮配方参考NRC(1982)营养标准设计,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
膨化玉米 Extruded corn	46.0
豆粕 Soybean meal	10.0
鱼粉 Fish meal	10.0
玉米蛋白粉 Corn protein meal	3.0
膨化大豆 Extruded soybean	12.5
牛肉骨粉 Beef bone meal	9.5
鸡油 Chicken fat	4.0
预混料 Premix¹⁾	5.0
合计 Total	100.0
营养水平 Nutrient levels²⁾
代谢能 ME/(MJ/kg)	14.63
水分 Moisture	10.12
粗蛋白质 CP	25.91
粗纤维 CF	2.542
粗脂肪EE	10.33
粗灰分 Ash	6.07
钙 Ca	1.51
总磷 TP	1.13
蛋氨酸 Met	0.83
赖氨酸 Lys	1.65

1)预混料为每千克饲粮提供The premix provided the following per kg of the diet:VA 10 500 IU,VD 1 650 IU,VE 250 IU,VB₁ 15 mg,VB₂ 15 mg,VB₆ 4 mg,VK 31.5 mg,VC 250 mg,烟酸 nicotinic acid 30 mg,泛酸 pantothenic acid 3.0 mg,胆碱 choline 750 mg,Fe (as ferrous sulfate) 80 mg,Mn (as manganese sulfate) 45 mg,Zn (as zinc sulfate) 60 mg,I (as potassium iodide) 0.5 mg,Se (as sodium selenite) 0.3 mg,Cu (as copper sulfate) 0.35 mg。

2)代谢能和氨基酸为参照《中国饲料成分及营养价值表(2021年第32版)》所得计算值,其他营养成分为实测值。ME and amino acids were calculated values, which were calculated according to Tables of Feed Composition and Nutritive Values in China (32nd ed, 2021), while the other nutrients were measured values.

1.2 试验方法

1.2.1 试验设计

将选取的160只乌苏里貉随机分为10组,每组4个重复,每个重复4只,公母比例1∶1。试验采用3×3双因素随机试验设计,牛至香酚的添加水平分别为100、200、300 mg/kg,枯草芽孢杆菌的添加水平分别为150、300、450 mg/kg,具体试验设计见表2。试验从2022年7月3日开始,试验期89 d,其中预试期7 d,正试期82 d。

表2 试验设计

Table 2 Experimental designmg/kg

项目 Items	试验组 Test groups									对照组 Control group
项目 Items	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Ⅴ	Ⅵ	Ⅶ	Ⅷ	Ⅸ	对照组 Control group
喹烯酮 Quinocetone	0	0	0	0	0	0	0	0	0	100
金霉素 Aureomycin	0	0	0	0	0	0	0	0	0	500
牛至香酚 Oregano carvacrol	100	100	100	200	200	200	300	300	300	0
枯草芽孢杆菌 Bacillus subtilis	150	300	450	150	300	450	150	300	450	0

1.2.2 饲养管理

试验乌苏里貉均为单笼饲养,自由饮水,充足光照的条件下进行饲养管理,固定人员于每日08:00与17:00各饲喂1次。饲养期间,保持正常驱虫及免疫,定期对料盆进行消毒和清洁。

1.2.3 测定指标

1.2.3.1 生长性能

在2022年7月10日与2022年9月30日早晨测量空腹状态下乌苏里貉体重分别作为初重和末重,进行详细记录。正试期内,准确无误测量每只仔貉早晚的饲喂量及剩料量,由此计算平均日采食量、平均日增重及料重比,计算公式如下:

平均日采食量=总采食量/(试验天数×试验仔貉数);

平均日增重=(末重-初重)/试验天数;

料重比=平均日采食量/平均日增重;

1.2.3.2 营养物质表观消化率

消化代谢试验于正试期第57~59天进行,为期3 d。采用全收粪尿法,每组选取3只公貉、3只母貉进行样品收集。使用粪尿分离的集粪盘进行消化代谢试验并记录每天的采食量。参照对应的国标方法对饲粮和粪样中营养物质(水分:GB/T 6435—2014;粗蛋白质:GB/T 6432—2018;粗脂肪:GB/T 6433—2006;粗纤维:GB/T 6434—2022;粗灰分:GB/T 6438—2007;钙:GB/T 6436—2018;总磷:GB/T 6437—2018)含量进行测定,按照以下公式计算营养物质表观消化率:

某营养物质表观消化率(%)=[(食入该营养物质质量-粪中该营养物质质量)/食入该营养物质质量]×100。

1.2.3.3 氮代谢指标测定

通过全自动凯氏定氮仪测定饲粮、粪样和尿样中的氮含量,利用以下公式计算貉氮沉积、净蛋白质利用率及蛋白质生物学价值:

氮沉积(g/d)=食入氮-尿氮-粪氮;

净蛋白质利用率(%)=(氮沉积/食入氮)×100;

蛋白质生物学价值(%)=[氮沉积/(食入氮-粪氮)]×100。

1.2.3.4 血清抗氧化指标测定

饲养试验结束时,每组随机选取6只乌苏里貉(公母比例为1∶1),于早晨空腹条件下在其后肢静脉处采血5 mL,分离血清。血清样品采用南京建成生物工程研究所生产的试剂盒测定抗氧化指标,包括超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性以及丙二醛(MDA)含量和总抗氧化能力(T-AOC)。

