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Novel duck reovirus resulting arthritis of chicken for meat and application of novel duck reovirus

A technology for reovirus and orphan intestinal disease, applied in the field of isolation and application of poultry reovirus strains, can solve problems such as increased feed-to-meat ratio, ineffective prevention and control, and lower pass rate of meat ducks for slaughter , to achieve good immunogenicity, complete protection, and good safety

Active Publication Date: 2018-10-16
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease mainly leads to a decrease in the egg production of breeding ducks and a decrease in the weight of meat ducks, an increase in the feed-to-meat ratio, and a significant reduction in the qualified rate of meat ducks for slaughter, causing serious economic losses to the meat ducks and breeding duck breeding industry.
The commercially available duck reovirus vaccine has been unable to effectively prevent and control the prevalence of the disease, so it is inferred that the disease may be caused by a new type of reovirus

Method used

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  • Novel duck reovirus resulting arthritis of chicken for meat and application of novel duck reovirus
  • Novel duck reovirus resulting arthritis of chicken for meat and application of novel duck reovirus
  • Novel duck reovirus resulting arthritis of chicken for meat and application of novel duck reovirus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Isolation and identification of duck reovirus strain

[0031] 1. Virus isolation:

[0032] (1) The arthritic ducks were clinically identified as positive by PCR reaction, and the swollen tendon tissue was collected by necropsy and placed in a 15mL centrifuge tube, and 5 times the volume of serum-free DMEM medium was added, homogenized and repeatedly frozen and thawed 3 times, shake on the shaker for 1-2min after each thawing, and then 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, and use 0.22μm micropore After membrane filtration, set aside;

[0033] (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 cultu...

Embodiment 2

[0057] Example 2: TCID 50 Potency determination:

[0058] 1. Test method:

[0059] The 5th generation cell culture of virus strain N-DAV-SD16 (DMEM medium containing 2% fetal calf serum) is done doubling dilution (10 -1 -10 -6 ). After the LMH cells in the 96-well cell culture plate are fully covered in each well, add 25 μL of virus solution of each dilution gradient in sequence, and add 25 μL of DMEM medium containing 2% fetal bovine serum to the last two wells as a blank control. Do 8 replicates on the plate at 37°C with 5% CO 2 After 30 minutes in the incubator, 100 μL of DMEM medium containing 2% fetal bovine serum was added to each well to continue culturing and observing, observing and recording the occurrence of cell lesions every day, and interpreting the results on the 5th day. Calculate isolate TCID by Reed-Muench method 50 potency.

[0060] 2. Test results:

[0061] After determination, the TCID of strain N-DAV-SD16 50 The potency determination result is: 1...

Embodiment 3

[0062] Embodiment 3: animal regression test

[0063] 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. Among them, the first group is the experimental group, and the experimental group adopts the 5th generation cell culture of the virus strain N-DAV-SD16 to inoculate through the claw pad, 0.5mL / only, and 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 hock joint swelling of each group of animals were observed and recorded every day.

[0064] The experimental group began to develop tarsal joint swelling and lesions 7 days after inoculation. On the 9th day after inoculation, all 20 experimental groups developed tarsal joint swelling and lesions. The symptoms were the same as those of natural infection cases. The control group was in go...

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Abstract

The invention discloses a novel duck reovirus which is preserved in the China Center for Type Culture Collection on April 26, 2018, and the preservation number is CCTCC NO: V201818. The novel duck reovirus disclosed by the invention has good immunogenicity upon popular duck reoviruses with a main symptom of hock arthrocele at present. An inactivated vaccine prepared from a novel duck reovirus strain disclosed by the invention is good in security, has a protection rate up to 100%, and is capable of providing complete protection on newly separated variants.

Description

technical field [0001] The invention relates to the field of isolation and application of avian reovirus strains, in particular to a novel duck reovirus causing duck arthritis and its application. Background technique [0002] Avian reovirus (ARV) belongs to the genus Orthoreovirus of the Reovirdae family and can cause a variety of diseases in poultry. vary by host. At present, the main means to control the incidence of ARV in poultry flocks is to control typical avian reovirus infection through immunization with live attenuated vaccine or inactivated whole virus vaccine. However, because avian reovirus is an RNA virus with multiple segments, there are certain differences in antigen structure, pathogenicity, cell culture characteristics and host specificity among different strains. It is easy to mutate during genetic evolution, which leads to the inability of vaccines prepared from traditional strains to prevent and control the current popular avian reovirus infection. ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/15A61P31/14C07K16/10
CPCA61K39/12A61K2039/5252A61K2039/5254A61K2039/552A61P31/14C07K16/10C12N7/00C12N2720/12221C12N2720/12234
Inventor 唐熠刁有祥王鸿志
Owner SHANDONG AGRICULTURAL UNIVERSITY
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