Method for improving shelf life of saltwater-freshwater bdellovibrio sp. microbial preparation

A technology of microbial preparations and Bdellovibrio, which is applied in the field of improving the shelf life of Bdellovibrio microbial preparations in salty and fresh water, can solve problems such as shortage, short shelf life of microbial preparations, and difficulty in lysing bacteria by Bdellovibrio microbial preparations, so as to maintain vitality, The effect of improving the shelf life

Inactive Publication Date: 2014-07-30
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At home and abroad, the research on the quality assurance technology of microbial preparations mainly focuses on the freeze-dried powder of bifidobacterium, microencapsulated beneficial bacteria microbial preparations, etc., but the technology for extending the shelf life of microbial preparations made by Bdellovibrio is still lacking. And mainly focus on methods such as freeze-drying or embedding, but the shelf life of the microbial preparations prepared by these methods is short, and the immobilized Bdellovibrio microbial preparations will be difficult to exert their cracking effect on bacteria, so there are certain limitations

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  • Method for improving shelf life of saltwater-freshwater bdellovibrio sp. microbial preparation

Examples

Experimental program
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Effect test

Embodiment 1

[0062] 1. Preparation of Bdellovibrio microbial preparations

[0063] (1) Preparation of host bacteria concentrate: Pick a single colony of Escherichia coli (E. coli, purchased from the Culture Collection Center of Guangdong Institute of Microbiology, No. GIM1.355), and inoculate it in the nutrient broth liquid medium , cultured at 200rpm and 28°C for 18 hours to obtain a culture solution, then centrifuged the culture solution at 4°C and 6000rpm for 20min, discarded the supernatant, suspended the precipitate collected per 100mL of the culture solution with 2-3mL DNB liquid medium, and obtained the concentrated solution of the host bacteria. Store at 4°C for later use;

[0064] (2) Preparation of Bdellovibrio microbial preparations: inoculate 1mL of Bdellovibrio BDS02 stored at -40℃ in the laboratory into 50mL of DNB liquid medium, add 1mL of E. coli host concentrate in step (1), and shake on a constant temperature shaker Cultivate at 230rpm and 28°C, add host concentrated sol...

Embodiment 2

[0075] 1. Preparation of Bdellovibrio microbial preparations. See Example 1 for specific steps, and the concentration obtained by fermentation is 6.4×10 10 pfu / mL of Bdellovibrio microbial preparations.

[0076] 2. Accelerated test of shelf life of Bdellovibrio microbial preparations

[0077] (1) Optimize the optimum concentration of protective agent

[0078] The protective agent used in the experiment of the present invention is 5%-25% sucrose in the final mass volume ratio (g / mL). Within this concentration range, 5 concentration points were taken, and the final mass-to-volume ratios (g / mL) were: 5%, 10%, 15%, 20% and 25% sucrose.

[0079] Mix 1 mL of Bdellovibrio microbial preparation fermented in step 1 with 1 mL of sucrose protectant of different concentrations, place in a 2.5 mL centrifuge tube, seal it, and place it at 25°C, 37°C, 45°C, 55°C, 65°C , 75°C, 85°C, and 95°C for constant temperature acceleration experiments. The mixture is equipped with sea crystals with...

Embodiment 3

[0086] 1. Preparation of Bdellovibrio microbial preparations. See Example 1 for specific steps, and the concentration obtained by fermentation is 7.2×10 10 pfu / mL of Bdellovibrio microbial preparations.

[0087] 2. Accelerated test of shelf life of Bdellovibrio microbial preparations

[0088] (1) Optimize the optimum concentration of protective agent

[0089] The protective agent used in the experiment of the present invention is 5%-25% glycerin in the final volume percentage. Within this concentration range, 5 concentration points are taken, and the final volume percentages are respectively: 5%, 10%, 15%, 20% and 25% glycerol.

[0090] Mix 1 mL of Bdellovibrio microbial preparation fermented in step 1 with 1 mL of glycerol protectant of different concentrations, place in a 2.5 mL centrifuge tube, seal it, and place it at 25°C, 37°C, 45°C, 55°C, 65°C , 75°C, 85°C, and 95°C for constant temperature acceleration experiments. The mixture is equipped with sea crystals with a ...

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Abstract

The invention discloses a method for improving shelf life of a saltwater-freshwater bdellovibrio sp. microbial preparation. The method comprises the following steps: preparing the bdellovibrio sp. microbial preparation; mixing the bdellovibrio sp. microbial preparation with a protection agent belonging to different kinds and / or with different concentration, so as to obtain a mixed solution; placing the mixed solution at different temperature to perform accelerated testing; sampling regularly, and detecting the live bacteria concentration of the bdellovibrio sp. microbial preparation in the mixed solution; and utilizing Arrhenius equation and recorded data to calculate out the shelf life of the bdellovibrio sp. microbial preparation, and determining the effect of the protection agent by comparing the shelf life. According to the method, by comparing the quality-guaranteeing effect of different protection agents on the bdellovibrio sp. microbial preparation, a protection agent with best quality-guaranteeing effect is screened out, and the shelf life of the bdellovibrio sp. microbial preparation is obviously prolonged; and also the activity of thalline in the bdellovibrio sp. microbial preparation can be effectively kept for a long time. The provided protection agent is wide in raw material selection scope and cheap in price, and is suitable for popularization and usage.

Description

technical field [0001] The invention belongs to the technical field of Bdellovibrio, and in particular relates to a method for improving the shelf life of salty and fresh Bdellovibrio microbial preparations. Background technique [0002] At present, for the sake of environmental friendliness, many bactericidal and disease-preventing agents used in animal husbandry and aquaculture are mostly microbial. However, most microbial preparations have disadvantages such as easy loss or decline of activity and short shelf life. At the same time, in our actual breeding environment, we need microbial preparations with higher concentration and activity so that the bacteria can be better put into production applications. At home and abroad, the research on the quality assurance technology of microbial preparations mainly focuses on the freeze-dried powder of bifidobacterium, microencapsulated beneficial bacteria microbial preparations, etc., but the technology for extending the shelf lif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12Q1/06C12R1/01
Inventor 蔡俊鹏孙秋平
Owner SOUTH CHINA UNIV OF TECH
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