1.3 数据处理

试验数据用Excel 2016进行整理,使用SPSS 27.0 软件生成的箱线图对数据进行筛选,将筛选后数据进行单因素方差(one-way ANOVA),并采用Duncan氏法进行多重比较;所有结果均以“平均值±标准误”表示,以P<0.05表示差异显著,P>0.05表示差异不显著。

2 结果

2.1 牛至香酚和枯草芽孢杆菌对乌苏里貉生长性能的影响

如表3所示,各试验组的初重、末重、平均日增重、平均日采食量及料重比与对照组相比均无显著差异(P>0.05),但试验Ⅲ组的末重及料重比均优于对照组。

表3 牛至香酚和枯草芽孢杆菌对乌苏里貉生长性能的影响

Table 3 Effects of oregano carvacrol and Bacillus subtilis on growth performance of Wusuli raccoon dogs

项目 Items	初重 IBW/kg	末重 FBW/kg	平均日采食量 ADFI/(g/d)	平均日增重 ADG/(g/d)	料重比 F/G
对照组 Control group	3.21±0.14	7.80±0.12	272.19±1.53	55.98±1.86	5.41±0.16
试验Ⅰ组 Test group Ⅰ	3.21±0.15	7.44±0.26	263.74±5.41	51.64±3.11	5.83±0.86
试验Ⅱ组 Test group Ⅱ	3.23±0.24	7.63±0.17	271.54±3.05	53.62±2.12	5.18±0.19
试验Ⅲ组 Test group Ⅲ	3.28±0.18	7.85±0.18	271.55±6.59	55.68±1.94	4.97±0.18
试验Ⅳ组 Test group Ⅳ	3.38±0.16	7.48±0.20	266.30±9.24	50.00±2.59	5.54±0.29
试验Ⅴ组 Test group Ⅴ	3.27±0.20	7.66±0.14	274.44±0.54	53.51±1.35	5.18±0.13
试验Ⅵ组 Test group Ⅵ	3.33±0.18	7.34±0.33	267.13±7.75	48.93±3.02	6.53±1.25
试验Ⅶ组 Test group Ⅶ	3.25±0.16	7.37±0.14	265.90±9.25	50.19±2.27	5.49±0.30
试验Ⅷ组 Test group Ⅷ	3.15±0.10	7.53±0.13	275.74±1.59	53.32±1.69	5.25±0.16
试验Ⅸ组 Test group Ⅸ	3.26±0.12	7.68±0.14	274.26±2.24	53.85±1.59	5.16±0.17
P值 P-value	0.998	0.615	0.833	0.326	0.559

	同行数据肩标无字母或相同字母表示差异不显著(P>0.05),不同小写字母表示显著差异(P<0.05)。下表同。
	Values in the same row 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 牛至香酚和枯草芽孢杆菌对乌苏里貉营养物质表观消化率的影响

如表4所示,试验Ⅰ、Ⅲ组的干物质表观消化率显著高于对照组(P<0.05),而试验Ⅳ组的干物质表观消化率则显著低于对照组(P<0.05),其余试验组与对照组无显著差异(P>0.05)。试验Ⅳ、Ⅷ及Ⅸ组的粗蛋白质表观消化率显著高于对照组(P<0.05),其余试验组与对照组无显著差异(P>0.05)。各组的粗脂肪表观消化率无显著差异(P>0.05),但以试验Ⅷ组的粗脂肪表观消化率最高。

表4 牛至香酚和枯草芽孢杆菌对乌苏里貉营养物质表观消化率的影响

Table 4 Effects of oregano carvacrol and Bacillus subtilis on apparent digestibility of nutrients of Wusuli raccoon dogs%

项目 Items	干物质表观消化率 DM apparent digestibility	粗蛋白质表观消化率 CP apparent digestibility	粗脂肪表观消化率 EE apparent digestibility
对照组 Control group	74.61±0.61^bc	66.00±0.52^bcd	87.67±0.92
试验Ⅰ组 Test group Ⅰ	81.00±1.97^a	67.67±0.56^ab	86.17±1.66
试验Ⅱ组 Test group Ⅱ	78.67±1.38^ab	64.83±0.87^cd	86.50±1.28
试验Ⅲ组 Test group Ⅲ	80.83±1.78^a	67.67±0.42^ab	87.50±1.34
试验Ⅳ组 Test group Ⅳ	68.50±2.64^d	68.67±0.84^a	88.33±1.61
试验Ⅴ组 Test group Ⅴ	73.17±0.87^c	64.00±1.48^d	86.67±2.09
试验Ⅵ组 Test group Ⅵ	75.50±0.85^bc	67.33±0.33^ab	87.33±0.67
试验Ⅶ组 Test group Ⅶ	76.67±1.50^abc	67.00±0.58^abc	87.50±0.76
试验Ⅷ组 Test group Ⅷ	75.83±0.65^bc	68.67±0.76^a	91.00±0.45
试验Ⅸ组 Test group Ⅸ	74.00±0.58^c	68.83±0.91^a	88.67±1.17
P值 P-value	<0.001	<0.001	0.342

