CN102847147A - Combined live vaccine for muscovy duck parvovirus disease and muscovy duck reovirus disease - Google Patents
Combined live vaccine for muscovy duck parvovirus disease and muscovy duck reovirus disease Download PDFInfo
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- CN102847147A CN102847147A CN2012100461390A CN201210046139A CN102847147A CN 102847147 A CN102847147 A CN 102847147A CN 2012100461390 A CN2012100461390 A CN 2012100461390A CN 201210046139 A CN201210046139 A CN 201210046139A CN 102847147 A CN102847147 A CN 102847147A
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Abstract
The invention discloses preparation and application of a combined vaccine for a muscovy duck parvovirus disease and a muscovy duck reovirus disease; the combined vaccine adopts a muscovy duck parvovirus attenuated P1 strain (preservation number: CCTCC V201013) and a muscovy duck liver white-spot disease (muscovy duck reovirus disease) attenuated MWCA strain (preservation number: CGMCC 0667) as seed viruses; the seed viruses are proliferated by using muscovy duck embryo fibroblasts and SPF chicken embryo fibroblasts through culture in spinner flasks; the cell viral liquid is gained, mixed according to a proper proportion, and freeze-dried by adding a freeze-drying protective agent to obtain the combined live vaccine; the combined live vaccine is applicable to muscovy duck breeding areas with epidemic muscovy duck three-week diseases and liver white-spot diseases.
Description
Technical field
The present invention relates to the sick bigeminal live vaccine of Muscovy duck parvovirus infection and muscovy duck reovirus, belong to the Preventive Veterinary Medicine field.
Background technology
Muscovy duck parvovirus infection (being commonly called as " three week sick ") be 1987 our province occurs first 1-3 age in week young Muscovy duck to breathe heavily a kind of new epidemic disease of rushing down as clinical symptoms.This disease is mainly encroached on the young Muscovy duck of 7-21 age in days, sickness rate 27-62% and case fatality rate 22-43%, and the duck major part of recovering becomes stiff duck, already causes very large economy loss for Muscovy duck.Muscovy duck reovirus sick (being commonly called as " liver ichthyophthiriasis ", " flower hepatopathy ", " ichthyophthiriasis ", " liver spleen ichthyophthiriasis " and " Muscovy duck hepatitis sequestrans " etc.) has been since 1997, in the Putian, Fujian Province, Fuqing, Foochow, permanently happy and ground Muscovy duck such as Jinhua, Zhejiang, Foshan raise the district and occur a kind ofly clinically take weak foot as cardinal symptom, have volume white point, kidney enlargement, hemorrhage, surface yellow streak to be arranged as the infectious disease of main pathological change take liver, spleen surface.This disease only betides Muscovy duck, and the morbidity age in days is 7～45 ages in days, with the young Muscovy duck of 10～30 ages in days for the most very.Sickness rate is 30～90%, and case fatality rate is 60～80%, becomes stiff duck after the dead duck of morbidity is anti-, already causes very large economy loss for Muscovy duck.
At present, Muscovy duck parvovirus infection and muscovy duck reovirus disease are still the Important Infectious Diseases of serious harm Muscovy duck aquaculture, and the immunity inoculation of vaccine can effectively be controlled this two kinds of new epidemic diseases, and repeatedly immunity easily causes bad stress to the Muscovy duck group.For this reason, the present invention develops muscovy duck reovirus disease and Muscovy duck parvovirus infection bigeminal live vaccine at home and abroad first.
Summary of the invention
The purpose of this invention is to provide the sick bigeminal live vaccine of Muscovy duck parvovirus infection and muscovy duck reovirus, the present invention is achieved by the following technical solution:
A, derive from the weak malicious P1 strain (MPV-P1 strain) of the exquisite weak Muscovy duck parvovirus of people, be deposited in Chinese Typical Representative culture collection center (China Center for Type Culture Collection, be called for short: CCTCC), depositary institution CCTCC address is in Wuhan, China city Wuhan University, June 2 2010 preservation day, deposit number: CCTCCNO:V201013;
B, derive from the weak malicious MWCA strain of the exquisite weak foreign duck liver ichthyophthiriasis of people (muscovy duck reovirus is sick), this strain is deposited at China Committee for Culture Collection of Microorganisms common micro-organisms center and (is called for short: CGMCC), the depositary institution address is in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, calendar year 2001 preservation day December 18 days, deposit number: CGMCC0667;
Malicious P1 strain a little less than malicious MWCA strain and the Muscovy duck parvovirus a little less than the above-mentioned foreign duck liver ichthyophthiriasis is applied to prepare Muscovy duck parvovirus infection and the sick bigeminal live vaccine of muscovy duck reovirus.
Compared with prior art, the present invention a little less than the Muscovy duck parvovirus malicious P1 strain (MPV-P1 strain) and the weak malicious MWCA strain of foreign duck liver ichthyophthiriasis (that is: the weak malicious CA strain of muscovy duck reovirus) for kind malicious, use respectively Muscovy duck embryo fibroblast and SPF chick embryo fibroblast spinner culture technology virus of proliferation, the harvesting venom, by adding the freeze drying protectant lyophilization after the proper proportion mixing, be developed into genetic stability, safety is good, the bigeminal live vaccine that immunogenicity is strong and cost is low, use in Muscovy duck cultivation area at popular Muscovy duck three all sick regulating liver-QI ichthyophthiriasises, can reach the purpose that one shot immunity could prevent and control these two kinds of epidemic diseases simultaneously, the young Muscovy duck immunity of 1-2 age in days once can effectively prevent Muscovy duck parvovirus infection and muscovy duck reovirus sick, not only reduced manpower and materials, also reduced because of inoculation repeatedly the distress that the duck group causes is reacted; The present invention is suitable for the large-scale production under the GMP condition.
The specific embodiment
The present invention is further described below in conjunction with specific embodiment.(but not being limitation of the present invention).
A, derive from the weak malicious P1 strain (MPV-P1 strain) of the exquisite weak Muscovy duck parvovirus of people, to utilize Muscovy duck embryo and Muscovy duck embryo fibroblast alternately to go down to posterity etc. to form a little less than the biotechnology artificially breeding causes, lost pathogenic to the young Muscovy duck of an age in days, and had following characteristic (1) virus titer: the MPV-P1 strain is to Muscovy duck embryo fibroblast cell monolayer TCID
50〉=10
-50/ 0.1ml; (2) safety: with young Muscovy duck 10 plumages of the healthy susceptible of 1 age in days, 0.2 milliliter of every plumage leg muscle injection seed culture of viruses stock solution was observed 20 days, should all survive; (3) immunogenicity: the healthy responsive young Muscovy duck of an age in days, 0.2 milliliter of leg muscle injection, its minimum immunity amount is 100TCID
50The MPV-P1 strain is deposited in CCTCC, June 2 2010 preservation day, deposit number CCTCCV201013;
B derives from the weak malicious MWCA strain of the exquisite weak foreign duck liver ichthyophthiriasis of people (muscovy duck reovirus is sick), to utilize Muscovy duck embryo fibroblast and chick embryo fibroblast to go down to posterity to form a little less than mutagenic and breeding causes, lost pathogenic to the young Muscovy duck of an age in days, and had following characteristic (1) virus titer: the MWCA strain is to chick embryo fibroblast cell monolayer TCID
50〉=10
-50/ 0.1ml; (2) safety: with young Muscovy duck 10 plumages of the healthy susceptible of 1 age in days, 0.2 milliliter of every plumage leg muscle injection seed culture of viruses stock solution was observed 20 days, should all survive; (3) immunogenicity: the young Muscovy duck of the healthy susceptible of an age in days, the lower limb intramuscular injection is penetrated 0.2 milliliter, and its minimum immunity amount is 300TCID
50This strain is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, (existing name: Chinese common micro-organisms culture presevation administrative center, China General Microbiological Culture Collection Center, be called for short CGMCC), the address is in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, calendar year 2001 preservation day December 18 days, deposit number: CGMCC0667.(see and authorize China Patent No. 01144485.1, Granted publication CN1179751C; The weak malicious MWCA strain of foreign duck liver ichthyophthiriasis (muscovy duck reovirus is sick) also claims in this patent documentation: the CGMCC0667 strain);
With the weak malicious MWCA strain of malicious P1 strain and foreign duck liver ichthyophthiriasis a little less than the above-mentioned Muscovy duck parvovirus (muscovy duck reovirus sick) for the preparation of Muscovy duck parvovirus infection and the sick bigeminal live vaccine of muscovy duck reovirus.
The concrete preparation method of the sick bigeminal live vaccine of above-mentioned Muscovy duck parvovirus infection and muscovy duck reovirus is as follows:
(1) MPV-P1 strain propagation, by the 1% synchronous inoculation Muscovy duck embryo fibroblast (MDEF) of cultivating liquid measure, 37 ℃ of rotating and culturing to cytopathys reach more than 75%, results MPV-P1 cell toxicant with MPV-P1 strain kind poison.
(2) MDRV-CA strain propagation connects the 1% synchronous inoculation SPF chick embryo fibroblast (CEF) of cultivating liquid measure with MDRV-CA strain kind poison, and 37 ℃ of rotating and culturing to cytopathys are stated more than 75%, results MDRV-CA cell toxicant.
(3) viral level TCID
50Measure the TCID50 difference 〉=10 of MDRV-CA and MPV-P1
-50/ 0.1ml and 〉=10
-4.5When/0.1ml is above, can be used for preparing vaccine.
(5) vaccine preparation adds the freeze drying protectant lyophilization after MPV-P1 and MDRV-CA mixed by proper proportion and forms.Make the viral level of various compositions in every plumage part vaccine reach respectively MDRVCA10
3.5TCID
50And MPV-P110
30TCID
50
Vaccine of the present invention is applied to prevent and treat the Muscovy duck parvovirus infection and muscovy duck reovirus is sick, and its amynologic index is determined as follows:
1. steriling test: carry out asepsis growth by existing " People's Republic of China's veterinary drug allusion quotation " appendix.
2. Security test: with young Muscovy duck 10 plumages of the healthy susceptible of 1 age in days, every plumage leg muscle is injected 10 multiple dose triple vaccinies, observes 20 days, and the inoculation duck is showed no diet wants and psychological problem, and growth promoter is good, all survives.
3. immunity is measured: get at random young Muscovy duck 20 plumages of an age in days susceptible, lower limb flesh is inoculated 1 plumage part bigeminal live vaccine, wherein 10 plumage immune ducks are measured for anti-MPV-LPAI antibody titer in inoculating 0 day, 7 days, 14 days, 21 days, 28 days, 60 days, 90 days, 150 days, 180 days together with not immune contrast duck blood sampling separation of serum; 10 plumage immune ducks are attacked by force poison of MDRV in inoculating rear 7 days simultaneously together with not immune matched group in addition, observe record Mortality situation 20 days.The result: 1. the immunity of the young Muscovy duck bigeminy of age in days Seedling produced antibody in rear 5 days, and the anti-MPV-LPAI antibody in rear 7 days serum of immunity is 2
2.1Above, be highly resistant to the MPV strong virus attack, extremely the rear antibody horizontal that peaked in 28 days of immunity is respectively 2
6.0, subsequently antibody horizontal still young bird be held in higher level, and continue to rear 180 days of immunity always, surpassed Muscovy duck this moment and delivered age in days for sale.2. rear 7 days of the young Muscovy duck bigeminy of age in days Seedling immunity is attacked the strong malicious 0.2ml/ plumage of MDRV, sickness rate 80% (8/10), the mortality rate 30% (3/10) of contrast counteracting toxic substances duck simultaneously together with not immune matched group; And sickness rate 10% (1/10), the mortality rate 0% of Immunization duck; Show the attack that was highly resistant to the strong poison of MDRV behind the young Muscovy duck immunity of the age in days bigeminy vaccine in 7 days.3. the immunity measurement result of bigeminy vaccine shows, vaccine immunity power is good, and 7 days immune generation phases, immune duration is more than 180 days.Primary immune response can protect young Muscovy duck to ride out vulnerable period; reach 1 inoculation and can prevent simultaneously Muscovy duck parvovirus infection (three weeks of Muscovy duck are sick) and muscovy duck reovirus sick (foreign duck liver ichthyophthiriasis); not only reduce manpower, material resources and financial resources, also reduce the stress that causes because of inoculation repeatedly.
Effect evaluation of the present invention: be total to the young Muscovy duck of inoculation plumage more than 6500 part one age in days in Ceng Liuhang Muscovy duck three all test duck fields sick and the foreign duck liver ichthyophthiriasis, tracing observation 20d has no immune duck and untoward reaction occurs; And the Muscovy duck parvovirus infection does not occur in the immune group Muscovy duck and muscovy duck reovirus is sick, and survival rate reaches more than 97.5%; And the foreign duck liver ichthyophthiriasis all occurs in the matched group Muscovy duck of 650 plumages in various degree and three weeks are sick, survival rate only 73%.Further proof bigeminal live vaccine safety of the present invention, effective can be used for preventing clinically muscovy duck reovirus sick (foreign duck liver ichthyophthiriasis) and Muscovy duck parvovirus infection (three weeks of Muscovy duck are sick).
Claims (3)
1. the sick bigeminal live vaccine of Muscovy duck parvovirus infection and muscovy duck reovirus is characterized in that, the sick bigeminal live vaccine of described Muscovy duck parvovirus infection and muscovy duck reovirus is to adopt:
A, derive from the weak malicious P1 strain of the exquisite weak Muscovy duck parvovirus of people, be deposited in Chinese Typical Representative culture collection center, June 2 2010 preservation day, deposit number: CCTCCNO:V201013;
B, derive from the weak malicious MWCA strain of the exquisite weak foreign duck liver ichthyophthiriasis of people, this strain is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, calendar year 2001 preservation day December 18 days, deposit number: CGMCC0667;
With malicious MWCA strain a little less than malicious P1 strain and the foreign duck liver ichthyophthiriasis a little less than the above-mentioned Muscovy duck parvovirus for the preparation of the sick bigeminal live vaccine of Muscovy duck parvovirus infection and muscovy duck reovirus.
2. the sick bigeminal live vaccine of Muscovy duck parvovirus infection according to claim 1 and muscovy duck reovirus is characterized in that, the sick bigeminal live vaccine preparation method of described Muscovy duck parvovirus infection and muscovy duck reovirus is as follows:
A.MPV-P1 strain propagation, by the 1% synchronous inoculation Muscovy duck embryo fibroblast of cultivating liquid measure, 37 ℃ of rotating and culturing to cytopathys reach more than 75%, results MPV-P1 cell toxicant with MPV-P1 strain kind poison;
B MDRV-CA strain propagation, by the 1% synchronous inoculation SPF chick embryo fibroblast of cultivating liquid measure, 37 ℃ of rotating and culturing to cytopathys reach more than 75%, results MDRV-CA cell toxicant with MDRV-CA strain kind poison;
C. viral level TCID50 measures, the TCID50 difference 〉=10 of MPV-P1 and MDRV-CA
-4.5/ 0.1ml and 〉=10
-5.0When/0.1ml is above, be used for the preparation vaccine;
E. vaccine preparation adds the freeze drying protectant lyophilization after MPV-P1 and MDRV-CA mixed by proper proportion and forms, and makes the viral level of various compositions in every plumage part vaccine reach respectively MDRV CA 10
3.5TCID
50And MPV-P110
3.0TCID
50
3. muscovy duck reovirus disease according to claim 1 and 2 and the application of Muscovy duck parvovirus infection bigeminal live vaccine on control Muscovy duck parvovirus infection and muscovy duck reovirus disease.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103833848A (en) * | 2014-03-07 | 2014-06-04 | 青岛易邦生物工程有限公司 | Egg yolk antibody for preventing and curing muscovy duck parvovirus diseases |
| CN108660117A (en) * | 2018-05-22 | 2018-10-16 | 山东农业大学 | One kind leading to the arthritic novel avian reovirus of broiler chicken and its application |
| CN111053897A (en) * | 2019-12-19 | 2020-04-24 | 广州渔跃生物技术有限公司 | Duck reovirus and duck adenovirus bivalent inactivated vaccine and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1426814A (en) * | 2001-12-19 | 2003-07-02 | 福建省农业科学院畜牧兽医研究所 | Foreign duck liver ichthyophthirius active vaccine and its preparation method |
| CN101880651A (en) * | 2010-07-01 | 2010-11-10 | 福建省农业科学院畜牧兽医研究所 | Preparation method of Muscovy duck parvo novel vaccines |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1426814A (en) * | 2001-12-19 | 2003-07-02 | 福建省农业科学院畜牧兽医研究所 | Foreign duck liver ichthyophthirius active vaccine and its preparation method |
| CN101880651A (en) * | 2010-07-01 | 2010-11-10 | 福建省农业科学院畜牧兽医研究所 | Preparation method of Muscovy duck parvo novel vaccines |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103833848A (en) * | 2014-03-07 | 2014-06-04 | 青岛易邦生物工程有限公司 | Egg yolk antibody for preventing and curing muscovy duck parvovirus diseases |
| CN103833848B (en) * | 2014-03-07 | 2016-05-04 | 青岛易邦生物工程有限公司 | A kind of Yolk antibody for prophylactic treatment kind Duck parvovirus disease |
| CN108660117A (en) * | 2018-05-22 | 2018-10-16 | 山东农业大学 | One kind leading to the arthritic novel avian reovirus of broiler chicken and its application |
| CN111053897A (en) * | 2019-12-19 | 2020-04-24 | 广州渔跃生物技术有限公司 | Duck reovirus and duck adenovirus bivalent inactivated vaccine and preparation method thereof |
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