CN107236715B - Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine - Google Patents

Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine Download PDF

Info

Publication number
CN107236715B
CN107236715B CN201710347027.1A CN201710347027A CN107236715B CN 107236715 B CN107236715 B CN 107236715B CN 201710347027 A CN201710347027 A CN 201710347027A CN 107236715 B CN107236715 B CN 107236715B
Authority
CN
China
Prior art keywords
raccoon
canine distemper
virus
strain
raccoon dog
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201710347027.1A
Other languages
Chinese (zh)
Other versions
CN107236715A (en
Inventor
闫喜军
胡博
吕爽
王洋
廉士珍
刘昊
张蕾
白雪
鲁荣光
张海玲
赵建军
朱言柱
史宁
薛向红
邓效宇
徐淑娟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute Special Animal and Plant Sciences CAAS
Original Assignee
Institute Special Animal and Plant Sciences CAAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute Special Animal and Plant Sciences CAAS filed Critical Institute Special Animal and Plant Sciences CAAS
Priority to CN201710347027.1A priority Critical patent/CN107236715B/en
Publication of CN107236715A publication Critical patent/CN107236715A/en
Application granted granted Critical
Publication of CN107236715B publication Critical patent/CN107236715B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • 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
    • 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
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
    • C12N2760/18421—Viruses as such, e.g. new isolates, mutants or their genomic sequences
    • 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
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
    • C12N2760/18434—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The invention discloses a raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccines, and belongs to the technical field of veterinary biological products. According to the invention, through a raccoon body continuous passage mode, the canine distemper vaccine strain CDV-O2 is artificially domesticated, and a canine distemper virus attenuated vaccine strain which is safe to raccoon dogs, good in immunogenicity and stable in titer is obtained and named as CDV-OR12 strain, and is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms (CGMCC NO. 13793). The invention provides a new technical means for preventing and controlling raccoon dog distemper.

