改良Frey氏液体培养基质控菌株菌种代次与生长曲线研究
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摘要
为探究改良Frey氏液体培养基质控菌CVCC2960滑液支原体(以下简称MS)生长规律,本试验首次同时采用CFU计数、CCU测定和p H值测定的方法,比较不同接种浓度、接种体积和不同代次MS生长情况。结果显示:5%与10%接种量的CCU测定结果峰值差异不明显,但5%接种的CFU计数峰值更高,pH下降更慢;1~5代菌在生长曲线各阶段的测定结果均无明显差别;3种不同接种方式的测定结果显示,用0.5 m L菌液接种9.5 m L培养基的方式更利于MS生长。本试验为改良Frey氏液体培养基的质控标准化提供依据。
To explore the growth law of a quality control strain for modified Frey liquid medium,CVCC2960 Mycoplasma Synoviae( MS),the colony counting method,CCU assay and pH determination were combined to compare MS growth in different inoculation concentrations,inoculation volumes and generations. The results indicated that there was no significant difference in growing titer peak,while 5% strains had a higher living cell concentration peak and a slower decrease pH value. The generation and growth value of 1 ~ 5 generations didn't have significant difference. That mix 0. 5 m L bacteria suspension with 9. 5 m L medium was proved to be more suitable than other two for MS. This study provided a basis evidence of standardization for modified Frey liquid medium.
To explore the growth law of a quality control strain for modified Frey liquid medium,CVCC2960 Mycoplasma Synoviae( MS),the colony counting method,CCU assay and pH determination were combined to compare MS growth in different inoculation concentrations,inoculation volumes and generations. The results indicated that there was no significant difference in growing titer peak,while 5% strains had a higher living cell concentration peak and a slower decrease pH value. The generation and growth value of 1 ~ 5 generations didn't have significant difference. That mix 0. 5 m L bacteria suspension with 9. 5 m L medium was proved to be more suitable than other two for MS. This study provided a basis evidence of standardization for modified Frey liquid medium.
引文
[1]The United States Phamacopeial Convention,Inc.USP36NF31,General Chapter 63,Mycoplasma Tests[S].
[2]European Directorate for the Quality of Medicines&Healthcare,Councile of Europe.European Phamacopoeia 5.0,2.6.7 Mycoplasmas[S].
[3]中国兽药典委员会.中国人民共和国兽药典二〇一〇年版三部[S].
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[2]European Directorate for the Quality of Medicines&Healthcare,Councile of Europe.European Phamacopoeia 5.0,2.6.7 Mycoplasmas[S].
[3]中国兽药典委员会.中国人民共和国兽药典二〇一〇年版三部[S].
[4]刘轶秋,丁家波,李蓓蓓,等.支原体检验用培养基质控菌株的生长曲线与世代时间测定[J].中国兽药杂志,2012,46(12):13-15.
[5]周锦龙,封柯宇,吴波良,等.禽滑液支原体液体培养中p H与活菌浓度关系研究[A].动物微生态学分会第四届第十一次全国学术研讨会论文集[C].2014.
[6]魏顺.衣阿华支原体与滑液支原体的全基因组测序及分析[D].武汉:华中农业大学,2012.
[7]吴移谋,叶元康.支原体学[M].北京:人民卫生出版社,2008.
[8]杜德燕,张洪,姜来生,等.不同容积的培养瓶对猪肺炎支原体培养滴度的影响[J].四川畜牧兽医,2013,(6):29-31.