Fluid medium for culturing mycoplasma synoviae (MS)

A liquid medium and solution technology, applied in the field of pathogenic culture, can solve the problems of strict pH value, high nutritional requirements, high cost of medium, etc., and achieve the effect of good safety and good immune efficacy

Pending Publication Date: 2016-06-22
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many problems in the culture of chicken synovial branch, such as high cost o

Method used

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  • Fluid medium for culturing mycoplasma synoviae (MS)
  • Fluid medium for culturing mycoplasma synoviae (MS)
  • Fluid medium for culturing mycoplasma synoviae (MS)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1 prepares culture medium of the present invention according to different formulations

[0014] 1 Culture medium preparation

[0015] 1.1 Preparation of base liquid Take 21.0g of PPLO broth, 5.0g of glucose, add to 1000ml of deionized water, 1.0ml of 1% phenol red solution, mix and dissolve the above ingredients, autoclave at 116°C for 30 minutes, cool and set aside for 2-8 Store at ℃.

[0016] 1.2 Culture medium preparation After cooling the base solution, aseptically add different amounts of inactivated horse serum, 25% yeast extract, 1% coenzyme I solution, 1% L-cysteine ​​solution, and 800,000 units / ml penicillin solution in 1.2 ml, adjust the pH value to 7.6 with 1mol / L sodium hydroxide solution, and prepare 6 kinds of liquid culture medium. The specific formula is shown in Table 1.

[0017] Table 1: Liquid medium preparation methods

[0018]

Embodiment 2

[0019] The screening of embodiment 2 different formula liquid culture medium:

[0020] Mycoplasmasynoviae YBF-MS1 strain (MycoplasmasynoviaeYBF-MS1) (Mycoplasmasynoviae YBF-MS1) has been preserved in the Chinese Type Culture Collection Center located in Wuhan and Wuhan University, the preservation date is March 3, 2016, and the preservation number is : CCTCCNO:M2016080.), inoculate the above-mentioned 6 kinds of liquid culture medium by 10% respectively, put 37 ℃, 150r / min shaking culture, the culture time is 36 hours, take samples respectively and carry out live bacteria count at different culture time. The screening test results of liquid culture media with different formulas show that the liquid culture media prepared according to formula 2, formula 3, formula 5 and formula 6 have better culture effects, and the number of viable bacteria in 24 to 30 hours is not less than 10 9 CCU / ml, formula 3 and formula 6, due to the high amount of supplementary liquid components such as...

Embodiment 3

[0023] Embodiment 3: Culture medium comparison of the present invention and improved Frey's liquid medium

[0024] After recovering the freeze-dried seeds of the YBF-MS1 strain, inoculate the seed liquid with 10% of the improved Frey's liquid medium and the liquid medium of the formula five of the present invention respectively, place them at 37°C and vibrate at 150r / min for 36 hours. Samples were taken at different times of culture for counting viable bacteria. The test results show that the amount of viable bacteria at different culture times is higher than that of the improved Frey's liquid medium with different culture times when the culture medium of the present invention is used. The results are shown in Table 3.

[0025] Table 3: Selection test results of medium

[0026]

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Abstract

The invention provides a fluid medium for culturing mycoplasma synoviae (MS), being prepared from the following components: pleuropneumonia-like organism (PPLO) broth, glucose, inactivated horse serum, yeast leachate, a phenol red solution, a coenzyme I solution and an L-cysteine solution; the pH value of the fluid medium is 7.6-7.8. The fluid medium is beneficial to the growth of the MS by supplementing various nutritional ingredients needed by microorganisms and effectively controlling the pH value of the fluid medium by sodium hydroxide; viable counts are not less than 109CCU/ml; an inactivated vaccine prepared by a bacteria solution of the MS is good in safety and immunization efficacy.

Description

technical field [0001] The invention belongs to the technical field of pathogen cultivation, and in particular relates to a liquid culture medium for cultivating Mycoplasma gallinarum. Background technique [0002] Mycoplasmasynoviae (Mycoplasmasynoviae, MS), also known as chicken infectious synovitis, is an acute or chronic infectious disease caused by Mycoplasmasynoviae in chickens and turkeys. Infectious synovitis was first reported by Olson et al (1964). The disease mainly affects joint synovial fluid and tendon sheath, which is characterized by swelling of joints, tendon sheaths and soles. Infection of flocks with the disease can result in significant lameness, stunted growth and carcass degradation. If the pathogen exists in chicken flocks, it is prone to mixed infection, which will aggravate the disease, increase the mortality rate, and cause serious economic losses. The pathogen of Mycoplasma synovialum is Mycoplasma synovialum, and the bacterial form of this bact...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20
Inventor 马爽宋新宇张志鸿窦小龙郭玉广王红朱艳梅申洪银刘新文
Owner YEBIO BIOENG OF QINGDAO
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