Mycoplasma bovis and application thereof
A technology of Mycoplasma bovis and inactivated vaccine, applied in veterinary vaccines, bacterial antigen components, antibody medical components, etc., can solve the problems of ineffective vaccine prevention, poor curative effect, economic loss of cattle industry, etc., and achieve good protection effect , Good immunogenicity and strong pathogenicity
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Embodiment 1
[0019] Embodiment 1, the separation and identification of Mycoplasma bovis
[0020] 1.1 Disease data collection
[0021] The disease material came from a 2-month-old black-and-white cow that died of severe pneumonia and dyspnea in a large dairy farm in Jiyuan City, Henan Province in November 2016.
[0022] 1.2 Preparation of medium
[0023] PPLO liquid medium: PPLO broth powder 10.5g, glucose 2.5g, yeast powder 2.5g, dissolved in 440mL ultrapure water, sterilized at 115°C for 15min, added MEM medium 5mL, horse serum 50mL, penicillin 80,000 units, Sterile 10% arginine 10 mL and 1% (w / v) phenol red 500 μL. After the culture was sterilized at 115°C for 15 minutes, it was stored at 4°C for later use.
[0024] PPLO solid medium: PPLO liquid medium with 1.5% (w / v) agar powder. After the culture was sterilized at 115°C for 15 minutes, it was stored at 4°C for later use.
[0025] 1.3 Isolation and culture of mycoplasma
[0026] The collected diseased lung tissue was cut into sma...
Embodiment 2
[0046] Embodiment 2, the preparation of vaccine, safety and efficacy test
[0047] 2.1 Preparation of vaccine
[0048] The Mycoplasma bovis strain HNMB1 was inoculated into PPLO liquid medium and placed at 37°C, 5% CO 2 Cultivate in an incubator, collect the culture after 3 days, measure the concentration, and adjust the number of bacteria in the culture to 2×10 9 CFU / mL, add formaldehyde to a final concentration of 0.2%, inactivate at 37°C for 12h. The inactivated culture is added with an aluminum hydroxide adjuvant at a volume ratio of 1:1, and mixed to prepare an inactivated Mycoplasma bovis vaccine.
[0049] 2.2 Vaccine safety test:
[0050] Ten healthy black-and-white dairy cows aged 60 days were selected and divided into 2 groups with 5 cows in each group. Vaccine group: intramuscular injection of 10 mL of this vaccine in the neck; control group: intramuscular injection of 10 mL of sterilized normal saline in the neck. Observation for 30 days. During the observatio...
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