鸡滑液囊支原体疫苗株的培养工艺研究

Culture Process of Mycoplasma Synoviae Vaccine Strain

为研究不同培养工艺对鸡滑液囊支原体(Mycoplasma synoviae,MS)疫苗株(QD株)活菌数的影响,试验通过对培养基、血清、培养方式、培养体积等参数的筛选,优化培养工艺,提升培养物的活菌含量。结果显示:与改良Frey氏培养基相比,大溪生物培养基能够使QD株培养时间缩短为16~19 h,活菌数稳定在109 CCU/mL;草原绿野猪血清的添加能够使QD株培养16~17 h,活菌数稳定在109~1010 CCU/mL;相比静置培养,100 r/min振荡培养使QD株平均培养时间缩短了2.5 h;50%培养体积的活菌数以及平均培养时间优于其他组。研究表明,鸡滑液囊支原体QD株发酵培养工艺确定为按10%比例接种于添加10%草原绿野猪血清的大溪生物培养基中,培养体积为50%,37℃恒温培养,转速为100 r/min,通气量为2 L/min,培养时间为12.5 h,10L发酵罐扩大培养能够使活菌数达到1010 CCU/mL。

In order to study the effect of different culture processes on viable bacteria count of Mycoplasma synoviae vaccine strain(QD strain),culture mediums,serum,culture methods,culture volumes and other parameters were screened to further optimize the culture process and increase viable bacteria content.The results showed that compared with the modified Frey medium,Daxi biological medium could shorten the culture time of QD strain to 16-19 h,and the number of viable bacteria was stable at 109 CCU/mL.The addition of Caoyuanlvye serum could make QD strain concentrated at 16-17 h,the number of viable bacteria was stable at 109 to 1010 CCU/mL.Compared with the static culture method,the average culture time of QD strain was shortened by 2.5 h by 100 r/min oscillation culture.The number of viable bacteria and average culture time of 50%culture medium volume group were better than other groups.The results showed that 10%QD strain was inoculated in Daxi biological medium supplemented with 10%Caoyuanlvye serum and 50%culture volume,incubated at 37℃with 100 r/min for rotational speed and 2 L/min for aeration rate,cultured for 12.5 h in a 10 L fermenter and the number of viable bacteria could reach 1010 CCU/mL.