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A kind of duck reovirus causing duck spleen necrosis and its inactivated vaccine and application

A technology of reovirus and inactivated vaccine, which is applied in the field of duck reovirus and its inactivated vaccine, can solve the economic loss of meat duck breeding industry, the egg production of breeding ducks and the weight loss of meat ducks, and the Problems such as the lower qualified rate of slaughtering have achieved the effect of preventing infection and outbreaks, good commercial development prospects, and good safety.

Active Publication Date: 2020-10-16
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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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  • A kind of duck reovirus causing duck spleen necrosis and its inactivated vaccine and application
  • A kind of duck reovirus causing duck spleen necrosis and its inactivated vaccine and application
  • A kind of duck reovirus causing duck spleen necrosis and its inactivated vaccine and application

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preparation example Construction

[0042] In one embodiment of the present invention, the preparation method of the inactivated vaccine of prevention or treatment duck spleen necrosis that provides, comprises the following steps:

[0043] (1) Inoculate the duck reovirus with the preservation number CCTCC NO: V201843 into the chicken liver cancer cell (LMH) cell line, proliferate and culture the duck reovirus, break the cells by freezing and thawing, and collect the supernatant , and purified to obtain duck reovirus strain virus liquid;

[0044](2) Inactivate the virus liquid of duck reovirus strain, add Tween-80 to mix as the water phase, mix white oil, aluminum stearate and Span-80 as the oil phase, and mix the water phase and the oil phase by volume 1:2 mixed evenly and emulsified to obtain an inactivated vaccine for preventing or treating duck spleen necrosis.

[0045] In the preparation method of above-mentioned inactivated vaccine, adopt LMH cell line to cultivate virus, this cell line has very good susce...

Embodiment 1

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

[0050] 1. Virus isolation:

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

[0052] (2) According to the proliferation characteristics of reovirus disease, 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 are covered with a monolayer, discard the culture medium in the flask and wash the cells twice ...

Embodiment 2

[0064] Embodiment 2: animal regression test

[0065] 40 healthy 1-day-old ducklings that did not carry novel reovirus and virus antibodies after pathogen and antibody detection were randomly divided into 2 groups, 20 in each group. Wherein the first group is the experimental group, and the experimental group adopts the 5th generation cell culture of the virus strain N-DRV-XT18 to inoculate through the claw pad, 0.5mL / only, the second group is the control group, and each claw pad is inoculated with DMEM medium as The control was kept in isolation. After inoculation, the mental state and spleen lesions of the animals in each group were observed and recorded every day.

[0066] The experimental group began to develop spleen necrosis 4 days after virus inoculation, and on the 7th day after inoculation, all 20 experimental groups developed spleen necrosis, which was the same as that of natural infection cases, and 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 causing duck spleen necrosis, an inactivated vaccine and application thereof. Background technique [0002] The genome of reovirus is a double-strand segmented RNA, which was designated as Reoviridae by the International Committee for Isolation of Viruses (ICTV) in 1978, including the genus Reovirus, Circovirus and Rotavirus. Symptovirus. Reovirus (Avian reovirus) was isolated from the respiratory tract of chickens for the first time. After infecting susceptible chickens, symptoms such as tendinitis, synovitis and respiratory diseases can be manifested. ARV can infect chickens, turkeys, ducks, geese, parrots, pigeons and other poultry. It can cause viral arthritis and tenosynovitis in chickens. It is also the cause of other diseases, including myocarditis, pericarditis, hydropericardium, hepatitis, and thymus atrophy. , respiratory disease...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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