紫色达利菊缩合单宁对大肠杆菌的抑制作用

刘秀丽; 王玉玺; 李元恒; 许中军; 金龙; 侯勇跃

doi:10.3969/j.issn.1006-267x.2015.11.036

动物营养学报 >

2015 , Vol. 27 >Issue 11: 3620 - 3627

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.11.036

研究简报 Short Communications

紫色达利菊缩合单宁对大肠杆菌的抑制作用

刘秀丽 ,
王玉玺 ,
李元恒 ,
许中军 ,
金龙 ,
侯勇跃

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1. 内蒙古自治区农牧业科学院, 呼和浩特 010031;
2. 加拿大农业与农业食品部列桥研究中心, 列桥 T1J4B1, 加拿大;
3. 中国农业科学院草原研究所, 呼和浩特 010010;
4. 东北农业大学动物科学与技术学院, 哈尔滨 150030

收稿日期: 2015-05-27

网络出版日期: 2015-11-21

基金资助

国家自然科学基金项目(31460652);公益性行业(农业)科研专项课题(201303040-11)

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Inhibiting Effects of Condensed Tannins from Purple Prairie Clover on Escherichia coli

LIU Xiuli ,
WANG Yuxi ,
LI Yuanheng ,
XU Zhongjun ,
JIN Long ,
HOU Yongyue

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1. Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010031, China;
2. Lethbridge Research Centre, Agriculture and Agri-Food Canada, Lethbridge T1J 4B1, Canada;
3. Institute of Grassland Research of CAAS, Hohhot 010010, China;
4. College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China

Received date: 2015-05-27

Online published: 2015-11-21

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摘要

本文旨在研究紫色达利菊(purple prairie clover,PPC)对肉牛瘤胃和大肠中大肠杆菌生长的体外抑制作用及其缩合单宁(condensed tannins,CT)对大肠杆菌的抑制机理。体外模拟肉牛瘤胃和大肠环境,以非致病性大肠杆菌ATCC 25922和致病性大肠杆菌O157:H7作为试验菌株,在分别以PPC和苜蓿(无CT,对照)为底物培养,在培养0、1.5、3.0、6.0、24.0 h测定菌落数;以大肠杆菌ATCC 25922为研究模型,分别以添加PPC的CT和红豆草的CT的菌悬液与邻-硝基苯-β-D-半乳糖苷溶液混合,同时以无CT添加的菌悬液和双蒸水混合作为对照,分别在0、40、80、120、160 min时测定β-D-半乳糖苷酶水解产物邻-硝基苯酚的量,以判断细胞内膜通透性的变化;同时观测不同pH(5.3、6.8、7.4、8.0、9.0)和环境温度(4、22℃)对CT作用效果的影响。结果表明:1)与苜蓿相比,PPC对瘤胃(6.0、24.0 h)和大肠中(1.5~24.0 h)的大肠杆菌生长均有显著抑制作用(P<0.05)。2)CT能够提高邻-硝基苯酚的量;与红豆草CT的作用规律有所不同,PPC的CT的作用会受到温度和pH的影响。结果提示,PPC在瘤胃及大肠环境下均具有抑制大肠杆菌生长的作用,其所含CT抑制大肠杆菌生长的机理部分与其增加细胞内膜的通透性有关,且细胞内膜通透性受到生长环境,如温度和pH的影响。

关键词： 紫色达利菊; 缩合单宁; 大肠杆菌; 抑制

本文引用格式

刘秀丽 , 王玉玺 , 李元恒 , 许中军 , 金龙 , 侯勇跃 . 紫色达利菊缩合单宁对大肠杆菌的抑制作用[J]. 动物营养学报, 2015 , 27(11) : 3620 -3627 . DOI: 10.3969/j.issn.1006-267x.2015.11.036

Abstract

This experiment was conducted to evaluate inhibiting effects of purple prairie clover (PPC, Dalea purpurea Vent.) on Escherichia coli (E. coli) growth prevalence in the rumen and large intestine and its condensed tannins (CT) antibacterial mechanism. Ruminal and large intestinal environments were simulated in vitro, non-pathogenic E. coli ATCC 25922 and pathogenic E. coli O157:H7 were used as test strains, and the strains were cultured with PPC and alfalfa (no CT, control) as substrates, respectively. Bacteria number was determined after cultured for 0, 1.5, 3.0, 6.0 and 24.0 h, respectively. With the E. coli ATCC 25922 as test model, the bacteria suspensions supplemented with CT from PPC and sainfoin were mixed with o-nitrophenyl-β-D-galactoside (ONPG) solution, meanwhile, the mixture of bacteria suspension without CT and ddH₂O was used as control. O-nitrophenyl amount, the hydrolysis product of β-D-galactosidase, was determined after cultured for 0, 40, 80, 120 and 160 min, respectively, to analysis the change of cell inner-membrane permeability. The effects of different pH and temperatures on CT actions were also observed. The results revealed as follows:1) compared with alfalfa, PPC had significant inhibiting effect on growth of E. coli from rumen (6.0 and 24.0 h) and large intestine (1.5 to 24.0 h) (P<0.05). 2) CT significantly increased o-nitrophenyl amount (P<0.05); differently from actions of CT from sainfoin, actions of CT from PPC were affected by pH and temperature. The results indicate that PPC showed inhibiting effect on E. coli growth in bovine rumen and large intestine in vitro incubation; its CT can increase inner membrane permeability of E. coli, however, action of inhibition associated with temperature and pH.

Key words： PPC; condensed tannins; E. coli; inhibition

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