Description

Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine
Technical Field
The invention relates to a raccoon canine distemper virus attenuated vaccine strain and application thereof in preparation of raccoon canine distemper live vaccines, and belongs to the technical field of veterinary biological products.
Background
Canine distemper is an acute, thermal and highly contact infectious disease caused by the canine distemper virus of measles virus of paramyxoviridae, young animals mostly pass through the acute and lethal way, adult animals can be infected chronically and persistently, clinical symptoms are mainly characterized by biphasic body temperature rise and accompanied by catarrhal pneumonia and gastroenteritis, occasionally neurological symptoms occur, mixed infection and secondary infection of other bacterial viruses are easy to secondary occur, the death rate can reach 80 percent, and the canine distemper is one of the most serious epidemic diseases harming fur-bearing animal breeding industry in China.
The raccoon dog distemper can occur in any season all the year around, and especially, the raccoon dogs are more sensitive before and after lactation interruption and during the breeding period, so that the raccoon dogs have very high morbidity and mortality, and have certain resistance for old raccoon dogs. The virus-carrying animals are the main infection source of raccoon dog distemper, and the virus is discharged through eye and nose secretion, saliva, urine and excrement and is mainly infected through digestive tract and respiratory tract. Because raccoon canine distemper has the characteristics of acute morbidity and lack of a specific treatment method, raccoon canine distemper still takes vaccination as a main prevention and control means, however, no raccoon canine distemper vaccine is sold in the market at present, and a plurality of farmers use canine distemper vaccines of animals such as dogs, minks and the like to prevent raccoon canine distemper and cannot obtain a good immune protection effect. Therefore, the development of safe raccoon canine distemper vaccines with good immune effect is not only an urgent need of the industry and the market, but also has wide market application prospect and has important significance for preventing and controlling raccoon canine distemper.
Disclosure of Invention
The invention aims to provide a raccoon dog pestivirus attenuated vaccine strain and application thereof in preparation of raccoon dog pestivirus live-heat vaccines.
In order to achieve the purpose, the invention adopts the following technical means:
the invention artificially domesticates the canine distemper vaccine strain CDV-O2 in a raccoon dog continuous passage way, and obtains the canine distemper virus attenuated vaccine strain which is safe to raccoon dogs, has good immunogenicity and stable titer.
The raccoon Canine Distemper virus attenuated vaccine strain obtained by the invention is named as CDV-OR12 strain and classified as Canine Distemper virus, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, is preserved in the microorganism research institute of China academy of sciences No. 3 of West Lu No.1 of North West Chen in the sunny district of Beijing, has the preservation number of CGMCC NO.13793 and the preservation time of 2017, 04 months and 21 days.
Furthermore, the invention also provides application of the raccoon dog distemper virus attenuated vaccine strain in preparation of raccoon dog distemper live vaccines.
Experiments prove that the raccoon canine distemper live vaccine prepared from the raccoon canine distemper virus attenuated vaccine strain can effectively protect raccoon dogs from being attacked by canine distemper virus virulent virus, the protection rate is 100% after 3 weeks and 3 months of immunization, and the protection rate is 90% after 6 months of immunization. Therefore, the vaccine can effectively prevent the raccoon dog distemper from occurring, the immune period is 6 months, and the invention provides a new technical means for preventing and controlling the raccoon dog distemper.
Detailed Description
The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
Example 1 domestication and identification of Canine distemper Virus vaccine Strain CDV-O2
1. Domestication of canine distemper virus vaccine strain CDV-O2
(1) Inoculating the preserved canine distemper virus vaccine strain CDV-O2 on African green monkey kidney cells (Vero) in an adsorption and virus inoculation mode for virus amplification culture, observing cell virus conditions every day, harvesting virus liquid by a repeated freeze thawing method when cytopathic effect reaches about 80%, and performing virus content (TCID)50≥105.0Ml), sterility testing. Subpackaging the qualified virus liquid and storing at-20 ℃.
