Mycoplasma hyopneumoniae dj-166 strain and its application
A technology of Mycoplasma hyopneumoniae, DJ-166, applied in bacterial antigen components, biochemical equipment and methods, instruments, etc., can solve problems such as the antigenic difference of isolates, achieve the effect of good immunogenicity and reduce production costs
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Embodiment 1
[0023] Embodiment 1 Mycoplasma hyopneumoniae DJ-166 strain isolation and identification of the present invention
[0024] 1 Medium and its preparation
[0025] Liquid medium:
[0026] Liquid A: 2.0g brain heart extract, 5.0g PPLO broth, 300ml deionized water
[0027] Mix and stir the above ingredients to dissolve them completely, autoclave at 116°C for 20 minutes, and cool for later use;
[0028] 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.
[0029] 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;
[0030] Solution C: healthy horse serum 140ml
[0031] 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
[0070] Embodiment 2 Mycoplasma hyopneumoniae DJ-166 strain biological characteristics of the present invention
[0071] 1 Culture characteristics
[0072] Inoculate the DJ-166 strain of Mycoplasma hyopneumoniae in liquid culture medium, and culture it for 5-9 days under the condition of 37±1℃5%CO2, the bacterial liquid is slightly turbid, and the pH drops from 7.6 to 6.5-7.0; On the base, the growth is slow, cultured at 5% carbon dioxide, 37±1°C for 7-10 days, needle-sized colonies can be seen, observed under a microscope, the colonies can be seen to have neat edges, loose texture around the periphery, and dense colonies in the center; Mycoplasma hyopneumoniae DJ-166 strain The growth of the bacterial liquid was not inhibited when the bacterial liquid was inoculated in the medium containing penicillin and thallium acetate.
[0073] 2 Morphological and biochemical properties
[0074] After Giemsa staining of the bacterial liquid smear, the oil microscope observation showed pl...
Embodiment 3
[0090] Embodiment 3 Mycoplasma hyopneumoniae DJ-166 strain living bacteria titer determination
[0091] In the prepared cell-free medium of Mycoplasma hyopneumoniae DJ-166 strain (see Example 1 for the preparation of raw materials and methods), the assay result of viable bacterial titer was 1010-1011 CCU / ml, as shown in Table 1.
[0092] Table 1 The results of three live bacterial titer determinations of Mycoplasma hyopneumoniae DJ-166 strain
[0093]
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