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Uses of Aspergillus brasiliensis as cigarette anti-mildew agent antibacterial effect indicating strains, and method for using Aspergillus brasiliensis as cigarette anti-mildew agent antibacterial effect indicating strain

A mold inhibitor and indicator bacteria technology, applied in the field of microorganisms, can solve problems such as threatening the health and environmental safety of experimenters, affecting the accuracy of experimental data, and time-consuming and laborious separation processes, so as to save experimental costs, shorten the cultivation period, and speed up the process. The effect of the R&D cycle

Inactive Publication Date: 2019-02-01
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method needs to screen and isolate multiple moldy microorganisms from moldy cigarettes, but the separation process is time-consuming and laborious, and operational errors are prone to occur when making spore suspensions. It is difficult to compare between different genera, and it is not easy to control variables, which will affect the experimental data. the accuracy of
In addition, there are literatures that single fungi such as Aspergillus flavus are used as indicator bacteria for the antifungal effect of antifungal agents, but the metabolites of most fungi are mycotoxins such as aflatoxin, which may have carcinogenic, teratogenic and mutagenic effects. Exposure to a large number of medium and high doses seriously threatens the health and environmental safety of experimenters
None of the above indicator bacteria are suitable for use as indicator bacteria for the antibacterial effect of cigarette antifungal agents

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: The mildew-causing performance test of mildew microorganisms.

[0034] To prepare tobacco juice culture medium, the method is as follows: take 50.0g tobacco leaves, add 1000mL distilled water to boil, filter twice with 2 layers of gauze, add 18.0g agar, set the volume to 1000mL, autoclave at 121°C for 15 minutes, cool to about 50°C , add 20mg chloramphenicol to every 200ml culture medium, shake evenly and cool for later use, and obtain the tobacco juice culture medium plate. Pick the marginal spores of cigarette mildew microorganisms isolated from 15 kinds of mildewed cigarettes stored on the slant medium, plant them in the center of the above-mentioned tobacco juice medium, and place them upside down for constant temperature and humidity culture under the culture conditions of 28°C and 85%RH Cultivate in the box for 5 days, measure the diameter of the colony by the cross method, do 3 parallels for each group, and calculate the average value, and use the u...

Embodiment 2

[0035] Embodiment 2: Aspergillus brasiliensis is used as the method of cigarette antifungal agent cinnamaldehyde antibacterial effect indicator bacteria

[0036] After activating the commercially available ATCC16404 standard strain under the specified conditions (the conditions are provided by the merchant), streak inoculate it in a slant medium test tube, and cultivate it in a static constant temperature and humidity incubator at 28°C and 85% RH for 2 days ; The medium formula includes the following components: potato flour 6.0g / L; glucose 20.0g / L; chloramphenicol 0.1g / L; agar 20.0g / L. Aspergillus brasiliensis spore suspension was prepared by culturing the resulting colony on the slant medium: wash the surface of the slant medium containing the colony with 85mg / ml sterile saline, and vortex for 30s to obtain the spore suspension; after filtering twice through 4 layers of sterile gauze, Count by hemocytometer, dilute to 1 x 10 5CFU / ml is enough. The same spore suspension pre...

Embodiment 3

[0037] Embodiment 3: Aspergillus brasiliensis is used as the method for the antibacterial effect indicator bacterium of cigarette antifungal agent eugenol

[0038] According to the method of Example 2, the antifungal agent of cigarettes is replaced by eugenol of equal concentration by cinnamaldehyde. The results showed that the zone of inhibition of 2.50mg / ml eugenol on the indicator bacteria of Aspergillus brasiliensis was 22.03±0.82mm, and the zone of inhibition on the mixture of cigarette mildew bacteria was 22.52±0.52mm. It shows that Aspergillus brasiliensis and cigarette mildew mixed bacteria are highly sensitive to cigarette antifungal agent eugenol (+++).

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Abstract

The invention discloses uses of Aspergillus brasiliensis as cigarette anti-mildew agent antibacterial effect indicating strains, and a method for using Aspergillus brasiliensis as a cigarette anti-mildew agent antibacterial effect indicating strain, wherein the ATCC standard strain number of the Aspergillus brasiliensis is No.16404. The method for using Aspergillus brasiliensis as the cigarette antifungal agent antibacterial effect indicating strain comprises: (1) activating Aspergillus brasiliensis; (2) culturing Aspergillus brasiliensis; (3) preparing an Aspergillus brasiliensis spore suspension; (4) preparing a bacterial-containing flat plate; (5) placing a sterile Oxford cup; (6) preparing an anti-mildew agent dilution liquid; (7) adding the anti-mildew agent to the Oxford cup; (8) culturing an experimental group and a control group; (9) measuring the diameter of the zones of inhibition by using a crossing method, and recording data; and (10) grading the sensitivity according to the average diameter of the zones of inhibition. According to the present invention, with the method, the problems of cumbersome screening and potential safety hazard of the cigarette anti-mildew agentantibacterial effect indicating strain are solved, and the method has advantages of short culture period, high safety, strong representativeness, high accuracy and the like.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to an application and a method of using Aspergillus brasiliensis as an indicator bacterium for the antibacterial effect of cigarette antifungal agents. Background technique [0002] my country is the world's largest producer of tobacco leaves, accounting for about one-third of the world's total output. The tobacco industry is one of the pillar industries that cannot be replaced in our country, and it is also one of the important sources of national fiscal revenue. With the acceleration of the joint reorganization of my country's cigarette industry enterprises and the continuous improvement of the structure, the cigarette industry enterprises have more urgent demand for high-quality tobacco. However, during the sale and storage of cigarettes, mildew is prone to occur due to the influence of ambient humidity, resulting in reduced quality or even scrapping of cigarettes. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/66
CPCC12Q1/04
Inventor 雷声杨乾栩师艳萍蒋举兴刘秀明段焰青张玲张晓鸣夏书芹陈文怡
Owner CHINA TOBACCO YUNNAN IND
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