为了研究双歧杆菌生物膜的成膜规律及其特性,利用微孔板实验对长双歧杆菌、短双歧杆菌、动物双歧杆菌、青春双歧杆菌、两歧双歧杆菌、假小链双歧杆6个种114株双歧杆菌的成膜能力进行分析。结果显示,基于微孔板吸光值,双歧杆菌分为不成膜(OD600 nm≤1)、弱成膜(1<OD600 nm<3)及强成膜(OD600 nm≥3)3类菌株;其中两歧双歧杆菌所有菌株为强成膜菌株,长双歧杆菌的成膜能力较弱,其中57.90%为非成膜株;而其余双歧杆菌的成膜能力存在菌株差异性,包含不成膜、弱成膜及强成膜3类菌株。进一步通过扫描电子显微镜发现,动物双歧杆菌和两歧双歧杆菌形成蘑菇状生物膜结构,而其余双歧杆菌形成扁平状生物膜;激光共聚焦显微镜显示长双歧杆菌和两歧双歧杆菌生物膜活菌比例较高。利用细菌粘附溶剂法研究双歧杆菌表面性质对其成膜的影响,结果发现短双歧杆菌、假小链双歧杆菌的成膜能力只与其路易斯酸特性有关,动物双歧杆菌和青春双歧杆菌表面疏水性和路易斯酸特性与其成膜能力呈正相关,长双歧杆菌表面性质与其成膜无相关性,而表现为高疏水性和路易斯酸特性的强成膜性两歧双歧杆菌,其成膜能力也与表面性质无关。综上,双歧杆菌生物膜成膜特性存在菌种和菌株特异性,并与菌株表面性质相关。
To explore the regularity and characteristics of biofilm formation of Bifidobacterium, the biofilm-forming ability of 114 strains distributing in six species of Bifidobacterium, including B. longum, B. breve, B. animalis, B. bifidum, B. adolescentis, and B. pseudocatenulatum, was analyzed by microplate assay. The results showed that according to the absorbance, Bifidobacterium was divided into three types: non-biofilm formers (OD600 nm≤1), weak biofilm formers (1<OD600 nm<3) and strong biofilm formers (OD600 nm≥ 3). All strains of B. bifidum were strong biofilm formers, while B.longum strains were not strong biofilm formers, 57.90% of which were non-biofilm formers. The biofilm-forming ability of other Bifidobacterium was different, including all three types of strains. It was found that the biofilm structures of B.animalis and B.bifidum were mushroom-shaped and all of other Bifidobacterium were flat using scanning electron microscopy, and the biofilms of B.longum and B.bifidum contained a higher proportion of viable bacteria using confocal laser scanning microscopy. The effect of surface properties of Bifidobacterium on the biofilm formation was performed using the adhesion of bacteria to the solvent. The results showed that the biofilm-forming ability of B. pseudocatenulatum was only related to the Lewis acid properties, B. breve, B. animalis and B. adolescentis were positively correlated with the surface hydrophobicity and Lewis acid properties, but there was no correlation between surface properties and biofilm formation for B. longum. The strains of B. bifidum with strong biofilm-forming ability, which exhibited high hydrophobicity and Lewis acid properties, had no relation between its surface properties to its biofilm-forming ability. In summary, the biofilm-forming characteristics of Bifidobacterium exerted species-specific and strain-specific effects and related to the surface characteristics.
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