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Cold-water soluble gel substrate resistant to decomposition of mesophilic bacteria and application thereof

A gel matrix and soluble technology, applied in the cold water soluble gel matrix and its application fields that are resistant to the decomposition of mesophilic bacteria, can solve the problems of inhibiting microbial growth, low strength, unfavorable observation, etc., to avoid adverse effects and ensure normal The effect of growth

Active Publication Date: 2018-08-28
GUANGDONG HUANKAI MICROBIAL SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This greatly limits the range of options
At the same time, in order to ensure the growth of microorganisms, the strength of the gel matrix is ​​required to be appropriate. If the strength is too high, it will inhibit the growth of microorganisms and affect the test results; if the strength is too low, it is not conducive to observation.

Method used

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  • Cold-water soluble gel substrate resistant to decomposition of mesophilic bacteria and application thereof
  • Cold-water soluble gel substrate resistant to decomposition of mesophilic bacteria and application thereof
  • Cold-water soluble gel substrate resistant to decomposition of mesophilic bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 The impact of different gel strength additives on gel strength

[0043] Carrageenan (100%) is mixed with 5% water-soluble starch, 5% urea, 5% sodium chloride, 2.5% calcium chloride, 1% sodium acetate respectively, and 1.5g is suspended in 20ml 10% (w / v) Polyethylene glycol-6000 methanol solution, take 2ml and add to the center of the test piece to dry. After adding 1ml of sterile deionized water to redissolve overnight, observe and test the gel strength by hand pressing. The results are shown in Table 1.

[0044] Table 1 The impact of gel strength additives on gel strength

[0045]

[0046]

[0047] Remarks: "+" stands for gel strength.

[0048] The gel strength of adding sodium chloride or sodium acetate is not significantly different from that of blank carrageenan. Calcium chloride, urea and starch can significantly enhance the gel strength, which may be caused by Ca 2+ The association with groups such as sulfate esters in carrageenan improves t...

Embodiment 2

[0049] The influence of embodiment 2 different factors on carrageenase enzymatic activity

[0050] Weigh 45.04mg of D-galactose, dissolve it with double distilled water to 50ml to form a standard solution of 5μmol / ml, add reagents and mix well, then develop color in a boiling water bath for 10min, cool to room temperature, and make up to 5ml with double distilled water . The absorbance value was measured at 520nm, and the standard curve was drawn with the absorbance value as the abscissa and the D-galactose content as the ordinate.

[0051] The enzyme activity was determined by 3,5-dinitrosalicylic acid (DNS) method. Take 50 μl carrageenan enzyme solution, add it to 550 μl 0.05M phosphate (PBS) buffer (pH7.0) of 0.5% carrageenan solution, add sodium chloride, potassium chloride, calcium chloride, magnesium chloride, EDTA respectively , SDS, β-mercaptoethanol, and urea in a 60°C water bath for 20 minutes, quickly add 900 μl DNS to a boiling water bath for 10 minutes, cool to ...

Embodiment 3

[0059] The effect of the total number of colonies detection sheet prepared by embodiment 3 enzyme inhibitor EDTA

[0060] 5g of tryptone, 2.5g of yeast extract powder, 1g of glucose, 0.1g of TTC, respectively mixed with 34g of carrageenan, 2g of water-soluble starch, and 0.015g of EDTA by ball milling, and fixed the dry powder on the culture device to make a test piece. Sterilized by irradiation with cobalt 60 at a dose of 15KGy. Escherichia coli ATCC25922, Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC27853, and Bacillus subtilis ATCC6633 were cultured in tryptone soybean broth at 36±1°C for 16-18 hours, and the enrichment solution was shaken with sterile physiological Saline gradient dilution to about 100 CFU / ml. Take 1ml and add it to the center of the sterilized test piece, cover with polyethylene film to spread the sample liquid to fill the whole area. Incubate at 36±1°C for 24 hours, and observe the results. The results are shown in Table 4.

[0061] The ...

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Abstract

The invention discloses a cold-water soluble gel substrate resistant to decomposition of mesophilic bacteria and application thereof. The gel of a substrate is at least one of carrageenan or a modifier thereof, and a gel strength adjusting agent and a carrageenan enzyme inhibitor are added in the gel substrate. According to a microbial testing piece, the normal growing of microorganisms is ensuredto avoid adverse effects due to the fact that cold-water gel which is not easily decomposed by the mesophilic bacteria directly is chosen and the special enzyme inhibitor and gel strength adjusting agent capable of inhibiting the decomposition of gel enzyme are added, so that samples containing bacillus are not liquefied and dispersed and single colony is clear, easy to observe and count during the cultivation process. Transparent gel with a certain degree of gel strength with the shape of agar is formed, so that colony edges after Escherichia coli, pseudomonas aeruginosa and the like are cultivated are relatively more regular, the color is brighter, and the characteristics are closer to traditional agar culture mediums and more typical.

Description

technical field [0001] The invention relates to a gel matrix for microbial culture medium and its application, in particular to a cold-water soluble gel matrix resistant to decomposition by mesophilic bacteria and its application. Background technique [0002] Usually the total number of colonies is counted using the pour method, surface coating method, spiral inoculation method or membrane filtration method, all of which without exception use agar medium and glass or plastic Petri dishes. Agar is a gelatin derived from reddish-purple marine algae such as Geliflower. Agar was first applied to microbiology in 1881 by Fanny Hesse. By the beginning of the 20th century, agar began to replace gelatin as the first choice of coagulant because it still maintained gel strength in the process of cultivating certain pathogenic bacteria at high temperature. Agar is in a gel state at room temperature, and remains a hard gel at temperatures as high as 65°C. Agar melts at about 85°C and...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C08J3/075C08L5/00C08L3/02
CPCC08J3/075C08J2305/00C08J2403/02C12Q1/045
Inventor 蔡芷荷滕昆仑陈博张建明卢勉飞吴清平熊争
Owner GUANGDONG HUANKAI MICROBIAL SCI & TECH