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Bdellovibrio bacteriovorus preparation for preventing grouper ulcer and application thereof

A technology for phagocytosing leech vibrio and grouper, which can be applied to medical preparations, bacteria, and microorganism-based methods without effective ingredients, etc. It is safe and efficient to use, beneficial to growth and reproduction, and has the effect of low cost of use

Inactive Publication Date: 2019-08-30
XIAMEN HUIYING ANIMAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In China, there are basically no reports on related articles and patents on the use of bacteriophage Bdellovibrio to prevent and treat grouper ulcers.
[0005] Through the investigation of domestic patent documents related to Bdellovibrio phage, it is found that most Bdellovibrio phage preparations are generally cultured with live host bacteria Escherichia coli, which has several defects. On the one hand, it is necessary to separate unlysed E. coli after the cultivation is completed, which is cumbersome to operate. On the other hand, if the separation of live host bacteria is not clean, there will be potential safety hazards, which may damage the environment and introduce pathogenic bacteria, which is not conducive to the large-scale use of Bdellovibrio preparations; Bacterial function is lost, and the ability to lyse the infection of other pathogenic bacteria is reduced or lost

Method used

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  • Bdellovibrio bacteriovorus preparation for preventing grouper ulcer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 effective phage Bdellovibrio isolation test

[0038] This example is used to illustrate the source of Bdellovibrio and its acquisition method of the present invention.

[0039] In a grouper breeding base, the cultured seawater was taken and centrifuged at 3000r / min for 5min, the supernatant was taken, and 10mL was divided into 50mL triangular flasks, and 4 vials were added with a concentration of 10 8 CFU / mL Vibrio harveii, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio vulnificus were enriched and cultured at 30°C, 100r / min for 48h in aerobic culture. After the enrichment culture, seawater agar double-layer plates were used for separation and detection, and Vibrio harvelii, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio vulnificus were used as host bacteria respectively. The plate was cultured at 30°C for 2 days, and multiple a transparent plaque. Pick multiple upper layer of phage plaque medium and add a small amount of sterilized se...

Embodiment 2

[0040] Embodiment 2 preparation of phage Bdellovibrio bacterial liquid

[0041] This example is used to illustrate the bacteriophage Bdellovibrio liquid provided by the present invention and its preparation method.

[0042] The Bdellovibrio phage described in this example is provided by Example 1.

[0043] 1. Preparation of inactivated host bacteria concentrate: inoculate Vibrio harveyi, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio vulnificus respectively in 10g / L peptone, 3g / L beef extract, 10g / L sodium chloride , pH7.0-7.5 culture medium, after aerobic culture at 30°C for 24 hours, the bacteria sludge was collected by centrifugation, mixed and placed in a water bath at 60°C for 30 minutes, and cooled for later use.

[0044] 2. Preparation of bacteriophage Bdellovibrio bacteria liquid: Take the uninactivated mixed host bacteria in the above step 1 and dilute them with sterilized seawater to make the bacteria concentration at 10 9 CFU / mL was inoculated with bacte...

Embodiment 3

[0045] Preparation of Example 3 Bdellovibrio Phage Preparation 1

[0046] This example is used to illustrate a bacteriophage Bdellovibrio preparation for preventing grouper ulcer provided by the present invention and its preparation method.

[0047] The phage Bdellovibrio bacterial liquid obtained in Example 2 was inserted into the inactivated host bacterium, and the bacterial concentration was about 10 6 CFU / mL, after aerobic culture for 2 hours, add 2% (V / V) glycerol, stir evenly, and then mix with freeze-dried carrier (sucrose: skimmed milk powder = 6%: 94%) at a liquid-solid mass ratio of 1:0.5 Proportionally mixed to obtain a mixture to be freeze-dried.

[0048] The above-mentioned mixed solution to be freeze-dried was divided into stainless steel shallow pans, placed in a vacuum freeze dryer at -40°C, pre-frozen for 3 hours, and then started to vacuumize. First raise the temperature to 0°C within 24 hours, then gradually raise the temperature to 10°C in the next 24 hou...

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Abstract

The invention discloses a Bdellovibrio bacteriovorus preparation for preventing grouper ulcer. A variety of pathogenic bacteria which can cause grouper ulcer are used as various mixed living host bacteria to culture the Bdellovibrio bacteriovorus, host pathogenic bacteria failed to be cracked are removed by filtering to obtain Bdellovibrio bacteriovorus seed liquid, the Bdellovibrio bacteriovorusseed liquid is co-cultured with a variety of mixed inactivated host bacteria, and finally, the Bdellovibrio bacteriovorus preparation is obtained by freeze-drying. The invention also discloses a preparation method and an application of the Bdellovibrio bacteriovorus preparation. The Bdellovibrio bacteriovorus preparation can be used by the way of diluting and sprinkling, has a cracking effect on avariety of pathogenic bacteria causing grouper ulcer, plays a role in effectively preventing the occurrence of grouper ulcer, and significantly reduces the incidence. The Bdellovibrio bacteriovorus preparation can be used for mariculture animals, and has the advantages that the preparation is green and environmentally friendly, safe in use and high in efficiency.

Description

technical field [0001] The invention relates to the technical field of seawater culture, in particular to a bacteriophage Bdellovibrio preparation for preventing grouper ulcers and its application, which is mainly used to prevent the occurrence of body surface ulcers in the seawater grouper culture process. Background technique [0002] The grouper belongs to the order Perciformes, and the grouper belongs to the genus Grouper. It is a rare and valuable fish at the bottom of warm water offshore. Its growth rate is fast, the meat is plump and tender, rich in nutrition, and the grouper has medicinal value of invigorating the spleen and replenishing qi. Due to the high economic value of grouper, the scale of artificially cultured grouper is also increasing. Wherein the grouper artificial breeding process disease problem is just becoming more and more serious. According to reports, the current grouper breeding process diseases mainly include bacterial, parasitic and viral disea...

Claims

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Application Information

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IPC IPC(8): A61K35/74A61K9/19A61K47/46A61P17/02C12N1/20C12N1/04C12R1/01
CPCA61K9/19A61K35/74A61K47/46A61P17/02C12N1/04C12N1/20
Inventor 乔欣君林松泉庄若飞李惠静曾海燕
Owner XIAMEN HUIYING ANIMAL TECH
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