(2) And inoculating 3 antibody-negative healthy raccoon dogs 5ml each to the amplified CDV-O2 virus liquid stock solution of the hind limb muscle, inoculating the raccoon dogs at three points, separately feeding the raccoon dogs and observing various physiological indexes of the inoculated raccoon dogs every day. 1 raccoon dog is killed by dissecting 3 days and 5 days after inoculation, spleen is collected under aseptic condition, virus is separated by using Vero cells, and identification is carried out by using canine distemper virus specific primers and canine distemper virus positive serum.
(3) Marking the separated canine distemper virus as CDV-OR3, namely domesticating the raccoon dog in the 1 st generation (F1), inoculating CDV-OR3 on Vero cells in an adsorption virus inoculation mode for virus amplification culture, observing the condition of cell virus every day, harvesting virus liquid by a repeated freeze thawing method when the cytopathic effect reaches about 80%, and performing virus content (TCID)50≥105.0Ml), sterility testing. Subpackaging the qualified virus liquid and storing at-20 ℃.
(4) And (3) repeating the steps (2) and (3), continuously domesticating the canine distemper virus in the raccoon dog for 9 times to obtain a canine distemper virus attenuated vaccine strain which is a 10 th generation (F10) strain domesticated in the raccoon dog, naming the strain as CDV-OR12 strain and being preserved in the common microorganism center of China Committee for culture Collection of microorganisms, wherein the preservation address is the institute of microorganisms of China academy of sciences No. 3, North Cheng Wen Lu 1 institute of China, North Yang district, Beijing city, the preservation number of the microorganism is CGMCC NO.13793, and the preservation time is 2017, 04 and 21 days.
2. Determination of immunogenicity of racoon dog by canine distemper virus attenuated vaccine strain CDV-OR12
Adjusting virus content to 10 by taking the preserved canine distemper virus attenuated vaccine strain CDV-OR124.5TCID50And (2) inoculating 5 healthy raccoon dogs which are negative in antibody to the muscle of the hind limb, wherein each dog is 1ml, measuring the titer of the neutralizing antibody of the canine distemper virus in serum after 21 days of inoculation, and using the canine distemper virus virulent HBF-1 strain to challenge, and measuring the protection rate. The result shows that the titer of the canine distemper virus neutralizing antibody in the raccoon serum inoculated by the canine distemper virus attenuated vaccine strain CDV-OR12 is respectively as follows: 1:256, 1:362, 1:256, 1:181, 1:362, the protection against toxic attack is 100% (5/5).
3. Return-to-strong toxicity test of canine distemper virus attenuated vaccine strain CDV-OR12 on raccoon dogs
And continuously inoculating the preserved canine distemper virus attenuated vaccine strain CDV-OR12 into antibody-negative raccoon dogs for 5 generations of muscles, and continuously observing for 14 days each generation. The result shows that physiological indexes of body temperature, appetite, mental state, excrement and the like of the raccoon dog inoculated in each generation are normal, and no canine distemper symptom appears, so that the avirulent return phenomenon of the canine distemper attenuated vaccine strain CDV-OR12 is proved, and the method can be used for researching the raccoon dog distemper vaccine.
Example 2 preparation of raccoon dog distemper live vaccine (CDV-OR12 strain)
1. Preparation of canine distemper virus vaccine preparation liquid
Inoculating the preserved canine distemper virus attenuated vaccine strain CDV-OR12 strain to Vero cells in an adsorption and virus inoculation mode for virus amplification culture, observing the condition of cell viruses every day, harvesting virus liquid by a repeated freeze thawing method when the cytopathic effect reaches about 80%, and carrying out virus content and purity test to obtain the content TCID of the raccoon canine distemper virus attenuated vaccine strain in the obtained virus liquid50≥104.5And/ml. According to the measured virus content data, the qualified virus liquid is testedAfter being diluted properly with serum-free MEM, the suspension was quantitatively dispensed into 90ml vaccine bottles and stored at-20 ℃.
2. Determination of canine distemper virus content in vaccine finished product
The virus content of the vaccine finished product is measured, and the content of the canine distemper virus TCID is measured50Is 104.5-105.5First part.
Example 3 safety test of raccoon dog distemper live vaccine (CDV-OR12 strain)
