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CN-121991820-A - Liquid culture medium of mycoplasma synoviae and fermentation method

CN121991820ACN 121991820 ACN121991820 ACN 121991820ACN-121991820-A

Abstract

The application relates to the technical field of microbial fermentation, and particularly provides a liquid culture medium of mycoplasma synoviae and a fermentation method. The liquid culture medium consists of a basic culture medium containing 1-20 g/L pplo of broth and 0.2-3 g/L of poloxamer. According to the application, poloxamer is added into the culture medium of mycoplasma synoviae for the first time, so that not only is the culture density of mycoplasma synoviae thalli improved, but also the immunogenicity of MS is improved by the poloxamer unexpectedly found. In addition, the liquid culture medium is used for fermenting chicken bursa of mycoplasma synoviae, so that the fermentation time is shortened to 12-14h.

Inventors

  • PANG WENQIANG
  • GENG XIAOLIN
  • XU JINGLONG
  • HUANG YUXIN
  • TIAN KEGONG
  • ZHANG XUKE

Assignees

  • 普莱柯生物工程股份有限公司
  • 洛阳惠中生物技术有限公司

Dates

Publication Date
20260508
Application Date
20241107

Claims (10)

  1. 1. The poloxamer is used for improving the culture density of the mycoplasma synoviae or the immunogenicity of the mycoplasma synoviae.
  2. 2. The use according to claim 1, wherein the poloxamer is poloxamer 188.
  3. 3. The chicken bursa mycoplasma liquid culture medium is characterized by comprising a basic culture medium containing 1-20 g/L pplo of broth and 0.2-3 g/L of poloxamer, wherein the basic culture medium is CM2 culture medium or modified Frey culture medium.
  4. 4. The liquid medium according to claim 3, wherein the poloxamer content is 1g/L, preferably wherein the poloxamer is poloxamer 188; Preferably, the pplo broth content is 1-10 g/L, preferably 5g/L; preferably, the pH of the liquid culture medium is 7.8-8.0.
  5. 5. A solid culture medium of mycoplasma synoviae, characterized in that the solid culture medium is the liquid culture medium of claim 3 or 4 containing 1.5% agar powder or agarose.
  6. 6. A fermentation method of mycoplasma synoviae, characterized in that the liquid culture medium of claim 3 or 4 is used for culturing mycoplasma synoviae.
  7. 7. The fermentation method according to claim 6, wherein the activated mycoplasma synoviae is inoculated into a fermentation tank containing the liquid culture medium for culturing, when DO is reduced to below 10%, the rotation speed and aeration of the fermentation tank are increased to make DO be more than 10%, and the culture is carried out until the pH is 6.6-6.7, namely, the culture is carried out for 12-14 h.
  8. 8. The fermentation method according to claim 7, comprising the steps of inoculating MS freeze-dried bacteria into the liquid culture medium, standing and culturing at 37 ℃ until the pH is 6.8-6.9 to obtain primary seeds, inoculating the primary seeds into the liquid culture medium at 10%, culturing at 37 ℃ until the pH is 6.8-6.9 to obtain secondary seeds, inoculating the secondary seeds into the liquid culture medium of a large-volume fermentation tank, and when DO is reduced to below 10%, increasing the rotating speed and ventilation of the fermentation tank to enable DO to be more than 10%, and culturing until the pH is 6.6-6.7 to obtain the fermentation tank, namely culturing for 12-14h; preferably, the large-volume fermentation tank is a 15L or 150L fermentation tank, wherein the stirring speed of the 15L fermentation tank is 50-150 rpm, the ventilation amount is 1-2L/min, the stirring speed of the 150L fermentation tank is 30-50 rpm, and the ventilation amount is 5-12L/min; preferably, the fermenter inoculum size is 2% -10%, preferably 5%.
  9. 9. A mycoplasma synoviae obtainable by the fermentation process of any one of claims 6 to 8.
  10. 10. Use of mycoplasma synoviae according to claim 9 in inactivated vaccines against mycoplasma synoviae.

