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Mycoplasma ovipneumoniae strain and application thereof in screening of anti-mycoplasma ovipneumoniae preparation

A technology of mycoplasma pneumoniae and mycoplasma, applied in the field of microorganisms, can solve the problems that hinder the rapid and efficient screening of anti-mycoplasma drugs and preparations, slow growth and discoloration, and strict nutritional requirements, so as to achieve easy and rapid screening and application, fast growth and low cost Effect

Active Publication Date: 2021-06-25
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] It can be seen from the above reports that because Mycoplasma ovis pneumoniae has strict nutritional requirements, it is difficult to cultivate, and the growth and discoloration speed is slow, which takes 3-10 days, which hinders the rapid and efficient screening of anti-mycoplasma drugs and preparations

Method used

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  • Mycoplasma ovipneumoniae strain and application thereof in screening of anti-mycoplasma ovipneumoniae preparation
  • Mycoplasma ovipneumoniae strain and application thereof in screening of anti-mycoplasma ovipneumoniae preparation
  • Mycoplasma ovipneumoniae strain and application thereof in screening of anti-mycoplasma ovipneumoniae preparation

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Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1: Obtaining and culturing identification of Ovine Mycoplasma pneumoniae NJ01 strain

[0036] 1. Strain isolation and culture

[0037] Select sheep with suspected characteristics of Mycoplasma ovis pneumonia from a sheep farm in Nanjing, collect typical diseased sheep with Mycoplasma ovis pneumonia characteristics, collect their lungs aseptically, inject the mycoplasma liquid medium directly into the bronchiole, and then suck it to obtain the bronchi Lavage fluid. The lavage solution can be directly filtered with a filter with a pore size of 0.45 μm, and then diluted with 1:1 and 1:10 to inoculate the mycoplasma liquid medium, and cultured at 37°C. When the color of the culture changes from red to orange or yellow, Subculture was carried out in this way for 5 consecutive generations. After the growth was stable, the mycoplasma solid medium was inoculated for purification. After the clone was purified, the mycoplasma liquid medium was inoculated, and the cult...

Embodiment 2

[0046] Embodiment 2: culture test of ovine mycoplasma pneumoniae

[0047] 1. Materials

[0048] Ovis Mycoplasma pneumoniae NJ01 strain; the deposit number is CCTCC NO: M 2020907. Ovine Mycoplasma pneumoniae Y98 strain, reference strain.

[0049] Mycoplasma medium formula and preparation method are: Eagles solution 50% (v / v), hydrolyzed milk protein 0.02g / mL, phosphate buffer (pH 7.4) 28.8% (v / v), 0.04g / mL phenol red 0.2 % (v / v); use NaOH to adjust the pH value to 7.4-7.6, after autoclaving, add 20% (v / v) of sterile horse serum and 1% (v / v) of fresh yeast extract. Wherein, the fresh yeast extract juice is prepared with reference to the method of "Zhang Ruiting et al. Improvement of Extraction Method of Yeast Extract. Chinese Journal of Veterinary Medicine. Issue 02, 1998".

[0050] 2. Method

[0051] (1) Take out the strains (Mycoplasma NJ01 strain and Mycoplasma Y98 strain) frozen at -20°C respectively, inoculate them in the Mycoplasma liquid culture medium at a volume rat...

Embodiment 3

[0064] Embodiment 3: the determination of drug susceptibility test of Mycoplasma pneumoniae

[0065] 1. Materials

[0066] Ovis Mycoplasma pneumoniae NJ01 strain; the deposit number is CCTCC NO: M 2020907. Ovine Mycoplasma pneumoniae Y98 strain, reference strain.

[0067] Mycoplasma medium formula and preparation method are: Eagles solution 50% (v / v), hydrolyzed milk protein 0.02g / mL, phosphate buffer (pH 7.4) 28.8% (v / v), 0.04g / mL phenol red 0.2 % (v / v); use NaOH to adjust the pH value to 7.4-7.6, after autoclaving, add 20% (v / v) of sterile horse serum and 1% (v / v) of fresh yeast extract. Wherein, the fresh yeast extract juice is prepared with reference to the method of "Zhang Ruiting et al. Improvement of Extraction Method of Yeast Extract. Chinese Journal of Veterinary Medicine. Issue 02, 1998".

[0068] Antibacterial drugs: Tylosin and doxycycline were purchased from Mdbio.

[0069] 2. Culture test

[0070] (1) Take Mycoplasma pneumoniae NJ01 and Y98 strains stored at...

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Abstract

The invention discloses a mycoplasma ovipneumoniae strain and application thereof in screening of an anti-mycoplasma ovipneumoniae preparation. The classification name of the mycoplasma ovipneumoniae strain is Mycoplasma ovipneumoniae NJ01 strain, the mycoplasma ovipneumoniae strain is preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: M 2020907, and the preservation date is December 14, 2020. The strain NJ01 provided by the invention is high in culture growth speed and high in titer which can reach 10<10> CCU / mL, and the color can be changed in 10-72 hours after the strain NJ01 is subjected to recovery subculture in a specific culture medium, so that time is saved for determination of an anti-mycoplasma ovipneumoniae preparation, the efficiency is improved, and the strain NJ01 can be used for rapid screening of the anti-mycoplasma ovipneumoniae preparation and the like.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of Mycoplasma pneumoniae and its application in screening anti-Mycoplasma pneumoniae preparations. [0002] technical background [0003] Mycoplasma pneumoniae of sheep and goat (MPSG) is a chronic respiratory infectious disease of sheep and goats caused by Mycoplasma ovipneumoniae (Mo). The disease is mainly clinically characterized by asthma, and it is widely prevalent in the world. Goats and sheep are more susceptible and almost universal, causing heavy losses to the sheep industry. Due to the poor immune effect of the Mycoplasma pneumoniae vaccine in sheep, the use of anti-Mycoplasma drugs and other preparations has become an important means of prevention and treatment of the disease. [0004] At present, the phenomenon of bacterial drug resistance, poor curative effect or ineffectiveness caused by the extensive use of antibacterial drugs has become a thorny p...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/18C12Q1/04C12R1/35
CPCC12Q1/18C12Q1/045
Inventor 刘茂军毛立李文良杨蕾蕾张纹纹孙敏杜改梅袁万哲马玉忠程子龙
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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