CN107653231A - Duck-origin coronavirus low virulent strain IBVDCV35 and its application - Google Patents
Duck-origin coronavirus low virulent strain IBVDCV35 and its application Download PDFInfo
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- CN107653231A CN107653231A CN201710897708.5A CN201710897708A CN107653231A CN 107653231 A CN107653231 A CN 107653231A CN 201710897708 A CN201710897708 A CN 201710897708A CN 107653231 A CN107653231 A CN 107653231A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5252—Virus inactivated (killed)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5254—Virus avirulent or attenuated
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20021—Viruses as such, e.g. new isolates, mutants or their genomic sequences
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- C—CHEMISTRY; METALLURGY
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- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Abstract
The present invention relates to microorganism field, specifically provides one plant of duck-origin coronavirus low virulent strain IBV DCV35 (deposit number is CGMCC NO.13852) and its application.The duck-origin coronavirus low virulent strain IBV DCV35 are to screen to obtain by duck source coronavirus velogen strain IBV ZZ2004 (deposit number is CGMCC NO.3842) Attenuation, obvious variation occurs for the S1 gene orders of the low virulent strain IBV DCV35, and the sequence after variation can be in the stable heredity of the 22nd~35 generation.The low virulent strain IBV DCV35 that the present invention screens carry out SPF chicken infection by Experimental infection and horizontal transmission experiment, are showed no virulence and return strong phenomenon.Experiment shows, after attenuated IBDVs of the invention are inoculated in chicken, can effectively activate the immune system in chicken body and have good prevention effect to infectious bronchitis of chicken, can be used as preventing the vaccine virus of infectious bronchitis of chicken.
Description
Technical field
The present invention relates to microorganism field, specifically, is related to a kind of duck-origin coronavirus low virulent strain.
Background technology
Avian infectious bronchitis virus (Infectious bronchitis virus, IBV) seriously endangers supporting for China
Grow industry, the economic loss caused by infectious bronchitis of chicken is very serious every year, and prevent this disease mainly be weak poison
Vaccine, wherein the most widely used attenuated vaccine is H120 and H52.But because IBV serotypes are numerous, IBV genomes are again
Easily variation, new IB variants continuously emerge so that currently available vaccines, existing vaccines can not produce to the emerging popular strain of variation
The protective effect of effect, causes IBV preventing and treating to become difficult.
IBV ZZ2004 are a kind of sick for what is be separated in kind duck of cardinal symptom from immunosupress and growth inhibition is caused
Poison, the virus can cause Broiler chicks that similar symptom, the incidence of disease 80% occurs, and the death rate is more than 10%, morbidity chicken colony
Mitigate 37% or so again, immunity degradation, economic loss is up to more than 30%.Duck source property IBVZZ2004 on pathogenic with the country
There is obvious difference in the IBV reported, cause significant immunosupress and growth inhibition, and chicken and duck can be infected.
IBVZZ2004 strains and domestic popular IBV affinities are very low, also very low with the affinity of the main vaccine strain of the country, such as M41
Strain, H120, Beaudette, IBV4/91 homology are 85.2%~87.9%.Virus isolated strain ZZ2004 separation and
The correlation properties such as authentication method, pathogenic and genetic mutation have all been opened up in patent of invention ZL201010225577.4
Show, currently available vaccines, existing vaccines can not produce to duck source property IBVZZ2004 and be effectively protected power in production practices so that the disease does not stop
OK.From having obtained in terms of research data, the immunogenicity of duck source property IBVZZ2004 strains is good, possesses as the basic of vaccine virus
Key element.It is IBVZZ2004 attenuated vaccines to prepare one plant of good low virulent strain therefore, it is necessary to carry out causing weak research to it
Research and development lay the foundation.
The content of the invention
In order to solve problems of the prior art, it is an object of the invention to provide a kind of weak poison of duck-origin coronavirus
Strain, and its gene expression characteristics and the application in terms of infective bronchitis prevention are studied.
Technical scheme is as follows：
In a first aspect, the present invention provides a kind of duck-origin coronavirus low virulent strain IBV DCV35, duck source property hat is identified as
The virus attenuated strain of shape, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address：
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode：100101), preservation date
For on 05 27th, 2017, deposit number was CGMCC No.13852.
