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Duck reovirus causing duck spleen necrosis as well as inactivated vaccine and application thereof

A technology of reovirus and inactivated vaccine is applied in the field of duck reovirus and its inactivated vaccine, which can solve the economic loss of the breeding duck breeding industry, the egg production of breeding ducks and the weight loss of meat ducks, and the reduction of meat ducks. The problem of reducing the pass rate of slaughtering and other problems has achieved the effect of preventing and controlling infection and outbreak, good commercialization development prospects, and good safety.

Active Publication Date: 2018-11-30
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0003] Since 2017, a large-scale outbreak of infectious diseases with spleen necrosis as the main symptom has occurred in land ducks in Shandong, Hebei, Henan, Anhui and Jiangsu. The disease mainly causes spleen enlargement, hemorrhage and necrosis in ducks of all ages, resulting in duck breeding The weight of eggs and meat ducks decreases, the feed-to-meat ratio increases, and the pass rate of meat ducks drops significantly, causing serious economic losses to the meat duck and duck breeding industry
[0004] At present, there is no effective preventive or therapeutic measure for the prevalence of this infectious disease, and conventional antiviral and antibacterial treatments are ineffective; it is also commonly used to prevent the occurrence of this infectious disease through immunization with attenuated live vaccines or inactivated virus vaccines One of the treatment methods, but the premise is that pathogenic strains with good immunogenicity and stable titer need to be isolated

Method used

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  • Duck reovirus causing duck spleen necrosis as well as inactivated vaccine and application thereof
  • Duck reovirus causing duck spleen necrosis as well as inactivated vaccine and application thereof
  • Duck reovirus causing duck spleen necrosis as well as inactivated vaccine and application thereof

Examples

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Effect test

preparation example Construction

[0042] In one embodiment of the present invention, the method for preparing an inactivated vaccine for preventing or treating duck spleen necrosis is provided, including the following steps:

[0043] (1) Duck reovirus with the deposit number CCTCC NO: V201843 was inoculated into the chicken liver cancer cell (LMH) cell line. After the duck reovirus was proliferated and cultured, the cells were broken by freezing and thawing, and the supernatant was collected , And purify to obtain the virus liquid of duck reovirus strain;

[0044] (2) Inactivate the virus liquid of the duck reovirus strain, add Tween-80 and mix it as the water phase, and mix the white oil, aluminum stearate and Span-80 as the oil phase. The water phase and the oil phase are in a volume ratio 1:2 is evenly mixed and emulsified to obtain an inactivated vaccine to prevent or treat duck spleen necrosis.

[0045] In the preparation method of the above-mentioned inactivated vaccine, the LMH cell line is used to culture th...

Embodiment 1

[0049] Example 1: Isolation and identification of duck reovirus strain

[0050] 1. Virus isolation:

[0051] (1) Necrosis of the spleen tissue of diseased ducks in a duck farm in Tai'an City, Shandong Province was placed in a 15 mL centrifuge tube, and 5 times the volume of serum-free DMEM medium was added. After homogenization, it was repeatedly frozen and thawed 3 times, each time thawing After shaking on a shaker for 1-2 minutes, perform the next freeze-thaw; after freeze-thaw, centrifuge the 15mL centrifuge tube containing the sample in a centrifuge at 4000 rpm for 5 minutes; take the supernatant, filter it with a 0.22μm microporous membrane, and set aside ;

[0052] (2) According to the proliferation characteristics of reo, chicken liver cancer cells (LMH) were selected for virus isolation. According to the conventional cell culture method, wait for 25cm 2 When the cells in the cell flask cover the monolayer, aspirate the culture medium in the flask and wash the cells twice wi...

Embodiment 2

[0064] Example 2: Animal regression test

[0065] Take 40 healthy 1-day-old ducklings that have passed pathogen and antibody testing and do not carry new reovirus and virus antibodies. They are randomly divided into 2 groups with 20 in each group. Among them, the first group is the experimental group, the experimental group uses the fifth generation cell culture of the strain N-DRV-XT18 to be inoculated through the paw pad, 0.5mL / grass, the second group is the control group, each paw pad is inoculated with DMEM medium as Control, reared in isolation. After inoculation, the mental state of each group of animals and the spleen lesions were observed and recorded daily.

[0066] The experimental group began to develop splenic necrosis lesions 4 days after inoculation. On the 7th day after inoculation, 20 experimental groups all developed splenic necrosis lesions, which had the same symptoms as natural infection cases. The control group was in good health. The diseased spleen of the e...

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Abstract

The invention discloses a duck reovirus causing duck spleen necrosis as well as inactivated vaccine and an application thereof. The duck reovirus is preserved in China Center for Type Culture Collection on July 18, 2018, with preservation number of CCTCC NO: V201843. The duck reovirus provided by the invention shows excellent immunogenicity on duck reovirus taking the spleen necrosis as a major symptom. The inactivated vaccine, which is prepared by the duck reovirus strain, is good in safety; and a 100% protecting rate can be achieved, so that newly separated variant strains can be completelyprotected.

Description

Technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a duck reovirus that causes duck spleen necrosis and an inactivated vaccine and application thereof. Background technique [0002] The genome of reovirus is a double-stranded segmented RNA. In 1978, the International Commission for Virus Isolation (ICTV) designated it as the Reoviridae family (Reoviridae), including the genus Reoviridae, Circovirus and Rot Virus genus. Avian reovirus (Avian reovirus) was isolated in the respiratory tract of chickens for the first time. After infection with susceptible chickens, it can show symptoms such as tendinitis, synovitis and respiratory diseases. ARV can infect chickens, turkeys, ducks, geese, parrots, pigeons and other birds. It can cause viral arthritis and tenosynovitis in chickens. It is also the cause of other diseases, including myocarditis, pericarditis, pericardial effusion, hepatitis, and thymic atrophy. , Resp...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/15A61P31/14C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61P31/14C12N7/00C12N2720/12221C12N2720/12234
Inventor 刁有祥唐熠王鸿志
Owner SHANDONG AGRICULTURAL UNIVERSITY
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