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摘要[目的]观察大黄（Rheum officinale）和公丁香（Flos caryophylli）对哈氏弧菌（Vibrio harveyi）及其生物膜（Biofilm）的体外抑制作用。[方法]采用琼脂扩散法测定大黄和公丁香对哈氏弧菌的体外抑菌作用；采用倍比稀释法测定大黄和公丁香对哈氏弧菌最小抑菌浓度（MIC）和最小杀菌浓度（MBC）；中图分类号S941.43文献标识码A文章编号0517-6611（2014）24-08188-03
The Inhibitory Activity of Flos caryophylli and Rheum officinale on Vibrio harveyi and Its Biofilm
LI Jiaqin， PANG Huanying et al（Fisheries College of Guangdong Ocean University， Zhanjiang， Guangdong 524088； Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals， Zhanjiang， Guangdong 524088； Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education Institutions， Zhanjiang， Guangdong 524088）
Abstract[Objective] To observe the inhibitory effects of Rheum officinale and Flos caryophylli on Vibrio harveyi and its biofilm. [Method] The inhibitory effects of Rheum officinale and Flos caryophylli on Vibrio harveyi were tested by disc agar diffusion， and the MIC and MBC were detected by the double dilution； the effects of Rheum officinale and Flos caryophylli on the formation of Vibrio harveyi biofilm were measured by MTT assay. [Result] Rheum officinale had inhibitory effects with the diameter of inhibition zone （DIZ） of （15.31±0.4） mm， and the MIC and MBC were 7.183 mg/ml； Flos caryophylli had inhibition effects with DIZ of （11.53±0.6） mm， and the MIC and MBC were 15.625 mg/ml， 62.5 mg/ml. Both Rheum officinale and Flos caryophylli exhibited a potent inhibitory activity against Vibrio harveyi biofilm formation （P<0.01）. [Conclusion] Rheum officinale and Flos caryophylli could significantly inhibit Vibrio harveyi and its biofilm.
Key wordsRheum officinalekl； Flos caryophylli； Vibrio harveyi； Biofilm； In vitro inhibitory activity
哈氏弧菌（Vibrio harveyi）隸属弧菌科（Vibrionaceae）弧菌属（Vibrio），是一种革兰氏阴性、发光海洋细菌，广泛分布于海洋环境中[1]，是危害水产养殖动物的重要致病菌之一。该菌能够感染中国对虾（Penaeus chinensis）[2]、文蛤（Meretrix meretrix ）[3]和大黄鱼（Pseudosciaena crocea）[4]等多种海水养殖动物，给水产养殖业造成巨大的经济损失。哈氏弧菌在水体中主要以生物膜（Biofilm）的形式生存。生物膜是细菌聚集粘结在一起分泌的黏性物质，其主要成分为多糖、核酸、脂类和蛋白等[5]。生物膜能保护细菌对抗外界危险，增强其对抗菌药物的耐受能力[6]。目前，水产养殖中主要采用抗生素等化学药物防治弧菌病，但抗生素的长期使用会使病原产生耐药性，会严重破坏养殖水域的生态环境，并且对人体健康构成潜在的威胁[7]。因此，寻找安全有效的抗菌药物是目前研究的热点之一。
中草药是我国医学宝库中最珍贵的财富之一，含有很多抗菌活性强、毒性低的有效成分。大黄和公丁香是常用的中草药，它们在防治水产动物疾病中有着广泛的应用。曹红峰等[8]证实大黄对致病性弧菌、鱼害粘球菌（Myxococcuspiscolasp.nov.）和假单胞菌（Pseudomonadaceae）等常见水产动物的病原菌有明显的抑制作用。王玉娥等[9]证实哈氏弧菌对大黄和公丁香具有较高的敏感性。但是，关于这2种中药对哈氏弧菌生物膜生成的抑制作用尚未见报道。笔者通过测定大黄和公丁香对哈氏弧菌及其生物膜的体外抑制作用，探索这2种中药的抑菌作用和抗生物膜功能，以期为今后应用中药防治由该菌引起的弧菌病提供理论依据。
The Inhibitory Activity of Flos caryophylli and Rheum officinale on Vibrio harveyi and Its Biofilm
LI Jiaqin， PANG Huanying et al（Fisheries College of Guangdong Ocean University， Zhanjiang， Guangdong 524088； Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals， Zhanjiang， Guangdong 524088； Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education Institutions， Zhanjiang， Guangdong 524088）
Abstract[Objective] To observe the inhibitory effects of Rheum officinale and Flos caryophylli on Vibrio harveyi and its biofilm. [Method] The inhibitory effects of Rheum officinale and Flos caryophylli on Vibrio harveyi were tested by disc agar diffusion， and the MIC and MBC were detected by the double dilution； the effects of Rheum officinale and Flos caryophylli on the formation of Vibrio harveyi biofilm were measured by MTT assay. [Result] Rheum officinale had inhibitory effects with the diameter of inhibition zone （DIZ） of （15.31±0.4） mm， and the MIC and MBC were 7.183 mg/ml； Flos caryophylli had inhibition effects with DIZ of （11.53±0.6） mm， and the MIC and MBC were 15.625 mg/ml， 62.5 mg/ml. Both Rheum officinale and Flos caryophylli exhibited a potent inhibitory activity against Vibrio harveyi biofilm formation （P<0.01）. [Conclusion] Rheum officinale and Flos caryophylli could significantly inhibit Vibrio harveyi and its biofilm.
Key wordsRheum officinalekl； Flos caryophylli； Vibrio harveyi； Biofilm； In vitro inhibitory activity
哈氏弧菌（Vibrio harveyi）隸属弧菌科（Vibrionaceae）弧菌属（Vibrio），是一种革兰氏阴性、发光海洋细菌，广泛分布于海洋环境中[1]，是危害水产养殖动物的重要致病菌之一。该菌能够感染中国对虾（Penaeus chinensis）[2]、文蛤（Meretrix meretrix ）[3]和大黄鱼（Pseudosciaena crocea）[4]等多种海水养殖动物，给水产养殖业造成巨大的经济损失。哈氏弧菌在水体中主要以生物膜（Biofilm）的形式生存。生物膜是细菌聚集粘结在一起分泌的黏性物质，其主要成分为多糖、核酸、脂类和蛋白等[5]。生物膜能保护细菌对抗外界危险，增强其对抗菌药物的耐受能力[6]。目前，水产养殖中主要采用抗生素等化学药物防治弧菌病，但抗生素的长期使用会使病原产生耐药性，会严重破坏养殖水域的生态环境，并且对人体健康构成潜在的威胁[7]。因此，寻找安全有效的抗菌药物是目前研究的热点之一。
中草药是我国医学宝库中最珍贵的财富之一，含有很多抗菌活性强、毒性低的有效成分。大黄和公丁香是常用的中草药，它们在防治水产动物疾病中有着广泛的应用。曹红峰等[8]证实大黄对致病性弧菌、鱼害粘球菌（Myxococcuspiscolasp.nov.）和假单胞菌（Pseudomonadaceae）等常见水产动物的病原菌有明显的抑制作用。王玉娥等[9]证实哈氏弧菌对大黄和公丁香具有较高的敏感性。但是，关于这2种中药对哈氏弧菌生物膜生成的抑制作用尚未见报道。笔者通过测定大黄和公丁香对哈氏弧菌及其生物膜的体外抑制作用，探索这2种中药的抑菌作用和抗生物膜功能，以期为今后应用中药防治由该菌引起的弧菌病提供理论依据。