Method for inducing bacteria to enter viable but nonculturable state

A technology of bacteria and state, which is applied in the field of biomedicine, can solve the problems of restricting the research progress of living non-culturable bacteria and long induction time, and achieve the effect of speeding up the preparation and improving the research progress

Active Publication Date: 2013-01-30
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the induction time required by this method is long, usually more than one month, which limits the progress of research on live non-culturable bacteria

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, induce bacterium to enter the non-culturable state of living

[0024] 1. High pressure carbon dioxide induction

[0025] 1. Bacterial suspension preparation

[0026] Wash the cells of E.coli O157:H7 (strain number NCTC12900) in the logarithmic growth phase twice with 0.85% (mass percentage) NaCl aqueous solution, and then resuspend the cells with 0.85% (mass percentage) NaCl aqueous solution, Make the cell concentration about 10 8 cfu / mL, as the bacterial solution to be induced (bacterial suspension);

[0027] 2. High pressure carbon dioxide treatment

[0028] Experimental group: use a self-designed high-pressure carbon dioxide sterilizer (model CAU-HPCD-1, high-density carbon dioxide sterilizer, disclosed in patent ZL200520132590.X) to treat the bacterial liquid with high-pressure carbon dioxide; specific steps: 20mL of the bacterial liquid to be induced (Bacterial suspension) was put into a glass bottle and sealed with a sealing film; the bacterial l...

Embodiment 2

[0038] Embodiment 2, induce bacterium to enter the non-culturable state of living

[0039] 1. High pressure carbon dioxide induction

[0040] 1. Bacterial suspension preparation

[0041] It is the same as the method of step 1 in Example 1.

[0042] 2. High pressure carbon dioxide induction

[0043] The method is basically the same as step 2 in Example 1, except that the treatment temperature is changed to 31° C., and the holding time is changed to 30 minutes; the induced bacterial liquid is obtained.

[0044] 2. Detection

[0045] Same as the method of two in embodiment 1.

[0046] The results are as follows: the number of viable bacteria in the bacterial solution after the experimental group was induced was 10 6.89 cfu / mL; the number of cultivable bacteria in the bacterial solution after induction is 0; therefore, the number of bacteria that have entered a viable non-culturable state is 10 6.89 cfu / mL.

[0047] The number of cultureable bacteria in the control group wa...

Embodiment 3

[0050] Embodiment 3, induce bacterium to enter live non-culturable state rapidly

[0051] 1. High pressure carbon dioxide induction

[0052] 1. Bacterial suspension preparation

[0053] It is the same as the method of step 1 in Example 1.

[0054] 2. High pressure carbon dioxide induction

[0055] The method is basically the same as step 1 in Example 1, only the treatment temperature is changed to 37° C., and the holding time is changed to 25 minutes; the induced bacterial liquid is obtained.

[0056] The results are as follows: the number of viable bacteria in the bacterial solution after the experimental group was induced was 10 5.72 cfu / mL; the number of cultivable bacteria in the bacterial solution after induction is 0; therefore, the number of bacteria that have entered a viable non-culturable state is 10 5.72 cfu / mL.

[0057] The number of cultureable bacteria in the control group was still about 10 8 cfu / mL, it is basically considered that there are no viable non-cu...

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PUM

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Abstract

The invention discloses a method for inducing bacteria to enter a viable but nonculturable state. The method provided by the invention comprises the step of treating target bacteria through high-pressure carbon dioxide to enable the bacteria to enter the viable but nonculturable state. According to an experiment, the method for inducing bacteria to enter the viable but nonculturable state provided by the invention can treat the bacteria by using a high-pressure carbon dioxide technique to enable the bacteria to rapidly enter the viable but nonculturable state within 1 hour. By adopting the method provided by the invention, the preparation of bacteria in the viable but nonculturable state is accelerated and the progress of relevant researches on the bacteria in the viable but nonculturable state is improved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for inducing bacteria to enter a living non-culturable state. Background technique [0002] Under unfavorable circumstances, a variety of bacteria enter a viable but nonculturable (VBNC) state. This state is a form of dormancy for non-spore-forming bacteria that improves the bacteria's ability to survive in adverse environments. At present, more than 60 kinds of bacteria are known to enter the living non-culturable state, most of which are pathogenic bacteria. Although live non-culturable bacteria are still metabolically active, they cannot grow or form colonies on the non-selective medium commonly used for the bacteria, so routine bacterial detection methods such as plate counts cannot detect live non-culturable bacteria In this way, the number of bacteria in the test sample may be underestimated, which will bring potential safety hazards to people. Carrying out ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/04C12R1/19
CPCC12N1/20C12N1/04C12N1/36
Inventor 廖小军赵凤
Owner CHINA AGRI UNIV
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