2.3 牛至香酚和枯草芽孢杆菌对乌苏里貉氮代谢的影响

如表5所示,各组的食入氮无显著差异(P>0.05)。试验Ⅳ组的粪氮显著高于其余各组(P<0.05),其余组间无显著差异(P>0.05)。试验Ⅶ组的尿氮显著高于对照组与试验Ⅴ、Ⅵ、Ⅷ及Ⅸ组(P<0.05),其余试验组与对照组无显著差异(P>0.05)。试验Ⅳ组的氮沉积、净蛋白质利用率及蛋白质生物学价值均显著低于对照组(P<0.05),其余试验组与对照组无显著差异(P>0.05)。

表5 牛至香酚和枯草芽孢杆菌对乌苏里貉氮代谢的影响

Table 5 Effects of oregano carvacrol and Bacillus subtilis on nitrogen metabolism of Wusuli raccoon dogs

项目 Items	食入氮 Nitrogen intake/ (g/d)	粪氮 Fecal nitrogen/ (g/d)	尿氮 Urine nitrogen/ (g/d)	氮沉积 Nitrogen retention/ (g/d)	净蛋白质利用率 Net protein utilization rate/%	蛋白质生物学价值 Biological value of protein/%
对照组 Control group	11.13±0.11	2.86±0.06^b	4.61±0.19^bcd	3.77±0.23^abc	33.52±2.05^abc	44.90±2.54^abc
试验Ⅰ组 Test group Ⅰ	11.48±0.00	2.76±0.21^b	5.76±0.67^abcd	2.96±0.60^abcd	25.75±5.21^abcd	34.09±7.34^abcd
试验Ⅱ组 Test group Ⅱ	11.27±0.21	3.02±0.08^b	6.03±1.59^abcd	2.44±1.67^bcd	21.21±14.5^bcd	28.6±19.42^bcd
试验Ⅲ组 Test group Ⅲ	11.48±0.00	2.75±0.24^b	6.29±0.76^abc	2.44±0.67^bcd	21.28±5.88^bcd	28.19±8.09^bcd
试验Ⅳ组 Test group Ⅳ	11.50±0.14	3.72±0.33^a	6.54±0.59^ab	1.21±0.30^d	10.56±2.56^d	16.07±3.96^d
试验Ⅴ组 Test group Ⅴ	11.09±0.39	3.07±0.14^b	4.18±0.34^cd	3.84±0.54^abc	34.41±4.22^abc	47.25±5.17^ab
试验Ⅵ组 Test group Ⅵ	11.66±0.00	2.83±0.11^b	4.81±0.43^bcd	4.02±0.39^abc	34.47±3.31^abc	45.64±4.49^abc
试验Ⅶ组 Test group Ⅶ	11.61±0.00	2.52±0.22^b	6.95±1.05^a	2.14±1.08^cd	18.41±9.29^cd	23.37±11.59^cd
试验Ⅷ组 Test group Ⅷ	11.61±0.00	2.69±0.12^b	4.42±0.85^bcd	4.50±0.81^ab	38.73±7.00^ab	50.59±9.19^ab
试验Ⅸ组 Test group Ⅸ	11.61±0.00	2.87±0.08^b	4.09±0.23^d	4.65±0.28^a	40.05±2.45^a	53.11±2.89^a
P值 P-value	0.072	0.005	0.012	0.003	0.005	0.005

2.4 牛至香酚和枯草芽孢杆菌对乌苏里貉血清抗氧化指标的影响

如表6所示,各组间血清SOD活性与T-AOC无显著差异(P>0.05)。试验Ⅷ组的血清GSH-Px活性最高,除与对照组及试验Ⅳ、Ⅸ组差异不显著外(P>0.05),显著高于其余各组(P<0.05)。各试验组血清MDA含量与对照组相比均无显著差异(P>0.05),但试验Ⅱ、Ⅶ组显著高于试验Ⅳ、Ⅵ及Ⅸ组(P<0.05)。

表6 牛至香酚和枯草芽孢杆菌对乌苏里貉血清抗氧化指标的影响

Table 6 Effects of oregano carvacrol and Bacillus subtilis on serum antioxidant indexes of Wusuli raccoon dogs

项目 Items	谷胱甘肽过氧化物酶 GSH-Px/(U/mL)	超氧化物歧化酶 SOD/(U/mL)	丙二醛 MDA/(nmol/mL)	总抗氧化能力 T-AOC/(mmol/mL)
对照组 Control group	1 596.55±39.81^ab	143.51±4.19	18.92±0.88^ab	0.97±0.02
试验Ⅰ组 Test group Ⅰ	1 269.36±57.41^d	126.59±5.19	19.44±1.52^ab	0.91±0.02
试验Ⅱ组 Test group Ⅱ	1 376.84±62.66^cd	137.14±5.14	22.58±1.75^a	0.97±0.03
试验Ⅲ组 Test group Ⅲ	1 290.90±57.75^d	144.59±4.60	19.75±1.82^ab	0.95±0.03
试验Ⅳ组 Test group Ⅳ	1 442.74±73.15^abc	144.66±4.84	16.88±1.48^b	0.95±0.02
试验Ⅴ组 Test group Ⅴ	1 290.90±57.01^d	142.43±6.10	17.80±1.53^ab	0.93±0.01
试验Ⅵ组 Test group Ⅵ	1 395.16±78.11^cd	132.75±7.56	16.67±1.29^b	0.93±0.01
试验Ⅶ组 Test group Ⅶ	1 375.35±70.87^cd	137.30±5.68	22.38±1.04^a	0.93±0.03
试验Ⅷ组 Test group Ⅷ	1 734.68±42.22^a	139.49±3.35	19.48±1.33^ab	0.92±0.02
试验Ⅸ组 Test group Ⅸ	1 539.86±38.95^ab	151.79±3.64	15.05±1.94^b	0.89±0.03
P值 P-value	<0.001	0.079	0.025	0.226

