CN113832113A

CN113832113A - Novel duck reovirus attenuated strain and application thereof

Info

Publication number: CN113832113A
Application number: CN202111076984.8A
Authority: CN
Inventors: 李泽君; 李雪松
Original assignee: Shanghai Veteromaru Research Institute Caas China Animal Health And Epidemiology Center Shanghan Branch Center
Current assignee: Shanghai Veteromaru Research Institute Caas China Animal Health And Epidemiology Center Shanghan Branch Center
Priority date: 2021-09-14
Filing date: 2021-09-14
Publication date: 2021-12-24
Anticipated expiration: 2041-09-14
Also published as: CN113832113B

Abstract

The invention discloses a novel duck reovirus attenuated strain which has a sigma C structural protein with an amino acid sequence shown in SEQ ID No. 1. The invention also discloses a vaccine containing the novel duck reovirus attenuated strain. The invention also discloses application of the novel duck reovirus attenuated strain in preparation of a product for preventing the novel duck reovirus disease. After the novel duck reovirus attenuated strain is used for immunizing healthy and susceptible ducklings at the age of 1 day, the protection rate of the novel live vaccine for the duck reovirus disease prepared by the novel duck reovirus attenuated strain on virulent attack reaches 100% after 6 days, the novel live vaccine has good safety and effectiveness, the toxicity is not returned to the strong state after artificial passage in the susceptible ducklings, and the novel live vaccine for the duck reovirus disease has a very good application prospect in prevention and treatment of the novel duck reovirus disease of the ducklings.

Description

Novel duck reovirus attenuated strain and application thereof

Technical Field

The invention belongs to the technical field of veterinary biological products, and particularly relates to a novel duck reovirus attenuated strain and application thereof as a novel live vaccine for duck reovirus diseases.

Background

Diseases caused by Novel Duck Reovirus (NDRV) infection are duck viral infectious diseases which are mainly characterized by irregular bleeding, swelling, necrosis and the like of spleen. The virus is sequentially separated from waterfowls such as geese, Muscovy ducks, cherry valley ducks, sheldrakes and the like, the NDRV can infect ducklings of different varieties, the host range of the NDRV infection is wider than that of the typical Muscovy Duck Reovirus (MDRV), and pathogenicity and clinical pathological changes caused by the two viruses are greatly different. Since 2017, the morbidity and mortality of the novel duck reovirus disease gradually increase, and the novel duck reovirus disease becomes one of important viral infectious diseases affecting the duck breeding industry in China.

NDRV can infect Muscovy ducks, semi-Muscovy ducks, cherry valley meat ducks, sheldrakes, geese and the like and cause morbidity. The virus has serious harm to ducklings, the disease days of Muscovy ducks and Muscovy ducks are mainly 6-25 days old, wherein the disease course is 5-7 days when the disease days of Muscovy ducks and Muscovy ducks are more than 7 days old; the disease age of the cherry valley duck is mainly 10-30 days. The morbidity of the novel reovirus is 5-30%, and the fatality rate is 4-20%.

Vaccination is one of the keys to the prevention of the novel duck reovirus disease. The existing vaccines for preventing the disease comprise inactivated vaccines and attenuated live vaccines, wherein the generation time of immunity of the inactivated vaccines is slow, multiple times of immunity are usually needed to achieve a good protection effect, the capability of causing cell-mediated immunity is weak, the attenuated live vaccines stimulate organisms to generate comprehensive immune response, the generation time of the immune efficacy is fast, and the sufficient protection effect can be generated once. Therefore, the novel duck reovirus attenuated live vaccine has more advantages in clinical application compared with an inactivated vaccine in the aspect of protecting ducklings.

Disclosure of Invention

The invention aims to solve the technical problem that no novel duck reovirus disease attenuated live vaccine for effective prevention exists clinically at present, and provides a novel duck reovirus attenuated strain.

In order to solve the technical problems, the invention is realized by the following technical scheme:

the invention provides a novel duck reovirus attenuated strain which has a sigma C structural protein with an amino acid sequence shown in SEQ ID No. 1.

Preferably, the virus attenuated strain further comprises a sigma A structural protein having an amino acid sequence shown in SEQ ID No. 2.

Preferably, the virus attenuated strain further comprises a sigma B structural protein having an amino acid sequence shown in SEQ ID No. 3.

The invention provides a novel duck reovirus attenuated strain JS01-105P with the preservation number of CCTCC NO: v202168.

