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Resuscitation medium capable of promoting growth of VBNC (viable but non-culture) bacteria and improving separation abundance, and preparation method and application thereof

A medium and liquid culture medium technology, applied in the field of microorganism screening and culture, can solve the problems of not publicly using a recovery medium, recovery growth and the effect of separation abundance is not very significant, etc., to improve the VBNC bacterial abundance, promote The effect of resuscitating growth and simple operation

Active Publication Date: 2014-07-23
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this document does not disclose what kind of recovery medium to use, and in practical applications, the effect of simple recovery-promoting factors on promoting the recovery and growth of non-culturable (VBNC) state bacteria and the isolation abundance has not been very significant.

Method used

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  • Resuscitation medium capable of promoting growth of VBNC (viable but non-culture) bacteria and improving separation abundance, and preparation method and application thereof
  • Resuscitation medium capable of promoting growth of VBNC (viable but non-culture) bacteria and improving separation abundance, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] A recovery medium that can promote the growth of VBNC bacteria and increase the abundance of isolation. The composition of the medium is composed of components A, B, and C (abbreviation: ABC recovery medium).

[0083] A component:

[0084] Bacterial solution: Micrococcus luteus IAM14879 was cultured on LMMD medium to the late logarithmic phase, and the protein was extracted from the centrifuged medium, containing the revival-promoting factor Rpf, and the dosage was 5% of the volume of component C.

[0085] B component:

[0086] Soil solution: take 1kg of garden soil that has been air-dried, sieved, and remove impurities such as stones, grass roots, etc., add 1L of tap water, and sterilize in a high-pressure sterilizer at 121°C for 30 minutes; after overnight at room temperature, sterilize again and repeat three times; 2000r / Centrifuge at a low speed for 1 min to take the supernatant (or filter with filter paper to obtain the filtrate), and the dosage is 2% of the volu...

Embodiment 2

[0123] A resuscitation medium that can promote the growth of VBNC bacteria and increase the abundance of isolation. The composition of the medium: it is composed of components A, B, and C (abbreviation: ABC resuscitation medium), suitable for Gram-positive bacteria Amycobacterium Genus ( Amycolatopsis genus).

[0124] A component:

[0125] Bacterial solution: Micrococcus luteus IAM14879 was cultured on LMMD medium to the late logarithmic phase, and the protein was extracted from the centrifuged medium, containing the revival-promoting factor Rpf, and the dosage was 1% of the volume of component C.

[0126] B component:

[0127] Soil solution: take 1kg of garden soil that has been air-dried, sieved, and remove impurities such as stones, grass roots, etc., add 1L of tap water, and sterilize in a high-pressure sterilizer at 121°C for 30 minutes; after overnight at room temperature, sterilize again and repeat three times; 2000r / Centrifuge at a low speed for 1 min to take the ...

Embodiment 3

[0136] A resuscitation medium that can promote the growth of VBNC bacteria and increase the abundance of isolation. The composition of the medium is composed of components A, B, and C (abbreviation: ABC resuscitation medium), suitable for Gram-positive bacteria Arthrobacter ( Arthrobacter genus), Rhodococcus ( Rhodococcus genus), Leif's genus ( Leifsonia genus), Nocardia ( Nocardia genus), Bilesporium ( Kitasatospora genus), Streptomyces ( Streptomyces genus), Bacillus subtilis ( Bacillus genus), Paenibacillus ( Paenibacillus genus).

[0137] A component:

[0138] Bacterial solution: Micrococcus luteus IAM14879 was cultured on LMMD medium to the late logarithmic phase, and the protein was extracted from the centrifuged medium, which contained the revival promoting factor Rpf, and the dosage was 8% of the volume of component C.

[0139] B component:

[0140] Soil solution: take 1kg of garden soil that has been air-dried, sieved, and remove impurities such as sto...

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Abstract

The invention relates to the field of microbial screening and cultivation, in particular to a recovery medium for viable non-culturable (VBNC) strains and a preparation method and application thereof. A kind of recovery medium that can promote the growth of VBNC bacteria and improve the separation abundance, the recovery medium is composed of culture medium base liquid, bacteria liquid and soil liquid; Cultivate on the culture medium to the late logarithmic phase, remove the bacteria by centrifugation and extract the liquid culture medium to obtain protein, which contains the revival promoting factor Rpf, and the bacterial liquid is 0.5-30% of the medium base liquid by volume; the soil liquid is composed of After air-drying, sieving, and removing impurities from the soil, add water, sterilize for many times, filter to get the filtrate or centrifuge to get the supernatant, the soil liquid is 0.1-20% of the medium base liquid by volume. The invention utilizes the revival promoting factor Rpf and adds soil liquid at the same time to construct a revival medium, which can further promote the revival and growth of VBNC bacteria, and increase the abundance of VBNC bacteria isolated from the ecological environment. The invention has the characteristics of simple operation and fast recovery.

Description

technical field [0001] The invention relates to the field of microbial screening and cultivation, in particular to a recovery medium for viable non-culturable (VBNC) strains and a preparation method and application thereof. Background technique [0002] VBNC (viable but non-culture) refers to the fact that some microorganisms can be detected in their habitat, but cannot be cultured artificially in the laboratory. In 1982, Xu Huaishu et al. first discovered and proposed the "viable but non-culturable state" (VBNC) of bacteria through the study of the survival rules of V.cholerae and E.coli, that is, the bacteria are under adverse environmental conditions. , its cells are usually shrunk into a spherical shape (recently many studies have found that some bacteria show increased volume and cell elongation), and cannot be grown and reproduced by conventional methods, but they still have metabolic activity and act as DNA synthesis inhibitors Under certain conditions, when cultured...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/04
Inventor 丁林贤陈建荣金夷林冬珍
Owner ZHEJIANG NORMAL UNIVERSITY
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