3 讨论

3.1 牛至香酚和枯草芽孢杆菌对乌苏里貉生长性能的影响

Ruan等^[15]的研究发现,饲粮中添加150~300 mg/kg的牛至精油对清远麻鸡的生长性能有积极影响。Pirgozliev等^[16]的研究发现,饲粮中添加包含5%香芹酚、3%肉桂醛和2%辣椒精油的复合物改善了罗斯(Ross)308雏鸡的平均日采食量,增加了肉鸡的体重。Lee等^[17]的研究发现,60~120 mg/kg的牛至精油(含有14%的相等水平的香芹酚和百里香酚)可显著缓解球虫病引起的肉鸡平均日采食量降低。以上研究结果显示牛至精油可作为生长促进剂替代抗生素的在畜禽生产中应用。益生菌也已经成为一种可以部分替代抗生素的安全替代品,且芽孢杆菌由于芽孢对极端条件的强抗性和能够长期储存的性能,被公认为是最合适的益生菌。研究表明,饲粮中添加枯草芽孢杆菌能显著提高肉鸡^[18]、肉兔^[19]、断奶仔猪^[20-21]等动物的生长性能。研究发现,在北极狐饲粮中添加250 mg/kg牛至提取物和300 mg/kg枯草芽孢杆菌能够提高其末重与平均日增重^[22]。宗文丽等^[23]的研究表明,在饲粮中添加牛至香酚和枯草芽孢杆菌组合能够提高育成期蓝狐的平均日增重,降低料重比。在本试验条件下,饲粮中添加不同水平牛至香酚和枯草芽孢杆菌的各试验组与添加抗生素的对照组相比,在末重、平均日增重、平均日采食量及料重比等方面均无显著差异,部分试验组的末重和料重比还优于对照组,说明牛至香酚和枯草芽孢杆菌组合对乌苏里貉生长性能与抗生素效果相似甚至更优。当牛至香酚添加水平为100 mg/kg,枯草芽孢杆菌添加水平450 mg/kg时,育成期乌苏里貉的末重最大,料重比最低。

3.2 牛至香酚和枯草芽孢杆菌对乌苏里貉营养物质表观消化率的影响

牛至香酚可以改善动物肠道菌群结构和小肠形态,保护肠道屏障功能,提高营养物质的消化利用率^[24-25]。枯草芽孢杆菌具有良好的分泌淀粉酶、蛋白酶、脂肪酶等消化酶的能力,且枯草芽孢杆菌还能够促进肠道绒毛发育,增加肠道与营养物质的接触面积,提高动物肠道的消化吸收能力^[26]。赵天成等^[27]的研究表明,在冬毛期蓝狐饲粮中添加百里香酚对其粗蛋白质和粗脂肪表观消化率有提升趋势。周立强等^[28]的研究表明,在黄羽肉鸡饲粮中添加150 mg/kg枯草芽孢杆菌制剂可极显著提高粗蛋白质表观消化率。在本试验条件下,饲粮中添加适宜水平的牛至香酚和枯草芽孢杆菌在一定程度上能够提高育成期乌苏里貉的干物质、粗蛋白质及粗脂肪的表观消化率,且效果优于抗生素。

牛至精油能够提高动物回肠胰凝乳蛋白酶和脂肪酶的活性以及盲肠中乳酸菌的丰度,且能抑制肠道内大肠杆菌的增殖^[29]。此外,牛至精油还能增加十二指肠、空肠和回肠绒毛高度增加,降低十二指肠隐窝深度,改善动物肠道形态^[30]。研究表明,枯草芽孢杆菌丰富了与外源生物降解和代谢、碳水化合物代谢、能量代谢、信号分子和相互作用、消化系统以及运输和分解代谢相关的途径,进而提高饲料转化率^[31]。枯草芽孢杆菌还能够通过上调紧密连接蛋白的表达显著增强基础条件下的肠道屏障完整性^[32],改善动物肠道形态^[33-34]。Kim等^[35]的研究结果表明,饲粮中补充枯草芽孢杆菌和牛至精油可以改善动物肠道微生物群的组成,对肠道健康有积极作用。在本试验条件下,乌苏里貉对饲粮中营养物质表观消化率提高可能是由于牛至香酚和枯草芽孢杆菌改善了其肠道形态,调节了肠道菌群,但具体的作用机制还需进一步通过分子生物学方法进行验证。

