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目的:研究阴道加德纳菌(GV)生物膜体外生长过程及阴道来源乳杆菌对GV生物膜的影响。方法:选取GV标准菌株(ATCC14018)体外培养生物膜,在成膜的不同时期检测生物膜生成量及观察生物膜生成形态。选取阴道来源乳杆菌1#(鼠李糖乳杆菌)、2#(乳酪乳杆菌),分别在GV生物膜生成后的0h、24h、48h加等量乳杆菌共培养24h,用Live/Dead荧光染色剂染色,观察GV生物膜生成情况。结果:(1)同一培养条件下,GV生物膜生成不同时期的OD值有明显差异(P<0.05)。电镜扫描图片显示,培养4~20h时见散落的单菌落,24h时单独菌落开始连接,48h时形成连接的菌落外面包裹一层胞外聚合物,48h后形态无变化。(2)GV生物膜的荧光染色显示,阴道内乳杆菌对抗生长后0h、24h时,GV生物膜厚度减少,形成松散的菌落,以乳杆菌为主;对抗生长后48h,菌落密度无变化,死菌面积增大,活菌以乳杆菌为主。结论:(1)阴道GV生物膜形成较缓慢,开始成膜时间约为培养后24h,约在48h后形成成熟的生物膜。(2)阴道内乳杆菌可干扰GV生物膜的形成,不同乳杆菌对不同时期GV生物膜的作用不同。
Objective: To study the in vitro growth of GV biofilm and the effect of vaginal Lactobacillus on GV biofilm. Methods: The biofilm was cultured in vitro with GV standard strain (ATCC14018). The biofilm production and the formation of biofilm were observed at different stages of biofilm formation. Lactobacillus rhamnosus 1 # (Lactobacillus rhamnosus) and 2 # (Lactobacillus casei) were selected and incubated with the same amount of Lactobacillus at 0h, 24h and 48h after GV biofilm formation, respectively. The cells were incubated with Live / Dead fluorescence Staining, observe the formation of GV biofilm. Results: (1) Under the same culture conditions, the OD values of GV biofilms at different stages were significantly different (P <0.05). Scanning electron micrographs showed that when colonies were cultured for 4 ~ 20h, single colonies scattered and scattered were observed. After 24 hours, the colonies started to connect with each other, and the colonies that formed after 48h were covered with an extracellular layer of polymer. (2) Fluorescence staining of GV biofilms showed that the thickness of GV biofilm decreased after 0 and 24 h of Lactobacillus inoculation to form loose colonies with Lactobacillus as the dominant ones. After 48 h of growth, the density of colonies did not change, Dead area increases, live bacteria to Lactobacillus. Conclusions: (1) The formation of vaginal GV biofilm is relatively slow, and the time of starting membrane formation is about 24 h after culture, forming a mature biofilm about 48 h later. (2) Lactobacillus intragastrically could interfere with the formation of GV biofilm, and different lactobacilli had different effects on GV biofilm in different periods.