Mycoplasma hyopneumoniae DJ-166 strain and application thereof

A technology of mycoplasma hyopneumoniae and DJ-166, which is applied in the direction of bacterial antigen components, biochemical equipment and methods, instruments, etc., can solve the problems of antigenic differences of isolates, achieve good immunogenicity and reduce production costs

Active Publication Date: 2013-07-03
兆丰华生物科技(南京)有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to data reports, Mycoplasma hyopneumoniae isolated from all over the world belong to the same serotype, but the antigenicity of the isolates is very different

Method used

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  • Mycoplasma hyopneumoniae DJ-166 strain and application thereof
  • Mycoplasma hyopneumoniae DJ-166 strain and application thereof
  • Mycoplasma hyopneumoniae DJ-166 strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 Mycoplasma hyopneumoniae DJ-166 strain isolation and identification of the present invention

[0023] 1 Medium and its preparation

[0024] Liquid medium:

[0025] Liquid A: 2.0g brain heart extract, 5.0g PPLO broth, 300ml deionized water

[0026] Mix and stir the above ingredients to dissolve them completely, autoclave at 116°C for 20 minutes, and cool for later use;

[0027] Liquid B: 5.0ml of 10× Hank’s liquid, 1.0g of whey protein hydrolyzate, 5.0g of yeast extract, 0.8g of sodium pyruvate, Peptone 3.0g, sodium thiosulfate 0.1g, 0.1% phenol red 10ml, penicillin 400U / ml, deionized water 545ml.

[0028] Mix and stir the above ingredients to dissolve them completely, filter and sterilize with a 0.22 μm filter membrane, and store at 4°C for later use;

[0029] Solution C: healthy horse serum 140ml

[0030] Thoroughly mix liquid A, liquid B and liquid C evenly, and adjust the pH to 7.6 with 1mol / L sodium hydroxide solution to prepare a liquid culture m...

Embodiment 2

[0068] Embodiment 2 Mycoplasma hyopneumoniae DJ-166 strain biological characteristics of the present invention

[0069] 1 Culture characteristics

[0070] Inoculate Mycoplasma hyopneumoniae DJ-166 strain in liquid medium, 37±1°C 5% CO 2 Under the same conditions, after 5-9 days of cultivation, the bacterial liquid was slightly turbid, and the pH dropped from 7.6 to 6.5-7.0; on solid medium, the growth was slow, and it was cultivated under 5% carbon dioxide and 37±1°C for 7-10 days. Needle-sized colonies can be seen. Under the microscope, the colonies can be seen to have neat edges, loose outer texture, and dense central colonies; the bacterial liquid of Mycoplasma hyopneumoniae DJ-166 was inoculated into the medium containing penicillin and thallium acetate, and the growth of the bacterial liquid was not inhibited.

[0071] 2 Morphological and biochemical properties

[0072] After Giemsa staining of the bacterial liquid smear, the oil microscope observation showed pleomorphi...

Embodiment 3

[0088] Embodiment 3 Mycoplasma hyopneumoniae DJ-166 strain living bacteria titer determination

[0089] Mycoplasma hyopneumoniae DJ-166 strain is in the cell-free culture medium of preparation (preparation raw material and method see embodiment 1), viable bacterial titer measurement result is 10 10 -10 11 CCU / ml, see Table 1.

[0090] Table 1 The results of three live bacteria concentration determinations of Mycoplasma hyopneumoniae DJ-166 strain

[0091]

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Abstract

The invention belongs to the field of veterinary microbial technology, and particularly relates to a mycoplasma hyopneumoniae DJ-166 strain and an application thereof. The mycoplasma hyopneumoniae can be used for preparing veterinary biological products or veterinary drugs for prevention. Experiments demonstrate that the mycoplasma hyopneumoniae DJ-166 strain has high active bacterium titer reaching 10<10-11> CCU / mL in a prepared cell-free culture medium, can greatly reduce production cost, and has good immunogenicity with an average pneumonia pathology reduction ratio reaching over 80%.

Description

technical field [0001] The invention belongs to the technical field of veterinary microorganisms, in particular to mycoplasma hyopneumoniae DJ-166 strain and application thereof. Background technique [0002] Mycoplasma swine pneumonia, also known as swine panting disease, is a contact chronic respiratory disease of pigs caused by Mycoplasma hyopneumoniae, which is prevalent in the world. According to reports, Mycoplasma hyopneumoniae isolated from all over the world belong to the same serotype, but the antigenicity of the isolates is very different. In 1992, Frey first confirmed the antigenic difference between isolates of Mycoplasma hyopneumoniae; in 1996, Artiushin and Minion further confirmed Frey's point of view; in 1999, Kokotovic also confirmed this conclusion. Therefore, it is very important to isolate a Mycoplasma hyopneumoniae strain with better antigenicity for vaccine and diagnostic kit application, especially to isolate a strain suitable for the preparation of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04G01N33/569C12R1/35
Inventor 车艳杰王海燕王勇鹣张锋高玉梅赵亚荣
Owner 兆丰华生物科技(南京)有限公司
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