3.3 牛至香酚和枯草芽孢杆菌对乌苏里貉氮代谢的影响

氮代谢可以反映体内蛋白质的代谢情况。枯草芽孢杆菌可以通过提高动物对氮的利用率来减少氮的排放,降低养殖环境中氨气的浓度^[36]。Zhang等^[37]的研究表明,种鹅饲粮中添加60或80 mg/kg铁(Fe)和5×10⁹ CFU/kg的枯草芽孢杆菌可显著降低血清尿素氮含量。Jeong等^[38]的研究表明,在肉鸡饲粮中添加枯草芽孢杆菌可显著降低粪便中氨气的排放量。在本试验条件下,当牛至香酚和枯草芽孢杆菌添加水平分别为200和450 mg/kg时,乌苏里貉的食入氮要高于对照组。乌苏里貉的食入氮主要来源于饲料中的蛋白质,说明适量的牛至香酚和枯草芽孢杆菌能够提高动物的采食量。在本试验条件下,牛至香酚和枯草芽孢杆菌能够在一定程度上提高乌苏里貉对氮的摄入量,侧面反映了两者联用能够增加动物的采食量,猜测其原因可能是牛至香酚所特有的气味刺激了乌苏里貉消化道感受器,从而增加其采食欲望。牛至香酚和枯草芽孢杆菌还提高了育成期乌苏里貉的净蛋白质利用率与蛋白质生物学价值,减少了粪氮、尿氮排放量。推测其原因可能是枯草芽孢杆菌在乌苏里貉生长过程中产生了多种消化酶,其中蛋白酶能够与内源酶协同作用,促进肠道消化吸收^[39-40]。枯草芽孢杆菌定植在肠道内后,促进有益菌的增殖,而乳酸菌能够将蛋白质类的大分子物质分解为游离的氨基酸,直接被机体吸收,减少粪氮含量^[41]。研究表明,枯草芽孢杆菌能显著增加仔猪粪便中乳酸菌数量,显著减少血清中尿素氮含量,减少氨气的排放量^[42]。

3.4 牛至香酚和枯草芽孢杆菌对乌苏里貉抗氧化能力的影响

牛至香酚含有非常丰富的酚类成分,能够调节抗氧化酶清除自由基,具有抗氧化的特性^[43-44]。枯草芽孢杆菌能够增加谷胱甘肽(GSH)基因的表达量。研究表明,枯草芽孢杆菌的孢子能够增加GSH含量和过氧化氢酶活性^[45]。枯草芽孢杆菌还能够通过激活核糖体和氧化磷酸化信号调节抗氧化和能量代谢来减轻大肠杆菌感染造成的氧化应激^[46]。Zhang等^[47]的研究表明,在肉鸡饲粮中添加牛至精油可以提高血清GSH-Px和SOD活性,降低MDA含量,达到与抗生素相似的效果。刘福鑫等^[48]的研究表明,香芹酚能够显著提高肉兔空肠GSH-Px活性和T-AOC,显著降低MDA含量,说明香芹酚能够增强机体的抗氧化能力。Xu等^[49]的研究表明,在饲粮中添加枯草芽孢杆菌能够提高肉鸡的抗氧化能力。研究发现,枯草芽孢杆菌能够显著提高采食受黄曲霉毒素污染的发霉玉米的鸭的抗氧化酶活性,并减少鸭肝中黄曲霉毒素的积累^[50]。Liu等^[51]的研究发现,饲粮中添加900 mg/kg枯草芽孢杆菌和植物精油的混合物可增强肉种鸡的抗氧化能力。在本试验条件下,当牛至香酚添加水平为300 mg/kg、枯草芽孢杆菌添加水平为300 mg/kg时,乌苏里貉血清中GSH-Px活性最高;当牛至香酚添加水平为300 mg/kg、枯草芽孢杆菌添加水平为450 mg/kg时,乌苏里貉血清中SOD活性最高,MDA含量最低。此外,牛至香酚和枯草芽孢杆菌对乌苏里貉血清T-AOC也有积极影响。上述结果说明适量的牛至香酚和枯草芽孢杆菌能够改善乌苏里貉的抗氧化能力。

4 结论

饲粮中添加适宜水平的牛至香酚和枯草芽孢杆菌在一定程度上能够改善育成期乌苏里貉的生长性能,促进营养物质的消化代谢,提高机体的抗氧化能力,且效果与抗生素相当甚至有些指标还优于抗生素。本试验条件下,育成期乌苏里貉饲粮中牛至香酚和枯草芽孢杆菌的适宜添加水平分别为100和450 mg/kg。

References

Publishing order | Descend order by publishing year | Descend order by cited within

[1]	王永颖, 唐佩娟, 窦新艳, 等. 饲用抗生素替代技术应用前景分析[J]. 天津农林科技, 2022(5):39-42. WANG Y Y, TANG P J, DOU X Y, et al. Analysis on the application prospect of alternative technology of feeding antibiotics[J]. Science and Technology of Tianjin Agriculture and Forestry, 2022(5):39-42. (in Chinese)

[2]	AL-KHALAIFA H, AL-NASSER A, AL-SURAYEE T, et al. Effect of dietary probiotics and prebiotics on the performance of broiler chickens[J]. Poultry Science, 2021, 100(2):1329.

[3]	ANDREW SELALEDI L, MOHAMMED HASSAN Z, MANYELO T G, et al. The current status of the alternative use to antibiotics in poultry production:an African perspective[J]. Antibiotics, 2020, 9(9):594.