The duck-origin coronavirus low virulent strain IBV DCV35 are by the coronavirus velogen strain IBV ZZ2004 (preservations of duck source
Numbering is CGMCC NO.3842, and preservation date is on 04 29th, 2010) seed virus is used as, carry out Attenuation and obtain,
Detailed process is：
First after being inoculated with SPF chicken embryos, 31~33 DEG C of low temperature trainings are carried out before seed virus inoculated into chick embryo through 56 DEG C of water-bath 20min
Support, then same processing is done with preceding method, continuously reached for the 25th generation, using 37 DEG C of cultures, then continuous passage since the 26th generation
To the 35th generation, attenuated IBDVs IBV DCV35 are obtained.
The variation situation of the present invention each generation virus S1 gene orders of comparative analysis in succeeding generations, discovery reach the 22nd
For when S1 genes there are 12 bases to be made a variation, this variation still keeps stable reaching for the 35th generation.12 changes of S1 genes
Ectopic sites are respectively：188ntT → C (Val → Ala), 191ntA → G (Asn → Ser), 228ntT → G (Asn → Lys),
232ntA→C(Ser→Arg)、347ntT→A(Phe→Try)、351ntG→T(Arg→Ser)、360nt T→G(Ser→
Arg)、454ntG→A(Valr→Ile)、471ntT→G(Ser→Arg)、560ntC→A(Thr→Lys)、1477ntT→C
(Ser→Pro)、1497ntA→C(Gln→His)。
Further, the present invention is determined by EID50, strong experiment is returned in the experiment of SPF chickens Virulence detection, cohabitation infection and virulence
To determine the pathogenic of DCV35；SPF chickens are inoculated with by the strain, inactivation strain is inoculated with rabbit and protest test to determine
DCV35 immunogenicity.Result of the test shows that DCV35 strains strengthen chicken embryo adaptability, does not have lethal to SPF chickens, together
Occupy only infection not fall ill, virulence is not returned by force；After target animals are immunized in DCV35 simultaneously, body antibody level rise, chicken attacks malicious protection
Rate also raises.
Accordingly, low virulent strain IBV DCV35 of the present invention are obtained by special cause incompetent person's section, and S1 genes have bright
Aobvious genetic stability feature.
Second aspect, it is excellent based on the duck-origin coronavirus low virulent strain IBVDCV35 of the invention for screening to obtain and its virulence
Gesture, the present invention further provides its application in terms of immunoreagent is prepared.
It is embodied as, a kind of attenuated vaccine or a kind of inactivated vaccine.Research is found, by low virulent strain IBV of the present invention
DCV35 direct immunizations animal either mixes the attenuated vaccine or inactivated vaccine so as to obtain with immune acceptable carrier,
The immunocompetence of animal body can be significantly improved.
Brief description of the drawings
Fig. 1 is former sequence, 7,15,20,22~25 generation base comparison results.
Fig. 2 is former sequence, 7,15,20,22~25 generation amino acid alignment results.
Embodiment
Following examples further illustrate present disclosure, but should not be construed limitation of the present invention, to this hair
The modification or replacement of bright method, step either condition, belong to the scope of the present invention.
The Low- temperature culture of embodiment 1 induces low virulent strain
By ZZV6 seeds poison (EID50For 10-4.88/0.2mL) first through 56 DEG C of water-bath 20min, with 0.22um bacterial filter
After filtering, 9 age in days SPF chicken embryos are inoculated with, per 5 pieces of pickup kind, per embryo 0.2mL, in 31~32 DEG C of hatchings.Daily observation, 24 hours
Within dead chicken embryo abandon it, dead chicken embryo temporarily puts 4 DEG C of refrigerators, 72 hours sterile lesions collected virus, while observe chicken embryo
Situation, select the allantoic fluid of typical cytopathic chicken embryo to be passed on, same processing and culture done per Dai Jun, continuously reached for the 25th generation,
Using 37 DEG C of cultures since the 26th generation, then continuously reach the 35th generation acquisition DCV35.
