Biological control method of crucian hemorrhagic disease and application thereof

A technology of hemorrhagic disease and crucian carp, which is applied in the field of biological control of crucian carp hemorrhagic disease, can solve the problems of crucian carp breeding industry impact, prevention and control difficulties, vaccines are difficult to work, etc., and achieve the effect of high-efficiency immunization and broad commercial development prospects

Active Publication Date: 2013-09-04
马悦 +1
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since 2009, the virus has successively caused large-scale diseases in crucian carp breeding areas in many provinces and cities in my country, which has brought a huge impact on crucian carp breeding industry
In addition, Aeromonas hydrophila has also been isolated and purified from the diseased crucian carp, and back-staining experiments have proved that crucian carp has bleeding symptoms (Liu Yanan et al., 2012, Journal of Aquatic Ecology 05; Lu Wenhao et al., 2009, Journal of Guangdong Ocean University, Issue 01), the pathogen often occurs simultaneously with viral diseases in the crucian carp culture process, and the developed monovalent vaccines are often difficult to be effective, which brings difficulties to the prevention and treatment

Method used

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  • Biological control method of crucian hemorrhagic disease and application thereof
  • Biological control method of crucian hemorrhagic disease and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Preparation of crucian carp haemorrhagic disease polyvalent carrier vaccine

[0027] 1. Culture medium preparation

[0028] (1) DSM liquid medium: bacterial nutrient broth (Difco) 8g, 10% (w / v) KCl 10ml, 1.2% (w / v) MgSO 4 ·7H 2 O 10ml, 1M NaOH 1.5ml, adjust pH to 7.6, ddH 2 O was adjusted to 1000ml. After autoclaving, cool to 50°C and add sterilized 1M Ca(NO 3 ) 2 , 0.01MMnCl 2 , 1mM FeSO 4 1ml each.

[0029] (2) PBS buffer: Na 2 HPO 4 2H 2 O 2.74g / L, NaH 2 PO 4 ·H 2 O 0.63g / L, the ingredients were dissolved in 1000ml of distilled water, and sterilized at 121°C for 20 minutes.

[0030] (3) LB medium: tryptone (Tryptone) 10g / L, yeast extract (Yeast extract) 5g / L, sodium chloride (NaCl) 10g / L, adjust the pH to 7.0.

[0031] 2. Recovery of the plate medium: take the seed of the recombinant Bacillus subtilis strain preserved at -80°C with the preservation number CCTCCNo: M2013029 and inoculate it on the LB solid plate by streaking and inoculat...

Embodiment 2

[0037] Example 2: Evaluation of the immune effect of carp herpesvirus II (CyHV-2) on crucian carp immunized with multivalent carrier vaccine orally.

[0038] Breeding of experimental fish: The experimental fish (crucian carp) purchased from Jiangsu was temporarily raised in a 2000L aquarium after being tested for the absence of CyHV-2, equipped with a flow circulation fresh water treatment and purification system. After raising for 1 week, remove unhealthy individuals. Select healthy crucian carp with a weight of 350±50g and randomly group them into 200L experimental aquariums, 30 in each aquarium, change the water twice a day in the experimental aquarium, and change the water volume to 1 / 3, maintain the water temperature at 20±2°C, and continue to raise temporarily 2 weeks.

[0039] Preparation of CyHV-2 virus suspension: dissect diseased fish, select gills, kidney and spleen, cut them into pieces and grind them under liquid nitrogen, add TN buffer (50 mmol / L Tris -HCl, ...

Embodiment 3

[0046] Example 3: Evaluation of the immune effect of crucian carp immunized orally with multivalent carrier vaccine against Aeromonas hydrophila.

[0047] Breeding of experimental fish: The experimental fish (crucian carp) purchased from Jiangsu was temporarily raised in a 2000L aquarium after being tested for the absence of CyHV-2, equipped with a flow circulation fresh water treatment and purification system. After raising for 1 week, remove unhealthy individuals. Select healthy crucian carp with a weight of 350±50g and randomly group them into 200L experimental aquariums, 30 in each aquarium, change the water twice a day in the experimental aquarium, and change the water volume to 1 / 3, maintain the water temperature at 20±2°C, and continue to raise temporarily 2 weeks.

[0048] Preparation of pathogenic bacteria suspension: culture Aeromonas hydrophila to OD in LB liquid medium 600 was 0.6, then centrifuged at 5000rpm for 10 minutes at room temperature, collected the pr...

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Abstract

The invention relates to a biological control method of crucian hemorrhagic disease, which is characterized in that: immunizing and inoculating a multivalent vector vaccine for cultured crucian; The multivalent vector vaccine is obtained from Bacillus subtilis recombination strain by a gene engineering method, and the classification and naming of the recombination strain is Bacillussubtilis HT5304, and the product is preserved in the China center for type culture collection, and the preservation number is CCTCCNo: M2013029. The invention also provides an application of Bacillussubtilis HT5304 in crucian culture and disease control. The biological prevention method of the present invention is used for immunizing and inoculating a multivalent vector vaccine for cultured crucian, thereby realizing multiple-effect immunization control for hemorrhagic disease with crucian viral disease cause and bacterial disease casue.

Description

Technical field [0001] The present invention involves a method and application of catfish hemorrhagic biological control methods and applications for catfish hemorrhage and immune prevention. Background technique [0002] Catfish, belongs to the family of carp, is one of the major freshwater fish breeding in my country.However, in the process of catfish breeding, it is not only threatened by the virus infection, but also facing potential risks of bacterial infections in the environment.At present, the major risk of epidemic disease in the process of catfish in my country mainly comes from the hemorrhage caused by carp virus II (CYHV-2) and gas monolithic (mainly hydrophobic bacteria). These diseases are given to the catfish breeding industry. Healthy development has brought destructive impact and serious economic losses. [0003] Herpesviral Haematopoietic Necrosis (HVHN) is a high pathogenic virus disease. At first, the pathogen of the disease was defined as a Goldfish Haematopo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61K39/245A61K48/00A61P31/04A61P31/22
Inventor 不公告发明人
Owner 马悦
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