Bacillus subtilis viable count medium, diluent and viable count method

A technology for counting Bacillus subtilis and viable bacteria, applied in the direction of microorganism-based methods, biochemical equipment and methods, instruments, etc., can solve the problems of high bacterial cell death rate, excessive foam, and excessive colonies on the counting plate, and reduce the Mortality, reduced foam formation, and improved counting accuracy

Active Publication Date: 2022-05-27
GUANGZHOU AONONG BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, although the above methods can solve the problems of some bacterial cell aggregation and counting colony spread, they cannot solve the problems of excessive foaming, high bacterial cell death rate and excessive colony counting plate during the shaking dilution process.

Method used

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  • Bacillus subtilis viable count medium, diluent and viable count method
  • Bacillus subtilis viable count medium, diluent and viable count method
  • Bacillus subtilis viable count medium, diluent and viable count method

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Effect test

Embodiment 1

[0050] Embodiment 1, Bacillus subtilis viable count solid culture medium of the present invention

[0051] The solid medium of the present invention (formula 1): 4g of peeled soybean meal powder (crushed to 100 mesh), 3g of corn flour (crushed to 100 mesh), 5g of sodium chloride, 3g of xanthan gum, 13g of agar, 1000mL of distilled water, pH value is 7.3.

[0052] The solid medium of the present invention (formulation 2): 6g of peeled soybean meal powder (crushed to 100 mesh), 5g of corn flour (crushed to 100 mesh), 10g of sodium chloride, 5g of xanthan gum, 11g of agar, 1000mL of distilled water, pH value is 7.5.

Embodiment 2

[0053] Embodiment 2, Bacillus subtilis live bacteria count sterilization diluent of the present invention

[0054] The sterilizing diluent of the present invention (Formulation 1): 8 g of sodium chloride, 2 g of potassium dihydrogen phosphate, 0.3 g of anhydrous glucose, 0.3 g of peptone, 0.1 g of polyoxypropylene oxide glycerol ether defoamer, and 1000 mL of distilled water.

[0055] The sterilizing diluent of the present invention (formula 2): 9 g of sodium chloride, 2 g of potassium dihydrogen phosphate, 0.5 g of anhydrous glucose, 0.5 g of peptone, 0.2 g of polyoxypropylene oxide glycerol ether defoamer, and 1000 mL of distilled water.

Embodiment 3

[0056] Embodiment 3, Bacillus subtilis viable count method of the present invention

[0057] Sample: Bacillus subtilis sample, the number of viable bacteria is about 5.0×10 10 CFU / g.

[0058] S1: Preparation of initial suspension:

[0059] Weigh 25g of the sample by aseptic operation, add 225mL of sterilized diluent, add 50 sterilized glass beads, shake for 30min, and make an initial suspension of 1:10.

[0060] S2: ten-fold serial dilution:

[0061] Prepare a vial with a capacity of 15 mL, add 9 mL of sterilized diluent and 10 sterilized glass beads into it, draw 1 mL of the initial suspension obtained in step S1 into the vial, vibrate with a vortex oscillator for 20-30 s, and make 1 : 100 dilution I, and the resulting dilution I is then subjected to ten-fold serial dilutions, respectively gradient dilution to a dilution of 10 7 , 10 8 , 10 9 ;

[0062] S3: Spread plate culture:

[0063] The dilution obtained in step S2 is 10 7 , 10 8 , 10 9 The diluted solution w...

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Abstract

The invention belongs to the technical field of microbial strains, and in particular relates to a culture medium for counting live bacteria of Bacillus subtilis, a sterilizing diluent and a method for counting live bacteria. Among them, the solid medium formula: peeled soybean meal powder (crushed to 100 mesh) 4‑6g, corn flour (ground to 100 mesh) 3‑5g, sodium chloride 5‑10g, xanthan gum 3‑5g, agar 11‑ 13g, distilled water 1000mL, pH value 7.3±0.2; sterile diluent: sodium chloride 8‑9g, potassium dihydrogen phosphate 1‑2g, anhydrous glucose 0.3‑0.5g, peptone 0.3‑0.5g, defoamer 0.1 ‑0.2g, 1000mL of distilled water; and the culture method of first adapting to temperature and then cooling down reduces the secretion of antibacterial substances of bacterial cells, reduces the speed of colony expansion, and reduces the competitive inhibition of bacterial cell growth. The method for counting viable Bacillus subtilis of the present invention improves the accuracy of dilution, reduces the death rate of bacterial cells in the oscillation dilution process, reduces the aggregation of bacterial cells, controls the spread of bacterial colonies, reduces the size of bacterial colonies, and improves the efficiency of Bacillus subtilis. Accuracy and repeatability of counting viable bacilli.

Description

technical field [0001] The invention belongs to the technical field of microbial strains, and in particular relates to a solid medium for counting viable bacteria of Bacillus subtilis, a sterilizing diluent and a method for counting viable bacteria. Background technique [0002] Bacillus subtilis exists in the form of endospores in the preparation. After the spores enter the intestinal tract of animals, they can quickly revive in the upper intestinal tract and secrete highly active proteases, lipases, and amylases, which help to degrade complex carbohydrates and produce Antibacterial and preventive effects by antagonizing the polypeptide substances of intestinal pathogenic bacteria. In addition, Bacillus subtilis is an aerobic bacterium, which can create an anaerobic environment by consuming oxygen in the intestinal tract, promote the reproduction of dominant anaerobic bacteria in the intestinal tract, and maintain the ecological balance of the intestinal tract. [0003] Th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/06C12Q1/04C12R1/125
CPCC12Q1/06C12Q1/045G01N2333/32
Inventor 赖水明肖俊峰曾诚张志榕杨伟春朱德钧李本松吴有林
Owner GUANGZHOU AONONG BIOTECH
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