黄连水提物对卵形鲳鲹源溶藻弧菌的抑菌作用
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摘要
卵形鲳鲹(Trachinotus ovatus)是一种生长快速且肉质鲜美的经济型海水养殖鱼类,但是近些年为满足人们的日常需求,在不断扩大养殖的同时病菌也逐渐泛滥。溶藻弧菌(Vibrio alginolyticus)是华南沿海地区海水养殖鱼类细菌性鱼病的主要致病菌之一,给水产养殖业带来巨大损失。本研究对黄连(Coptis chinensis Franch)的抗菌作用进行系统研究,目的是为由溶藻弧菌引起的爆发性细菌性鱼病的高效防治提供科学的用药依据。本研究采用二倍稀释法测定黄连水提物对卵形鲳鲹源溶藻弧菌的最低抑菌浓度(MIC)、最低杀菌浓度(MBC)和半数致死量(LD50),并使用BCA蛋白浓度测定试剂盒测定细菌超声破碎上清液中可溶性蛋白含量。结果表明,黄连水提物对溶藻弧菌具有明显的抑制作用,其对溶藻弧菌的MIC、MBC和LD50的值分别为7.800mg/mL、31.250mg/mL和15.625mg/mL。胞内可溶性蛋白含量变化的结果提示:黄连发挥抑菌作用的机制可能是通过损伤细菌的细胞壁导致细菌内容物的释放并引起菌体裂解死亡。黄连具有发展成为一种高效抗水产病害中草药制剂的潜力。
Trachonotus ovatus is an economical marine fish that grows fast and has a delicious taste.However,in recent years,in order to meet people's daily needs,the germs are gradually spreading under the continuous expansion of breeding.Vibrio alginolyticusis one of the main pathogens of bacterial fish disease in marine fish in coastal areas of South China,causing huge losses to the aquaculture industry.In this paper,the antimicrobial effects of Coptis chinensis Franch was systematically studied to provide a scientific basis for the effective prevention and treatment of explosive bacterial fish disease caused by Vibrio alginolyticus.In this study,the minimum inhibitory concentration(MIC),the minimum bactericidal concentration(MBC)and the median lethal dose(LD50)of the aqueous extract of Cc.hinensis against V.alginolyticus fromTo.vatus were determined by two-fold dilution method.After sonic disruption,the soluble protein contents in V.alginolyticus were determined by BCA protein concentration assay kit.The experiment showed that the water extracts of Cc.hinensis had obvious inhibitory effects on V.alginolyticus,and the MIC,MBC and LD_(50) values of V.alginolyticus were 7.800 mg/mL,31.250 mg/mL and 15.625 mg/mL,respectively.The changes of soluble protein contents in supernatants suggest that Cc.hinensis can play the inhibitory role by damaging the bacterial cell wall and causing the release of bacterial contents,which results in the lysis and death of bacteria.Cc.hinensis has the potential to develop a highly effective Chinese herbal preparation against aquatic diseases.
Trachonotus ovatus is an economical marine fish that grows fast and has a delicious taste.However,in recent years,in order to meet people's daily needs,the germs are gradually spreading under the continuous expansion of breeding.Vibrio alginolyticusis one of the main pathogens of bacterial fish disease in marine fish in coastal areas of South China,causing huge losses to the aquaculture industry.In this paper,the antimicrobial effects of Coptis chinensis Franch was systematically studied to provide a scientific basis for the effective prevention and treatment of explosive bacterial fish disease caused by Vibrio alginolyticus.In this study,the minimum inhibitory concentration(MIC),the minimum bactericidal concentration(MBC)and the median lethal dose(LD50)of the aqueous extract of Cc.hinensis against V.alginolyticus fromTo.vatus were determined by two-fold dilution method.After sonic disruption,the soluble protein contents in V.alginolyticus were determined by BCA protein concentration assay kit.The experiment showed that the water extracts of Cc.hinensis had obvious inhibitory effects on V.alginolyticus,and the MIC,MBC and LD_(50) values of V.alginolyticus were 7.800 mg/mL,31.250 mg/mL and 15.625 mg/mL,respectively.The changes of soluble protein contents in supernatants suggest that Cc.hinensis can play the inhibitory role by damaging the bacterial cell wall and causing the release of bacterial contents,which results in the lysis and death of bacteria.Cc.hinensis has the potential to develop a highly effective Chinese herbal preparation against aquatic diseases.
