25种中草药及其联合用药配伍对5种水产养殖常见致病菌的抑菌作用
|
摘要
为探究25种中草药及其联合用药配伍对水产养殖中常见5种细菌致病菌的抑制作用,采用琼脂扩散法(纸片法)测定了夏枯草、石榴皮、黄连、白头翁、乌梅等25种中草药对嗜水气单胞菌Aeromonas hydrophila、副溶血弧菌Vibrio parahemolyticus、蜡样芽孢杆菌Bacillus cereus、荧光假单胞菌Pseudomonas fluorescens和弗氏柠檬酸杆菌Citrobacter freundii的抑制作用,用二倍稀释法测定了抑菌效果较强的4种中草药对5种致病菌的最小抑菌(MIC)和最小杀菌浓度(MBC),采用正交试验法对这5种致病菌具有较强抑制作用的中草药配伍进行了研究。结果表明:25种中草药对这5种致病菌均有抑制作用,乌梅、石榴皮、黄连、白头翁的抑菌作用最强,其抑菌圈直径达10～30 mm,MIC<62.5 mg/mL,MBC<125 mg/mL,其中对嗜水气单胞菌、荧光假单胞菌、弗氏柠檬酸杆菌抑菌作用最明显的是乌梅,MIC均为15.63 mg/mL,对副溶血弧菌抑菌作用最明显的是黄连,MIC为15.63 mg/mL,对蜡样芽孢杆菌抑菌作用最明显的是乌梅,MIC为7.81 mg/mL;正交试验结果表明,对嗜水气单胞菌、副溶血弧菌具有最佳抑菌作用的组合为乌梅+石榴皮,对蜡样芽孢杆菌具有最佳抑菌作用的组合为乌梅+白头翁、石榴皮+白头翁,对荧光假单胞菌具有最佳抑菌作用的组合为乌梅+白头翁,对弗氏柠檬酸杆菌具有最佳抑菌作用的组合为乌梅+石榴皮、乌梅+白头翁。研究表明,针对水产养殖中常见的5种致病菌,乌梅+石榴皮、乌梅+白头翁为最佳抑菌中药配伍组合,中草药配伍效果好于单一用药。
Antimicrobial effects of 25 kinds of Chinese herbal medicines on Aeromonas hydrophila, Vibrio parahemolyticus, Bacillus cereus, Pseudomonas fluorescens and Citrobacter freundii were examined by means of the disc diffusion method, and the minimal inhibitory concentration(MIC) and minimal bactericidal concentration(MBC) of four kinds of Chinese herbal medicines with significantly antibacterial effect against the five pathogenic bacteria were determined with the double dilution method. Then the combinations of Chinese herbal medicines with better antibacterial effect against the five pathogenic bacteria were optimized through an orthogonal test. All of the 25 kinds of Chinese herbal medicines were found to have to some extent antimicrobial effects, with the best antibacterial role against the five pathogenic bacteria in the four kinds including Prunus mume, Punica granatum, Coptis chinensis Franch and Pulsatilla chinensis, which had diameter of inhibition zone over 10-30 mm, MIC<62.5 mg/mL, and MBC<125 mg/mL. P. mume had the best antimicrobial effect on A.hydrophila, P.fluorescens and C.freundii, with MIC of 15.63 mg/mL. P.mume and C.chinensis Franch had most effect against V.parahemolyticus, with MIC of 15.63 mg/mL. P.mume was found to have significant antimicrobial effect on B.cereus with MIC of 7.81 mg/mL. The orthogonal tests revealed that the best MIC on A.hydrophila was observed in the combination of P.mume+P.granatum and V.parahemolyticus. P.mume+P.chinensis and P.granatum+P.chinensis were the best combination on B.cereus. For P.fluorescens, P.mume+P.chinensis were the best combinations. P.mume+P.granatum and P.mume+P.chinensis had best antibacterial combination against C.freundii. All the test results indicated that P.mume+P.granatum, and P.mume+P.chinensis were the best herbal medicine combination to fight against the five pathogenic bacteria, and that herbal medicine combination was better than single herbal medicine.
