Attenuated method for fowl infectious bronchitis viruses

A bronchitis and infectious technology, which is applied in the attenuation field of chicken infectious bronchitis virus, can solve the problems of slow attenuation speed, achieve the effects of reducing loss, saving passage time, and shortening attenuation time

Inactive Publication Date: 2013-11-20
WENS FOOD GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, it is necessary to provide a weakening method of avian infectious bronchitis virus in view of the slow attenuation rate of avian infectious bronchitis virus in conventional temperature continuous subculture

Method used

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  • Attenuated method for fowl infectious bronchitis viruses
  • Attenuated method for fowl infectious bronchitis viruses
  • Attenuated method for fowl infectious bronchitis viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 continuous passage weakens

[0026] A method for attenuating avian infectious bronchitis virus. After diluting the virus strain YX10 by 10 times by 100 times, inoculate 3 SPF chicken embryos that have been incubated at the corresponding culture temperature for 10 hours, and inoculate each embryo with 103.5-4EID50 / 0.2 mL, after culturing for a certain period of time, put the chicken embryos at -20°C for half an hour, collect the allantoic fluid aseptically, and carry out the next generation and sub-passage; the culture temperature from the first generation to the tenth generation is 37°C, and the culture time is 48 hours ;The culture temperature of the 11th generation to the 14th generation is reduced by 1°C for each generation, that is, the culture temperature of the 11th generation is 36°C, the culture temperature of the 12th generation is 35°C, the culture temperature of the 13th generation is 34°C, and the culture temperature of the 14th generation is 33...

Embodiment 2

[0027] Embodiment 2 continuous passage weakens

[0028] A method for attenuating avian infectious bronchitis virus. After diluting the virus strain YX10 by 10 times to 100 times, inoculate 3 SPF chicken embryos that have been incubated at the corresponding culture temperature for 12 hours, and each embryo is inoculated with 10 3.5-4 EID 50 / 0.2mL, after culturing for a certain period of time, put the chicken embryo at -20°C for half an hour, collect the allantoic fluid aseptically, and carry out the next generation and subculture; 48 hours; from the 11th generation to the 14th generation, the culture temperature is reduced by 1°C for each generation, and the culture time is 48 hours; from the 15th generation onwards, the culture temperature is 32°C, and the culture time from the 15th generation to the 60th generation is 40 hours; From generation 61 onwards, the culture time is 32 hours.

Embodiment 3

[0029] Embodiment 3 continuous passage weakens

[0030] A method for attenuating avian infectious bronchitis virus. After diluting the virus strain YX10 by 10 times to 100 times, inoculate 3 SPF chicken embryos that have been incubated at the corresponding culture temperature for 8 hours, and each embryo is inoculated with 10 3.5-4 EID50 / 0.2mL, after culturing for a certain period of time, put the chicken embryo at -20°C for half an hour, collect the allantoic fluid aseptically, and carry out the next generation and subculture; 48 hours; from the 11th generation to the 14th generation, the culture temperature is reduced by 1°C for each generation, and the culture time is 48 hours; from the 15th generation onwards, the culture temperature is 32°C, and the culture time from the 15th generation to the 60th generation is 44 hours; From generation 61 onwards, the culture time is 36 hours.

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Abstract

The invention discloses an attenuated method for fowl infectious bronchitis viruses. The method comprises the following steps: the toxic strain YX10 is inoculated on SPF chick embryos, 103.5-4EID50 / 0.2mL of the toxic strain is inoculated on every embryo; after cultured for certain time, the chick embryos are placed at the temperature of minus 20 DEG C for half an hour; allantoic fluids are collected in sterile conditions; the passage of the next generation is carried out; the culture temperature of the first generation to the tenth generation is 37 DEG C and the culture time is 48 h; the culture temperature of the eleventh generation to the fourteenth generation is lowered by 1 degree per generation and the culture time is 48 h; from the fifteenth generation, the culture temperature is 32 DEG C, and the culture time is 32-48 h. The method can save the passage time effectively, and reduces the loss of immunogenicity. The method is beneficial to develop novel vaccines in time and effectively.

Description

technical field [0001] The invention relates to a method for attenuating virus, in particular to a method for attenuating chicken infectious bronchitis virus. Background technique [0002] Avian "kidney type" infectious bronchitis is a pathological form of avian infectious bronchitis (Avian Infectious Bronchitis), an acute, highly contagious infectious disease that combines respiratory tract and kidney lesions; it is characterized by Respiratory symptoms in sick chickens, decreased egg production and quality, nephropathy, and infection in young hens can cause permanent degeneration of oviducts. [0003] Avian infectious bronchitis virus (Infectious Bronchitis Virus, IBV) can infect chickens of all ages, resulting in growth retardation, death, weight gain and reduced feed remuneration; it also often causes mycoplasma mixed infection and secondary infections such as Escherichia coli to increase The mortality rate of chicken flock brings huge loss to chicken production. At pr...

Claims

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Application Information

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IPC IPC(8): C12N7/08C12R1/93
Inventor 陈峰薛瑜严专强曾凡桂覃健萍李海燕鲁俊鹏刘闯
Owner WENS FOOD GRP CO LTD
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