Preparation method and application of sea bdellovibrio bacteriovorus ecological preparation

The technology of a microecological preparation and leech vibrio is applied in the field of preparation of seawater microecological preparations, and can solve the problems of being unsuitable for the process of marine aquaculture, the titer needs to be further improved, etc. The effect of avoiding environmental pollution

Inactive Publication Date: 2010-07-21
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although there are some products related to Bdellovibrio preparations at home and abroad, they basically come from fresh water and are not suitable for mariculture, and the potency needs to be further improved

Method used

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  • Preparation method and application of sea bdellovibrio bacteriovorus ecological preparation
  • Preparation method and application of sea bdellovibrio bacteriovorus ecological preparation
  • Preparation method and application of sea bdellovibrio bacteriovorus ecological preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Isolation, purification and preparation of concentrated solution of Bdellovibrio bacillus:

[0010] 1. Preparation of host bacterial suspension

[0011] Pseudomonas stutzeri (provided by the Institute of Microbiology, Chinese Academy of Sciences) was activated on a slant, and cultured for 18-24 hours, and about 2ml of normal saline was added to the test tube to prepare a bacterial suspension, and then placed in a nutrient broth medium, at 30°C Under the shaker liquid culture 18 ~ 24h. Add 1% chitosan oligosaccharide to the obtained bacterial liquid as a flocculant, stir evenly at a speed of 150 r / min, and then filter with a multi-layer polyester high-efficiency filter bag to collect the precipitate. Finally add normal saline to get 10 12 The cfu / mL host bacterial suspension can be stored at 4°C for 10 days.

[0012] 2. Isolation and purification of Bdellovibrio

[0013] The double-layer agar plate method was used to separate and purify the collected seawater and sed...

Embodiment 2

[0022] Preservation of Bdellovibrio:

[0023] The Bdellovibrio concentrated solution and the milk sucrose stabilizer are uniformly mixed at a ratio of 1:5 to obtain a mixed solution to be freeze-dried. Milk sucrose stabilizer composition: 5% sucrose + 10% skim milk powder + 85% deionized water. The stabilizer needs to be sterilized by autoclaving, and it is ready for use after passing the sterility test.

[0024] Divide the above mixed solution, pre-freeze at -40°C for 3 hours, and then freeze-dry it with a freeze dryer at -80°C and 2-7mTorr vacuum for 18-24h to obtain Bdellovibrio Freeze-dried formulations.

Embodiment 3

[0026] Application of Bdellovibrio in purifying mariculture pond water:

[0027] In order to study the application of Bdellovibrio in purifying the water in seawater culture ponds, the present invention uses Bdellovibrio Bdh5221 to treat the water in the culture ponds of Penaeus vannamei. The test container was a 125mL aquarium, the water temperature was 25°C, and the test water was untreated prawn breeding pond water. Ten vannamei shrimp were placed in each aquarium, and the concentration of Bdellovibrio was 1.5×10 1 pfu / mL (test group 1), 1.5×10 2 pfu / mL (test group 2), 1.5×10 3 pfu / mL (test group 3), and set up a control group without adding Bdellovibrio Bdh5221, adopt 24h aeration, and cultivate continuously for one week. During the experiment, the total number of bacteria, the number of Vibrio and the number of Bdellovibrio were recorded, and the survival of the prawns was observed from time to time. The results are shown in Table 1.

[0028] Table 1. The detection res...

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Abstract

The invention provides a preparation method and application of a sea bdellovibrio bacteriovorus ecological preparation, and relates to a sea microbial ecological preparation and a preparation method of a bdellovibrio bacteriovorus preparation for mariculture. The preparation method comprises the following steps of: preparing a host bacteria suspension; separating and purifying bdellovibrio bacteriovorus; and propagating and culturing bdellovibrio bacteriovorus. The preparation method is characterized in that after pseudomonas stutzeri is connected with an incline and is activated, adding physiological saline in a test tube to prepare the bacterial suspension; then propagating and culturing with nutrient bouillon; adding a flocculating agent in the obtained bacterial suspension; standing and precipitating after uniformly shaking; adding the physiological saline to obtain 1012 cfu / mL of host bacteria suspension; separating and purifying the bdellovibrio bacteriovorus of collected sea and bottom sediment samples with a double agar plate method; pouring culture media containing the host bacteria suspension and the collected samples into sea agar in the lower layer for culture after uniformly mixing; and adding the host bacteria and the bdellovibrio bacteriovorus obtained by separation into sterilized natural sea, and culturing until the concentration of the bdellovibrio bacteriovorus is 108 to 109 pfu / mL for future use.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the preparation of a seawater micro-ecological preparation, which is used for water quality purification and prevention and treatment of bacterial diseases in the seawater culture process. Background technique [0002] Since the 1980s, my country's marine aquaculture industry has developed rapidly, not only aquaculture species are rich, but also have high economic value. However, some phenomena that are not conducive to the sustainable and healthy development of the aquaculture industry have also followed, such as the disorderly development of a single species, blindly increasing the breeding density, etc., all of which lead to deterioration of water quality and outbreaks of aquaculture diseases. Since 1993, the frequent occurrence of bacterial diseases in aquaculture species such as prawns and large yellow croakers has caused huge economic losses to the mariculture industry in my coun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/63
Inventor 房文红周帅胡琳琳谢群英
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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