生孢梭菌半固体培养基菌落计数方法的研究  

Study Sporogenes Semi-Solid Medium Colony Counting Method

作　　者：刘晓菲 

机构地区：[1]华北制药河北华民药业有限责任公司,河北藁城052165

出　　处：《化工设计通讯》2016年第3期173-173,179,共2页Chemical Engineering Design Communications

摘　　要：目的 ：建立使用半固体培养基进行生孢梭菌菌落计数的方法,使菌落计数结果能够接近最大活菌数。方法 ：接种生孢梭菌的新鲜培养物至硫乙醇酸盐培养基中,30~35℃培养18~24小时,用0.9%无菌氯化钠溶液稀释生孢梭菌菌液至与标准比浊管相同的浓度,用10倍稀释法分别稀释至1×10-6、1×10-7、1×10-8三个稀释级备用。分别配制琼脂浓度为0.2%、0.3%、0.4%、0.5%、0.6%、0.7%的6种硫乙醇酸盐培养基,分别取三个稀释级的菌液1m L接种至配制好的上述6种硫乙醇酸盐培养基中,同时在胰酪大豆胨琼脂培养基上接种等量菌液,并放入厌氧袋中,将接种好的上述培养基至35℃培养18~24小时,进行菌落计数并比较结果。结果 ：半固体培养基计数法比厌氧袋计数法效果好,使用琼脂浓度为0.5%的硫乙醇酸盐培养基,30~35℃培养18小时,菌落计数最多,最接近最大活菌数。结论 ：生孢梭菌半固体培养基计数法适用于生孢梭菌计数。Objective ： To establish a semi-solid media were sporogenes colony counting method, so that the results can be close to the maximum colony count the number of viable cells.Methods ： Fresh culture was inoculated sporogenes to thioglycollate medium, 30 - 35 ℃ for 18 to 24 hours, with sterile 0.9% sodium chloride solution was diluted sporogenes bacteria to the standard ratio the same concentration turbidity tube with a 10-fold dilution were diluted to 1 × 10 -6, 1 × 10-7, 1 ×10 -8 diluted three-level backup.Agar were prepared at a concentration of 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7% of the six kinds of thioglycollate medium were taken three stages diluted broth was inoculated into lmL above six kinds of the prepared thioglycollate medium, while in the pancreatic casein peptone soy broth agar medium was inoculated the same amount and placed in an anaerobic bag the inoculated medium to above 35 ℃ cultured 18 to 24 hours.colony count and compare the results.Results ： The semi-solid medium count better than anaerobic bags counting results, use agar concentration of 0.5% thioglycoUate medium, 30 - 35 ℃ 18 hours, colony counts most, closest to the maximum number of viable cells.Conclusion ： sporogenes semi-solid medium suitable for counting sporogenes count.

关 键 词：生孢梭菌 半固体培养基 菌落计数 

分 类 号：R927[医药卫生—药学]