引文
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[14]吴华彰,费鸿君,赵云利,等.萝卜硫素对大肠杆菌抑菌机制的研究[J].四川大学学报(医学版),2012,43(3):386-390.
[15]YU Q,LIU M Z,SU H F,et al.Selection and characterization of ssDNA-aptamers specifically recognized pathogenic Vibrio alginolyticus[J].Journal of Fish Diseases,2019.DOI:10.1111/jfd.12985.[Epub ahead of print].
[16]YU Q,LIU M Z,XIAO H H,et al.The inhibitory activities and antiviral mechanism of Viola philippica aqueous extracts against grouper iridovirus infection in vitro and in vivo[J].Journal of Fish Diseases,2019.DOI:10.1111/jfd.12987.[Epub ahead of print].
[2]余庆,李菲,覃仙玲,等.广西卵形鲳鲹小脑来源细胞系的建立及特征分析[J].广西科学,2018,25(1):74-79.
[3]余庆,李菲,王一兵,等.广西北部湾大宗海水养殖鱼类卵形鲳鲹感染溶藻弧菌及其致病性研究[J].广西科学,2018,25(1):68-73.
[4]陈昌福.我国水产养殖动物病害防治研究的主要成就与当前存在的问题[J].饲料工业,2007,28(10):1-3.
[5]PARK Y H,HWANG S Y,HONG M K,et al.Use of antimicrobial agents in aquaculture[J].Revue Scientifique et Technique,2012,31(1):189-197.
[6]HAI N V.The use of probiotics in aquaculture[J].Journal of Applied Microbiology,2015,119(4):917-935.
[7]CARBONE D,FAGGIO C.Importance of prebiotics in aquaculture as immunostimulants.Effects on immune system of Sparus aurataand Dicentrarchus labrax[J].Fish&Shellfish Immunology,2016,54:172-178.DOI:10.1016/j.fsi.2016.04.011.
[8]SONG S K,BECK B R,KIM D,et al.Prebiotics as immunostimulants in aquaculture:A review[J].Fish&Shellfish Immunology,2014,40(1):40-48.
[9]李鑫.黄连、姜黄连抗菌谱及其物质基础比较研究[D].武汉:湖北中医药大学,2015.
[10]王维,李丹,刘洋,等.黄连解毒汤的体外抑菌试验及拆方研究[J].动物医学进展,2016,37(11):65-69.
[11]王凯,于永翔,张正,等.黄连素对海水病原菌的杀灭效果及药效稳定性[J].水产科学,2019,38(1):67-72.
[12]张文青,龚一富,金思,等.中草药及其配伍对嗜水气单胞菌的抑菌作用[J].渔业科学进展,2012,33(1):114-121.
[13]蒋启欢,叶应旺,胡王,等.草鱼嗜水气单胞菌L1灭活疫苗培养条件的优化[J].生物技术进展,2012,2(3):206-211.
[14]吴华彰,费鸿君,赵云利,等.萝卜硫素对大肠杆菌抑菌机制的研究[J].四川大学学报(医学版),2012,43(3):386-390.
[15]YU Q,LIU M Z,SU H F,et al.Selection and characterization of ssDNA-aptamers specifically recognized pathogenic Vibrio alginolyticus[J].Journal of Fish Diseases,2019.DOI:10.1111/jfd.12985.[Epub ahead of print].
[16]YU Q,LIU M Z,XIAO H H,et al.The inhibitory activities and antiviral mechanism of Viola philippica aqueous extracts against grouper iridovirus infection in vitro and in vivo[J].Journal of Fish Diseases,2019.DOI:10.1111/jfd.12987.[Epub ahead of print].