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Production method of bdellovibrio bacteriovorus preparation

The invention relates to a technology for the production of leech vibrio and a production method, which can be applied to the field of microorganisms, and can solve the problems of insufficient bacterial count, reduced lysis ability of leech vibrio, and small number of phage plaques, so as to achieve enhanced lysis activity and stability, Enhance the effect of disease control and reduce the effect of chemical oxygen demand

Pending Publication Date: 2016-10-12
南京阡晟源生物科技有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Practice has proved that the bacteriophage Bdellovibrio preparations produced by this method will reduce the ability of Bdellovibrio phage to lyse live host bacteria, or even reduce the lysis ability to no lysis ability. Therefore, the effect on the prevention and treatment of animal bacterial diseases is unstable. At the same time, the preparation has a short storage time, which limits the development and application of the preparation, and is even more restricted in aquaculture
In the patent ZL200610039346.8, the applicant successfully developed a production method using Rhodopseudomonas sphaeroides as the host. The bacteriophage Bdellovibrio preparations produced in the past ten years have been welcomed by users, but the bacteriophage Bdellovibrio There are still some defects in the preparation, such as the number of plaques produced is less, and the number of bacteria is not stable enough. In order to overcome these shortcomings, we improved the single host bacteria

Method used

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  • Production method of bdellovibrio bacteriovorus preparation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Preparation of the host bacteria suspension: the photosynthetic bacterium Rhodopseudomonas sphaeroides P4 suspension cultured to the stationary phase, let stand for 24 hours, remove the supernatant which accounts for 1 / 2 of the total solution volume; add and remove the above The mass fraction of 0.2% NaCl solution equal to the volume of the supernatant, shake well, and then stand still for 24 hours, remove the supernatant that accounts for 1 / 2 of the total solution volume, and then add the mass fraction of 0.2% NaCl solution to prepare the original photosynthetic bacteria mixture. Suspension volume 1 / 2 of the host bacteria suspension A, with a bacterial content of 3 billion / ml, and then adding 5% of the volume of the host bacteria suspension A to the inactivated E.Coli600 bacterial suspension cultivated to the stable phase, the obtained Host bacteria suspension B, spare.

[0023] 2. Prepare culture medium: Dilute phosphate buffered saline (PBS) with pH 6.5 10 times w...

Embodiment 2

[0026] 1. Preparation of the host bacteria suspension: the photosynthetic bacterium Rhodopseudomonas sphaeroides P4 suspension cultured to the stationary phase was left to stand for 60 hours, and the supernatant accounting for 2 / 3 of the total solution volume was removed; adding and removing the above The mass fraction of 0.5% NaCl solution equal to the volume of the supernatant, shake well, and then stand still for 48 hours, remove the supernatant that accounts for 2 / 3 of the total solution volume, and then add the mass fraction of 0.5% NaCl solution to form the original photosynthetic bacteria mixture. Suspension volume 2 / 3 of the host bacteria suspension A, with a bacterial content of 17 billion / ml, and then adding 10% of the volume of the host bacteria suspension A to the inactivated E.Coli600 bacterial suspension cultivated to the stable phase, the obtained Host bacteria suspension B, spare.

[0027] 2. Preparation of culture medium: Dilute phosphate buffered saline (PBS)...

Embodiment 3

[0030] 1. Preparation of the host bacteria suspension: put the photosynthetic bacteria Rhodopseudomonas palustris 101 suspension cultured to the stationary phase, let it stand for 96 hours, remove the supernatant which accounts for 1 / 2 of the total solution volume; add and remove the above The mass fraction of 0.85% NaCl solution equal to the volume of the supernatant, shake well, and then stand still for 72 hours, remove the supernatant that accounts for 1 / 2 of the total solution volume, and then add the mass fraction of 0.85% NaCl solution to form the original photosynthetic bacteria mixture. Suspension volume 1 / 2 of the host bacteria suspension A, with a bacterial content of 30 billion / ml, and then adding 15% of the volume of the host bacteria suspension A to the inactivated E.Coli600 bacterial suspension cultivated to the stable phase, the obtained Host bacteria suspension B, set aside.

[0031] 2. Preparation of culture medium: Dilute phosphate buffered saline (PBS) with ...

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Abstract

The invention discloses a production method of a bdellovibrio bacteriovorus preparation. The production method comprises the following: (1) preparing host bacteria mixed suspension: taking rhodopseudomonas sphaeroides P4 or rhodopseudomonas palustris 101 in photosynthetic bacteria and E.Coli600 as mixed host bacteria for producing and culturing bdellovibrio bacteriovorus; (2) preparing a culture solution; (3) carrying out a fermentation process of the bdellovibrio bacteriovorus. The viable count in the prepared bdellovibrio bacteriovorus preparation is obviously improved and a lot of plaques can be produced, so that the use effect of the bdellovibrio bacteriovorus preparation is improved, and the cracking activity, stability and disease prevention and treatment effect of the bdellovibrio bacteriovorus are enhanced.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a production method of a phage Bdellovibrio preparation. Background technique [0002] In aquaculture and animal disease prevention and control at home and abroad, chemical preparations such as antibiotics are currently mainly used. The side effects of these preparations, such as drug residues, drug resistance of pathogenic bacteria, inhibition of beneficial bacteria, etc., bring adverse effects on human health and animal husbandry and aquatic production. The impact is becoming more and more obvious. In recent years, the research and development of microbial ecological preparations has attracted the attention of all parties, especially the research on Bdellovibrio bacteriophage has been favored by people. [0003] Since Bdellovibrio phage was discovered in 1963, because it is a parasitic bacterium, it can crack Escherichia coli, Salmonella, Aeromonas hydrophil...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34A61K35/74A61P1/12A61P31/04C12R1/01
CPCA61K2035/11C02F3/34C12N1/20Y02A50/30
Inventor 薛恒平薛彦青
Owner 南京阡晟源生物科技有限公司
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