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Bacterial Culture Preservation in Frozen and Dry-Film Methylcellulose

机译:冷冻和干膜甲基纤维素中的细菌培养保存

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摘要

Forty-seven of 61 bacterial cultures, including strains of Pseudomonas, Xanthomonas, Erwinia, Agrobacterium, Corynebacterium, Serratia, Klebsiella, and Escherichia, remained viable after storage in frozen methylcellulose or in dried methylcellulose for up to 38 months. Pathogenicity remained intact for those strains tested. Bacteria were grown on a solid medium and then removed and placed in 1.0% methylcellulose (cellulose methyl ether) to make a final suspension of 108 colony-forming units (CFU) per ml. For storage in dried form, the bacteria-methylcellulose suspension was placed in a petri dish and dried in a forced-air incubator. After 24 h of storage at 25°C, viable populations of 105 CFU/mg (equivalent to 106 CFU/ml) were recovered. Populations of 102 to 104 CFU/mg were recovered after storage of up to 38 months. Similar results were obtained in frozen methylcellulose. Survival was greatly enhanced when the growth medium for the bacteria was potato dextrose peptone rather than nutrient agar, yeast dextrose calcium carbonate peptone, or King's medium B. Addition of 0.1 M MgSO4 to the methylcellulose suspension and to the resuspending liquid also increased survival and recovery from storage for some strains. Methylcellulose storage should be a simple, inexpensive, and reliable method of maintaining cultures for short or long periods of time.
机译:61种细菌培养物中的47种,包括假单胞菌,黄单胞菌,欧文氏菌,农杆菌,棒状杆菌,沙雷氏菌,克雷伯菌和大肠埃希氏菌,在冷冻的甲基纤维素或干燥的甲基纤维素中保存长达38个月后仍能存活。这些测试菌株的致病性保持不变。将细菌在固体培养基上生长,然后移出并置于1.0%甲基纤维素(纤维素甲基醚)中,使最终悬浮液为每毫升10 sup 8菌落形成单位(CFU)。为了以干燥形式储存,将细菌-甲基纤维素悬浮液置于培养皿中,并在强制空气培养箱中干燥。在25°C下储存24小时后,回收到10 5 CFU / mg(相当于10 6 CFU / ml)的存活种群。储存长达38个月后,恢复出10 2 至10 4 CFU / mg的种群。在冷冻的甲基纤维素中获得了相似的结果。当细菌的生长培养基是马铃薯葡萄糖蛋白ept而不是营养琼脂,酵母葡萄糖碳酸钙蛋白ept或King's培养基B时,存活率大大提高。向甲基纤维素悬浮液和重悬液中添加0.1 M MgSO4也可以提高存活率和回收率。从存储一些应变。甲基纤维素的储存应该是一种简单,便宜且可靠的短期或长期维持培养物的方法。

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