The passaged virus S1 gene sequencings of embodiment 2
The 7th generation, the 15th generation, the 20th~35 generation virus, RT-PCR clone's S1 genes, measure are chosen during continuous passage
Its nucleotide sequence.S1 gene primers：Upstream (P1)：5'-AGTTATTGGTTAGAGATGTTGGGGA-3', downstream (P2)：5'-C
GTA TGG ACA GCT TGT GAC ATT TTC-3'。
Using DNASTAR analysis softwares by the 7th of sequencing the, 15,20, the base of 22~35 generations poison and the S1 genes of maternal poison
Sequence and the difference for deriving amino acid sequence, are shown in Table 1, respectively see appendix A and Appendix B for base and amino acid comparative analysis result.
S1 genes have 12 bases to be made a variation when as a result showing to reach for 22 generation, and this variation still keeps steady reaching for the 35th generation
It is fixed.12 variant sites of S1 genes are respectively：188ntT → C (Val → Ala), 191ntA → G (Asn → Ser), 228ntT
→ G (Asn → Lys), 232ntA → C (Ser → Arg), 347ntT → A (Phe → Try), 351ntG → T (Arg → Ser),
360nt T→G(Ser→Arg)、454ntG→A(Val r→Ile)、471ntT→G(Ser→Arg)、560ntC→A(Thr
→Lys)、1477ntT→C(Ser→Pro)、1497ntA→C(Gln→His)。
The base and amino acid of 1 former sequence of table and the 7th, 15,20,22~25 generation S1 genetic mutations
Pathogenicities of the embodiment 3DCV35 to SPF chick
(1)DCV35EID50Measure takes DCV35 virus liquids 1mL to be made with after 0.22um sterile filters filtering with physiological saline
10 times of incremental dilutions, each dilution factor are inoculated with the SPF chicken embryos of 5 piece of 9 age in days, and 0.2mL/ is per embryo, 37 DEG C of cultures, daily observational record
Dead germ number, dead chicken embryo abandons it within 24 hours, is incubated to 19 ages in days and dissects remaining embryo living, and sense is used as using dead and dwarf embryo
The criterion of dye.The SPF chicken embryos that 9 ages in days are inoculated with by the use of same dosage physiological saline are used as control.In terms of Reed-Muench Er Shi methods
Calculate EID50。
As a result show：As the increase of viral passages number, virus are more and more stronger to the adaptability of SPF chicken embryos, EID50
Increase, DCV35 EID is calculated through Reed-Muench methods50For 10-6.75/0.2mL。
(2) 39 5 age in days SPF chick of DCV35 artificial challenges SPF chick, 3 big groups are divided into using completely random method,
1st and 2 group every group 15 chickens, 9 chickens of control group, by the generations of IBV ZZ2004 the 6th poison (104.88EID50/ 0.2mL), 35 generations poison
(106.75EID50/ 0.2mL) filtered through 0.22 μm of sterilizing filter after, through eye droppings, collunarium approach be inoculated with respectively the 1st group, the
The physiological saline of 2 groups, every group of inoculation 2mL, the 3rd group chicken injection Isodoses.From inoculation, morbidity feelings are observed and recorded daily
Condition, cut open inspection is carried out to the chicken of dead chicken lesion, observes dead chicken lesion.
As a result as shown in table 2, DCV35 strains inoculation SPF chickens do not fall ill, also not lethal SPF chickens.
Pathogenicity of table 2 IBV ZZ2004ZZV6 and the DCV35 virus to SPF chick
(3) experiment of DCV35 infected chickens cohabitation infection is inoculated with 5 age in days SPF with DCV35 viral allantoic fluid through collunarium, eye droppings
Chicken 4, every dosage are not less than 106.75EID50/ 0.2mL, 4 same group feedings of healthy SPF chickens with the non-virus inoculation of same age in days
Support in same SPF isolators, observe 7 days, record the clinical manifestation situation of chicken；All chickens are put to death after 7 days, to DCV35
RT-PCR detections are carried out with the chicken internal organs without inoculation poison that group feeding is supported.
As a result show：5 inoculated DCV35 SPF chick is supported 7 days with 5 with age in days SPF chickens with group feeding, and none is only sent out
Disease.It is the positive to carry out RT-PCR testing results to the chicken internal organs lived together.