The invention also provides a nucleic acid encoding the following structural proteins or fragments thereof:

sigma C structural protein of an amino acid sequence shown in SEQ ID NO. 1;

or sigma A structural protein of an amino acid sequence shown in SEQ ID NO. 2;

or sigma B structural protein of an amino acid sequence shown in SEQ ID NO. 3.

Preferably, the nucleic acid is selected from the group consisting of:

the nucleotide sequence of sigma C structural protein shown in SEQ ID NO. 4;

or a nucleotide sequence of the sigma A structural protein shown in SEQ ID NO. 5;

or the nucleotide sequence of sigma B structural protein shown in SEQ ID NO. 6.

The invention also provides a vaccine containing the novel duck reovirus attenuated strain.

The vaccine has good safety and effectiveness on ducklings, and the virus content of each feather is not less than 10^3.5TCID₅₀。

The invention also provides application of the novel duck reovirus attenuated strain in preparation of a product for preventing novel duck reovirus diseases.

The novel duck reovirus attenuated strain JS01-105P is used as a live vaccine of the novel duck reovirus, after 1 day-old healthy susceptible ducklings are immunized, the protection rate of virulent attack reaches 100% after 6 days, the novel duck reovirus attenuated strain JS01-105P has good safety and effectiveness, the virulent return is not generated in the susceptible duck body after artificial passage, and the novel duck reovirus attenuated strain JS has a very good application prospect in prevention and treatment of the novel duck reovirus disease of the ducklings.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

FIG. 1 is a graph showing the survival rate of ducks inoculated with NDRV JS01 strain virulent strain and subcultured virulent strain thereof in example 2 of the present invention;

FIG. 2 is a graph showing the measurement of the virus titer of the NDRV JS01 strain virulent virus and the passage virus thereof inoculated to the viscera of a 1-day-old duck in example 2 of the present invention;

figure 3 is a schematic representation of the novel reovirus attenuated live vaccine final product of example 4 of the present invention.

The optimized novel duck reovirus attenuated strain is preserved in China center for type culture Collection (CCTCC for short, the address: preservation center of Wuchang Lodojia mountain Wuhan university, Wuhan City, Hubei province) at 9 months and 1 day 2021, and the preservation number is CCTCC NO: v202168, which is classified and named as novel duck reovirus attenuated strain JS 01-105P.

Detailed Description

In order to develop a safe and effective novel duck reovirus attenuated live vaccine, the separated and identified novel duck reovirus virulent strain JS01 is continuously passaged and purified on SPF chick embryos for 105 generations to obtain a novel duck reovirus attenuated strain JS 01-105P. The novel duck reovirus attenuated strain (JS01-105P) disclosed by the invention is used as a live vaccine to immunize healthy and susceptible ducklings of 1 day age, the protection rate of virulent attack reaches 100% after 6 days, the novel duck reovirus attenuated strain has good safety and effectiveness, and the virulence return does not occur in artificial passage in the susceptible ducks, so that the novel duck reovirus attenuated strain disclosed by the invention has a very good application prospect in the aspect of prevention and control of novel duck reovirus diseases of the ducklings

EXAMPLE 1 cultivation and sequence analysis of novel attenuated reovirus vaccine Strain

A novel duck reovirus strain is separated and identified from a case causing spleen necrosis of a duckling, named as a novel duck reovirus strain JS01, and inoculated to an allantoic cavity of an SPF chick embryo of 9-10 days old and continuously cultured in an incubator. Discarding the dead chick embryos within 24 hours of inoculation, taking the rest chick embryos once within 24 hours of embryo culture, taking out the dead chick embryos in 2-6 days, storing at the temperature below-15 ℃, melting at room temperature, grinding, collecting supernatant, centrifuging at 3000 r/min for 10 minutes, and taking the supernatant for packaging. And subcultured continuously in the same manner to 100 generations, and then purified 5 times by limiting dilution to 105 generations. The JS01 and the virus of the 105 generation are subjected to whole-gene sequencing, and the result shows that the virus of the 105 generation has 4 differences compared with JS01 in amino acid mutation sites of main structural proteins sigma C, sigma A and sigma B of the virus of the 105 generation. Of the 5 amino acid mutation points, the sigma C (sigma C) protein has 1 mutation point (T120P), the sigma A (sigma A) protein has 2 mutation points (T44P, E120G), and the sigma B (sigma B) protein has 1 mutation point (L260F), and the amino acid mutations of the main structural proteins of different subvirals and the conditions are shown in the following table 1. The amino acid sequence of the JS01-105P strain structural protein sigma C is shown as SEQ ID NO.1, and the nucleotide sequence is shown as SEQ ID NO. 4; the amino acid sequence of the structural protein sigma A is shown as SEQ ID NO.2, and the nucleotide sequence is shown as SEQ ID NO. 5; the amino acid sequence of the structural protein sigma B is shown as SEQ ID NO.3, and the nucleotide sequence is shown as SEQ ID NO. 6.

