Charles University, Faculty of Pharmacy in Hradec Králové, Teaching and Research Centre, Czech Republic.
J Med Microbiol. 2021 May;70(5). doi: 10.1099/jmm.0.001371.
(SA) and (SE) are the most common pathogens from the genus causing biofilm-associated infections. Generally, biofilm-associated infections represent a clinical challenge. Bacteria in biofilms are difficult to eradicate due to their resistance and serve as a reservoir for recurring persistent infections. A variety of protocols for drug activity testing against staphylococcal biofilms have been introduced. However, there are often fundamental differences. All these differences in methodical approaches can then be reflected in the form of discrepancies between results. In this study, we aimed to develop optimal conditions for staphylococcal biofilm formation on pegs. The impact of peg surface modification was also studied. The impact of tryptic soy broth alone or supplemented with foetal bovine serum (FBS) or human plasma (HP), together with the impact of the inoculum density of bacterial suspensions and the shaking versus the static mode of cultivation, on total biofilm biomass production in SA and SE reference strains was studied. The surface of pegs was modified with FBS, HP, or poly-l-lysine (PLL). The impact on total biofilm biomass was evaluated using the crystal violet staining method and statistical data analysis. Tryptic soy broth supplemented with HP together with the shaking mode led to crucial potentiation of biofilm formation on pegs in SA strains. The SE strain did not produce biofilm biomass under the same conditions on pegs. Preconditioning of peg surfaces with FBS and HP led to a statistically significant increase in biofilm biomass formation in the SE strain. Optimal cultivation conditions for robust staphylococcal biofilm formation might differ among different bacterial strains and methodical approaches. The shaking mode and supplementation of cultivation medium with HP was beneficial for biofilm formation on pegs for SA (ATCC 29213) and methicillin-resistant SA (ATCC 43300). Peg conditioning with HP and PLL had no impact on biofilm formation in either of these strains. Peg coating with FBS showed an adverse effect on the biofilm formation of these strains. By contrast, there was a statistically significant increase in biofilm biomass production on pegs coated with FBS and HP for SE (ATCC 35983).
（SA）和（SE）是导致生物膜相关感染的最常见病原体。通常，生物膜相关感染是一个临床挑战。由于生物膜中的细菌具有耐药性，因此很难将其消灭，并成为反复持续感染的储库。已经引入了各种针对葡萄球菌生物膜的药物活性测试方案。但是，通常存在根本差异。所有这些方法上的差异都会以结果之间的差异形式反映出来。在这项研究中，我们旨在为葡萄球菌在 peg 上形成生物膜开发最佳条件。还研究了 peg 表面改性的影响。单独使用胰蛋白酶大豆肉汤或补充胎牛血清（FBS）或人血浆（HP），以及细菌悬浮液接种密度的影响，以及摇动与静态培养方式，对 SA 和 SE 参考菌株的总生物膜生物量产生的影响。 peg 的表面用 FBS、HP 或聚-l-赖氨酸（PLL）改性。使用结晶紫染色法和统计数据分析评估对总生物膜生物量的影响。补充 HP 的胰蛋白酶大豆肉汤与摇动模式相结合，可显著增强 SA 菌株 peg 上的生物膜形成。在相同条件下，SE 菌株不能在 peg 上产生生物膜生物量。peg 表面用 FBS 和 HP 预处理可使 SE 菌株的生物膜生物量形成显著增加。不同细菌菌株和方法学方法之间可能存在差异，从而导致稳健的葡萄球菌生物膜形成的最佳培养条件也不同。摇动模式和补充 HP 的培养介质有利于 SA（ATCC 29213）和耐甲氧西林的 SA（ATCC 43300）在 peg 上的生物膜形成。在这两种菌株中，peg 用 HP 和 PLL 预处理对生物膜形成没有影响。peg 用 FBS 包被对这些菌株的生物膜形成有不利影响。相比之下，在 peg 上包被 FBS 和 HP 会使 SE（ATCC 35983）的生物膜生物量产生显著增加。
