Method for improving shelf life of bdellovibrio sp. microbial prepartion

A technology of microbial preparations and Bdellovibrio, which is applied in the field of microbiology, can solve the problems of prolonging the shelf life of Bdellovibrio microbial preparations, and achieve the effects of shortening the shelf life, improving the shelf life, and prolonging the decay time

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

AI Technical Summary

Problems solved by technology

At present, no feasible plan has been reported for the long-term shelf life of Bdellovibrio preparations. The exploration of this research method is of great significance for prolonging the shelf life of Bdellovibrio microbial preparations

Method used

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  • Method for improving shelf life of bdellovibrio sp. microbial prepartion
  • Method for improving shelf life of bdellovibrio sp. microbial prepartion
  • Method for improving shelf life of bdellovibrio sp. microbial prepartion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] 1. Preparation of Bdellovibrio microbial preparations

[0061] (1) Preparation of DNB medium: Weigh 0.8g / L nutrient broth, 0.5g / L casein hydrolyzate, 0.1g / L yeast extract, 30g / L sea crystal, dissolve and mix with distilled water Mix well, adjust the pH to 7.5, and sterilize by steam at 115°C for 20 minutes.

[0062] (2) Preparation of host bacteria suspension: in 100mL LB liquid medium (tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, pH7.4, autoclaved at 121°C for 15min) Inoculate Aeromonas hydrophila (Ameromonas hydrophila) (purchased from Guangdong Microbial Culture Collection Center, No. GIM1.172), cultured on a shaker at 250 rpm and 30 °C for 24 h, and centrifuged the culture solution at 4 °C and 6000 rpm for 15 min. Discard the supernatant to collect the bacteria, suspend in 2-3 mL of DNB medium to obtain the host bacteria suspension, and store in a 4°C refrigerator for later use.

[0063] (3) Activation of Bdellovibrio: Take one tube (2 mL) of Bdellov...

Embodiment 2

[0075] 1. Preparation of Bdellovibrio microbial preparations. See Example 1 for specific steps, and the concentration obtained by fermentation is 5.74×10 12 pfu / mL of Bdellovibrio microbial preparations.

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

[0077] The optimal concentration of the protective agent obtained according to Example 1 is respectively final concentration (mass volume fraction g / mL) 5% sodium glutamate (MSG), final concentration (mass volume fraction g / mL) 5% sucrose+final concentration (mass volume fraction g / mL) 5% sodium glutamate, final concentration (mass volume fraction g / mL) 5% meso-inositol, final concentration (mass volume fraction g / mL) 5% raffinose, final Concentration (mass volume fraction g / mL) 10% glucose, final volume percentage 15% glycerin 6 kinds of protective agents.

[0078] ① Screening of the optimal group: Take 100 μL of the Bdellovibrio microbial preparation cultured in step 1, mix it with fresh D...

Embodiment 3

[0085] Embodiment 3 confirmatory experiments

[0086] 1. Preparation of Bdellovibrio microbial preparations. See Example 1 for specific steps, and the concentration obtained by fermentation is 5.74×10 12 pfu / mL of Bdellovibrio microbial preparations.

[0087] 2. Confirmatory experiment of shelf life

[0088] In the confirmatory experiment, take the Bdellovibrio microbial preparation cultured in step 1, add fresh DNB medium and the best protective agent group obtained in Example 2, and place it at 4°C for testing, and prepare 3 parallel samples . The mixed mixtures were placed under the same conditions for experiments. The variation of the concentration of Bdellovibrio in each sample is detected once a week, and the statistical data is used to evaluate the experimental effect of the accelerated test.

[0089] Effect evaluation: draw the statistical data of the confirmatory experiment for 8 weeks as Figure 7 of the graph. From Figure 7 We can see that the concentration...

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Abstract

The invention discloses a method for improving shelf life of a 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, then filling with a DNB medium, so as to obtain a mixed solution; placing the mixed solution at 25 DEG C, 37 DEG C, 45 DEG C, 55 DEG C and 65 DEG C to perform accelerated testing; sampling regularly, and detecting the bdellovibrio sp. concentration in the mixed solution; utilizing Arrhenius equation and recorded data to calculated out the shelf life of the bdellovibrio sp. microbial preparation; and determining the effect of the protection agent by comparing the shelf life. The protection agent is at least one of monosodium glutamate, cane sugar, internal-compensation inositol, melitose, glucose and glycerin. By employing the method, the attenuation time of a high-concentration bdellovibrio sp. microbial preparation is obviously prolonged, and the shelf life of the bdellovibrio sp. microbial preparation is improved; and by comparing the quality-guaranteeing effect of different protection agents, an optimum quality-guaranteeing method is screened out.

Description

technical field [0001] The invention belongs to the field of microbiology, in particular to a method for improving the shelf life of Bdellovibrio microbial preparations. Background technique [0002] At present, the shelf life of Bdellovibrio microbial preparations generally has the shortcoming of a short shelf life, and a large number of Bdellovibrio will die as time goes by during the shelf life. However, in actual experiments and applications, we need to be able to provide a certain concentration and activity of Bdellovibrio so that these strains can be better put into the preliminary experiments and applications. Therefore the present invention intends to study a method that can improve the shelf life of Bdellovibrio microbial preparations. At present, no feasible solution has been reported for the long-term shelf life of Bdellovibrio preparations. The exploration of this research method is of great significance for prolonging the shelf life of Bdellovibrio microbial pr...

Claims

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Application Information

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