A kind of high-density cultivation method of Haemophilus parasuis type 5 500l fermenter

Abstract

The invention relates to a high-density culture method of a fermentation tank of 500 L for model-5 haemophilus parasuis. The method includes the following steps that A, the freeze-dried model-5 haemophilus parasuis is activated for 18-24 h through a TSA flat plate till single colonies grow out; B, 4-8 single colonies are selected and put into a TSB shake flask first-grade seed culture medium to be cultured for 6-14 h till the OD600 value reaches 0.8-1.6; C, according to the inoculum size of 1-5% V / V, a first-grade seed solution is transferred into a self-prepared second-grade shake flask seed culture medium to be cultured for 8-14 h till the OD600 value reaches 2.0-3.5; D, according to the inoculum size of 1-5% V / V, a second-grade seed solution is inoculated into a sterilizing culture medium of the fermentation tank of 500 L, a fermentation tank culture medium is contained inside the fermentation tank, culture conditions include that temperature ranges from 35 DEG C to 39 DEG C, tank pressure ranges from 0.02 Mpa to 0.1 Mpa, the percentage of dissolved oxygen is 20-50%, and pH value ranges from 7.3 to 7.8, and the tank is discharged to collect the haemophilus parasuis after culturing is carried out till the pH value is stable, and the dissolved oxygen rises again. In the method, preparation of raw materials is simple, cost is low, it can be guaranteed that the haemophilus parasuis grows powerfully, and metabolism is normal, growth speed can be increased, the production period is short, the viable count is high and is generally 72-80 hundred millions / milliliter, and the method can completely adapt to production of model-5 haemophilus parasuis vaccines.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a high-density culture method and application of a 500L fermentation tank of Haemophilus parasuis type 5. Background technique [0002] Haemophilus parasuis (HPs) is a common bacterium in the upper respiratory tract of conventionally reared pigs, but under certain conditions it can invade the body and cause severe systemic diseases, such as fibrinous polyserositis, It is characterized by arthritis and meningitis, also known as Glasser's disease (Oliverira et al., 2004). Antibiotics were used for treatment in the early stage, but the phenomenon of drug resistance of the bacteria is becoming more and more common. The state restricts the application of antibiotics in animal feed is becoming more and more strict, and the application of antibiotics in the prevention and control of this disease will be strictly restricted and gradually banned. Therefore...

AI Technical Summary

Problems solved by technology

However, the cost is high, and the number of viable bacteria obtained in the end is small. Generally, the number of viable bacteria is 1.5 to 2 billion, and the production cycle is long, which is not suitable for large-scale production by companies.

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