Method for quickly finding early bacterium infection in microorganism fermentation process

A technology for fermentation broth and seeds, which is applied in the fields of biochemical equipment and methods, and the determination/inspection of microorganisms, can solve the problems of time-consuming, expensive, cumbersome operation of checking miscellaneous bacteria, etc., and achieves the effect of low price of instruments and reduced loss.

Pending Publication Date: 2020-03-27
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there are still problems of cumbersome, time-consuming and expensive inspection of miscellaneous bacteria

Method used

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  • Method for quickly finding early bacterium infection in microorganism fermentation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Hyaluronic Acid Fermentation Process Seed Liquid Miscellaneous Bacteria Detection Before Transplantation

[0067] (1) Calculate the bacterial concentration of the seed solution

[0068] Take the seed liquid whose lot number is 181203, and use a hemocytometer (model: XB.K.25, 0.1mm, 1 / 400mm 2 , manufacturer: Shanghai Qiujing Biochemical Reagent Instrument Co., Ltd.) According to the instructions, calculate the bacterial concentration of the required test sample: 1.88×10 10 (RBCs) / L, repeat the counting results twice according to the above method: 1.89×10 10 (RBCs) / L, 1.90×10 10 (RBCs) / L, it is further determined that the minimum concentration factor of the sample is 16-21 times, preferably 20 times.

[0069] Small square position upper left lower left upper right bottom right middle Total bacteria count (N) The number of bacteria for the first time (a) 78 79 70 73 75 375 The second bacteria count (a) 80 74 77 76 70 3...

Embodiment 2

[0074] Example 2 Detection of miscellaneous bacteria in the fermentation broth during the fermentation of hyaluronic acid for 3 hours

[0075] (1) Calculate the bacterial concentration of the fermentation broth

[0076] Take the fermentation broth with batch number 181203 and cultivate it for 3 hours. According to the microscopic examination experience, the cocci are still in the lagging phase at this time. The cocci basically do not proliferate, and the amount of cocci during microscopic examination is very small. Therefore, the first step of concentration is carried out according to the amount of transplantation and fermentation volume. The concentration ratio is: fermentation volume / seed volume=4000L / 400L=10. Take the fermented liquid that has been concentrated 10 times with a hemocytometer (model: XB.K.25, 0.1mm, 1 / 400mm 2 , manufacturer: Shanghai Qiujing Biochemical Reagent Instrument Co., Ltd.) According to the instructions, the bacterial concentration of the fermented ...

Embodiment 3

[0082] Example 3 Detection of miscellaneous bacteria in the fermentation broth during the fermentation of hyaluronic acid for 7 hours

[0083] (1) Calculate the bacterial concentration of the fermentation broth

[0084] Take the fermentation broth whose batch number is 181203 and which has been cultured for 7 hours. According to the microscopic examination experience, the cocci are in the early logarithmic phase at this time, and the cocci have multiplied to a certain extent. The same large amount of bacteria is not suitable for direct counting on a hemocytometer. The fermentation broth can be diluted, and the dilution factor should be diluted to a suitable count, which can be adjusted by oneself. The batch number is 181203 and the fermented liquid whose cultivation time is 7 hours is diluted 5 times, and the bacteria concentration of the fermented liquid after counting three times with a hemocytometer is as follows: 1.79×10 10 (RBCs) / L, 1.81×10 10 (RBCs) / L, 1.75×10 10 (RBC...

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Abstract

The invention relates to a method for finding early bacterium infection of fermentation liquor. The method comprises the following steps of preparing culture mediums, performing inoculation into a strain to be fermented, and beginning fermentation; performing fermentation for some time, and before the fermentation liquor reaches a given OD value, taking out fermentation liquor samples; calculatingthe bacterium concentration of the taken-out fermentation liquor samples with a blood cell counting chamber, and according to the bacterium concentration of the fermentation liquor samples, determining the multiple to be concentrated of the fermentation liquor samples, wherein the concentration multiple is large enough to concentrate the bacterium concentration of the sampled fermentation liquorsamples to the concentration of given microscope inspection bacteria which are suitable for observation under a microscope; according to the determined concentration multiple, concentrating the fermentation liquor samples to the concentration of the microscope inspection bacteria; observing the concentrated samples having given area under the microscope, and determining whether infectious microbesexist or not and the number of the infectious microbes; and based on the existence and the number of the infectious microbes contained in the concentrated samples having the given area, judging wither the fermentation liquor has a phenomenon of bacterium infection or not.

Description

technical field [0001] The invention relates to the field of microbial fermentation process detection, in particular to a method for quickly discovering early bacterial contamination in the microbial fermentation process. Background technique [0002] Microorganisms are tiny organisms that exist in nature, are small in size, simple in structure, invisible to the naked eye, and must be magnified with the help of special instruments to observe tiny organisms. The microbial growth curve includes four periods: lag period, logarithmic period, stable period, and decay period. [0003] The characteristics of the lag phase: the growth rate constant is zero, the bacteria are bulky, the RNA content increases, the metabolic activity is strong, and the resistance to adverse environments decreases. [0004] The characteristics of the logarithmic phase: the fastest growth rate, strong metabolism, active enzyme system, the number of viable bacteria and the total number of bacteria are rou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06C12Q1/04
CPCC12Q1/06C12Q1/04
Inventor 刘文超郭学平栾贻宏穆惠军任福群穆淑娥王玉盈张宁宁杨健民王岩
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD
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