福赛斯坦纳菌体外培养实验技术的研究

目的 探索福赛斯坦纳菌生长繁殖的基本条件，比较N-乙酰胞壁酸及不同营养因子对其生长繁殖的影响，筛选该菌体外培养的最佳营养条件，建立其体外培养的技术方法。方法 （1）培养基制备：在血平板中加0.005% N-乙酰胞壁酸形成改良培养基；制备血平板与不同浓度（0.5、3、10、50、100 mg/L）的N-乙酰胞壁酸形成的培养基；制备10 mg/L N-乙酰胞壁酸与氯化血红素、酵母提取物、维生素K₃、脱纤维羊血组合形成不同组别的培养基。（2）接种培养后，用磷酸盐缓冲液将细菌洗脱，并用酶标仪（OD₆₀₀）测定其吸光度。结果 福赛斯坦纳菌在含N-乙酰胞壁酸的培养基中生长更好，浓度为10 mg/L时促进作用最明显且细菌形态更为典型。在组3培养基中生长最快。结论 N-乙酰胞壁酸能促进福赛斯坦纳菌的生长繁殖，浓度为10 mg/L时促进作用最明显；N-乙酰胞壁酸及氯化血红素、维生素K₃、酵母提取物和脱纤维羊血组合能使其生长周期明显缩短。

Investigation of Experimental Technique of Tannerella forsythia Cultured in Vitro

Objective To investigate the basic requirements for the growth and proliferation of Tannerella forsythia and compare the growth-promoting effects with different nutritional factors and screen the optimum culture in vitro conditions and establish the technique of culturing it.Methods (1)Preparation of culture medium:0.005% N-acetylmuramic acid was added to the blood agar medium,forming the modified medium.Different concentration of N-acetylmuramic acid(0.5,3,10,50,100 mg/L)was added to the blood agar medium.The different mediums of N-acetylmuramic acid(10 mg/L)combined with other nutritional factors (chlorhematin,yeast extract,Vitamin K₃,off fiber sheep blood) were formed.(2)After culturing,use PBS to elute the bacterial and measure its absorbance with enzyme-labelled instrument (OD₆₀₀).Results Tannerella forsythia grew more better in the culture medium containing N-acetylmuramic acid,and the concentration at 10 mg/L was most obvious in growth-promoting effect and can lead to the change of bacterial morphology.The medium which consisted of hemin,yeast extract,menadione and sheep blood was the most suitable condition for its growth.Conclusion N-acetylmuramic acid can promote the growth of Tannerella forsythia,and the combination of N-acetylmuramic acid,hemin,menadione,yeast extract and sheep blood can shorten the growth cycle of it dramatically.