Antimicrobial effects of 25 kinds of Chinese herbal medicines on Aeromonas hydrophila, Vibrio parahemolyticus, Bacillus cereus, Pseudomonas fluorescens and Citrobacter freundii were examined by means of the disc diffusion method, and the minimal inhibitory concentration(MIC) and minimal bactericidal concentration(MBC) of four kinds of Chinese herbal medicines with significantly antibacterial effect against the five pathogenic bacteria were determined with the double dilution method. Then the combinations of Chinese herbal medicines with better antibacterial effect against the five pathogenic bacteria were optimized through an orthogonal test. All of the 25 kinds of Chinese herbal medicines were found to have to some extent antimicrobial effects, with the best antibacterial role against the five pathogenic bacteria in the four kinds including Prunus mume, Punica granatum, Coptis chinensis Franch and Pulsatilla chinensis, which had diameter of inhibition zone over 10-30 mm, MIC<62.5 mg/mL, and MBC<125 mg/mL. P. mume had the best antimicrobial effect on A.hydrophila, P.fluorescens and C.freundii, with MIC of 15.63 mg/mL. P.mume and C.chinensis Franch had most effect against V.parahemolyticus, with MIC of 15.63 mg/mL. P.mume was found to have significant antimicrobial effect on B.cereus with MIC of 7.81 mg/mL. The orthogonal tests revealed that the best MIC on A.hydrophila was observed in the combination of P.mume+P.granatum and V.parahemolyticus. P.mume+P.chinensis and P.granatum+P.chinensis were the best combination on B.cereus. For P.fluorescens, P.mume+P.chinensis were the best combinations. P.mume+P.granatum and P.mume+P.chinensis had best antibacterial combination against C.freundii. All the test results indicated that P.mume+P.granatum, and P.mume+P.chinensis were the best herbal medicine combination to fight against the five pathogenic bacteria, and that herbal medicine combination was better than single herbal medicine.
引文
[1] Zhang Dunhua,Xu Dehai,Shoemaker C.Experimental induction of motile Aeromonas septicemia in channel catfish (Ictalurus punctatus) by waterborne challenge with virulent Aeromonas hydrophila[J].Aquaculture Reports,2016,3(C):18-23.
[2] 卢静,王振宁,陈锐,等.几种中药单体和抗生素对嗜水气单胞菌及温和气单胞菌的体外抑菌活性研究[J].水生生物学报,2013,37(6):1128-1132.
[3] 张金金,扈庆华,李连青.副溶血弧菌的致病性及检测方法[J].分子诊断与治疗杂志,2014,6(4):274-278.
[4] 刘芳,罗臻,黄静敏,等.致病性蜡样芽孢杆菌的研究进展[J].检验检疫学刊,2016,26(1):68-71.
[5] 唐金凤,李莉,左国营.铜绿假单胞菌致病机理及中药抗感染的研究现状[J].中华中医药学刊,2016,34(5):1108-1111.
[6] 薛巧,赵战勤,刘会胜,等.弗氏柠檬酸杆菌对动物和人致病性研究进展[J].动物医学进展,2015,36(7):81-85.
[7] Shakir Z,Khan S,Sung K,et al.Molecular characterization of fluoroquinolone-resistant Aeromonas spp.isolated from imported shrimp[J].Applied and Environmental Microbiology,2012,78(22):8137-8141.
[8] Choi W M,Mo W Y,Wu S C,et al.Effects of traditional Chinese medicines (TCM) on the immune response of grass carp (Ctenopharyngodon idellus)[J].Aquaculture International,2014,22(2):361-377.
[9] 李国,闺茂仓,常维山,等.我国海水养殖贝类弧菌病研究进展[J].浙江海洋学院学报:自然科学版,2008,27(3):327-334.
[10] 杨移斌,余琳雪,刘永涛,等.罗非鱼源蜡样芽孢杆菌分离、鉴定及药敏特性研究[J].淡水渔业,2017,47(4):51-56.
