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Production of extra-cellular slime by Staphylococcus epidermidis during stationary phase of growth: its association with adherence to implantable devices.

机译:表皮葡萄球菌在生长的稳定阶段产生细胞外粘液:其与对可植入装置的粘附有关。

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摘要

A method of optimising slime production produced by Staphylococcus epidermis and its quantitative assay was developed, which gave a preliminary indication of its identity and an assessment of the correlation between slime production and adherence of the organism to implants. After inducing vigorous growth in brain heart infusion broth to stationary phase, all nutrients were removed by washing and the organisms resuspended in sterile deionised water with added magnesium. After further incubation the culture was centrifuged and the supernatant reacted with alcian blue in 50 mM magnesium chloride/sodium acetate solution, and the amount of bound dye was measured spectrophotometrically at 620 nm after its resolubilisation using sodium dodecyl sulphate. Large quantities of slime were produced by some, but not all, strains. Preliminary electrophoresis of the slime showed mobility and staining similar to that of the glycosaminoglycans. Adherence was tested by growing strains in wells of tissue culture plates and aspirating the supernatant after incubation. After fixation and staining of adherent growth the amount of bound stain was determined spectrophotometrically after its elution with ethanol. In this series of organisms there was no correlation between the result of tests for adherence or production of extracellular slime, and no correlation between either of these and the clinical source of the organisms.
机译:开发了一种优化表皮葡萄球菌产生的粘液产生的方法及其定量测定方法,该方法初步表明了其身份,并评估了粘液产生与生物对植入物的粘附之间的相关性。在使脑心输液肉汤剧烈生长至固定相后,通过洗涤除去所有营养,并将生物体重新悬浮在添加了镁的无菌去离子水中。进一步温育后,将培养物离心,使上清液与阿尔辛蓝在50mM氯化镁/乙酸钠溶液中反应,在使用十二烷基硫酸钠将其再溶解后,在620nm下用分光光度法测定结合染料的量。一些(但不是全部)菌株产生了大量的煤泥。粘液的初步电泳显示出与糖胺聚糖相似的迁移率和染色。通过在组织培养板的孔中生长菌株并在孵育后吸出上清液来测试粘附性。固定和粘附生长的染色后,用乙醇洗脱后用分光光度法测定结合的染色量。在这一系列生物中,粘附性或细胞外粘液产生的测试结果之间没有相关性,并且这两者与生物体的临床来源之间也没有相关性。

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