Description

Liquid culture medium of mycoplasma synoviae and fermentation method Technical Field The application relates to the technical field of microbial fermentation, and particularly provides a liquid culture medium of mycoplasma synoviae and a fermentation method. Background Mycoplasma synoviae of chickens is an infectious disease of chickens and turkeys caused by Mycoplasma synoviae (Mycoplasma Synoviae, MS), also known as infectious synovitis (avian onfectious synovitis), characterized by arthrocele, bursa and tendon inflammation, and enlargement of the parenchymal organs. The disease has slow development and long disease course, once the disease is infected in chicken flock, the disease is difficult to eradicate, so the disease spreads in chicken flock for a long time, and the problems of low feed utilization rate, slow growth and development, high elimination rate, reduced egg yield and the like are caused. When the pathogen exists in the chicken flock, mixed infection is easy to occur, the illness state is aggravated, the death rate is increased, and serious economic loss is caused. MS is pear-shaped or elliptic, has a diameter of about 0.2 μm, is facultative anaerobic, and has no cell wall. There are two current methods for controlling chicken bursal mycoplasmosis, antibiotics or immunization. MS is sensitive to multiple antibiotics, but is prone to developing drug resistance, which causes severe mycoplasma infection while the expected therapeutic effect is inadequate, and is the causative agent of the transition to chronic disease. And the inoculation of the inactivated vaccine is an ideal control method. In the manufacturing process of inactivated vaccines, MS high-density culture is an important link. However, in large-scale high-density culture of MS, the number of living bacteria in stationary culture is low and the fermentation time is long, and the use of a mechanical stirring ventilation fermentation tank improves the culture density and shortens the fermentation time, but the shearing force generated by MS does not have cell walls, so that MS bacteria are damaged, and the number of living bacteria is not high. In view of this, the present application has been made. Disclosure of Invention The application aims to provide a liquid culture medium of mycoplasma synoviae and a fermentation method thereof, so as to improve the culture density and immunogenicity of the mycoplasma synoviae. In order to achieve the above purpose, the present application adopts the following technical scheme: the poloxamer is used for improving the culture density of the mycoplasma synoviae or the immunogenicity of the mycoplasma synoviae. Further, the poloxamer is poloxamer 188. The chicken bursa mycoplasma liquid culture medium consists of a basic culture medium containing 1-20 g/L pplo of broth and 0.2-3 g/L of poloxamer, and the basic culture medium is CM2 culture medium or modified Frey culture medium. Further, the poloxamer content is 1g/L, preferably, the poloxamer is poloxamer 188; Preferably, the pplo broth content is 1-10 g/L, preferably 5g/L; preferably, the pH of the liquid culture medium is 7.8-8.0. A solid culture medium of mycoplasma synoviae is prepared from the liquid culture medium containing 1.5% of agar powder or agarose. A fermentation method of mycoplasma synoviae utilizes the liquid culture medium to culture mycoplasma synoviae. Further, inoculating the activated mycoplasma synoviae to the liquid culture medium for culture, when DO is reduced to below 10%, increasing the rotating speed and ventilation to enable DO to be more than 10%, and harvesting when the pH is 6.6-6.7, namely culturing for 12-14 h. The fermentation method comprises the steps of inoculating MS freeze-dried bacteria into a liquid culture medium, standing and culturing at 37 ℃ until the pH value is 6.8-6.9 to obtain first-stage seeds, inoculating the first-stage seeds into the liquid culture medium according to 10%, culturing at 37 ℃ until the pH value is 6.8-6.9 to obtain second-stage seeds, inoculating the second-stage seeds into the liquid culture medium of a large-volume fermentation tank, and culturing for 12-14h after DO is reduced to below 10%, and increasing the rotating speed and ventilation to enable DO to be greater than 10%, and culturing until the pH value is 6.6-6.7; preferably, the large-volume fermentation tank is a 15L or 150L fermentation tank, wherein the stirring speed of the 15L fermentation tank is 50-150 rpm, the ventilation amount is 1-2L/min, the stirring speed of the 150L fermentation tank is 30-50 rpm, and the ventilation amount is 5-12L/min; preferably, the fermenter inoculum size is 2% -10%, preferably 5%. The chicken bursa mycoplasma obtained by the fermentation method. The application of the mycoplasma synoviae in the inactivated vaccine of mycoplasma synoviae is provided. Compared with the prior art, the application has the technical effects that: According to the application, poloxamer is added into th