[11] Fu Bu,Wu Qiaolian,Dang Minyan,et al.Inhibition of Pseudomonas aeruginosa biofilm formation by traditional Chinese medicinal Herb Herba patriniae[J].Bio Med Research International,2017, doi: 10.1155/2017/9584703.
[12] 吴秋云,黄琳,皮真,等.中草药抑菌作用及其机制研究进展[J].中兽医医药杂志,2018,37(1):25-29.
[13] 张传亮,李槿年,汪兴生,等.水产动物常见病原菌的耐药性分析及中草药体外抑菌活性的检测[J].中国微生态学杂志,2007,19(2):152-155,157.
[14] 温晓鹿,杨雪芬,胡友军,等.猪饲用抗生素替代技术的研究进展[C]//第十二届(2014)中国猪业发展大会论文集.成都:中国畜牧业协会,2014:106-109.
[15] 金建玲,徐轲,高培基.中西药联合使用在应对细菌抗药性方面的优势[J].中医药学报,2011,39(4):95-98.
[16] 彭金菊,马驿,梁淑鋆,等.抗菌中药及复方对嗜水气单胞菌的体外抑制效果[J].安徽农业科学,2009,37(28):13623-13625.
[17] 曹良,李英伦.斑点叉尾鮰“腹水症”病原的体外中药药敏实验[J].水生态学杂志,2009,2(1):95-97.
[18] 肖辉,苏振霞,单娟娟,等.16种中草药提取物对嗜水气单胞菌的体外抑菌实验[J].水生态学杂志,2009,2(3):53-56.
[19] 罗新,张其中.42种中草药对嗜水气单胞菌的体外抑菌试验[J].淡水渔业,2011,41(3):61-65.
[20] 帅丽华,姜登钊,刘怀,等.多重耐药大肠埃希菌的黄连-左氧氟沙星联合用药体外活性研究[J].中国医院药学杂志,2017,37(5):1-4.
[21] 黄宇星,刘二伟.联合用药的药物相互作用及研究方法[J].药物评价研究,2014,37(3):276-279.
[22] 邓家刚,覃骊兰.试论中药配伍中的中医“和谐”观[J].时珍国医国药,2009,20(2):503-504.
[23] 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.
[24] 陈霞,张其中,李春涛.47种中草药体外抑杀嗜水气单胞菌的药效研究[J].水产科学,2012,31(7):387-391.
[25] 李春涛,陈霞,张其中,等.100种中草药体外抑杀嗜水气单胞菌的药效研究[J].淡水渔业,2012,42(2):27-34.
[26] 聂鲡蓉,郑曙明,仇登高.12种中药有效成分对3株水产致病菌体外抑菌试验[J].水生态学杂志,2010,31(6):141-144.
[27] 俞树荣.微生物学和微生物学检验[M].2版.北京:人民卫生出版社,1997:452-458.
[28] 张海宾,杨桂芳.12种中草药对嗜水气单胞菌杀伤能力的研究[J].水产科学,2006,25(1):16-18.
[29] 李茜,张懿瑾,华汝泉,等.23种中草药及复方对鲫肠道3种细菌的体外抑菌试验[J].淡水渔业,2007,37(4):7-11.
[30] 曹红峰,宋靖芳,李国庆,等.中草药对嗜水气单胞菌ST-3-3抑菌作用的研究[J].中医药导报,2007,13(5):86-88.
[31] 田甜.鱼源嗜水气单胞菌的分离鉴定及耐药性研究[D].武汉:华中农业大学,2010.
[32] 韦嫔,谭艾娟,黄弘宇,等.五倍子降低大肠杆菌耐药性的效果[J].贵州农业科学,2015,43(7):137-139,143.
[33] 赵建荣,李晓玫.中药复方物质基础研究的现状与进展[J].中草药,2003,34(11):963-966,1010.