Example 2 evaluation of the pathogenicity of NDRV JS01 strain subculture virus to 1 day old ducklings

In order to evaluate the virus pathogenicity change conditions of NDRV JS01 virulent strain 5 th generation (5P), subculture virus 75 th generation (75P) and subculture virus 105 th generation (105P), the purchased SPF duck embryos are incubated to 1 day of age for animal experiments, 40 susceptible ducklings are randomly divided into 4 groups, 10 ducklings are divided into 4 groups, the 1 group is NDRV JS01-5P group, the 2 group is NDRV JS01-75P group, the 3 group is NDRV JS01-105P group, and the 4 group is PBS control group. The 1 st to 3 th groups of 1-day-old ducklings are subjected to leg intramuscular injection passage virus, and the virus dose is 10^5.0TCID₅₀0.1mL, 0.1mL each. The group 4 is a blank control group, 0.1mLPBS buffer solution is injected into legs, and the continuous observation is carried out for 14 days to observe whether the inoculated ducks have clinical symptoms caused by virus or not and whether local and systemic adverse reactions caused by the inoculation occur or not.

Test for isolation of visceral viruses: the purchased SPF duck embryos are incubated to 1 day of age and then animal experiments are performed,40 ducklings are randomly divided into 4 groups, 10 ducklings are arranged in each group, the 1 st group is NDRV JS01-5P generation virus group, the 2nd group is NDRV JS01-75P generation virus group, the 3 rd group is NDRV JS01-105P generation virus group, and the 4 th group is PBS control group. In groups 1-3, each 1-day-old duckling is injected with passage virus at leg part with virus dose of 10^5.0TCID₅₀0.1mL, 0.1mL each. Group 4 was a blank control group, and each leg was injected with 0.1ml of pbs buffer. 4 days after virus inoculation, all ducklings are dissected and killed to carry out the dissection examination and the measurement of the content of the visceral virus, and collected visceral organs comprise heart, liver, spleen, kidney, duodenum, brain, bursa of fabricius and other visceral tissues.

As a result:

clinical symptoms and mortality within 14 days after different NDRV JS01 strains of passage virus infect 1-day-old ducklings: after the NDRV JS01-5P strain is inoculated to the muscle for strong toxicity, two ducklings die within 14 days, the survival rate is 80 percent, and the NDRV JS01-75P and NDRV JS01-105P inoculated ducks do not die, and the survival rate is 100 percent (figure 1). NDRV JS01-5P strain inoculation group part of ducklings after 3 days of toxin attack have symptoms of listlessness, decreased feed intake, diarrhea and the like; the NDRV JS01-75P strain inoculated duckling group part has a diarrhea symptom; NDRV JS01-105P vaccinated groups and PBS vaccinated groups did not have any adverse clinical symptoms.

Different NDRV JS01 strains are subcultured to ducklings of 1 day old after 4 days, and the pathological observation is shown in figure 2, the livers of the NDRV JS01-5P inoculated ducks have severe bleeding points, and the livers of the subcultured ducks 75P and 105P and the control ducks have no obvious change; spleen of the duck inoculated with NDRV JS01-5P shows severe hemorrhage, enlargement and necrosis, spleen of the duck inoculated with subculture virus 75P shows hemorrhage, slight enlargement and necrosis point, and spleen of the duck of 105P and a control group has no obvious change. NDRV JS01 strain virulent virus and passage virus thereof are inoculated on healthy susceptible ducklings of 1 day age, the ducklings are killed after 4 days, the heart, the liver, the spleen, the lung, the kidney, the duodenum, the bursa of fabricius and the brain of each group of inoculated ducks are subjected to virus titration, NDRV sensitive LMH cells are used for titration, and the titration results of the organ viruses of NDRV JS01-5P and the passage virus (75P and 105P) of the NDRV inoculated ducks are shown in figure 2. After NDRV JS01-5P infects ducks, viruses can be detected in heart, liver, spleen, lung, kidney, duodenum, bursa of Fabricius and brain, and systemic infection is shown, wherein the virus content in lung, spleen, lung and kidney is the highest; NDRV JS01-75P inoculated ducks mainly infect spleen and bursa of fabricius; the NDRV JS01 strain passage virus 105P inoculated duck can only detect low-level virus in the spleen of part of ducks.