[4]	SWEENEY M T, LUBBERS B V, SCHWARZ S, et al. Applying definitions for multidrug resistance,extensive drug resistance and pandrug resistance to clinically significant livestock and companion animal bacterial pathogens-authors’ response[J]. Journal of Antimicrobial Chemotherapy, 2019, 74(2):536-537.

[5]	佚名. 中华人民共和国农业农村部公告第194号[J]. 浙江畜牧兽医, 2020, 45(1):28. Anon. Announcement No. 194 of the Ministry of Agriculture and Rural Affairs of the People’s Republic of China[J]. Zhejiang Journal Animal Science and Veterinary Medicine, 2020, 45(1):28. (in Chinese)

[6]	GADDE U, KIM W H, OH S T, et al. Alternatives to antibiotics for maximizing growth performance and feed efficiency in poultry:a review[J]. Animal Health Research Reviews, 2017, 18(1):26-45.

[7]	吴樟强. 牛至香酚在禽呼吸道病上的应用[J]. 畜牧兽医科技信息, 2019(6):129-130. WU Z Q. Application of oregano in avian respiratory diseases[J]. Chinese Journal of Animal Husbandry and Veterinary Medicine, 2019(6):129-130. (in Chinese)

[8]	LUO K Y, ZHAO P Y, HE Y F, et al. Antibacterial effect of oregano essential oil against Vibrio vulnificus and its mechanism[J]. Foods, 2022, 11(3):403.

[9]	KACHUR K, SUNTRES Z. The antibacterial properties of phenolic isomers,carvacrol and thymol[J]. Critical Reviews in Food Science and Nutrition, 2020, 60(18):3042-3053.

[10]	LI X Y, WU S R, LI X Y, et al. Simultaneous supplementation of Bacillus subtilis and antibiotic growth promoters by stages improved intestinal function of pullets by altering gut microbiota[J]. Frontiers in Microbiology, 2018,9:2328.

[11]	刘帅, 熊云霞, 孙铝辉, 等. 枯草芽孢杆菌在猪生产中的应用及其作用机制的研究进展[J]. 动物营养学报, 2022, 34(4):2105-2113. DOI LIU S, XIONG Y X, SUN L H, et al. Research progress of application of Bacillus subtilis as feed additive in pigs production[J]. Chinese Journal of Animal Nutrition, 2022, 34(4):2105-2113. (in Chinese)

[12]	PU J N, CHEN D W, TIAN G, et al. Effects of benzoic acid,Bacillus coagulans and oregano oil combined supplementation on growth performance,immune status and intestinal barrier integrity of weaned piglets[J]. Animal Nutrition, 2020, 6(2):152-159.

[13]	TAN B F, LIM T, BOONTIAM W. Effect of dietary supplementation with essential oils and a Bacillus probiotic on growth performance,diarrhoea and blood metabolites in weaned pigs[J]. Animal Production Science, 2021,61:64-71.

[14]	WANG J, LI G Y, ZHONG W, et al. Effect of dietary Paeoniae Radix Alba extract on the growth performance,nutrient digestibility and metabolism,serum biochemistry,and small intestine histomorphology of raccoon dog during the growing period[J]. Frontiers in Veterinary Science, 2022,9:839450.

[15]	RUAN D, FAN Q L, FOUAD A M, et al. Effects of dietary oregano essential oil supplementation on growth performance,intestinal antioxidative capacity,immunity,and intestinal microbiota in yellow-feathered chickens[J]. Journal of Animal Science, 2021, 99(2):skab033.

[16]	PIRGOZLIEV V, MANSBRIDGE S C, ROSE S P, et al. Immune modulation, growth performance,and nutrient retention in broiler chickens fed a blend of phytogenic feed additives[J]. Poultry Science, 2019, 98(9):3443-3449.

[17]	LEE J W, KIM D H, KIM Y B, et al. Dietary encapsulated essential oils improve production performance of coccidiosis-vaccine-challenged broiler chickens[J]. Animals, 2020, 10(3):481.

[18]	李泽然. 枯草芽孢杆菌对肉鸡生长性能、屠宰性能、肉品质、免疫功能、抗氧化功能和肠道形态的影响[J]. 饲料研究, 2023, 46(11):51-55. LI Z R. Effect of Bacillus subtilis on growth performance,slaughter performance,meat quality,immune function,antioxidant function and intestinal morphology of broilers[J]. Feed Research, 2023, 46(11):51-55. (in Chinese)

[19]	冯宇, 崔萌. 枯草芽孢杆菌或凝结芽孢杆菌对肉兔生长性能、糖代谢及肠道菌群的影响[J]. 中国饲料, 2023(6):77-80. FENG Y, CUI M. Effects of Bacillus subtilis or Bacillus coagulans on growth performance,glucose metabolism and intestinal microflora of rabbit[J]. China Feed, 2023(6):77-80. (in Chinese)

[20]	朱五文, 施伟领, 陈晓峰. 不同剂量枯草芽孢杆菌制剂对断奶仔猪饲养效果试验[J]. 畜牧与兽医, 2007, 39(8):32-33. ZHU W W, SHI W L, CHEN X F. Feeding effect of different doses of Bacillus subtilis preparation on weaned piglets[J]. Animal Husbandry & Veterinary Medicine, 2007, 39(8):32-33. (in Chinese)

[21]	周映华, 周小玲, 吴胜莲, 等. 枯草芽孢杆菌对断奶仔猪生产性能的影响[J]. 饲料研究, 2012(5):71-73. ZHOU Y H, ZHOU X L, WU S L, et al. Effects of Bacillus subtilis on growth performance of weaned piglets[J]. Feed Research, 2012(5):71-73. (in Chinese)

[22]

宗文丽, 郭亮, 邓露芳, 等. 枯草芽孢杆菌和牛至提取物对北极狐生长性能和粪便中细菌数量的影响[J]. 特产研究, 2023, 45(5):116-120,125.