[34] 白峰,王文魁,王广彬,等.22味中草药对3种奶牛乳腺炎病原菌的体外抑菌试验[J].畜牧兽医杂志,2009,28(5):5-7.
[35] 陶健,李绍戊,刘红柏.21种中草药及复方制剂对嗜水气单胞菌的体外抑菌作用[J].水生态学杂志,2013,34(3):90-93.
[36] 王荻,刘红柏,罗添允.中草药与益生菌在水产养殖中协同作用的研究进展[J].大连海洋大学学报,2016,31(5):583-588.
[37] Akbary P.In vitro inhibitory activity of the leaf methanol extract of green tea (Camellia sinensis) against Lactococcus garvieae and Aeromonas hydrophila isolated of rainbow trout (Oncorhynchus mykiss)[J].Advances in Microbiology,2014,4(12):829-834.
[38] 王广军,谢骏,胡隐昌,等.中草药添加剂对日本鳗鲡生长和非特异性免疫效应的研究[J].淡水渔业,2008,38(6):38-41.
[39] 路晶晶,郭冉,齐国山,等.复方中草药对大菱鲆幼鱼生长性能及非特异性免疫指标的影响[J].大连海洋大学学报,2018,33(6):722-728.
[2] 卢静,王振宁,陈锐,等.几种中药单体和抗生素对嗜水气单胞菌及温和气单胞菌的体外抑菌活性研究[J].水生生物学报,2013,37(6):1128-1132.
[3] 张金金,扈庆华,李连青.副溶血弧菌的致病性及检测方法[J].分子诊断与治疗杂志,2014,6(4):274-278.
[4] 刘芳,罗臻,黄静敏,等.致病性蜡样芽孢杆菌的研究进展[J].检验检疫学刊,2016,26(1):68-71.
[5] 唐金凤,李莉,左国营.铜绿假单胞菌致病机理及中药抗感染的研究现状[J].中华中医药学刊,2016,34(5):1108-1111.
[6] 薛巧,赵战勤,刘会胜,等.弗氏柠檬酸杆菌对动物和人致病性研究进展[J].动物医学进展,2015,36(7):81-85.
[7] Shakir Z,Khan S,Sung K,et al.Molecular characterization of fluoroquinolone-resistant Aeromonas spp.isolated from imported shrimp[J].Applied and Environmental Microbiology,2012,78(22):8137-8141.
[8] Choi W M,Mo W Y,Wu S C,et al.Effects of traditional Chinese medicines (TCM) on the immune response of grass carp (Ctenopharyngodon idellus)[J].Aquaculture International,2014,22(2):361-377.
[9] 李国,闺茂仓,常维山,等.我国海水养殖贝类弧菌病研究进展[J].浙江海洋学院学报:自然科学版,2008,27(3):327-334.
[10] 杨移斌,余琳雪,刘永涛,等.罗非鱼源蜡样芽孢杆菌分离、鉴定及药敏特性研究[J].淡水渔业,2017,47(4):51-56.
[11] Fu Bu,Wu Qiaolian,Dang Minyan,et al.Inhibition of Pseudomonas aeruginosa biofilm formation by traditional Chinese medicinal Herb Herba patriniae[J].Bio Med Research International,2017, doi: 10.1155/2017/9584703.
[12] 吴秋云,黄琳,皮真,等.中草药抑菌作用及其机制研究进展[J].中兽医医药杂志,2018,37(1):25-29.
[13] 张传亮,李槿年,汪兴生,等.水产动物常见病原菌的耐药性分析及中草药体外抑菌活性的检测[J].中国微生态学杂志,2007,19(2):152-155,157.
[14] 温晓鹿,杨雪芬,胡友军,等.猪饲用抗生素替代技术的研究进展[C]//第十二届(2014)中国猪业发展大会论文集.成都:中国畜牧业协会,2014:106-109.
[15] 金建玲,徐轲,高培基.中西药联合使用在应对细菌抗药性方面的优势[J].中医药学报,2011,39(4):95-98.