Example 3 minimal immune dose study of novel reovirus strain JS01-105P

To determine the optimal immune dose of NDRV JS01 strain low virulent strain JS-105P to 1 day old duckling: the purchased SPF duck embryos are incubated to 1 day old and then subjected to animal experiments, and 40 healthy susceptible ducklings are randomly divided into 4 groups of 10 ducklings. Group 1: muscle inoculation of legs with NDRV JS01-105P at a dose of 10^2.0TCID₅₀0.1mL for each; group 2: muscle inoculation of legs with NDRV JS01-105P at a dose of 10^2.5TCID₅₀0.1mL for each; group 3: muscle inoculation of legs with NDRV JS01-105P at a dose of 10^3.0TCID₅₀0.1mL for each; group 4: PBS control group, leg muscle inoculated with PBS, 0.1mL each. When ducklings are 7 days old (6 days after immunization), all 4 groups of ducklings are detoxified, and muscle inoculation is carried out for 10^5.0TCID₅₀NDRV JS01 is virulent, and on day 4 after challenge, all ducks are subjected to autopsy and virus titration of organs. Caesarean display, 10^2.0TCID₅₀After the dose immunization group is detoxified, 3/10 spleen still has certain swelling and bleeding phenomena. And 10^2.5TCID₅₀And 10^3.0TCID₅₀After the duck is detoxified by the dose immunization group, no obvious visceral lesion appears. The titration results of the virus in different organs showed that the titer of the virus in different organs was 10 after the challenge^3.0TCID₅₀And 10^2.5TCID₅₀No virus was detected in the organs of the dose immunized group ducks, and 10^2.0TCID₅₀The virus still existed in spleen of 3/10 duck in dose immunization group. The above results show 10^2.5TCID₅₀The strain JS01-105P, the NDRV subculture virus, can provide complete protection. Determination of minimum immune dose of NDRV subculture virus JS01-105P strain is 10^2.5TCID₅₀。

Example 4 preparation and testing of novel attenuated live reovirus vaccines

Adding a freeze-drying protective agent into NDRV JS01-105P, 110P and 115P 3 virus liquid amplified by generations in an aseptic safety cabinet according to the proportion of 9:1, mixing uniformly, quantitatively subpackaging into aseptic ampoule bottles, carrying out freeze vacuum drying in a freeze dryer, capping after the freeze drying is finished, and storing in a refrigerator below-15 ℃, wherein the vaccine finished product is shown in figure 3.

And (3) checking a vaccine finished product: the freeze-dried vaccine is subjected to character inspection, the character of the vaccine is spongy loose lumps and is easy to separate from the wall of a vaccine bottle, and the vaccine is quickly dissolved after a diluent is added; and (4) performing sterile inspection, mycoplasma inspection, exogenous viruses inspection, residual water and vacuum degree inspection according to the appendix of the current Chinese veterinary pharmacopoeia. And (3) identification and inspection: diluting the prepared vaccine to 100TCID with virus maintenance solution₅₀0.1ml, mixing with NDRV hyperimmune serum inactivated at 56 ℃ for 30min in equal volume (the neutralization titer is not less than 1:10), neutralizing at 37 ℃ for 1h, inoculating a 96-well cell culture plate of well-grown LMH cells, wherein each batch of vaccine is 10 wells and 200 ul/well, and simultaneously setting normal control cells and virus control wells, and inoculating 10 wells each. Placing at 37 ℃ and 5% CO₂For 7 days, and observing and recording the pathological condition of the cells. Normally maintaining no cytopathy in the liquid cell hole and the neutralizing hole, and generating cytopathy in the virus control hole; the virus content of the vaccine was tested using LMH cells: randomly extracting one bottle of vaccine, diluting with virus maintaining liquid to 0.1ml of vaccine feather with 1 according to the vaccine feather marked by the label, diluting with 10 times of diluent, and taking 10^-2、10^-3、10^-4、10^-54 dilutions were added to well-grown LMH cells in 96-well plates, and each dilution was plated in 4 replicates, 0.1mL per well, at 37 ℃ in 5% CO₂Culturing for 7 days in a cell culture box, observing and recording the cytopathic condition, and calculating the virus content TCID according to a Reed-Muench method₅₀The virus content of each feather should be more than or equal to 10^3.5TCID₅₀。

