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Preparation method and application of specific yolk antibody resisting mycoplasma synoviae diseases

A technology of mycoplasma synovialum and yolk antibody, which is applied in the direction of antibodies, chemical instruments and methods, specific peptides, etc., can solve the problems of animal dependence, drug treatment restrictions, and clearing plan conflicts, and achieve easy large-scale production. Toxic and side effects, low cost effect

Inactive Publication Date: 2020-08-25
NINGXIA UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] MS is mainly transmitted through contact, and once the infection is life-long carrier, there is no effective treatment at present. Antibiotic treatment can significantly reduce the infection level of MS and recover the losses caused by the disease, but it cannot eradicate MS. Once the drug is stopped, the infection may immediately Exacerbation, and overdose will not only cause dependence in animals, but also increase the drug resistance of MS and cause drug residue problems, so the use of drug treatment in production is increasingly limited
Vaccination is a long-term solution to control mycoplasma infection. Unfortunately, only Australia currently has a mutation-selected attenuated vaccine. The immune effect of this vaccine has been confirmed by research. Conflicts have arisen and it has been banned in many countries, and there is no commercialized mycoplasma vaccine on the market in China

Method used

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  • Preparation method and application of specific yolk antibody resisting mycoplasma synoviae diseases
  • Preparation method and application of specific yolk antibody resisting mycoplasma synoviae diseases
  • Preparation method and application of specific yolk antibody resisting mycoplasma synoviae diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: the preparation of chicken synovial bursa mycoplasma antigen

[0025] 1. Materials and methods

[0026] 1.1 Strains and experimental animals

[0027] The strain of Mycoplasma synovialum was obtained from the Clinical Veterinary Laboratory of the Agricultural College of Ningxia University; 10 20-week-old sea blue and brown chicks were provided by Ningxia Xiaoming Poultry Co., Ltd.

[0028] 1.2 Media and reagents

[0029] Improved Frey's medium, purchased from Beijing Zhonghai Biotechnology Co., Ltd.; Marcol-52 white oil adjuvant, purchased from Nanjing Tianbang Biotechnology Co., Ltd.; common nutrient agar medium and blood agar medium; both purchased from Qingdao Haibo Biological Technology Co., Ltd.

[0030] 1.3 Determination of the concentration of mycoplasma in chicken synovial bursa

[0031] After recovery, the strain was inoculated in the modified Frey's liquid medium at a 10% inoculum size, and cultured at 37°C with constant temperature and shakin...

Embodiment 2

[0044] Embodiment 2: the preparation of anti-Mycoplasma chicken synovial bursa specific egg yolk antibody

[0045] 1. Materials and methods

[0046] 1.1 Main reagents and instruments

[0047] The mycoplasma strain of chicken synovial bursa was isolated and preserved by the clinical veterinary laboratory of Ningxia University; Marcol-52 white oil adjuvant was purchased from Nanjing Tianbang Biotechnology Co., Ltd.; anhydrous sodium sulfate (analytical pure); Cellulose phthalic acid (HPMCP); chicken mycoplasma synovial bursa antibody detection kit (code CK118), purchased from Beijing IDESI Yuanheng Biotechnology Co., Ltd.; rabbit anti-chicken IgY, protein markers were purchased from Pierce company ; Improved Frey's medium, purchased from Beijing Zhonghai Biotechnology Co., Ltd.;

[0048] 1.2 Experimental animals and immunization

[0049] Experimental animals: 30 20-week-old sea blue and brown laying hens (non-mycoplasma synovialum infection), provided by Ningxia Xiaoming Poul...

Embodiment 3

[0065] Example 3: Application Effect Evaluation of Anti-Mycoplasma Gallinarum-specific Egg Yolk Antibody Injection

[0066] 1. Test steps and methods

[0067] 1. Establishment of pathological model of Mycoplasma synovialis

[0068] 1.1 Preparation of bacterial suspension for challenge

[0069] Use the modified Frey's medium to revive and cultivate the Mycoplasma chicken synovial bursa strain, and expand the culture after three generations, stop the culture when the concentration of the bacteria solution reaches 106CCU / mL, centrifuge at 10000r / min for 25min, discard the supernatant and take the precipitate, and use PBS After washing with buffer three times, resuspend to the original volume as the bacterial suspension for challenge.

[0070] 1.2 Grouping and challenge of experimental animals

[0071] Fifty 5-week-old healthy chicks whose anti-Mycoplasma synovialum antibody test was negative were selected as experimental animals and divided into three groups, the experimental ...

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Abstract

The application discloses a preparation method of a specific yolk antibody resisting mycoplasma synoviae diseases. The preparation method comprises the following steps of recovering mycoplasma synoviae strains, performing propagation culture for three generations, performing sufficient inactivation, performing centrifuging, and taking precipitate so as to obtain a mycoplasma synoviae antigen; performing mixed emulsification on a mycoplasma synoviae antigen solution and a white oil adjuvant, performing basic immunization on each hen, and performing injection for 0.5ml; and after the basic immunization, performing strengthened immunization twice, wherein an immunization method and the dosage are the same as those of the basic immunization, after the first-time strengthened immunization, beginning to collect immunized eggs, disinfecting the obtained immunized eggs with alcohol, breaking shells, collecting yolk, and performing purifying to obtain the specific yolk antibody. The specific yolk antibody resisting mycoplasma synoviae diseases, prepared by the preparation method can effectively prevent and treat the mycoplasma synoviae diseases. The specific yolk antibody is easy in mass production and low in cost, and an obtained product is a safe preparation resisting the mycoplasma synoviae diseases.

Description

technical field [0001] The present application relates to the field of biotechnology, in particular to a preparation method and application of a specific egg yolk antibody against mycoplasma synovial bursa disease. Background technique [0002] Mycoplasma synoviae disease is an acute or chronic infectious disease of poultry caused by Mycoplasma synoviae (MS). It is characterized by joint exudative synovial meningitis and tenosynovitis, hence the name Mycoplasma synovialum. MS mainly infects commercial layer chickens, broiler chickens and breeder chickens, and chickens will cause obvious lameness, growth retardation and carcass degradation after infection. MS infection was first reported in the United States in the 1950s, followed by subsequent reports of the disease in Australia, Asia, Africa, and South America. MS infection can also cause subclinical respiratory symptoms, arthritis, synovitis, air sacculitis, genital tract inflammation, and egg tip syndrome in poultry, ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02A61K39/40A61P31/04
CPCA61K2039/505A61P31/04C07K16/1253C07K2317/11
Inventor 何生虎姜肖军郭磊郭亚男魏晓明李勇祁燕蓉
Owner NINGXIA UNIVERSITY
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