植物提取物对犬源大肠杆菌的体外抑菌效果及作用机制

doi:10.16431/j.cnki.1671-7236.2025.08.037

摘要/Abstract

摘要： 【目的】筛选对犬源大肠杆菌有抑制效果的植物提取物，为植物提取物在犬细菌性疾病防控领域的应用提供借鉴。【方法】选取莽草酸、甜茶苷、绿原酸等15种植物提取物，采取打孔法测定各提取物对犬源大肠杆菌的体外抑菌活性，筛选出具有明显抑菌作用的植物提取物。采用二倍稀释法测定植物提取物对犬源大肠杆菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。将MIC剂量的植物提取物与犬源大肠杆菌恒温摇床共培养，定时取样离心收集上清，以上清的D_{600 nm}、D_{260 nm}和D_{280 nm}值来判断不同植物提取物对大肠杆菌的生长曲线和胞外核酸及蛋白的影响。使用试剂盒测定菌液上清中碱性磷酸酶(AKP)和菌体中乳酸脱氢酶(LDH)活性。【结果】莽草酸、甜茶苷、绿原酸、根皮素、红景天苷对犬源大肠杆菌具有良好的抑菌效果，按照抑菌圈直径大小判断抑菌能力依次为：莽草酸＞绿原酸＞甜茶苷＞根皮素＞红景天苷。根皮素、莽草酸、甜茶苷、绿原酸和红景天苷对犬源大肠杆菌的MIC分别为0.48、15.625、62.5、31.25和125 mg/mL，MBC分别为0.48、31.25、125、62.5和250 mg/mL。生长曲线显示，MIC剂量的各植物提取物均对犬源大肠杆菌的生长起到了抑制作用。分析菌液上清D_{260 nm}和D_{280 nm}值可知，绿原酸、红景天苷、甜茶苷、根皮素对大肠杆菌的细胞膜具有显著的破坏作用。在甜茶苷、红景天苷作用下菌液上清AKP活性显著高于对照组(P＜0.05)，说明二者能破坏大肠杆菌的细胞壁从而发挥抑菌作用。在莽草酸、甜茶苷、绿原酸、根皮素、红景天苷5种植物提取物作用菌体LDH活性均显著低于对照组(P＜0.05)，说明这5种植物提取物均可抑制菌体内LDH的活性，影响细菌的代谢活动。【结论】莽草酸、甜茶苷、绿原酸、根皮素、红景天苷对犬源大肠杆菌具有明显抑制作用。

关键词: 植物提取物; 犬源大肠杆菌; 体外抑菌

Abstract: 【Objective】 The purpose of the experiment was to screen plant extracts that had inhibitory effects on canine Escherichia coli (E.coli),provide a reference for the application of plant extracts in the prevention and control of bacterial diseases in dogs. 【Method】 Fifteen plant extracts including shikimic acid,rubusoside,chlorogenic acid,etc.,were selected.The antibacterial activity of these extracts against E.coli was determined using the hole-punching method to identify those with significant antibacterial effects.The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of plant extracts to canine E.coli were measured using the double dilution method.The MIC dose of each plant extract was co-cultured with E.coli in a constant temperature shaker.Samples were taken periodically,centrifuged,and supernatants were collected.The absorbance at D_{600 nm},D_{260 nm} and D_{280 nm} values of the supernatant was used to assess the impact of different plant extracts on the growth curve and extracellular nucleoprotein of E.coli.Meanwhile,the activity of alkaline phosphatase (AKP) in the supernatant and lactate dehydrogenase (LDH) in the bacteria was measured using kits. 【Result】 Shikimic acid,rubusoside,chlorogenic acid,phloretin and salidroside showed good antibacterial effects against E.coli.Based on the size of the inhibition zone diameter,their antibacterial capacities ranked as follows:Shikimic acid＞chlorogenic acid＞rubusoside＞phloretin＞salidroside.The MICs of phloretin,shikimic acid,rubusoside,chlorogenic acid and salidroside to canine E.coli were 0.48,15.625,62.5,31.25 and 125 mg/mL respectively,while their MBCs were 0.48,31.25,125,62.5 and 250 mg/mL,respectively.Growth curve measurements indicated that all plant extracts at MIC doses inhibited the growth of E.coli.Analysis of the absorbance at D_{260 nm} and D_{280 nm} values revealed that chlorogenic acid,salidroside,rubusoside and phloretin significantly damaged the cell membrane of E.coli.AKP activity in the supernatant under the influence of rubusoside and salidroside was significantly higher than that of control group (P<0.05),indicating that they could destroy the cell wall of E.coli and exert antibacterial effects.LDH activity in bacteria treated with shikimic acid,rubusoside,chlorogenic acid,phloretin and salidroside was significantly lower than that of control group (P<0.05),suggesting that all five plant extracts could inhibit intracellular LDH activity and affect bacterial metabolism. 【Conclusion】 Shikimic acid,rubusoside,chlorogenic acid,phloretin and salidroside had significant inhibitory effects on canine E.coli.

Key words: plant extract; canine Escherichia coli; in vitro antibacterial activity

中图分类号:

S852.65^＋1

李梓豪, 郭兴扬, 卢军霞, 位治国, 丁家波, 秦彤, 李睿文. 植物提取物对犬源大肠杆菌的体外抑菌效果及作用机制[J]. 中国畜牧兽医, 2025, 52(8): 3888-3895.

LI Zihao, GUO Xingyang, LU Junxia, WEI Zhiguo, DING Jiabo, QIN Tong, LI Ruiwen. Antibacterial Effect and Mechanism of Plant Extracts on Canine Escherichia coli in vitro[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3888-3895.

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