3 batches of experimental lyophilized NDRV JS01-105P attenuated vaccine prepared in laboratory, the freeze-drying effect is good. And (3) displaying a finished product inspection result: the freeze-drying property is spongy loose lumps which are easy to separate from the vaccine bottle wall, and the vaccine can be quickly dissolved by adding diluent; the sterility test, mycoplasma test and exogenous virus test were performed according to the appendix of the current "Chinese veterinary pharmacopoeia", and all of the 3 batches of vaccines were free of bacteria, mycoplasma growth and exogenous virus contamination. The identification test results show that the virus control wells all generate cytopathic effect, and the neutralization wells and the maintenance solution control wells of 3 vaccine samples have no cytopathic effect.

Example 5 safety evaluation of novel reovirus attenuated live vaccines

After the novel duck reovirus disease attenuated vaccine JS01-105P is inoculated to healthy and susceptible ducklings of 1-day age in single dose, repeated single dose and super dose, no obvious adverse reaction is seen in the inoculated part and the whole body, the inoculated ducks are normal in ingestion, drinking, excretion and mental state, and no clinical reaction or pathological change is found (see tables 1-3).

TABLE 1 safety test of single-dose 1-time inoculation of 1-day-old ducklings by novel reovirus attenuated vaccine JS01-105P

TABLE 2 safety test of single-dose repeated inoculation of 1-day-old ducklings with novel reovirus attenuated vaccine JS01-105P

TABLE 3 safety test of 1 day old duckling inoculated with strain JS01-105P of novel reovirus attenuated vaccine

Example 6 evaluation of effectiveness of novel reovirus attenuated live vaccines

The novel attenuated reovirus live vaccine JS01-105P is inoculated to the virus attack protection condition of 1 day old ducklings: (1) the clinical manifestations, the 1-day-old ducklings are immunized with 3 batches of NDRV JS01-105P vaccine, the 7-day-old ducklings are attacked, the vaccinated group ducklings have no abnormal manifestations, and the contrast attacking group ducklings are manifested by reduced feed intake and diarrhea; (2) pathological changes are as follows: killing the duck in 4 days after challenge, checking pathological changes of the immune ducks, wherein each visceral organ of each batch of immune ducklings has no obvious pathological changes, spleen of the control challenge ducklings has different degrees of heaviness and necrosis, and other visceral organs have no obvious macroscopic lesions; (3) virus separation: after 4 days of challenge, the organs such as heart, liver, spleen, lung and kidney of the inoculated ducklings were collected and virus-separated by LMH cells, and the results show that no virus was separated from the organs of the ducklings immunized by 3 batches of vaccines, while viruses could be separated from the spleen of the control challenge group (Table 4).

TABLE 4 novel reovirus attenuated live vaccine JS01-105P Strain effectiveness test

The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Sequence listing

<110> Shanghai animal doctor institute of Chinese academy of agricultural sciences (Shanghai center of Chinese centers of animal health and epidemiology)

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aatagagacg cgtggcgcaa tttcaacgtc ttcggctttt catggacaga tgaaggtttg 300