ZONG

W L

, GUO

, DENG

L F

, et al. Effect of Bacillus subtilis and Origanum vulgare L. extracts on the growth performance and numbers of bacteria in the feces of arctic fox (Alopex lagopus)[J]. Special Wild Economic Animal and Plant Research, 2023, 45(5):116-120,125. (in Chinese)

[23]	宗文丽, 邓露芳, 辛娜, 等. 牛至香酚和枯草芽孢杆菌对育成期蓝狐生长性能和泪痕的影响[J]. 饲料研究, 2022, 45(8):63-66. ZONG W L, DENG L F, XIN N, et al. Effects of oregano carvacrol and Bacillus subtilis on growth performance and tear stains in breeding blue foxes[J]. Feed Research, 2022, 45(8):63-66. (in Chinese)

[24]	JIA L, WU J P, LEI Y, et al. Oregano essential oils mediated intestinal microbiota and metabolites and improved growth performance and intestinal barrier function in sheep[J]. Frontiers in Immunology, 2022,13:908015.

[25]	AMER H Y, HASSAN R I M, MUSTAFA F E Z A, et al. Modulation of immunity,antioxidant status,performance,blood hematology,and intestinal histomorphometry in response to dietary inclusion of Origanum majorana in domestic pigeons’ diet[J]. Life, 2023, 13(3):664.

[26]	AKHTAR N, CAI H Y, KIARIE E G, et al. A novel Bacillus sp. with rapid growth property and high enzyme activity that allows efficient fermentation of soybean meal for improving digestibility in growing pigs[J]. Journal of Applied Microbiology, 2022, 133(1):3-17.

[27]	赵天成, 刘钊, 臧晓, 等. 百里香酚对冬毛期蓝狐生产性能与生化指标的影响[J]. 饲料工业, 2022, 43(9):60-64. ZHAO T C, LIU Z, ZANG X, et al. Effect of thymol on production performance and biochemical indexes of blue fox in winter fur-growing period[J]. Feed Industry, 2022, 43(9):60-64. (in Chinese)

[28]	周立强, 兰丽燕, 徐春生, 等. 枯草芽孢杆菌对黄羽肉鸡生长性能及营养物质代谢率的影响[J]. 中国畜牧兽医, 2012, 39(2):71-74. ZHOU L Q, LAN L Y, XU C S, et al. Effects of Bacillus subtilis on growth performance and nutrient metabolic rate in yellow broilers[J]. China Animal Husbandry & Veterinary Medicine, 2012, 39(2):71-74. (in Chinese)

[29]

GIANNENAS

, PAPANEOPHYTOU

C P

, TSALIE

, et al. Dietary supplementation of benzoic acid and essential oil compounds affects buffering capacity of the feeds,performance of turkey poults and their antioxidant status,pH in the digestive tract,intestinal microbiota and morphology[J]. Asian-Australasian Journal of Animal Sciences, 2014, 27(2):225-236.

[30]	AMER S A, TOLBA S A, ALSADEK D M M, et al. Effect of supplemental glycerol monolaurate and oregano essential oil blend on the growth performance,intestinal morphology,and amino acid digestibility of broiler chickens[J]. BMC Veterinary Research, 2021, 17(1):312.

[31]	LIU S, XIAO G S, WANG Q, et al. Effects of dietary Bacillus subtilis HC6 on growth performance,antioxidant capacity,immunity,and intestinal health in broilers[J]. Animals, 2023, 13(18):2915.

[32]	RHAYAT L, MARESCA M, NICOLETTI C, et al. Effect of Bacillus subtilis strains on intestinal barrier function and inflammatory response[J]. Frontiers in Immunology, 2019,10:564.

[33]	MA Y B, WANG W W, ZHANG H J, et al. Supplemental Bacillus subtilis DSM 32315 manipulates intestinal structure and microbial composition in broiler chickens[J]. Scientific Reports, 2018, 8(1):15358.

[34]	ZHANG Q Q, LI J, WANG G X, et al. The replacement of bacitracin methylene disalicylate with Bacillus subtilis PB6 in the diet of male cherry valley ducks reduces the feed conversion ratio by improving intestinal health and modulating gut microbiota[J]. Poultry Science, 2022, 101(11):102155.

[35]	KIM H J, KIM H S, YUN Y S, et al. Effect of Bacillus subtilis and oregano oil on performance,gut microbiome,and intestinal morphology in pullets[J]. Animals, 2023, 13(16):2550.

