北冰洋来源白色念珠菌拮抗菌株的分离与鉴定
|
摘要
采用涂布平板法、对峙培养法及琼脂扩散法从北冰洋沉积物样品中分离、筛选对白色念珠菌(Canidia albicans)具有良好拮抗作用的菌株,通过形态观察、生理生化试验及分子生物学技术对其进行鉴定,并对其抑菌谱进行研究。结果表明,从北冰洋沉积物样品中共分离纯化出314株菌株,其中,4株菌株对白色念珠菌具有明显拮抗作用,且菌株56#拮抗效果最好,抑菌圈直径为(32.14±0.72)mm。菌株56#被鉴定为枯草芽孢杆菌(Bacillus subtilis),其对大肠杆菌、副溶血性弧菌、铜绿假单胞菌、鼠伤寒沙门氏菌、肺炎克雷伯氏菌、屎肠球菌、金黄色葡萄球菌和鲍曼不动杆菌都有一定的抑制作用,是一株广谱抗菌的海洋微生物。
The strain with good antagonistic effects on Canidia albicans was isolated and screened from the sediment samples in the Arctic Ocean by dilution-plate method, confrontation culture method and agar diffusion method. The strain was identified by morphological observation, physiological and biochemical tests and molecular biological technique, and its antimicrobial spectrum was studied. The results showed that a total of 314 strains were isolated and purified from the sediment samples in the Arctic Ocean. Among them, 4 strains had obvious antagonistic effect on C. albicans, and strain 56 had the optimal antagonistic effect. The diameter of the inhibition zone of the strain 56#was(32.14±0.72) mm. The strain was identified as Bacillus subtilis and was a broad-spectrum antibacterial marine microorganism, which had certain antagonistic effects on Escherichia coli, Vibrio parahaemolyticu, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella pneumoniae, Enterococcus faecium, Staphylococcus aureus and Acinetobacter baumannii.
The strain with good antagonistic effects on Canidia albicans was isolated and screened from the sediment samples in the Arctic Ocean by dilution-plate method, confrontation culture method and agar diffusion method. The strain was identified by morphological observation, physiological and biochemical tests and molecular biological technique, and its antimicrobial spectrum was studied. The results showed that a total of 314 strains were isolated and purified from the sediment samples in the Arctic Ocean. Among them, 4 strains had obvious antagonistic effect on C. albicans, and strain 56 had the optimal antagonistic effect. The diameter of the inhibition zone of the strain 56#was(32.14±0.72) mm. The strain was identified as Bacillus subtilis and was a broad-spectrum antibacterial marine microorganism, which had certain antagonistic effects on Escherichia coli, Vibrio parahaemolyticu, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella pneumoniae, Enterococcus faecium, Staphylococcus aureus and Acinetobacter baumannii.
引文
[1]阎力君,张多,卢文,等.正交试验优化白头翁汤抗白色念珠菌物质的提取工艺[J].黑龙江畜牧兽医,2017(12):137-139.
[2]沈洁,张欣,刘建钗,等.鸡白色念珠菌和烟曲霉双重SYBR GreenⅠ荧光定量PCR检测方法的建立[J].中国兽医科学,2018,48(4):484-487.
[3]房耀维,王淑军,刘姝,等.西沙群岛海域海洋放线菌的分离及其抗菌活性[J].微生物学杂志,2014,34(4):47-52.
[4]BULL A T,STACH,J E M.Marine actinobacteria:new opportunities for natural product search and discovery[J].Trends Microbiol,2007,15:491-499.
[5]CHEN G,WANG H,PEI Y.Secondary metabolites from marine-derived microorganisms[J].J Asian Nat Prod Res,2014,16(1):105-122.
[6]WANG K W,DING P.New bioactive metabolites from the marine-derived fungi Aspergillus[J].Mini Rev Med Chem,2018,18(13):1072-1094.
[7]WU B,WIESE J,LABES A,et al.Lindgomycin,an unusual antibiotic polyketide from a marine fungus of the Lindgomycetaceae[J].Mar Drugs,2015,13(8):4617-4632.
[8]IMHOFF J F.Natural products from marine fungi-still an underrepresented resource[J].Mar Drugs,2016,14(1):19.
[9]王碧霞,吴祺豪,李鑫,等.中国东海及东太平洋海山区来源抗菌活性海洋微生物的筛选研究[J].中国海洋药物,2017,36(5):9-15.
[10]金黎明,金鑫浩,侯熙彦,等.具有抗菌活性的海洋真菌天然产物研究进展[J].中国海洋药物,2017,36(4):66-74.
[11]TARMAN K,LINDEQUIST U,WENDE K,et al.Isolation of a new natural product and cytotoxic and antimicrobial activities of extracts from fungi of Indonesian marine habitats[J].Mar Drugs,2011,9(3):294-306.
[12]GEBREYOHANNES G,MOGES F,SAHILE S,et al.Isolation and characterization of potential antibiotic producing actinomycetes from water and sediments of Lake Tana,Ethiopia[J].Asian Pac J Trop B,2013,3(6):426-435.
