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Chicken infectious bronchitis divalent live vaccine and preparation method thereof

A bronchitis and chicken infectious technology, applied in the field of chicken infectious bronchitis bivalent live vaccine and its preparation, can solve the problem of no prevention and achieve the effect of reducing immune stress

Inactive Publication Date: 2014-02-26
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a kind of bivalent live vaccine of chicken infectious bronchitis, realize the effect that can effectively prevent respiratory type and kidney type chicken infectious bronchitis simultaneously, solve the above-mentioned two types of chickens that current domestic existing IBV vaccine Infectious bronchitis can only provide partial prevention or even no preventive effect, so as to avoid the widespread occurrence and prevalence of chicken infectious bronchitis to a greater extent

Method used

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  • Chicken infectious bronchitis divalent live vaccine and preparation method thereof
  • Chicken infectious bronchitis divalent live vaccine and preparation method thereof
  • Chicken infectious bronchitis divalent live vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Screening of Chicken Infectious Bronchitis Kidney Type K136 Strain

[0026] From 2003 to 2005, our company collected suspected chicken infectious bronchitis virus isolates from different chicken farms in my country. The sick chickens mainly manifested as lack of energy, coughing, and often heard grunting. Pathological autopsy found that there were obvious lesions in the kidneys in addition to small bleeding spots in the tracheal ring. After preliminary identification, 4 strains of renal-type infectious bronchitis virus were obtained. They were named CK / IBV / QD03 strain, CK / IBV / ZC03 strain, CK / IBV / YT04 strain and CK / IBV / SD05 strain respectively. Use 10-day-old SPF chicken embryos to carry out continuous subculture to attenuate the four strains. In each generation, 4 chicken embryos were inoculated through the allantoic cavity, incubated at 37°C, and the chicken embryos were frozen to death 36 hours after inoculation, and the allantoic fluid was harvested for th...

Embodiment 2

[0035] Embodiment 2: K136 strain safety detection

[0036] 1. Determination of toxicity of K136 strains of different generations on chicken embryos

[0037] Take the E2, E4, E7, E10, E15 generation virus seeds of the K136 strain, make appropriate multiple dilutions, inoculate 10-day-old SPF chicken embryos through the allantoic cavity, 0.1 mL per embryo, and set a blank control group at the same time, The cavity was inoculated with 0.1 mL of physiological saline, and incubated at 37°C. The chicken embryos that died within 24 hours were discarded, and after 144 hours of incubation, the embryos were taken out to observe the development of the chicken embryos. Calculation of EID according to the Reed-Muench method 50 , the poisonous price of E2, E4, E7, E10, E15 generation poison species is respectively: 10 6.7 、10 6.8 、10 6.5 、10 6.6 、10 6.8 EID 50 / 0.1mL. The test results showed that the K136 strain proliferated stably on chicken embryos, and there was no reversion of ...

Embodiment 3

[0044] Example 3: Evaluation of immune efficacy of different generations of K136 strain

[0045] Use the E2, E4, E7, E10, E15 generation K136 strains to breed chicken embryo allantoic fluid in 5×10 2.0 EID 50 Dosage, intranasal drops for 1-day-old SPF chickens, 14 days after immunization, together with control chickens, each chicken used 10 3.68 EID 50 / 0.1mL of the isolated virulent CK / IBV / SD05 strain 0.05mL for eye drops and nasal drops respectively. On the 5th day after the challenge, the tracheal swabs of each chicken were collected and inoculated into the 10-day-old SPF chicken embryo 5 through the allantoic cavity. Embryo, 0.1mL / embryo, incubated for 144 hours. The results showed that the toxicity rate of the immune group was between 0 / 10-1 / 10, and the protection rate was between 9 / 10-10 / 10; the toxicity rate of the control group was 10 / 10. It shows that the immunogenicity of IBV-K136 strain virus seeds is basically the same within 15 generations, and they all have g...

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Abstract

The invention discloses a chicken infectious bronchitis divalent live vaccine and a preparation method thereof. The divalent live vaccine comprises antigens and a freeze-drying protective agent. The antigens comprise a chicken kidney-type infectious bronchitis virus K136 strain having an accession number of CCTCC-V201320 and a chicken breath-type infectious bronchitis virus H120 strain. The preparation method includes respectively performing amplification culture to the IBV-K136 strain and the H120 strain through chick embryos; mixing culturing products uniformly to prepare the vaccine antigens; and mixing uniformly the vaccine antigens and the freeze-drying protective agent and performing freeze drying to prepare the vaccine. The divalent live vaccine can prevent the chicken breath-type infectious bronchitis and the chicken kidney-type infectious bronchitis simultaneously. The vaccine conforms to the epidemiological characteristics of the chicken infectious bronchitis in our country at present, and solves a problem that no vaccine for the chicken kidney-type epidemic infectious bronchitis is available in our country at present. The divalent live vaccine can be popularized and used in a large scale.

Description

technical field [0001] The invention belongs to the technical field of new biological medicines for livestock and poultry, and in particular relates to a bivalent live vaccine for chicken infectious bronchitis and a preparation method thereof. Background technique [0002] Avian Infectious Bronchitis (IB) is an acute and highly contagious infectious disease of chickens, which mainly affects the respiratory system, genitourinary system and digestive system. The disease was first reported by Schalk and Hawn in the U.S. Clinically, the respiratory symptoms of coughing, sneezing and tracheal rales in chickens were the main ones, and then it occurred in various countries in the world. In 1936, Beach and Schalm determined that the pathogen was a virus (respiratory IBV). Named as chicken infectious bronchitis virus (Infectious Bronchitis Virus, IBV), classified into Coronaviridae, is a representative strain of coronavirus. In 1972, Kuang Ronglu and others first confirmed in Guangd...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K9/19A61P31/14A61K39/215
Inventor 孙美玉何召庆杨保收陈冰邢婧珉李云霞梁武李守军
Owner TIANJIN RINGPU BIO TECH
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