摘要
为探讨黄连素对主要海水养殖病原菌的抑菌作用,采用二倍稀释法测定黄连素对迟缓爱德华氏菌、哈维氏弧菌等10种病原菌的最小抑菌质量浓度和最小杀菌质量浓度,并以美人鱼发光杆菌美人鱼亚种为指示菌,研究黄连素在不同温度、存储条件和金属离子影响下的药效稳定性。试验结果显示,黄连素对10种海水养殖病原菌均具有较好的抑菌及杀菌作用,最小抑菌质量浓度为64～256mg/L,最小杀菌质量浓度为256～1024mg/L。其中,黄连素对大菱鲆弧菌和溶藻弧菌抑菌作用最明显,最小抑菌质量浓度为64mg/L;对鳗弧菌、大菱鲆弧菌、迟缓爱德华氏菌、溶藻弧菌、鲨鱼弧菌和轮虫弧菌杀菌作用最强,最小杀菌质量浓度为256mg/L。质量浓度为512mg/L的黄连素溶液分别在20、40、60、80、100、121℃处理下作用30min,对美人鱼发光杆菌美人鱼亚种的杀菌率均超过99%,将20、60、100℃处理的黄连素溶液于室温下避光存储35d,对指示菌的杀菌率依然大于99%,显示其良好的药物稳定性。金属离子的影响试验也证实,水环境中的Na+、Mg~(2+)、Ca~(2+)、K~+等离子对黄连素的杀菌效果无显著影响。因此,黄连素适宜作为防治水产动物细菌性疾病的临床药物。
A double dilution method was used to determine minimal inhibitory concentration(MIC)and minimal bactericidal concentration(MBC)of berberine on 10 different pathogenic bacteria including Edwardsiella tarda,Vibrio harveyi,V.carchariae,V.alginolyticus,V.anguillarum,V.scophthalmi,V.rotiferianus,V.parahemolyticus,Pseudoalteromonas nigrifaciens,and Photobacterium damselae ssp.damselae to investigate the bacteriostatic and bactericidal effect of berberine against main pathogens in mariculture in vitro.Meanwhile,drug efficacy stability of berberine under various temperatures,storage conditions and different metal ions was studied using P.damselae ssp.damselae as a strain indicator.The results showed that berberine had both good bacteriostatic and bactericidal effects on all testing strains,with the MIC from 64mg/L to 256mg/L,and the MBC varying from 256mg/L to 1024mg/L.Especially,it had the best inhibitory effects on V.scophthalmi and V.alginolyticus at 64mg/L and the best bactericidal effect on V.anguillarum,V.scophthalmi,E.tarda,V.alginolyticus,V.carchariae and V.rotiferianus,with the MBC of 256mg/L.In the stability test,the berberine had sterilization rate of above 99%on indicator bacteria(P.damselae ssp.damselae)at concentration of 512mg/L at 20℃,40℃,60℃,80℃,100℃ or 121℃for 30 min,even when the berberine was stored at 20℃,60℃and 100℃in photophobic place under room temperature for 35 d.There was no variation in sterilization effect of the berberine solution treated with different metal ions including Na+,Mg~(2+),Ca~(2+),and K~+,indicating that berberine as a Chinese herb monomer was suitable for a clinical drug against bacterial diseases in mariculture.
引文
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