Inductive formation method for VBNC (viable but nonculturable) salmonella

A Salmonella, state technology, applied in the field of food microbial cultivation and detection, can solve the problems of food safety and public health safety, and achieve the effect of avoiding the risk of contamination

Active Publication Date: 2015-12-30
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Once food-borne pathogens and spoilage bacteria enter the VBNC state, it will be very harmful to food safety and public health safety

Method used

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  • Inductive formation method for VBNC (viable but nonculturable) salmonella
  • Inductive formation method for VBNC (viable but nonculturable) salmonella
  • Inductive formation method for VBNC (viable but nonculturable) salmonella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Ultrasonic heat treatment is used to induce Salmonella typhimurium in BPY medium to enter the VBNC state

[0024] Take the number of cultivable bacteria as 10 7 ~10 8 CFU / mL of Salmonella typhimurium (strain number CMCC50115) BPY culture medium suspension 60 mL was aseptically transferred to a jacketed beaker. Connect to the circulating water bath device and set the temperature at 52°C. Equilibrate for 5 minutes, so that the temperature of the BPY bacterial solution in the center of the jacketed beaker is 52°C. Immediately turn on the ultrasonic device, using 190W power. Ultrasonic heat treatment for 20min. Remove the sample and cool to 4 °C.

[0025] Mix the Salmonella suspension before and after treatment evenly with a vortex mixer, take a sample by sterility test, and evenly spread it on an SS agar plate, place it upside down in a biochemical incubator, and incubate at 37°C for 24 hours. Salmonella colony growth, it is preliminarily believed that it ...

Embodiment 2

[0026] Example 2: Ultrasonic heat treatment is used to induce Salmonella typhimurium in bayberry juice to enter the VBNC state

[0027] Take the number of cultivable bacteria as 10 7 ~10 8 CFU / mL of Salmonella typhimurium (strain number CMCC50115) BPY medium suspension 60mL, centrifuged at 6000rpm for 5min, collected the bacterial sediment and transferred it to a jacketed beaker filled with 60mL bayberry juice after sterilization . Connect to the circulating water bath device and set the temperature to 45°C. Balance treatment for 2 minutes, so that the temperature of the bayberry juice bacterial liquid in the center of the jacketed beaker is 45°C. Immediately turn on the ultrasonic device, using 190W power. Ultrasonic heat treatment for 28min. Remove the sample and cool to 4 °C.

[0028] Mix the Salmonella suspension before and after treatment evenly with a vortex mixer, take a sample by sterility test, and evenly spread it on an SS agar plate, place it upside down in a ...

Embodiment 3

[0029] Example 3: Resuscitation of VBNC Salmonella typhimurium

[0030] Take 10 mL of the BPY bacterial solution that entered the VBNC state in Example 1, and centrifuge at 8000 rpm for 5 min. Rinse the bacterial cells twice with sterilized physiological saline, and finally suspend the bacterial cell pellet in 10 mL of sterilized BPY medium, and incubate at a constant temperature of 37°C and 200 rpm for 36 hours. Take 100 μL of the resuscitated bacteria solution and spread it evenly on the SS agar plate, place it upside down in the incubator, incubate at 37°C for 24 hours, and record the number of viable Salmonella bacteria after the resuscitation according to the colony counting method. The results are shown in figure 1 .

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Abstract

The invention discloses an inductive formation method for VBNC (viable but nonculturable) salmonella. The method comprises the following steps: salmonella is suspended in a liquid and uniformly mixed, the concentration of a bacterial liquid is enabled to be 107-108 CFU / mL, the bacterial liquid is treated with ultrasonic waves of 190-570 W at the constant temperature of 42-55 DEG C for 5-25 min, and whether the salmonella is induced into the VBNC state is detected. According to the method, understanding about the VBNC salmonella is expanded, and theoretical basis and technical support are provided for better application of the ultrasonic heat technology to food processing, microorganism control and food safety control.

Description

technical field [0001] The invention belongs to the field of food microorganism culture and detection, and more specifically relates to a method for inducing the formation of Salmonella in VBNC state. Background technique [0002] The Viable but non-culturable (VBNC) state of microorganisms refers to a special dormant state in which they have lost the ability to grow and reproduce on conventional media, but can recover and restore cultivability under suitable conditions. Since 1982 when Xu Huaishu et al. first discovered Vibrio cholerae (Vibriocholerae) and Escherichia coli (Escherichia coli) which survived but could not be cultivated in estuarine and marine environments, until Colwell formally proposed the concept of VBNC in 1985, it has been used in more than 90 species of 20 species. VBNC status was found in several bacteria. In the process of food processing and storage, there are two main categories of physical stress factors and chemical induction factors to induce ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12Q1/10C12R1/42
Inventor 廖红梅蒋丽芬丁占生李群英
Owner JIANGNAN UNIV
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