Hussain M, Collins C, Hastings J G, White P J
Department of Molecular Biology and Biotechnology, The University, Western Bank, Sheffield.
J Med Microbiol. 1992 Jul;37(1):62-9. doi: 10.1099/00222615-37-1-62.
A firmly adherent mass of slime plus organisms (biofilm) accumulates on the sides of culture tubes when some strains of coagulase-negative staphylococci are grown in a chemically-defined medium containing [14C]glucose. This mass was washed (to remove labelled medium) and then counted after adding scintillation fluid. Organisms from the liquid culture were also washed and counted to check that [14C]glucose had been utilised to label the bacteria. Nine strains were examined in this way, and the results were compared with those obtained with four older techniques for recognising slime production or adherent bacteria. The new method is quick, and has advantages of reproducibility and good discrimination between strains; there was a 15-fold difference in counts in the biofilm between slime-producing and non-producing strains respectively. With the new radiolabel assay, the effects of several antibacterial compounds on the build-up of the biofilm were investigated with four slime-producing strains. Tunicamycin, chloramphenicol and 5-fluorouracil, at levels below their minimum growth-inhibitory concentrations, each greatly diminished biofilm formation; several other drugs had less effect.
当一些凝固酶阴性葡萄球菌菌株在含有[14C]葡萄糖的化学限定培养基中生长时，在培养管的管壁上会积聚一团牢固附着的黏液与微生物（生物膜）。将这团物质洗涤（以去除标记培养基），然后加入闪烁液后进行计数。对液体培养物中的微生物也进行洗涤和计数，以检查[14C]葡萄糖是否已被用于标记细菌。用这种方法检测了9个菌株，并将结果与另外4种用于识别黏液产生或黏附细菌的较老技术所获得的结果进行了比较。新方法快速，具有可重复性的优点，并且对不同菌株有良好的区分能力；产黏液菌株和不产黏液菌株的生物膜计数分别相差15倍。使用新的放射性标记测定法，用4个产黏液菌株研究了几种抗菌化合物对生物膜形成的影响。衣霉素、氯霉素和5-氟尿嘧啶在低于其最低生长抑制浓度时，均能大大减少生物膜的形成；其他几种药物的作用较小。