(4) DCV35 virulence returns strong experiment DCV35 for passaged virus liquid (106.75EID50/ 0.2mL) 5 age in days SPF of inoculation
Chicken 5, every from the inoculation from chicken incidence, inoculation slaughters after 7 days, takes its kidney and tracheae to add physiology
Salt solution makees 1:2 mixing.After grinding, centrifuging and taking supernatant, SPF chick 5 is inoculated after dual anti-processing, it is the second generation, such as
This continuously passed for 5 generations by susceptible chick.And it is determined whether with poison as RT-PCR to the virus in per generation.
As a result show：Virus returns the generation of chicken body 5, and it is positive that per generation, which returns and detects its M gene with RT-PCR, is inoculated with chick
But have no that inoculation chicken has any clinical manifestation, illustrate that the virulence of the attenuated virus is stable.
Embodiment 4DCV35 immunogenicity determining
(1) DCV35 it is weak poison inoculation the antibody test of SPF chicken serums by SPF fowl raisings in negative pressure isolator, free choice feeding with
Drinking-water, and first immunisation is carried out when 15 age in days, DCV35 dosages of inoculation are 106.75EID50/0.2mL.14 days after first immunisation,
Every group of chicken takes the blood of 4 chickens at random, and the blood sample of collection is placed in 37 DEG C of insulating box and is incubated 2h, then 1000r/min from
Heart 10min, serum is collected, be sub-packed in 200uL PCR pipe, -20 DEG C of preservations, indirect ELISA method to be used detects chicken group's serum
In antibody.
As a result it is as shown in table 3：Inoculated DCV35 strains chicken serum antibody is 1 in dilution ratio:When 800, P/N value
Reach 2.1, nonvaccinated control group chicken serum sample antibody detection is feminine gender.
The chicken group serum antibody of table 3 produces horizontal
(2) Serum Antibody Detection selection health after the weak immune rabbits of poison of DCV35, body weight 2.0kg or so White Rabbit 2,
Emulsion made of viral antigen-Freund's complete adjuvant (FAC) of first immunisation inactivation (inactivation antigen concentration is 2.4mg/mL,
Antigen：FAC volumes are 1:1), the subcutaneous multi-point injection of nape part, each positions of 0.2mL/, every rabbit inject 1mL；Two Zhou Houyong
Emulsion two made of Freund's incomplete adjuvant (FIC) is exempted from, dosage 1mL；Directly booster immunization is carried out after two weeks with the 35th generation virus.Three
7 days ear edge vein exploitating bloods after exempting from, its potency is detected with indirect ELISA.
As a result show：Rabbit blood sampling after the albumen wrapper sheet weak to DCV35 malicious 3 preserved using this laboratory is exempted from is carried out
ELISA is detected, and rabbit immune DCV35 can produce good antibody, and potency reaches 1:3200, it is shown in Table 4.
The rabbit anteserum antibody level of table 4
(3) 14 SPF chick of protest test, DCV35 is immunized respectively in 5 ages in days and 19 ages in days, dosage of inoculation is every
Only 106.75EID50/0.2mL.Two exempt from two weeks after using ZZV6 attack poison, every chicken is inoculated with 2mL through collunarium, eye droppings
(104.88EID50), while set and nonimmune attack malicious control group.From self tapping poison, observation daily, falling ill, being dead for chicken group is recorded
Situation, cut open inspection is carried out to dead chicken, observes pathological change, Continuous Observation 7 days.
As a result show：DCV35 strain group chickens do not fall ill, case fatality rate 0, rather than the morbidity of immune group chicken, and death occur.Its
As a result such as table 5.
Immune protective efficiencies of the DCV35 of table 5 to SPF chick
Claims (9)
1. a kind of duck-origin coronavirus low virulent strain IBV DCV35, it is characterised in that deposit number is：CGMCC NO.13852.
2. duck-origin coronavirus low virulent strain IBV DCV35 according to claim 1, it is characterised in that its to SPF chickens not
With lethal, virulence is not returned by force.
3. duck-origin coronavirus low virulent strain IBV DCV35 according to claim 1 or 2, it is characterised in that it is by duck source
Coronavirus velogen strain IBV ZZ2004 Attenuations and obtain.
4. duck-origin coronavirus low virulent strain IBV DCV35 according to claim 3, it is characterised in that its S1 gene with
Duck source coronavirus velogen strain IBV ZZ2004 S1 genes are compared, and 12 nucleotide variations be present.
