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Breeding method, application and screening method of mycoplasma synoviae biofilm

A technology of mycoplasma synovialis and mycoplasma organisms, which is applied in the field of pathogen isolation and identification and pathogen culture, can solve the problems such as the lack of formation of chicken mycoplasma synovial mycoplasma biofilm growth form, the inability of theoretical dosage to guide practical applications, and the like, achieving antibacterial Active Accurate Effects

Active Publication Date: 2022-07-15
FOSHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, for the in vitro antibacterial activity screening of Mycoplasma gallisovus of antibacterial drugs, the Mycoplasma gallisovesus used is generally prepared by simply suspension culture, and the growth form of Mycoplasma gallisovesus biofilm has not been formed.
However, it is generally believed that the Mycoplasma gallisyoviosus obtained from cultivation is the Mycoplasma gallisovus present in the environment of live chickens, which makes the results of the antibacterial activity of the screened drugs different from the actual application, and the theoretical dosage is not enough guide practical application

Method used

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  • Breeding method, application and screening method of mycoplasma synoviae biofilm
  • Breeding method, application and screening method of mycoplasma synoviae biofilm
  • Breeding method, application and screening method of mycoplasma synoviae biofilm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Separation of Mycoplasma gallis synovium and cultivation of Mycoplasma gallis synovium biofilm, comprising the following steps:

[0066] (1) Isolation and culture of MS: 148 sick chickens suspected of MS infection found in different regions were killed by bloodletting from the neck, amputated from the hip joint, sterilized with 75% alcohol and marked, and used in the ultra-clean workbench. Samples such as arthritic exudates were collected and inoculated into vials containing 3 mL of modified Frey's liquid medium. After the inoculation is completed, cover the bottle stopper, and set up a blank control group at the same time, seal the vial with parafilm, and place it at 37 °C for cultivation. When the color of the medium changes from red to yellow or orange, subculture immediately. A culture capable of stable passage is obtained.

[0067] (2) Purification of MS: Dilute the MS bacterial solution grown to the logarithmic growth phase by 1000 times. Take 50 μL of the dilut...

Embodiment 2

[0084] Isolation of Mycoplasma gallis synovium and cultivation of Mycoplasma gallis synovium biofilm

[0085] The difference between Example 2 and Example 1 is: Example 2 is also preserved by freeze-drying; specifically, the following steps are included:

[0086] (1) Isolation and culture of MS: 148 sick chickens suspected of MS infection found in different regions were sacrificed by bloodletting from the neck, amputated from the hip joint, sterilized with 75% alcohol and marked, and used in the ultra-clean workbench. Samples such as arthritic exudates were collected and inoculated into vials containing 3 mL of modified Frey's liquid medium. After the inoculation is completed, cap the bottle, and set up a blank control group. Seal the vial with parafilm and place it at 37 °C for cultivation. When the color of the medium changes from red to yellow or orange, subculture immediately. A culture capable of stable passage is obtained.

[0087] (2) Purification of MS: Dilute the MS...

Embodiment 3

[0092] The effect of temperature on the growth of MS biofilm, including the following steps:

[0093] (1) Cultivation of biofilm (BF) at different temperatures: add in 96-well plates and dilute to 10 with modified Frey's liquid medium 6 CFU / mL standard strain WVU185 bacterial liquid, and blank medium was used to set up a control group. After completion, they were cultured at 25°C, 30°C, 33°C, 37°C, and 40°C respectively. After the medium changed from red to yellow Take out, and each group of experiments is repeated 3 times.

[0094] (2) Counting of biofilm (BF) biomass: Take out the above-mentioned 96-well plate with well-grown BF, aspirate the medium in the ultra-clean bench and rinse with sterile 0.1M phosphate buffer (pH7.0) for 2- The residual medium and suspended bacteria were washed 3 times. After washing, 200 μL of fresh medium (modified Frey's liquid medium) was added to disperse MS in BF by repeated pipetting with a micropipette, and the dispersed bacterial liquid w...

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Abstract

The invention relates to the technical field of pathogen separation and identification and pathogen culture, in particular to a breeding method, application and screening method of a mycoplasma synoviae biofilm. The culture method of the mycoplasma synoviae biofilm comprises the following steps: diluting mycoplasma synoviae growing to a logarithmic phase to (0.9-1.5) * 10 < 6 > CFU / mL by using a liquid culture medium, carrying out first culture, growing to the logarithmic phase, and removing suspended thalli to prepare the mycoplasma synoviae biofilm; the liquid culture medium is an improved Frey liquid culture medium, and the judgment standard from the growth to the logarithmic phase is that the improved Frey liquid culture medium turns yellow from red; the cultivation temperature of the first cultivation is 30-37 DEG C. According to the method, the mycoplasma synoviae biofilm can be cultured in vitro, and data measured in a biological experiment can be more accurate.

Description

technical field [0001] The invention relates to the technical field of pathogen isolation and identification and pathogen culture, in particular to a method for cultivating a Mycoplasma synovialis biofilm and a method for its application and screening. Background technique [0002] Mycoplasma synoviae (MS) infection can cause infectious synovitis in chickens, ducks, geese, turkeys and other poultry, which is characterized by joint exudative synovial membrane and tenosynovitis synovitis . MS infection can also cause respiratory diseases, but it is mainly subclinical, so it is not easy to observe. If concurrent infection with Newcastle disease or infectious bronchitis will cause airsacculitis. In addition, MS infection can cause abnormalities at the top of the eggshell. Therefore, MS-infected chickens often exhibit symptoms such as lameness, swollen joints, and respiratory tract inflammation, which in turn lead to decreased egg production, stunted growth, and reduced feed co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C12Q1/18C12R1/35
CPCC12N1/20C12Q1/04C12Q1/18
Inventor 张楠康天灏周明虎严秀宋姝缇
Owner FOSHAN UNIVERSITY