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A Strain of Mycoplasma bovis and Its Application

A technology of mycoplasma bovis and system, applied in the field of cattle vaccines, can solve the problems of micro-effect, etc., and achieve the effect of simple preparation process, good application prospects, and reliable immune protection effect

Active Publication Date: 2019-01-01
福清市默克兽医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, conventional antibacterial drugs are generally used to treat M. bovis-related diseases, but the effect is minimal

Method used

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  • A Strain of Mycoplasma bovis and Its Application
  • A Strain of Mycoplasma bovis and Its Application
  • A Strain of Mycoplasma bovis and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Isolation and Identification of Mycoplasma bovis

[0020] Bacteria isolation and culture, animal regression test and 16S rRNA gene sequencing were carried out on the clinical disease materials, and three strains of Mycoplasma bovis (Mycoplasma bovis) causing bovine pneumonia were isolated, which were named Mycoplasma bovis FJ1201 (MbovFJ1201), Mycoplasma bovis FJ1301 (MbovFJ1301) and Mycoplasma bovis FJ1501 (MbovFJ1501). The results of 16S rRNA gene sequence comparison showed that the sequence similarity of MbovFJ1201, MbovFJ1301 and MbovFJ1501 was 100%, indicating that the three isolates were the same strain.

[0021] 1 Materials and methods

[0022] 1.1 Materials

[0023] 1.1.1 Disease materials: from multiple cattle farms in Fuzhou City, Changle City, Xiamen City, Nanping City and Sanming City in Fujian Province from 2012 to 2015. The clinical disease materials are lung tissues of cattle with severe pneumonia or dead cattle , mainly collect necrotic focus and surr...

Embodiment 2

[0047] Vaccine preparation and safety testing

[0048] 1. Preparation of the vaccine:

[0049] The Mycoplasma bovis MbovFJ1201 strain was inoculated into PPLO liquid medium and placed at 37°C in 5% CO 2 Cultivate in an incubator, collect the culture after 5 days, and use it as a seed solution. The seed solution was inoculated again into PPLO liquid medium at 3.0% (V / V) at 37°C in 5% CO 2 After cultivating for 5 days, the cultured vaccine bacteria liquid is obtained. Adjust the number of bacteria in the vaccine liquid to 2×10 8 ccu / mL, add formaldehyde at a final concentration of 0.2%, and inactivate at 4°C for 72 hours. After the vaccine bacteria liquid is inactivated, ISA-206 adjuvant is added at a volume ratio of 1:1 (V / V), and mixed to prepare the inactivated Mycoplasma bovis vaccine.

[0050] 2. Vaccine safety testing:

[0051] a. Sterility test: take 0.2mL of Mycoplasma bovis inactivated vaccine, spread on 3 defibrated sheep blood agar plates, incubate them upside d...

Embodiment 3

[0056] Vaccine immune effect evaluation

[0057] A total of 90 healthy Holstein bull calves with similar body weight were randomly selected and divided into three groups. Vaccine group: 30 calves were intramuscularly injected with 2 mL of the vaccine of the present invention at the age of 30 days in the neck triangle, and 2 mL of the vaccine of the present invention was injected again at the age of 60 days; Inject 2 mL of sterilized normal saline, and inject 2 mL of sterilized normal saline again at the age of 60 days; normal control group: 30 calves were used as the normal control group without injection. The experimental animals were observed for 5 months, and all experimental animals adopted the same feeding and management method during the observation period.

[0058] Calculate the morbidity and mortality rate of experimental animals caused by mycoplasma during the entire observation period; calculate the average daily weight gain at the age of 2 to 4 months and 4 to 6 mo...

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Abstract

The invention relates to a mycoplasma bovis and its application. The steps are to determine the pathogen as Mycoplasma bovis through pathogen isolation and culture, animal regression test and 16S rRNA gene sequence determination, named as Mycoplasma bovis MbovFJ1201 strain, and the preservation number is CCTCC NO: M2015772. Inoculate the bacterium into the culture medium to expand the culture, harvest the culture, inactivate the culture, add ISA‑206 adjuvant at a ratio of 1:1 (V / V) after inactivation and mix to obtain an inactivated vaccine. The vaccine of the invention has strong pertinence and good immune protection effect, can significantly reduce lung lesions caused by mycoplasma bovis infection, increase average daily weight gain, and achieve good effects of preventing and controlling diseases related to mycoplasma bovis.

Description

technical field [0001] The invention relates to the field of cattle vaccines, in particular to a strain of mycoplasma bovis and its application. Background technique [0002] Mycoplasma bovis-associated diseases are important diseases that endanger the cattle industry. Mycoplasma bovis infection can cause diseases such as pneumonia, arthritis, mastitis and conjunctivitis in cattle. Since the isolation of Mycoplasma bovis was first reported in the United States in 1961, there have been outbreaks in different countries, causing serious economic losses to the cattle industry. The losses caused by pneumonia in calves caused by Mycoplasma bovis infection in the United States and Europe amount to hundreds of millions of dollars. Shi Lei and others reported the prevalence of Mycoplasma bovis in my country for the first time in 2008, and subsequent investigations and studies have shown that Mycoplasma bovis is widespread in cattle herds in China. Mycoplasma bovis not only leads t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/35
CPCA61K39/02A61K2039/54A61K2039/545C12N1/205C12R2001/35
Inventor 张德明林燕花王锦祥李金凤车勇良张梦青
Owner 福清市默克兽医院
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