基于响应面法分析乌梅和抗生素对哈维氏弧菌的协同抑制作用  

Synergism between Fructus mume and antibiotics against Vibrio harveyi based on response surface methodology

作　　者：钟志鸿 王菲[1,2] 陈永贵 邓恒为 王世锋[2] 孙云 陈雪芬[1] 郭伟良[1,2] 周永灿[1,2] ZHONG Zhihong;WANG Fei;CHEN Yonggui;DENG Hengwei;WANG Shifeng;SUN Yun;CHEN Xuefen;GUO Weiliang;ZHOU Yongcan(State Key Laboratory of Marine Resource Utilization in South China Sea,Hainan University,Haikou 570228,China;Hainan Provincial Key Laboratory of Tropical Hydrobiology and Biotechnology,College of Marine Science,Hainan University,Haikou 570228,China;State Key Laboratory of Biocontrol,School of Life Sciences,Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]海南大学南海海洋资源利用国家重点实验室,海南海口570228 [2]海南大学海洋学院,海南省热带水生生物技术重点实验室,海南海口570228 [3]中山大学生命科学学院有害生物控制与资源利用国家重点实验室,广东广州510275

出　　处：《南方水产科学》2018年第6期81-88,共8页South China Fisheries Science

基　　金：国家自然科学基金项目(31560725);海洋公益性行业科研专项经费项目(201405020);海南省自然科学基金创新研究团队项目(2016CXTD005);南海海洋生物健康养殖团队(hdkytg201704)。

摘　　要：为了给哈维氏弧菌(Vibrio harveyi)疾病防治新手段的开发提供基础实验数据,筛选了协同抗哈维氏弧菌的乌梅(Fructus mume)水提液与抗生素复合物。首先绘制药物浓度对数对抑菌圈直径标准曲线,采用全组合法筛选与乌梅水提液具有协同抗菌活性的抗生素,然后采用中心组合设计法(CCD)结合响应面法(RSM)分析乌梅水提液与筛出抗生素复合后两两药物间的交互作用,并确定最佳复合配比。全组合法筛出的四环素、土霉素、金霉素和强力霉素分别与乌梅水提液具有协同抗菌作用,RSM分析显示将这4种药物复合后,乌梅水提液与土霉素对哈维氏弧菌存在显著协同抗菌作用;土霉素分别与金霉素和强力霉素存在交互抑制作用;乌梅水提液、土霉素、金霉素和强力霉素最佳复合配比为45∶0.12∶0.00∶0.00 (mg·mL^(–1))。模型预测最优复合物对哈维氏弧菌的抑菌圈直径为18.54 mm,琼脂扩散法平行3次测得该复合物的抑菌圈直径为(19.03±0.07) mm,相对误差为2.6%,表明该方法结果可靠,为考察药物间的交互作用提供了新思路。To provide basic experimental data for developing protocols to prevent and cure the disease caused by Vibrio harveyi,we collected the compound of Fructus mume water extract and antibiotics,which has synergism of antibacterial activity against V.harveyi.First,we plotted the standard curves of concentration logarithms versus inhibitory zone diameters of the test drugs,and used comprehensive combination experimental design method to screen the antibiotics which had synergism effect with F.mume water extract.Then,we applied central concentration design(CCD)with response surface methodology(RSM)to investigate the interaction among F.mume water extract and antibiotics against V.harveyi and to find their optimum combination.Results show that significant synergism existed between F.mume water extract and tetracycline,oxytetracycline,aureomycin and deoxytetracycline.RSM results indicate that the synergism between F.mume water extract and oxytetracycline contributed to the most significant effect on the antibacterial activity against V.harveyi.However,antagonism effect existed between oxytetracycline and aureomycin,and between oxytetracycline and deoxytetracycline.The optimum ratio of F.mume water extract,oxytetracycline,aureomycin and deoxytetracycline was 45∶0.12∶0.00∶0.00(mg·mL^–1).For this optimum combination,the predictive inhibitory zone diameter was 18.54 mm.According to cup method,agar diffusion method with three replicates,the practical maximum inhibitory zone diameter was(19.03±0.07)mm,and the relative error between predictive values and actual values was 2.6%.Thus,RSM is a reliable approach to describe the relationship among test drugs and inhibitory zone diameter.

关 键 词：哈维氏弧菌 乌梅 抗生素 响应面分析法 协同抗菌 

分 类 号：R931.71[医药卫生—生药学]