高级检索
刘优招, 冯梅, 王丽, 陈炜聪, 韩宇宇, 王祉蕴, 陈清, 刘志华, 胡静. 香港海鸥菌不同保存方法效果比较[J]. 中国公共卫生, 2017, 33(3): 471-474. DOI: 10.11847/zgggws2017-33-03-34
引用本文: 刘优招, 冯梅, 王丽, 陈炜聪, 韩宇宇, 王祉蕴, 陈清, 刘志华, 胡静. 香港海鸥菌不同保存方法效果比较[J]. 中国公共卫生, 2017, 33(3): 471-474. DOI: 10.11847/zgggws2017-33-03-34
LIU You-zhao, FENG Mei, WANG Li.et al, . Comparison of methods for Laribacter hongkongensis strain storage[J]. Chinese Journal of Public Health, 2017, 33(3): 471-474. DOI: 10.11847/zgggws2017-33-03-34
Citation: LIU You-zhao, FENG Mei, WANG Li.et al, . Comparison of methods for Laribacter hongkongensis strain storage[J]. Chinese Journal of Public Health, 2017, 33(3): 471-474. DOI: 10.11847/zgggws2017-33-03-34

香港海鸥菌不同保存方法效果比较

Comparison of methods for Laribacter hongkongensis strain storage

  • 摘要: 目的比较不同方法对香港海鸥菌的短期和长期保存效果,以期建立该菌的简单、实用和经济的短期和长期保存方法。方法采用半固体石蜡封存法,磁珠菌种保存管、10%甘油、15%甘油肉汤和30%甘油肉汤等5种方法保存香港海鸥菌,其中,半固体石蜡封存法的保存温度为室温和4 ℃,其他方法均在-20 ℃和-80 ℃进行菌种保存。定期进行菌种复苏,采用活菌菌落计数比较不同方法的保存效果,观察期为12个月。结果半固体石蜡封存法在室温环境下保存的活菌数在6个月内呈缓慢下降趋势(活菌数的对数值下降幅度<1),但6个月以后活菌数急速下降且菌落变小,至第9个月时为0 CFU/mL;而4 ℃保存的第1个月活菌数下降幅度小,但到第3个月时活菌数为0 CFU/mL;在12个月内,磁株及3种不同甘油浓度保存的活菌数在-80 ℃下保持相对稳定,但-20 ℃下保存时间短、效果差。结论室温条件下的半固体石蜡封存法适合短期保存香港海鸥菌,而-80 ℃低温条件下的磁株或甘油适合长期保存。

     

    Abstract: ObjectiveTo compare different methods for preserving Laribacter hongkongensis (LH) strains and to develop a simple and practicable preservation protocol for short- and long-term storage and reactivation LH strain.MethodsThe five methods for LH strain storage included:(i) semisolid with paraffin storage,(ii) Microbank vial,(iii) 10% glycerol,(iv) 15% glycerol/85% Luria-Bertani (LB) broth culture,and (v) 30% glycerol/70% LB broth culture.The LH strains were put at 4 ℃ and room temperature when using methods i,while the stains were disposed at conventional freezing temperatures of -20 ℃ and -80 ℃ when using other methods,respectively.All storage techniques were evaluated periodically by counting with colony-forming unit over 12 months.ResultsFor semisolid with paraffin storage,the viable strains were reduced gently up to 6 months at room temperature,then reduced rapidly and LH colonies became pinpoint size,contrasting with no colony growth on MacConkey agar at 3-month at 4 ℃.Preservation of LH strains in Microbank vial,10% glycerol,15% glycerol broth,and 30% glycerol broth stored at -80 ℃ were proved to be stable for up to 12 months.However,recovery of the strains from these methods stored at -20 ℃ showed poor growth after one month.ConclusionSemisolid with paraffin storage at room temperature is an effective way for short-term preservation of LH strain.Preservation in glycerol or Microbank vial at -80 ℃ is recommended for prolonged periods of LH strain storage.

     

/

返回文章
返回