[13]王勇勇,蒙春蕾,蔡爽,等.抗白色念珠菌海洋真菌的筛选及抑菌活性初探[J].中国酿造,2015,34(2):55-59.
[14]ZHANG Y,LI X M,WANG C Y,et al.A new naphthoquinoneimine derivative from the marine algal derived endophytic fungus Aspergillus niger EN-13[J].Chin Chem Lett,2007,18(8):951-953.
[15]WEN L,LIN Y C,SHE Z G,et al.Paeciloxanthone,a new cytotoxic xanthone from the marine mangrove fungus Paecilomyces sp.(Tree1-7)[J].J Asian Nat Prod Res,2008,10(2):133-137.
[16]张静楠,聂灿玉,赵胜楠,等.抗白色念珠菌的海洋微生物的分离与鉴定[J].天津农业科学,2016,22(1):79-81
[17]BUCHANAN R E,GIBBONS N E.伯杰细菌鉴定手册(第8版)[M].北京:科学出版社,1984:729-736.
[18]蔡妙英,东秀珠.常见细菌系统鉴定手册[M].北京:科学出版社,2001:349-386.
[19]杨耀刚,郜晋楠,田瑞华.自然发酵玉米秸秆中优良乳酸菌的筛选鉴定及混菌发酵试验[J].黑龙江畜牧兽医,2018(7):145-148.
[20]王硕,孟庆丽,武天娇,等.抗金葡菌深海真菌的分离鉴定及抑菌谱研究[J].生物技术,2015,25(2):169-172.
[2]沈洁,张欣,刘建钗,等.鸡白色念珠菌和烟曲霉双重SYBR GreenⅠ荧光定量PCR检测方法的建立[J].中国兽医科学,2018,48(4):484-487.
[3]房耀维,王淑军,刘姝,等.西沙群岛海域海洋放线菌的分离及其抗菌活性[J].微生物学杂志,2014,34(4):47-52.
[4]BULL A T,STACH,J E M.Marine actinobacteria:new opportunities for natural product search and discovery[J].Trends Microbiol,2007,15:491-499.
[5]CHEN G,WANG H,PEI Y.Secondary metabolites from marine-derived microorganisms[J].J Asian Nat Prod Res,2014,16(1):105-122.
[6]WANG K W,DING P.New bioactive metabolites from the marine-derived fungi Aspergillus[J].Mini Rev Med Chem,2018,18(13):1072-1094.
[7]WU B,WIESE J,LABES A,et al.Lindgomycin,an unusual antibiotic polyketide from a marine fungus of the Lindgomycetaceae[J].Mar Drugs,2015,13(8):4617-4632.
[8]IMHOFF J F.Natural products from marine fungi-still an underrepresented resource[J].Mar Drugs,2016,14(1):19.
[9]王碧霞,吴祺豪,李鑫,等.中国东海及东太平洋海山区来源抗菌活性海洋微生物的筛选研究[J].中国海洋药物,2017,36(5):9-15.
[10]金黎明,金鑫浩,侯熙彦,等.具有抗菌活性的海洋真菌天然产物研究进展[J].中国海洋药物,2017,36(4):66-74.
[11]TARMAN K,LINDEQUIST U,WENDE K,et al.Isolation of a new natural product and cytotoxic and antimicrobial activities of extracts from fungi of Indonesian marine habitats[J].Mar Drugs,2011,9(3):294-306.
[12]GEBREYOHANNES G,MOGES F,SAHILE S,et al.Isolation and characterization of potential antibiotic producing actinomycetes from water and sediments of Lake Tana,Ethiopia[J].Asian Pac J Trop B,2013,3(6):426-435.
[13]王勇勇,蒙春蕾,蔡爽,等.抗白色念珠菌海洋真菌的筛选及抑菌活性初探[J].中国酿造,2015,34(2):55-59.
[14]ZHANG Y,LI X M,WANG C Y,et al.A new naphthoquinoneimine derivative from the marine algal derived endophytic fungus Aspergillus niger EN-13[J].Chin Chem Lett,2007,18(8):951-953.
[15]WEN L,LIN Y C,SHE Z G,et al.Paeciloxanthone,a new cytotoxic xanthone from the marine mangrove fungus Paecilomyces sp.(Tree1-7)[J].J Asian Nat Prod Res,2008,10(2):133-137.
[16]张静楠,聂灿玉,赵胜楠,等.抗白色念珠菌的海洋微生物的分离与鉴定[J].天津农业科学,2016,22(1):79-81
[17]BUCHANAN R E,GIBBONS N E.伯杰细菌鉴定手册(第8版)[M].北京:科学出版社,1984:729-736.
[18]蔡妙英,东秀珠.常见细菌系统鉴定手册[M].北京:科学出版社,2001:349-386.
[19]杨耀刚,郜晋楠,田瑞华.自然发酵玉米秸秆中优良乳酸菌的筛选鉴定及混菌发酵试验[J].黑龙江畜牧兽医,2018(7):145-148.
[20]王硕,孟庆丽,武天娇,等.抗金葡菌深海真菌的分离鉴定及抑菌谱研究[J].生物技术,2015,25(2):169-172.