[36]	于洁, 赵敏孟, 龚道清. 枯草芽孢杆菌在畜禽生产中的研究进展[J]. 中国饲料, 2020(19):6-11. YU J, ZHAO M M, GONG D Q. Research progress in application of Bacillus subtilis on animal production[J]. China Feed, 2020(19):6-11. (in Chinese)

[37]	ZHANG B B, SUI F L, WANG B W, et al. Dietary combined supplementation of iron and Bacillus subtilis enhances reproductive performance,eggshell quality,nutrient digestibility,antioxidant capacity,and hematopoietic function in breeder geese[J]. Poultry Science, 2020, 99(11):6119-6127.

[38]	JEONG J S, KIM I H. Effect of Bacillus subtilis C-3102 spores as a probiotic feed supplement on growth performance,noxious gas emission,and intestinal microflora in broilers[J]. Poultry Science, 2014, 93(12):3097-3103.

[39]	刘兴, 姚晶, 夏元军. 枯草芽孢杆菌在固态发酵饲料中的应用研究进展[J]. 饲料研究, 2020, 43(2):113-116. LIU X, YAO J, XIA Y J. Research progress on the application of Bacillus subtilis in solid-state fermented feed[J]. Feed Research, 2020, 43(2):113-116. (in Chinese)

[40]	朱瑾, 朱红军. 枯草芽孢杆菌的作用机制及其在动物生产中的应用[J]. 粮食与饲料工业, 2019(8):47-51. ZHU J, ZHU H J. Mechanism of Bacillus subtilis and its application in animal production[J]. Cereal & Feed Industry, 2019(8):47-51. (in Chinese)

[41]	HIGGINS S E, HIGGINS J P, WOLFENDEN A D, et al. Evaluation of a lactobacillus-based probiotic culture for the reduction of Salmonella enteritidis in neonatal broiler chicks[J]. Poultry Science, 2008, 87(1):27-31.

[42]	LAN R X, TRAN H, KIM I. Effects of probiotic supplementation in different nutrient density diets on growth performance,nutrient digestibility,blood profiles,fecal microflora and noxious gas emission in weaning pig[J]. Journal of the Science of Food and Agriculture, 2017, 97(4):1335-1341.

[43]

黄其春, 吴樟强, 赖建彬, 等. 牛至提取物对生长育肥猪生长性能、胴体性状及肉品质的影响[J]. 中国畜牧杂志, 2020, 56(8):150-153.

HUANG

Q C

, WU

Z Q

, LAI

J B

, et al. Effects of Origanum vulgare L. extract on growth performance,carcass traits and meat quality in growing-finishing pigs[J]. Chinese Journal of Animal Science, 2020, 56(8):150-153. (in Chinese)

[44]	DING X M, WU X P, ZHANG K Y, et al. Dietary supplement of essential oil from oregano affects growth performance,nutrient utilization,intestinal morphology and antioxidant ability in Pekin ducks[J]. Journal of Animal Physiology and Animal Nutrition, 2020, 104(4):1067-1074.

[45]	ABDEL-MONEIM A M E, SELIM D A, BASUONY H A, et al. Effect of dietary supplementation of Bacillus subtilis spores on growth performance,oxidative status,and digestive enzyme activities in Japanese quail birds[J]. Tropical Animal Health and Production, 2020, 52(2):671-680.

[46]	LI C, LI Y, LI S Z, et al. Bacillus subtilis protects the ducks from oxidative stress induced by Escherichia coli:efficacy and molecular mechanism[J]. Antioxidants, 2022, 11(10):1951.

[47]	ZHANG L Y, PENG Q Y, LIU Y R, et al. Effects of oregano essential oil as an antibiotic growth promoter alternative on growth performance,antioxidant status,and intestinal health of broilers[J]. Poultry Science, 2021, 100(7):101163.

[48]

刘福鑫, 孔繁根, 王国洲, 等. 香芹酚对肉兔生长性能、养分表观消化率、肠道消化酶活性、免疫器官发育及抗氧化能力的影响[J]. 动物营养学报, 2022, 34(3):1875-1884.

DOI

LIU

F X

, KONG

F G

, WANG

G Z

, et al. Effects of carvacrol on growth performance,nutrient apparent digestibilities,intestinal digestive enzyme activities,immune organ development and antioxidant capacity of meat rabbits[J]. Chinese Journal of Animal Nutrition, 2022, 34(3):1875-1884. (in Chinese)

[49]	XU Y L, YU Y, SHEN Y Y, et al. Effects of Bacillus subtilis and Bacillus licheniformis on growth performance,immunity,short chain fatty acid production,antioxidant capacity,and cecal microflora in broilers[J]. Poultry Science, 2021, 100(9):101358.

[50]	ZHANG L Y, MA Q G, MA S S, et al. Ameliorating effects of Bacillus subtilis ANSB060 on growth performance,antioxidant functions,and aflatoxin residues in ducks fed diets contaminated with aflatoxins[J]. Toxins, 2016, 9(1):1.

[51]	LIU X, CHEN Y B, TANG S G, et al. Dietary encapsulated Bacillus subtilis and essential oil supplementation improves reproductive performance and hormone concentrations of broiler breeders during the late laying period[J]. Livestock Science, 2021,245:104422.

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