tcaggacttg tcgctgccca agaccctcct ccagctccac cgtaccaacc agcttcggga 360

cagtggtctg atctgcttca gtatcctcga tgggctaatc gtcaacgtga gttgcagtct 420

aaatatccta ttctgttgcg atctactttg ctttcagcta tgcgcgctgg accggtgttg 480

tacgttgaga cttggcccaa cacgatttct ggtcgacttg ctgactggtt catgtcgcag 540

tatgggaaca acttcgttga catgtgtgca cgattaactc agtcctgcat gaacatgcca 600

gttgagccgg atggtaatta tgaccagcag atgcgagcat taattagtct gtggctcctc 660

tcgtatattg gcgttgtgaa tcaatctaac actattaacg gcttctactt tgcttccaag 720

acgcgggggc aggctatgga taactggacg ctgttctatg ctaccaacac caatcgggtg 780

cagattactc agcgccactt tgcctacgtt tgcgctcgct cccccgactg gaatgtcgat 840

aagtcctgga tcagcgctgc caatttgact gccattgtca tggcttgccg ccagccgcca 900

gcctttgcca accagggggt aatcaaccag gcccagaatc gacctggatt ctcgatgaat 960

ggtggaacgc ctgtgcacga gctgaacatg ctgactaccg ctcaagcttg tatccagcag 1020

tgggtcgtag ctgatttgat ctcagcagct aagggtcagt ccatgacgca ggaggcgaat 1080

gacttctcga acctcatcca agccgatctt gggcggatca aagcgcaaga tgacgcgttg 1140

tacaaccaac aacctggtta cgctcgtcgc attaagccgt ttgccaacgg agattggact 1200

cctggaatga ctgcgcaagc attagctcta ctagccactt ttactgtcta g 1251

<210> 6

<211> 1104

<212> DNA

<213> Novel duck reovirus (Novel duck reovirus) attenuated strain JS01-105P

<400> 6

atggaggtgc gtgtgccaaa ctttcactcc tttatcgagg gtattactac tagttacttg 60

tgttctcctg cgagctggaa ttcgaagacg ttatgggata ttgaagaatt tcacacacct 120

gacgttatca gggtcggcaa tgcttattgt tgcactcagt gctgtggtgt tctgtactat 180

ggtgcccctc cctctgatgg aaactgtttt ccacatcaca agtgtcatca acagcaatat 240

cgtactgaga ctccgctcat gagatatatt aaggtgggtc gcactacaga gcaactgctt 300

gatcaatatg ccattgctct gcatgtcatt gcagattact atgacgaggc gagtaagcaa 360

cctcatgata tcgctgaaac tgagtcaatc gcaccatttg atatcgtaac caggactgaa 420

tctattcgca gtgaccgtgc cgttgacccg gaattctgga cttatccgtt agagagacga 480

ggatacgacg cgcgacatga gattgctaga gcgggttgga agatgatcga tgcttcatcg 540

cgaagtcaca ctcttcctga atgtctggtg tcaaatatgc tacatactag acatgtcttc 600

agccaaatgt tgaccacgac aaccatctat gatgtcgctg tcacgggtaa agctgttaca 660

ttcagcccga tggtagcaac catgccaact cgaggagatg gtgctgtggc tctgtcaaga 720

ggtaacttgg atcatgatgt cgaggactgt tggatggatg gttttgcatt ctcccccttc 780

atcggcggtg ttggcatcac tggtcaattt gagcgtggct cctgccataa ttttggacac 840

cccatgattg gaagcggtaa gaaggcttct cactaccgca atttattcat ggaatcctgg 900

cgtggatggt caaagtcgtg ctttacatgt gctgtaggga tggagcccgc ggagtgcgaa 960

tctaggctgc gaggccatgc cagaactatg ttcggacgtt ctcttccgga tatctgtgac 1020

ttcgaggaga ctacccacat tggccagtcg tccgcgccct taaagaaggc cacgaaattg 1080

tccttcctgg agtgtaggtg gtaa 1104

Claims

1. A novel duck reovirus attenuated strain has a sigma C structural protein with an amino acid sequence shown in SEQ ID NO. 1.

2. The novel attenuated strain of duck reovirus of claim 1, further comprising a σ a structural protein having the amino acid sequence shown in SEQ ID No. 2.

3. The novel attenuated strain of duck reovirus as claimed in claim 1 or 2, further comprising a sigma B structural protein having an amino acid sequence shown in SEQ ID No. 3.

4. A novel duck reovirus attenuated strain has a sigma A structural protein with an amino acid sequence shown in SEQ ID NO. 2.

5. A novel duck reovirus attenuated strain has a sigma B structural protein with an amino acid sequence shown in SEQ ID NO. 3.

6. A novel duck reovirus attenuated strain has a preservation number of CCTCC NO: v202168.

7. A nucleic acid encoding the following structural proteins or fragments thereof:

sigma C structural protein of an amino acid sequence shown in SEQ ID NO. 1;

or sigma A structural protein of an amino acid sequence shown in SEQ ID NO. 2;

or sigma B structural protein of an amino acid sequence shown in SEQ ID NO. 3.

8. A vaccine comprising the novel attenuated strain of duck reovirus as claimed in any one of claims 1 to 6.

9. The vaccine of claim 8, wherein the vaccine has a viral content of no less than 10 per plume^3.5TCID₅₀。

10. Use of the novel attenuated strain of duck reovirus as claimed in any one of claims 1 to 6 in the manufacture of a product for the prevention of novel duck reovirus diseases.