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目的建立和鉴定铜绿假单胞菌体外生物膜。方法采用改良平板法,用扫描电镜、Fontana镀银染色法和刚果红-阿利新蓝(AB)染液联合胞外多糖染色法观察体外生物膜型的铜绿假单胞菌和浮游型铜绿假单胞菌的形态,建立稳定、可靠的铜绿假单胞菌体外生物膜模型。结果刚果红-阿利新蓝(AB)染液联合细菌胞外糖染色法,细菌细胞呈淡红色,胞外糖为深紫红色,背景呈蓝色;Fontana镀银染色法细菌细胞呈绛红色,胞外糖为深黄色,背景呈桔黄色;扫描电镜下生物膜细菌呈短杆状,周围被黏稠状物质紧紧包绕,菌体间以黏稠的纤维状黏液丝相连。结论铜绿假单胞菌体外培养建立生物膜的方法简便易行,结果可靠,重复性好,为进一步开展铜绿假单胞菌临床治疗的研究提供了实验手段。
Objective To establish and identify in vitro biofilms of Pseudomonas aeruginosa. Methods In vitro biofilm-type Pseudomonas aeruginosa and planktonic pseudomonas aeruginosa were observed by modified plate method, scanning electron microscopy, Fontana silver staining and Congo red-alitake blue (AB) combined with exopolysaccharide staining To establish a stable and reliable biofilm model of Pseudomonas aeruginosa in vitro. RESULTS: Congo red-alitake blue (AB) stain combined with bacterial extracellular polysaccharide staining showed that the bacterial cells were reddish-colored, the extracellular sugar was dark purple and the background was blue. The Fontana silver-stained bacterial cells were magenta, The extracellular sugar was dark yellow and the background was orange. The biofilm bacteria under the scanning electron microscope showed a short rod shape, surrounded by viscous material and surrounded by viscous fibrous mucilage filaments. Conclusion The biofilm culture of Pseudomonas aeruginosa in vitro is simple, reliable, and reproducible. It provides an experimental method for further research on the clinical treatment of Pseudomonas aeruginosa.