畜牧与饲料科学 ›› 2018, Vol. 39 ›› Issue (11): 1-6.doi: 10.12160/j.issn.1672-5190.2018.11.001

• 著者文摘 •    下一篇

副猪嗜血杆菌高密度发酵工艺研究

耿笑林;张翼;刘鹏;张浩浩;翟路峰;李向东;黄玉欣;田克恭   

  1. 国家兽用药品工程技术研究中心,河南洛阳471003
  • 收稿日期:2018-09-20 出版日期:2018-11-30 发布日期:2019-08-19
  • 通讯作者: 耿笑林-国家兽用药品工程技术研究中心,河南洛阳471003
  • 作者简介:耿笑林(1985-),男,硕士,主要研究方向为微生物发酵工艺,共同第一作者;张翼(1990-),男,主要从事过程装备与控制相关工作,共同第一作者。;通讯作者:黄玉欣(1974-),女,主要从事微生物发酵技术研究工作。;通讯作者:田克恭(1964-),男,教授,主要从事动物疫病诊断与防控技术研究工作。
  • 基金资助:
    国家重点研发计划(2016YFD0500703);河南省重大科技专项(171100110200);河南省省级引智项目.

Eatablisment of a High-density Fermentation Process of Haemophilus parasuis

GENG Xiao-lin[1];ZHANG Yi[1];LIU Peng[1];ZHANG Hao-hao[1];ZHAI Lu-feng[1];LI Xiang-dong[1];HUANG Yu-xin[1];TIAN Ke-gong[1]   

  1. National Research Center of Veterinary Medicine,Luoyang 471003,China
  • Received:2018-09-20 Online:2018-11-30 Published:2019-08-19

摘要: 旨在建立副猪嗜血杆菌高密度发酵工艺。通过对TSB、BH1、LB培养基进行比较,选择TSB培养基进行优化,利用优化后的培养基对副猪嗜血杆菌在10L发酵罐中进行高密度发酵培养。结果表明,血清4型JS株的活菌数达到2.5×10^10CFU/mL,血清5型ZJ株的活菌数达到2.5×10^10CFU/mL,且通过批次补料与流加相结合的工艺使关键成本血清的使用量降低了1/2。在1000L发酵规模上进行连续3批的发酵试验.经验证该工艺可用于副猪嗜血杆菌灭活疫苗抗原的规模化生产。对3批发酵抗原利用替代动物豚鼠进行动物实验,结果显示抗原均合格。研究结果为国内副猪嗜血杆菌流行菌株血清4型和5型二价灭活疫苗的规模化生产提供了理论依据。

关键词: 副猪嗜血杆菌;高密度发酵;血清

Abstract: This experiment aims to establish a high-density fermentation process for Haemophilus parasuis. By comparing the culture effect of TSB, BHI, and LB medium, the TSB medium was selected for further optimization, and the optimized medium was used to carry out high-density fermentation culture of Haemophilus parasuis in a 10 L fermentation tank. The results showed that the number of viable bacteria of serum type 4 JS strain was 2.5×10^10 CFU/mL, and that of serum type 5 ZJ strain was 2.5×10^10 CFU/mL, and the addition of serum was reduced by half through the combination use of batch -supplementation and fed batch culture. Three consecutive batches of fermentation experiments at the 1 000 L fermentation scale identified that the process could be used for large-scale production of inactivated vaccine antigens of Haemophilus parasuis. In addition, animal experiments were carried out on three batches of fermentation antigens using replacement animal of guinea pigs, and the results showed that the antigens were all qualified. In summary, our results provide a theoretical basis for the large-scale production of bivalent inactivated vaccines of serum type 4 and type 5 of Haemophilus parasuis.

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