The qualified raccoon canine distemper live vaccine (prepared in example 2) is taken, the hind limb muscle is inoculated with the antibody-negative raccoon, and a single dose (1 part is inoculated and the content of the canine distemper virus TCID is determined)50＝104.5Per head), single dose repeat and overdose (10 times) to the immunized group, while setting the non-vaccinated raccoon dog as a negative control. After the vaccination, the physiological indexes of the raccoon dog, such as body temperature, appetite, mental state, excrement and the like, are observed for 14 consecutive days, and the pathological changes of the main organs of the vaccinated raccoon dog are observed by means of autopsy. Test results show that after single-dose, single-dose repeat and overdose inoculation, all physiological indexes of the raccoon dog are normal and do not show any clinical symptoms, and the raccoon dog inoculated with the raccoon dog main visceral organs is free of pathological changes through autopsy observation, which indicates that the raccoon dog distemper live vaccine is safe for the raccoon dog.
Example 4 efficacy and immunization period test of raccoon canine distemper live vaccine (CDV-OR12 strain)
The qualified raccoon canine distemper live vaccine (prepared in example 2) is taken, and the hind limb muscle is inoculated with 1ml of antibody-negative raccoon (inoculated with 1 part of vaccine with canine distemper virus content TCID)50＝104.5First part), and a control group inoculated with Vero cell culture solution is also arranged. Blood was collected 3 weeks, 3 months, 6 months and 9 months after inoculation, serum was separated, and neutralizing antibodies against canine distemper virus in serum were measured while using 100-half Lethal Dose (LD)50) The canine distemper test uses virulent HBF-1 to attack, and the physiological indexes of the attacked raccoon dog, such as body temperature, appetite, mental state, feces and the like, are observed for 14 days continuously.
The result shows that after 3 weeks and 3 months of inoculation, the titer of the neutralizing antibody of the canine distemper virus in the serum of the inoculated raccoon dog is more than 1:256, and the protection rate of the raccoon dog on the strong canine distemper virus is 100 percent (10/10); after 6 months of inoculation, the protection rate of the raccoon dog on the canine distemper virus is 90 percent (9/10); after 9 months of inoculation, the protection rate of the raccoon dog on the canine distemper virus is 70 percent (7/10). Thus, the immunization period of the vaccine was 6 months.
Example 5 comparison test of the immune efficacy of racoon dog Canine distemper live vaccine (CDV-OR12 strain) and similar vaccine (CDV3-CL strain) on racoon dog
The qualified raccoon dog canine distemper live vaccine (prepared in example 2) and the commercial mink canine distemper live vaccine (CDV3-CL strain) are respectively inoculated to the raccoon dogs with negative antibody at the hind limb muscles, and each raccoon dog is inoculated with 1 part (the content of the canine distemper virus is TCID)50＝104.5Per head), along with a control group that was not vaccinated. Blood was collected 3 weeks, 3 months, 6 months and 9 months after inoculation, serum was separated, and neutralizing antibodies against canine distemper virus in serum were measured while using 100-half Lethal Dose (LD)50) The canine distemper test uses virulent HBF-1 to attack, and the physiological indexes of the attacked raccoon dog, such as body temperature, appetite, mental state, feces and the like, are observed for 14 days continuously.
The result shows that after 3 weeks and 3 months of inoculation, the titer of the neutralizing antibody of the canine distemper virus in the serum of the raccoon canine distemper live vaccine immune group is 1:256, the protection rate of the inoculated raccoon dog against the canine distemper virus is 100% (10/10), the titer of the neutralizing antibody of the canine distemper virus in the serum of the mink canine distemper live vaccine immune group is more than 1:128, and the protection rate of the inoculated raccoon dog against the canine distemper virus is 100% (10/10); after 6 months of inoculation, the protection rate of the raccoon dog canine distemper live vaccine immunization group for inoculating raccoon dogs to the canine distemper virus is 100 percent (10/10), and the protection rate of the mink canine distemper live vaccine immunization group for inoculating raccoon dogs to the canine distemper virus is 80 percent (8/10); after 9 months of inoculation, the protection rate of the raccoon dog canine distemper live vaccine immunization group to the raccoon dog against the canine distemper virus is 70 percent (7/10), and the protection rate of the mink canine distemper live vaccine immunization group to the raccoon dog against the canine distemper virus is only 50 percent (5/10).
The above results show that: the immunization efficacy of the raccoon dog canine distemper live vaccine (CDV-OR12 strain) prepared by the invention is superior to that of the clinically applied mink canine distemper live vaccine.
The experiment results show that the raccoon dog canine distemper live vaccine prepared by the invention can effectively protect raccoon dogs from being attacked by strong canine distemper viruses and effectively prevent raccoon dog canine distemper.

Claims (2)

1. The racoon dog canine distemper virus attenuated vaccine strain is named as CDV-OR12 strain, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, is preserved in the microorganism institute of China academy of sciences No. 3 of Xilu No.1, North Chen, west, in the area facing the sun, in Beijing, has the preservation number of CGMCC NO.13793 and the preservation time of 2017, 04 months and 21 days.
2. Use of the raccoon canine distemper virus attenuated vaccine strain of claim 1 in preparation of raccoon canine distemper live vaccines.
CN201710347027.1A 2017-05-16 2017-05-16 Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine Active CN107236715B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710347027.1A CN107236715B (en) 2017-05-16 2017-05-16 Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710347027.1A CN107236715B (en) 2017-05-16 2017-05-16 Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine

Publications (2)

Publication Number Publication Date
CN107236715A CN107236715A (en) 2017-10-10
CN107236715B true CN107236715B (en) 2020-01-14

Family

ID=59984933

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710347027.1A Active CN107236715B (en) 2017-05-16 2017-05-16 Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine

Country Status (1)

Country Link
CN (1) CN107236715B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116064317B (en) * 2022-11-16 2023-10-17 石家庄市农林科学研究院 Raccoon dog-derived extra-intestinal pathogenic escherichia coli and application thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101612396B (en) * 2009-07-17 2011-12-07 齐鲁动物保健品有限公司 Canine distemper live vaccine and preparation method thereof
CN101914503B (en) * 2010-07-30 2012-04-18 中国农业科学院哈尔滨兽医研究所 Canine distemper attenuated vaccine strain and application thereof
CN106399260B (en) * 2015-07-31 2021-03-02 北京大北农科技集团股份有限公司动物医学研究中心 Canine distemper and parvovirus bivalent live vaccine and preparation method thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Isolation and sequence analysis of a canine distemper virus from a raccoon dog in jilin province,China;ChengＹ et al.;《Virus Genes》;20151031;第51卷(第2期);第298-301页 *

Also Published As

Publication number Publication date
CN107236715A (en) 2017-10-10

Similar Documents

Publication Publication Date Title
Matthijs et al. Ability of Massachusetts-type infectious bronchitis virus to increase colibacillosis susceptibility in commercial broilers: a comparison between vaccine and virulent field virus
CN104383530B (en) Mink canine distemper-Canine Parvovirus Enteritis bigeminal live vaccine and its production and use
WO2021103421A1 (en) Gene vii type newcastle disease virus attenuated strain and use thereof
CN102220287B (en) Avian infectious bronchitis cold adaptation attenuated vaccine strain and application thereof
CN101117627A (en) Pig epidemic diarrhea virus attenuated vaccine strain and uses thereof
CN110387354B (en) Pseudorabies virus passage attenuated strain and application thereof
CN112316128B (en) Canine distemper virus and canine parvovirus infectious hepatitis virus triple live vaccine
CN111748529B (en) Porcine pseudorabies virus strain and application thereof
WO2021217959A1 (en) Recombinant vector containing african swine fever virus immunogenic protein, recombinant bacteria, and application thereof
CN108066758B (en) Raccoon dog canine distemper-parvovirus enteritis bigeminal live vaccine and preparation method and application thereof
CN111073863B (en) Porcine epidemic diarrhea and porcine delta coronavirus bivalent attenuated vaccine and preparation method thereof
CN106754594A (en) A kind of Salmonella choleraesuls attenuated carrier bacterium and its construction method
CN107158370B (en) Raccoon dog canine distemper freeze-dried live vaccine and preparation method and application thereof
CN110302374B (en) Canine vaccine and preparation method and application thereof
CN107236715B (en) Raccoon dog distemper virus attenuated vaccine strain and application thereof in preparation of raccoon dog distemper live vaccine
CN101745106A (en) Porcine parvnvirus living vaccine and preparation method thereof
CN106190988B (en) Inactivated vaccine of feline calicivirus CH-JL5 strain
CN107338227B (en) Bovine parainfluenza virus PBIV3-B strain and application thereof
CN112402599B (en) Canine distemper and parvovirus disease bivalent inactivated vaccine for dogs and preparation method thereof
CN105039270B (en) H9N2 subtype avian influenza acclimatization to cold attenuated strains and its application
CN107881154B (en) Raccoon parvovirus attenuated vaccine strain and application thereof in preparation of raccoon parvovirus attenuated vaccine
CN107115525B (en) Racoon dog canine distemper live vaccine and its preparation method and application
CN112807424A (en) Bivalent live vaccine for bovine viral diarrhea and bovine infectious rhinotracheitis and preparation method thereof
CN104388394B (en) Mink Parvovirus attenuated vaccine strain and its purposes in Mink Parvovirus attenuated vaccine is prepared
CN107029230A (en) A kind of vaccine combination, kit and its preparation method and application

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant