海洋环境微生物的分离和抑菌活性菌株的筛选

doi:10.14012/j.cnki.fjsc.2023.01.007

摘要/Abstract

摘要：

海洋菌株,因其复杂的生存环境而蕴含着许多未知的活性物质,是探索水产养殖上的应用拮抗菌的良好资源。本研究通过分析从海洋生物与环境中分离得到的海洋微生物提取物对动物致病菌的抑菌谱,筛选获得具有拮抗活性的海洋菌株。以3株鳗鲡爱德华氏菌(Edwardsiella anguillarum)、3株迟缓爱德华氏菌(Edwardsiella tarda)、4株嗜水气单胞菌(Aeromonas hydrophila)、2株维氏气单胞菌(Aeromonas veronii)、8株副溶血弧菌(Vibrio parahemolyticus)、1株溶藻弧菌(Vibrio alginolyticus)、1株哈维氏弧菌(Vibrio harveyi)、3株大肠杆菌(Escherichia coli)、2株鸡白痢沙门氏菌(Salmonella pullorum)等共9种27株作为指示病原菌,采用滤纸片扩散法,测定74株海洋菌株对病原菌的抑菌谱。研究发现,在74株海洋菌株中,对爱德华氏菌有抑制作用的有14株;对气单胞菌有抑制作用的有19株;对副溶血弧菌有抑制作用的有14株;对溶藻弧菌有抑制作用的有12株;对大肠杆菌有抑制作用的有27株;对沙门氏菌有抑制作用的有24株;没有抑制哈维氏弧菌的菌株。其中,芽孢杆菌N-14对维氏气单胞菌E-12(MIC为10 240 μg/mL)、大肠杆菌和鸡白痢沙门氏菌均具有一定的抑制作用。另外,研究结果还发现共有9种海洋真菌属对除哈维氏弧菌外的其余8种指示病原菌均具有明显的抑制活性。

关键词: 海洋菌株, 致病菌, 抑菌谱, 拮抗菌

Abstract:

Marine microorganisms contain many unknown active substances due to their complex living environment, which is a good resource to explore the application of antagonism in aquaculture. In this study, the antimicrobial spectrum of marine microbial extracts isolated from marine organisms and environment against animal pathogenic bacteria was analyzed, and the strains with antagonistic activity were screened. 27 strains of 9 species including 3 strains of Edwardsiella anguillarum, 3 strains of E.tarda, 4 strains of Aeromonas hydrophila, 2 strains of A.veronii, 8 strains of Vibrio parahemolyticus, 1 strain of V.alginolyticus, 1 strains of V.harveyi, 3 strains of Escherichia coli, 2 strains of Salmonella pullorum were used as indicator pathogens. The antimicrobial spectrum of 74 marine strains was determined by filter paper diffusion method. It was found that among the 74 marine strains, 14 strains had inhibition on Edwardsiella, 19 strains of had inhibition on the Aeromonas, 14 strains had inhibitory effect on V.parahemolyticus, 12 strains had inhibition on V.alginolyticus, 27 strains had inhibitory effect on E.coli, 24 strains had inhibition on Salmonella, but no strain could inhibit the V.harveyi strains. Among them, N-14 (Bacillus) had certain inhibitory effects on A.veronii E-12 (MIC of 10 240 μg/mL), E.coli and S.pullorum. In addition, a total of 9 marine fungi genera were found to have significant inhibitory activities against the other 8 indicator pathogens except V.harveyi.

Key words: marine strains, pathogenic bacteria, antimicrobial spectrum, antagonistic strains

中图分类号:

Q939

程茜彤, 靳婷, 马楚雯, 陈永轩. 海洋环境微生物的分离和抑菌活性菌株的筛选[J]. 渔业研究, 2023, 45(1): 46-53.

CHENG Xitong, JIN Ting, MA Chuwen, CHEN Yunghusan. Isolation of marine microorganisms and screening of antimicrobial strains[J]. Journal of Fisheries Research, 2023, 45(1): 46-53.

图/表 4

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编号 Number	菌属 Genus	A-01	A-02	A-03	A-11	A-12	A-13	B-01	B-02	B-03	B-04	B-05	B-06	B-07	B-08	B-11	B-21	C-01	C-02	C-03	D-01	D-02	E-01	E-02	E-03	E-04	E-11	E-12
N-1	Cystobasidium	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	-	-	+	+	-	+	+	-	+
N-2	Hortaea	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	-	-	-	-	-	-	-	-	-
N-3	Aspergillus	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	++	-	+++	+++	+++	+++	+++	+++	+	+++	+++	+++	+++
N-4	Ochroconis	+++	+++	++	++	++	+	+	++	++	++	++	++	++	++	++	-	++	++	+	+	+++	++	-	+	++	+	+++
N-5	Roussoella	+++	+++	++	++	++	++	++	++	++	++	++	++	++	++	+++	-	++	++	++	++	+++	+++	+	++	+++	++	+++
N-6	Alfaria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	-	+	+	-	+	-	-	+
N-7	Alfaria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	++	++	+	+	++	++	-	+	+	-	+
N-8	Emericellopsis	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	+	++	+	-	-	+	-	+
N-9	Acremoyces	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	-	-	-	-	-	-	-	-	-
N-10	Hortaea	+++	+++	+++	+++	+++	+++	+++	++	+++	+++	+++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	+	++	+++	++	+++
N-11	Streptomyces	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	-	-	-	-	-	-	-	-
N-12	Emericellopsis	+++	+++	++	+++	+++	+++	+++	++	++	++	++	+++	+++	+++	++	-	+++	+++	+++	+++	+++	++	-	++	++	-	+++
N-13	Pseudocosmospora	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	-	++	+++	+	+++
N-14	Bacillus	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	++	++	++	++	++	++	-	+	++	-	++
N-15	Beauveria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	++	++	+	++	+	+	-	+	+	-	+
N-16	Alfaria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	-	-	-	-	-	-	-	-
N-17	Streptomyces	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	++	++	-	++	-	-	-	-	-	-
N-18	Acremoyces	-	-	-	-	-	-	-	+	+	+	-	+	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-
N-19	Penicillium	-	-	-	-	-	-	-	-	-	+	-	-	-	-	-	-	+	++	+	-	++	-	-	-	-	-	-
N-20	Emericellopsis	+++	+++	+++	+++	+++	+++	+++	++	++	++	++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	-	++	++	-	+++
N-21	Cystobasidium	+++	+++	+++	+++	+++	+++	+++	++	++	++	+++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	-	++	++	-	+++
N-22	Cystobasidium	+++	+++	+++	+++	+++	+++	+	++	++	++	++	++	+++	++	+++	-	+++	+++	+++	+++	+++	+++	-	++	+++	-	+++
N-23	Phaeophieospora	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	+	+	+	-	-	+	-	+
N-24	Cystobasidium	-	-	-	-	-	-	-	-	+	+	-	++	-	-	+	-	+	++	++	+	+	++	-	+	+	-	+
N-25	Acremonium	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	++	++	+	++	+	-	+	+	-	+
N-26	Hortaea	++	-	-	-	-	-	-	+	+	+	-	-	-	+	-	-	++	++	++	+	++	++	-	+	++	-	+
N-27	Trichosporon	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	-	-	-	-	-	-	-	-
N-28	Trichosporon	-	-	-	-	-	-	-	+	+	+	-	-	-	-	+	-	-	++	+	-	-	-	-	-	-	-	-

编号 Number	菌属 Genus	A-01	A-02	A-03	A-11	A-12	A-13	B-01	B-02	B-03	B-04	B-05	B-06	B-07	B-08	B-11	B-21	C-01	C-02	C-03	D-01	D-02	E-01	E-02	E-03	E-04	E-11	E-12
N-1	Cystobasidium	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	-	-	+	+	-	+	+	-	+
N-2	Hortaea	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	-	-	-	-	-	-	-	-	-
N-3	Aspergillus	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	++	-	+++	+++	+++	+++	+++	+++	+	+++	+++	+++	+++
N-4	Ochroconis	+++	+++	++	++	++	+	+	++	++	++	++	++	++	++	++	-	++	++	+	+	+++	++	-	+	++	+	+++
N-5	Roussoella	+++	+++	++	++	++	++	++	++	++	++	++	++	++	++	+++	-	++	++	++	++	+++	+++	+	++	+++	++	+++
N-6	Alfaria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	-	+	+	-	+	-	-	+
N-7	Alfaria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	++	++	+	+	++	++	-	+	+	-	+
N-8	Emericellopsis	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	+	++	+	-	-	+	-	+
N-9	Acremoyces	+	+	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	-	-	-	-	-	-	-	-	-
N-10	Hortaea	+++	+++	+++	+++	+++	+++	+++	++	+++	+++	+++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	+	++	+++	++	+++
N-11	Streptomyces	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	-	-	-	-	-	-	-	-
N-12	Emericellopsis	+++	+++	++	+++	+++	+++	+++	++	++	++	++	+++	+++	+++	++	-	+++	+++	+++	+++	+++	++	-	++	++	-	+++
N-13	Pseudocosmospora	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	-	++	+++	+	+++
N-14	Bacillus	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	++	++	++	++	++	++	-	+	++	-	++
N-15	Beauveria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	++	++	+	++	+	+	-	+	+	-	+
N-16	Alfaria	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	-	-	-	-	-	-	-	-
N-17	Streptomyces	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	++	++	-	++	-	-	-	-	-	-
N-18	Acremoyces	-	-	-	-	-	-	-	+	+	+	-	+	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-
N-19	Penicillium	-	-	-	-	-	-	-	-	-	+	-	-	-	-	-	-	+	++	+	-	++	-	-	-	-	-	-
N-20	Emericellopsis	+++	+++	+++	+++	+++	+++	+++	++	++	++	++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	-	++	++	-	+++
N-21	Cystobasidium	+++	+++	+++	+++	+++	+++	+++	++	++	++	+++	+++	+++	+++	+++	-	+++	+++	+++	+++	+++	+++	-	++	++	-	+++
N-22	Cystobasidium	+++	+++	+++	+++	+++	+++	+	++	++	++	++	++	+++	++	+++	-	+++	+++	+++	+++	+++	+++	-	++	+++	-	+++
N-23	Phaeophieospora	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	+	+	+	+	-	-	+	-	+
N-24	Cystobasidium	-	-	-	-	-	-	-	-	+	+	-	++	-	-	+	-	+	++	++	+	+	++	-	+	+	-	+
N-25	Acremonium	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	++	++	+	++	+	-	+	+	-	+
N-26	Hortaea	++	-	-	-	-	-	-	+	+	+	-	-	-	+	-	-	++	++	++	+	++	++	-	+	++	-	+
N-27	Trichosporon	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	+	+	-	-	-	-	-	-	-	-
N-28	Trichosporon	-	-	-	-	-	-	-	+	+	+	-	-	-	-	+	-	-	++	+	-	-	-	-	-	-	-	-