Quantification of the extracellular matrix of the Listeria monocytogenes biofilms of different phylogenic lineages with optimization of culture conditions
Quantification of the extracellular matrix of the Listeria monocytogenes biofilms of different phylogenic lineages with optimization of culture conditions
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DOI：
10.1111/jam.12127
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发表时间：
2013-04-01
影响因子：
4
通讯作者：
Midelet-Bourdin, G.
中科院分区：
文献类型：
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作者：
Combrouse, T.;Sadovskaya, I.;Midelet-Bourdin, G.
目的定量研究单核细胞增生李斯特菌生物膜的细胞外基质。初步研究了27株菌株的系统发育谱系与其在各种条件下形成生物膜的能力之间的关系。方法和结果使用结晶紫测定法在不同条件[两种温度（37和22 ℃）和两种培养基（胰蛋白胨大豆肉汤酵母提取物培养基（TSBYE）和MCDB 202限定培养基）]下评价属于谱系I或II的27株单核细胞增生李斯特菌菌株在微量滴定板上的生物膜形成。谱系II菌株比谱系I菌株产生显著更多的生物膜。在微量滴定板测定中，MCDB 202培养基中的生物膜量大于TSBYE培养基[通过扫描电子显微镜（SEM）分析证实]，并且在37 ℃下大于22 ℃下。在TSBYE中，培养皿上生物膜群体中的可培养细菌的数量比在MCDB 202培养基中更多。SEM研究证实，在37 ℃下，单核细胞增生李斯特菌生物膜在两种培养基中均产生细胞外基质。TSBYE中细胞外基质中外聚合物的量和pH值显著高于MCDB 202培养基。例外的是ScottA菌株，其在两种培养基中呈现相似的pH值和外聚合物含量。蛋白质是最丰富的外聚合物组分，其次是DNA和多糖。结论生物被膜定量结果的解释取决于细菌的生长条件、存活率和分析方法。蛋白质、DNA和多糖的含量因菌株和培养基的不同而不同。本研究筛选了一组广泛的单核细胞增生李斯特菌菌株在生物膜生长条件下合成外聚合物的潜力。对单核细胞增生李斯特菌生物膜组成的研究有助于开发新的策略来防止生物膜的形成和去除生物膜。
Aims The purpose of this study was to quantify the extracellular matrix of Listeria monocytogenes biofilm. A preliminary study was carried out to establish a relationship between phylogenetic lineage of 27 strains and their ability to form biofilm in various conditions. Methods and Results Biofilm formation on microtitre plates of 27 strains of L.monocytogenes belonging to lineages I or II was evaluated in different conditions [two temperatures (37 and 22 degrees C) and two media (tryptone soy broth yeast extract medium (TSBYE) and MCDB 202 defined medium)] using crystal violet assay. Lineage II strains produced significantly more biofilm than lineage I strains. In microtitre plates assay, biofilm quantities were greater in MCDB 202 vs TSBYE medium [confirmed by scanning electron microscopy (SEM) analysis] and at 37 vs 22 degrees C. Cultivable bacteria from biofilm population on Petri dishes were enumerated in greater quantities in TSBYE than in MCDB 202 medium. The SEM investigation established that L.monocytogenes biofilms produce extracellular matrix in both media at 37 degrees C. The amount of exopolymers in the extracellular matrix and the pH values were significantly higher in TSBYE than in MCDB 202 medium. The exception was the ScottA strain that presented similar pH values and exopolymer contents in both media. Proteins were the most abundant exopolymer components, followed by DNA and polysaccharides. Conclusions The interpretation of results of biofilm quantification was depending on the growth conditions, the viability of the bacteria and the analysis method. The quantities of proteins, DNA and polysaccharides were different according to the strains and the medium. Significance and Impact of the Study This study screened the potential of a wide panel of L.monocytogenes strains to synthesize exopolymers in biofilm growing condition. The characterization of L.monocytogenes biofilm composition may help to develop new strategies to prevent the formation of biofilms and to remove the biofilms.