5. the duck-origin coronavirus low virulent strain IBV DCV35 described in any one of Claims 1 to 4 are in immune drug is prepared
Using.
6. application according to claim 5, it is characterised in that the medicine is vaccine.
7. the application according to claim 5 or 6, it is characterised in that the medicine is used to prevent infectious bronchitis of chicken.
8. a kind of attenuated vaccine, it is characterised in that as the duck-origin coronavirus low virulent strain described in any one of Claims 1 to 4
It is prepared by IBV DCV35.
9. a kind of inactivated vaccine, it is characterised in that as the duck-origin coronavirus low virulent strain described in any one of Claims 1 to 4
Prepared after IBV DCV35 inactivations.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402065A (en) * | 2018-08-08 | 2019-03-01 | 温氏食品集团股份有限公司 | One kind kind duck C hypotype fowl metapneumovirus low virulent strain and its preparation and application |
| CN113786404A (en) * | 2021-10-13 | 2021-12-14 | 山东领海生物科技有限公司 | Application of bromophenol-pyrazoline compound in treating avian coronavirus diseases |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE60012042D1 (en) * | 1999-11-26 | 2004-08-12 | Merial Lyon | ENTEPNEUMOVIRUS AND APPROPRIATE VACCINE |
| CN1729996A (en) * | 2005-08-03 | 2006-02-08 | 中国人民解放军军事医学科学院军事兽医研究所 | The preparation technology and the preparation of the important eqpidemic disease hyper-immune serum of Canis animals anti-canidae animal |
| CN101993856A (en) * | 2010-11-24 | 2011-03-30 | 河南科技学院 | Duck-origin coronavirus ZZ2004-resisting monoclonal antibody, hybridoma cell strain and application thereof |
| CN104762271A (en) * | 2015-01-26 | 2015-07-08 | 河南科技学院 | Preparation method and use of duck-derived coronavirus attenuated strain DCV35 |
-
2017
- 2017-09-28 CN CN201710897708.5A patent/CN107653231B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE60012042D1 (en) * | 1999-11-26 | 2004-08-12 | Merial Lyon | ENTEPNEUMOVIRUS AND APPROPRIATE VACCINE |
| CN1729996A (en) * | 2005-08-03 | 2006-02-08 | 中国人民解放军军事医学科学院军事兽医研究所 | The preparation technology and the preparation of the important eqpidemic disease hyper-immune serum of Canis animals anti-canidae animal |
| CN101993856A (en) * | 2010-11-24 | 2011-03-30 | 河南科技学院 | Duck-origin coronavirus ZZ2004-resisting monoclonal antibody, hybridoma cell strain and application thereof |
| CN104762271A (en) * | 2015-01-26 | 2015-07-08 | 河南科技学院 | Preparation method and use of duck-derived coronavirus attenuated strain DCV35 |
Non-Patent Citations (4)
| Title |
|---|
| SIXIN YAO等: "Isolation of a novel serotype strain of infectious bronchitis virus ZZ2004 from ducks in China", 《VIRUS GENES》 * |
| 朱广蕊等: "鸡传染性支气管病毒ZZ2004株致SPF感染鸡小肠损伤的病理与超微病变研究", 《中国预防兽医学报》 * |
| 闫艺婷: "IBVZZ-V35弱毒株增殖特性与免疫原性研究及安全性评价", 《万方数据库》 * |
| 闫艺婷等: "鸡传染性支气管炎病毒弱毒株ＺＺ－Ｖ３５在鸡胚中的增殖规律", 《动物医学进展》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402065A (en) * | 2018-08-08 | 2019-03-01 | 温氏食品集团股份有限公司 | One kind kind duck C hypotype fowl metapneumovirus low virulent strain and its preparation and application |
| CN109402065B (en) * | 2018-08-08 | 2021-07-30 | 温氏食品集团股份有限公司 | Muscovy duck subtype C avian metapneumovirus attenuated strain and preparation and application thereof |
| CN113786404A (en) * | 2021-10-13 | 2021-12-14 | 山东领海生物科技有限公司 | Application of bromophenol-pyrazoline compound in treating avian coronavirus diseases |
| CN113786404B (en) * | 2021-10-13 | 2024-03-26 | 山东领海生物科技有限公司 | Use of bromophenol-pyrazoline compounds in treatment of avian coronavirus diseases |
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