Sakimura Toshiyuki, Kajiyama Shiro, Adachi Shinji, Chiba Ko, Yonekura Akihiko, Tomita Masato, Koseki Hironobu, Miyamoto Takashi, Tsurumoto Toshiyuki, Osaki Makoto
Department of Orthopaedic Surgery, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8501, Japan.
Department of Gross Anatomy, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8501, Japan.
Biomed Res Int. 2015;2015:943056. doi: 10.1155/2015/943056. Epub 2015 Jan 31.
We investigated biofilm formation and time of vancomycin (VCM) resistance expression after adhesion to a metal surface in Staphylococcus epidermidis. Biofilm-forming Staphylococcus epidermidis with a VCM MIC of 1 μg/mL was used. The bacteria were made to adhere to a stainless steel washer and treated with VCM at different times and concentrations. VCM was administered 0, 2, 4, and 8 hours after adhesion. The amount of biofilm formed was evaluated based on the biofilm coverage rates (BCRs) before and after VCM administration, bacterial viability in biofilm was visually observed using the fluorescence staining method, and the viable bacterial count in biofilm was measured. The VCM concentration required to decrease BCR significantly compared with that of VCM-untreated bacteria was 4 μg/mL, even in the 0 hr group. In the 4 and 8 hr groups, VCM could not inhibit biofilm growth even at 1,024 μg/mL. In the 8 hr group, viable bacteria remained in biofilm at a count of 10(4) CFU even at a high VCM concentration (1,024 μg/mL). It was suggested that biofilm-forming Staphylococcus epidermidis expresses resistance to VCM early after adhesion to a metal surface. Resistance increased over time after adhesion as the biofilm formed, and strong resistance was expressed 4-8 hours after adhesion.
我们研究了表皮葡萄球菌在粘附于金属表面后生物膜的形成以及对万古霉素(VCM)耐药性表达的时间。使用对VCM的最低抑菌浓度(MIC)为1μg/mL的产生物膜表皮葡萄球菌。使细菌粘附于不锈钢垫圈,并在不同时间和浓度下用VCM处理。在粘附后0、2、4和8小时给予VCM。根据给予VCM前后的生物膜覆盖率(BCR)评估生物膜形成量,使用荧光染色法肉眼观察生物膜中的细菌活力,并测量生物膜中的活菌数。与未用VCM处理的细菌相比,即使在0小时组中,显著降低BCR所需的VCM浓度为4μg/mL。在4小时和8小时组中,即使在1024μg/mL时,VCM也无法抑制生物膜生长。在8小时组中,即使在高VCM浓度(1024μg/mL)下,生物膜中仍有10⁴CFU的活菌。提示产生物膜的表皮葡萄球菌在粘附于金属表面后早期就对VCM产生耐药性。随着生物膜形成,粘附后耐药性随时间增加,在粘附后4 - 8小时表现出强耐药性。