[16] 彭金菊,马驿,梁淑鋆,等.抗菌中药及复方对嗜水气单胞菌的体外抑制效果[J].安徽农业科学,2009,37(28):13623-13625.
[17] 曹良,李英伦.斑点叉尾鮰“腹水症”病原的体外中药药敏实验[J].水生态学杂志,2009,2(1):95-97.
[18] 肖辉,苏振霞,单娟娟,等.16种中草药提取物对嗜水气单胞菌的体外抑菌实验[J].水生态学杂志,2009,2(3):53-56.
[19] 罗新,张其中.42种中草药对嗜水气单胞菌的体外抑菌试验[J].淡水渔业,2011,41(3):61-65.
[20] 帅丽华,姜登钊,刘怀,等.多重耐药大肠埃希菌的黄连-左氧氟沙星联合用药体外活性研究[J].中国医院药学杂志,2017,37(5):1-4.
[21] 黄宇星,刘二伟.联合用药的药物相互作用及研究方法[J].药物评价研究,2014,37(3):276-279.
[22] 邓家刚,覃骊兰.试论中药配伍中的中医“和谐”观[J].时珍国医国药,2009,20(2):503-504.
[23] 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.
[24] 陈霞,张其中,李春涛.47种中草药体外抑杀嗜水气单胞菌的药效研究[J].水产科学,2012,31(7):387-391.
[25] 李春涛,陈霞,张其中,等.100种中草药体外抑杀嗜水气单胞菌的药效研究[J].淡水渔业,2012,42(2):27-34.
[26] 聂鲡蓉,郑曙明,仇登高.12种中药有效成分对3株水产致病菌体外抑菌试验[J].水生态学杂志,2010,31(6):141-144.
[27] 俞树荣.微生物学和微生物学检验[M].2版.北京:人民卫生出版社,1997:452-458.
[28] 张海宾,杨桂芳.12种中草药对嗜水气单胞菌杀伤能力的研究[J].水产科学,2006,25(1):16-18.
[29] 李茜,张懿瑾,华汝泉,等.23种中草药及复方对鲫肠道3种细菌的体外抑菌试验[J].淡水渔业,2007,37(4):7-11.
[30] 曹红峰,宋靖芳,李国庆,等.中草药对嗜水气单胞菌ST-3-3抑菌作用的研究[J].中医药导报,2007,13(5):86-88.
[31] 田甜.鱼源嗜水气单胞菌的分离鉴定及耐药性研究[D].武汉:华中农业大学,2010.
[32] 韦嫔,谭艾娟,黄弘宇,等.五倍子降低大肠杆菌耐药性的效果[J].贵州农业科学,2015,43(7):137-139,143.
[33] 赵建荣,李晓玫.中药复方物质基础研究的现状与进展[J].中草药,2003,34(11):963-966,1010.
[34] 白峰,王文魁,王广彬,等.22味中草药对3种奶牛乳腺炎病原菌的体外抑菌试验[J].畜牧兽医杂志,2009,28(5):5-7.
[35] 陶健,李绍戊,刘红柏.21种中草药及复方制剂对嗜水气单胞菌的体外抑菌作用[J].水生态学杂志,2013,34(3):90-93.
[36] 王荻,刘红柏,罗添允.中草药与益生菌在水产养殖中协同作用的研究进展[J].大连海洋大学学报,2016,31(5):583-588.
[37] Akbary P.In vitro inhibitory activity of the leaf methanol extract of green tea (Camellia sinensis) against Lactococcus garvieae and Aeromonas hydrophila isolated of rainbow trout (Oncorhynchus mykiss)[J].Advances in Microbiology,2014,4(12):829-834.
[38] 王广军,谢骏,胡隐昌,等.中草药添加剂对日本鳗鲡生长和非特异性免疫效应的研究[J].淡水渔业,2008,38(6):38-41.
[39] 路晶晶,郭冉,齐国山,等.复方中草药对大菱鲆幼鱼生长性能及非特异性免疫指标的影响[J].大连海洋大学学报,2018,33(6):722-728.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |