CN102327603B - Preparation method and use method of loach aeromonas sobria vaccine - Google Patents
Preparation method and use method of loach aeromonas sobria vaccine Download PDFInfo
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- CN102327603B CN102327603B CN 201110040577 CN201110040577A CN102327603B CN 102327603 B CN102327603 B CN 102327603B CN 201110040577 CN201110040577 CN 201110040577 CN 201110040577 A CN201110040577 A CN 201110040577A CN 102327603 B CN102327603 B CN 102327603B
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- aeromonas sobria
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- sobria
- radix astragali
- loach
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Abstract
The invention discloses a preparation method of a loach aeromonas sobria vaccine, which is characterized by comprising the following step: preparing a water aqua type loach aeromonas sobria vaccine using radix astragali as an adjuvant by using loach pathogenic aeromonas sobria LD081008A-1 as immunogen and a radix astragali lixivium as the adjuvant, wherein the concentration of the pathogenic aeromonas sobria as the immunogen in the vaccine is 2.7*108 cells/mL, and the concentration of the radix astragali lixivium is 1 g/mL. The invention further discloses a use method of the loach aeromonas sobria vaccine. By using the loach aeromonas sobria vaccine prepared by the method, the capability of the resistance of loaches to aeromonas sobria infection can be obviously improved; the immune protecting capability of the loaches to the pathogenic aeromonas sobria is enhanced; the survival rate of cultivating loaches is improved; the yield of cultivating the loaches is increased; the export obstacle caused by drug residue is avoided; the economic benefit in the cultivation production of the loaches is improved; and the problem that diseases caused by the aeromonas sobria easily appearing in the current cultivation production process of the loaches is solved foundationally.
Description
Technical field
The present invention relates to a kind of preparation method of animal disease resistant vaccine, particularly relate to a kind of preparation method of Misgurni anguillicaudati Aeromonas sobria vaccine.The invention still further relates to the using method of this vaccine.
Background technology
Aeromonas sobria extensively is present in water environment, and distribution is all arranged all over the world, belongs to conditioned pathogen, can cause the various diseases of Fish and other aquatic animal, has caused serious economic loss for the Aquatic product aquaculture.Therefore, to the efficient prevention of cause of disease Aeromonas sobria, to reduce to greatest extent its extent of injury, can improve significantly the economic benefit that loach breeding is produced.Aspect the prevention and control of cultured fishes bacterial disease, the generation of most use chemical synthetic drug or antibiotics control disease and popular in breeding production at present, but in long-term drug use, it is a lot of not enough that people recognize that Drug therapy exists: the one, and the cause of disease is complicated, there is blindness in a large amount of medications, and a lot of diseases there is no active drug targetedly; The 2nd, some chemicals significant surfaces are narrow, and a little less than preventive effect, and sick fish often is difficult to picked-up to effective drug dose because of anorexia; The 3rd, medicine is residual in animal body, jeopardizes the mankind's health, and long-term prescription brings environmental pollution, and causes resistant micro-organism to occur in a large number; In addition, make for a long time the resistance against diseases decline that the weak individuality of premunition that should be eliminated in a large number survives and proportion increases and cause whole colony gradually in population under medicine " protective effect ", had a strong impact on the development of aquaculture.Therefore, the generation of applied immunology technical prevention fish bacterial disease has been subject to increasing attention, has become the important content that various countries' researcher is paid close attention to.
Summary of the invention
Technical problem to be solved by this invention is for the deficiencies in the prior art, provides a kind of and improve Misgurni anguillicaudati immunity, reduces the harm that caused by the cause of disease Aeromonas sobria in loach breeding production, with the preparation method of the Misgurni anguillicaudati Aeromonas sobria vaccine that satisfies the disease-resistant demand that loach breeding produces.
Another technology to be solved by this invention has been to provide the immune using method of several Misgurni anguillicaudati Aeromonas sobria vaccines as above.
Technical problem to be solved by this invention is to realize by following technical scheme.The present invention is a kind of preparation method of Misgurni anguillicaudati Aeromonas sobria vaccine, is characterized in: it is with Misgurni anguillicaudati cause of disease Aeromonas sobria LD081008A-1(
A.sobriaLD081008A-1) be immunogen, take Radix Astragali leachate as adjuvant; Its step body step is as follows:
(1) with Aeromonas sobria LD081008A-1(
A.sobriaLD081008A-1) inoculation on the ordinary nutrient agar broth bouillon, is cultivated 24h for 28 ℃, adds 28 ℃ of deactivation 48h of formaldehyde of final concentration 0.3%, gets the deactivation thalline;
(2) get the Radix Astragali and decoct after with distilled water immersion 30-60min, filter, being concentrated into final concentration is former medicine 1g/mL, gets Radix Astragali leachate;
(3) with the deactivation thalline with Radix Astragali leachate mix, make water aqua type Radix Astragali adjuvant Misgurni anguillicaudati Aeromonas sobria vaccine; In vaccine, cause of disease Aeromonas sobria immunogen is 2.7 * 10
8Cells/mL, Radix Astragali leachate are 1g/mL.
The Aeromonas sobria LD081008A-1(that relates in the inventive method
A.sobriaLD081008A-1) open in " Misgurni anguillicaudati Peptic Ulcers and cause of disease Aeromonas sobria biology and characterization of molecules research ", the publication of delivering of the disclosure document is: " aquatic science ", and 2010,29(12): 696-702.The isolate DNA sequence length of this bacterial strain and accession number such as following table:
Isolate DNA sequence length and accession number
The immune using method of the prepared Misgurni anguillicaudati Aeromonas sobria of above-described preparation method vaccine is:
1, it can adopt the method for immersion immunity, first will be exempted from fish and with mass concentration be 8% NaCl solution soaking 5min under venting condition, then 1:5 dilute with water Misgurni anguillicaudati Aeromonas sobria vaccine gets Misgurni anguillicaudati Aeromonas sobria vaccine solution by volume, to be exempted from again Misgurni anguillicaudati and be put in inflation immersion 1h in Misgurni anguillicaudati Aeromonas sobria vaccine solution, be pressed the preceding method booster immunization 1 time after the week.
2, it can adopt the method for injecting immune, gets body long for the Misgurni anguillicaudati of 10-15cm carries out the initial injection immunity by the immunizing dose of 0.1mL/ tail, after week about by aforementioned immunizing dose booster immunization 1 time, immunity is 4 times altogether.
3, it can adopt the oral immunity method, throws something and feeds after hard pellet diet is soaked with Misgurni anguillicaudati Aeromonas sobria vaccine, and the fish body absorption every day inactivated bacteria scale of construction is 2.7 * 10
6The cells/g body weight is thrown something and fed a week continuously, carries out one week of booster immunization by preceding method again after every other week.
The inventor carries out the experiment of Misgurni anguillicaudati immunoprotection 2010 March in year ~ June at vaccine and the immunization method of the preparation of Jiangsu Province's Huaihai Institute of Technology aquatic animal disease use for laboratory the inventive method, and after duration of immunity finished, three kinds of immunization methods were 3 * 10 with concentration respectively
8The Aeromonas sobria LD081008A-1(of CFU/mL
A.sobriaLD081008A-1) infectable infection is exempted from fish.Experimental result for use Aeromonas sobria vaccine of the present invention through soak, oral and inject three kinds of modes immunity Misgurni anguillicaudatis; all can produce certain immune protective efficiency; best with the injection system immune effect, soak, immune protective rate oral and that inject three kinds of modes is respectively 46.6%, 53.3%, 83.3%.See following table for details:
The immune protective efficiency table of immunity fish after the viable bacteria counteracting toxic substances
The Misgurni anguillicaudati Aeromonas sobria vaccine of the inventive method preparation can significantly improve the ability of Misgurni anguillicaudati temperature resistance and Aeromonas infection; strengthen Misgurni anguillicaudati to the immune protective efficiency of cause of disease Aeromonas sobria; improve the survival rate of loach breeding; increase the output of loach breeding; avoid the export obstacle that causes because of drug residue; improve the economic benefit that loach breeding is produced, fundamentally solved the problem of the disease that in present loach breeding production process, easy generation Aeromonas sobria causes.
The specific embodiment
Below further describe concrete technical scheme of the present utility model, so that those skilled in the art understands the present invention further, and do not consist of its Copyright law.
Embodiment 1.A kind of preparation method of Misgurni anguillicaudati Aeromonas sobria vaccine, it is with Misgurni anguillicaudati cause of disease Aeromonas sobria LD081008A-1(
A.sobriaLD081008A-1) be immunogen, take Radix Astragali leachate as adjuvant; Its step body step is as follows:
(1) with Aeromonas sobria LD081008A-1(
A.sobriaLD081008A-1) inoculation on the ordinary nutrient agar broth bouillon, is cultivated 24h for 28 ℃, adds 28 ℃ of deactivation 48h of formaldehyde of final concentration 0.3%, gets the deactivation thalline;
(2) get the Radix Astragali and decoct after with distilled water immersion 30-60min, filter, being concentrated into final concentration is former medicine 1g/mL, gets Radix Astragali leachate;
(3) with the deactivation thalline with Radix Astragali leachate mix, make water aqua type Radix Astragali adjuvant Misgurni anguillicaudati Aeromonas sobria vaccine; In vaccine, cause of disease Aeromonas sobria immunogen is 2.7 * 10
8Cells/mL, Radix Astragali leachate are 1g/mL.
Embodiment 2.The using method of the prepared Misgurni anguillicaudati Aeromonas sobria of a kind of preparation method as described in Example 1 vaccine, it adopts the method for immersion immunity, first will be exempted from fish and with mass concentration be 8% NaCl solution soaking 5min under venting condition, then 1:5 dilute with water Misgurni anguillicaudati Aeromonas sobria vaccine gets Misgurni anguillicaudati Aeromonas sobria vaccine solution by volume, to be exempted from again Misgurni anguillicaudati and be put in inflation immersion 1h in Misgurni anguillicaudati Aeromonas sobria vaccine solution, be pressed the preceding method booster immunization 1 time after the week.
Embodiment 3.The using method of the prepared Misgurni anguillicaudati Aeromonas sobria of a kind of preparation method as described in Example 1 vaccine, it adopts the method for injecting immune, get body length and carry out the initial injection immunity for the Misgurni anguillicaudati of 10-15cm by the immunizing dose of 0.1mL/ tail, after week about by aforementioned immunizing dose booster immunization 1 time, immunity is 4 times altogether.
Embodiment 4.The using method of the prepared Misgurni anguillicaudati Aeromonas sobria of a kind of preparation method as described in Example 1 vaccine, it is characterized in that: it adopts the oral immunity method, throw something and feed after hard pellet diet is soaked with Misgurni anguillicaudati Aeromonas sobria vaccine, the fish body absorption every day inactivated bacteria scale of construction is 2.7 * 10
6The cells/g body weight is thrown something and fed a week continuously, carries out one week of booster immunization by preceding method again after every other week.
Claims (1)
1. the preparation method of a Misgurni anguillicaudati Aeromonas sobria vaccine, it is characterized in that: it is take Misgurni anguillicaudati cause of disease Aeromonas sobria LD081008A-1(A.sobria LD081008A-1) as immunogen, take Radix Astragali leachate as adjuvant; Its concrete steps are as follows:
(1) with Aeromonas sobria LD081008A-1(A.sobria LD081008A-1) inoculation on the ordinary nutrient agar broth bouillon, cultivate 24h for 28 ℃, add 28 ℃ of deactivation 48h of formaldehyde of final concentration 0.3%, get the deactivation thalline;
(2) get the Radix Astragali and decoct after with distilled water immersion 30-60min, filter, being concentrated into final concentration is former medicine 1g/mL, gets Radix Astragali leachate;
(3) the deactivation thalline is mixed with Radix Astragali leachate, make water aqua type Radix Astragali adjuvant Misgurni anguillicaudati Aeromonas sobria vaccine; In vaccine, cause of disease Aeromonas sobria immunogen is 2.7 * 10
8Cells/mL, Radix Astragali leachate are 1g/mL.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110040577 CN102327603B (en) | 2011-02-18 | 2011-02-18 | Preparation method and use method of loach aeromonas sobria vaccine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110040577 CN102327603B (en) | 2011-02-18 | 2011-02-18 | Preparation method and use method of loach aeromonas sobria vaccine |
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| CN102327603B true CN102327603B (en) | 2013-05-08 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109847059A (en) * | 2017-11-30 | 2019-06-07 | 淮海工学院 | A kind of preparation and application of all grand Aeromonas vaccines of loach |
| CN109198268A (en) * | 2018-09-29 | 2019-01-15 | 扬州大学 | The disease-resistant Chinese herbal medicine of mandarin fish and Yolk antibody compound additive and feeding method |
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2011
- 2011-02-18 CN CN 201110040577 patent/CN102327603B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 仇登高等.复方乌梅添加免疫单体对草鱼免疫功能影响的研究.《中国南方十六省（市、区）水产学会渔业学术论坛第二十六次学术交流大会》.2010,522. * |
| 姚东瑞等.泥鳅(Misgurnus anguillicaudatus)病原温和气单胞菌(Aeromonas sobria)分子鉴定及耐药性研究.《海洋与湖泊》.2010,第41卷(第5期),756-762. * |
| 张晓君等.泥鳅溃疡病及病原温和气单胞菌生物学及分子特征研究.《水产科学》.2010,第29卷(第12期),676-702. * |
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