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Bdellovibrio bacteriovorus preparation for preventing bacterial diseases of anguilla japonica and application of preparation

A bacteriophage leech vibrio, bacterial technology, applied in the field of freshwater aquaculture, can solve problems such as unfavorable large-scale use of leech vibrio preparations, safety, lack of leech vibrio function, etc., to prevent the occurrence of eel bacterial diseases, Safe and efficient use, beneficial to growth and reproduction

Inactive Publication Date: 2019-09-10
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 Bdellovibrio to prevent and treat eel bacterial diseases.
[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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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] Take the intestinal contents of the sick eel and add an appropriate amount of sterile saline, dilute and shake, centrifuge at 3000r / min for 5min, take the supernatant, put 10mL in 50mL triangular flasks, and add 5 small bottles with a bacterial concentration of 10 8 CFU / mL of Vibrio anguillarum, Edwardsiella, Pseudomonas aeruginosa, Aeromonas hydrophila and Vibrio vulnificus were enriched and cultivated at 30°C, 100r / min for 48h aerobically. After the enrichment culture, the tap water agar double-layer plate was used for separation and detection, and Vibrio anguillarum, Edwardsiella, Pseudomonas aeruginosa, Aeromonas hydrophila and Vibrio vulnificus were used as host bacteria for testing respectively, and the plates were cultured at 30°C On 2d, multiple transparent phage plaques app...

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 anguillarum, Edwardsiella, Pseudomonas aeruginosa, Aeromonas hydrophila and Vibrio vulnificus respectively in 10g / L peptone, 3g / L beef extract, 5g / L sodium chloride, pH7.0-7.5 culture medium, after aerobic culture at 30°C for 24 hours, collect the sludge by centrifugation, mix and place in a water bath at 60°C for 30 minutes, and cool 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 physiological saline to make the bacterial concentration at 10 9 CFU / mL was inoculated w...

Embodiment 3

[0045] Preparation of Example 3 Bdellovibrio Phage Preparation 1

[0046] This example is used to illustrate a Bdellovibrio bacteriophage preparation for preventing eel bacterial diseases provided by the present invention and a preparation method thereof.

[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 2 hours of aerobic culture, add lyoprotectant 2% (V / V) glycerin, stir evenly, and then mix with lyophilized carrier (sucrose: skimmed milk powder = 6%: 94%) according to the liquid-solid mass ratio Mix at a ratio of 1:0.5 to obtain a mixture to be lyophilized.

[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 temper...

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PUM

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Abstract

The invention discloses a bdellovibrio bacteriovorus preparation for preventing bacterial diseases of anguilla japonica. Various pathogenic bacteria capable of causing the bacterial diseases of anguilla japonica are adopted as various mixed living host bacteria for culturing bdellovibrio bacteriovorus, then the host pathogenic bacteria which are not subjected to pyrolysis are filtered out and removed to obtain a bdellovibrio bacteriovorus seed solution, the bdellovibrio bacteriovorus seed solution and various mixed inactivated host bacteria are co-cultured, a lyoprotectant is added to an obtained bdellovibrio bacteriovorus culture solution for uniform mixing at first, then a lyophilization carrier is added for mixing to obtain a to-be-lyophilized preparation, and finally the bdellovibrio bacteriovorus preparation is prepared through freeze drying. The invention further discloses a preparation method and application of the preparation. The preparation can be used in the manners of material mixing for feeding and dilution for splashing, has a pyrolysis function on various pathogenic bacteria of the bacterial diseases of anguilla japonica, effectively prevents the occurrence of the bacterial diseases of anguilla japonica, significantly reduces the incidence rate and has the advantages of being environmentally friendly, safe to use and efficient.

Description

technical field [0001] The invention relates to the technical field of freshwater aquaculture, in particular to a bacteriophage Bdellovibrio preparation for preventing eel bacterial diseases and its application, which is mainly used in the freshwater eel breeding process to prevent the occurrence of bacterial diseases. Background technique [0002] Eel is an animal of the family Eelidae, which belongs to fish, resembles a snake, but has no scales, and is generally produced in sea areas at the junction of brackish and fresh water. Adult eels grow fast, round and cone-shaped in appearance, black in color and firm in body, and more artificially bred in recent years, with crispy meat. Eel is rich in nutrition, delicious in taste, less thorny and more fleshy, and has the functions of cooling and relieving heat, nourishing and strengthening the body. The meat, bone, blood and swim bladder of eel can be used as medicine. Its meat is sweet and flat, and has the effects of nourishi...

Claims

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Application Information

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IPC IPC(8): A61K35/74A61K9/19A61K47/26A61P31/04
CPCA61K9/19A61K35/74A61K47/26A61P31/04
Inventor 林章秀林松泉乔欣君曾海燕黄榕
Owner XIAMEN HUIYING ANIMAL TECH
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