Microbiology and Infection Research Group, Department of Biomedical Science, Cardiff School of Sport and Health Sciences, Cardiff Metropolitan University, Cardiff, UK.
Costerton Biofilm Center, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
J Appl Microbiol. 2021 Nov;131(5):2579-2585. doi: 10.1111/jam.15120. Epub 2021 May 8.
To use a flow-based method to establish, quantify and visualize biofilms of Ureaplasma parvum.
Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow-cell with a flow rate of 0·01 ml min and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High-resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm /µm ± 0·152 and 4 × 10 CCU per ml, respectively) compared with static conditions (0·008 µm /µm ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation.
Flow is an essential requirement for the establishment of U. parvum biofilms.
This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence-associated with adhesion.
建立、定量和可视化微小脲原体生物膜的流动方法。
为了建立生长曲线，每 3 小时在 550nm 处测量微小脲原体 HPA5 培养物的吸光度读数，共 30 小时，并通过颜色变化单位（CCU）的数量来确定活力。使用 DTU 流动池以 0.01ml/min 的流速建立生物膜，并与静态对照进行比较。通过 CCU 确定细菌浓度，通过 Syto9 染色和 COMSTAT 分析定量生物膜生物量。通过扫描电子显微镜（SEM）获得高分辨率图像。与静态条件相比（分别为 0.008µm/µm±0.010 和无可回收细胞），流动导致生物膜和细胞浓度显著增加（分别为 0.599µm/µm±0.152 和 4×10 CCU/ml）。SEM 显示多形细胞，有出芽和可能的膜囊泡形成的迹象。
流动是微小脲原体生物膜形成的必要条件。
这是首次对微小脲原体生物膜生物量进行定量。现在可以建立微小脲原体的存活生物膜，这将允许未来测试抗微生物剂并理解与粘附相关的毒力。
