TWI806393B - Method of coating pests and fungicides on a pest culture medium platform for direct drug efficacy evaluation - Google Patents

Abstract

A method of coating harmful organisms and fungicides on the medium platform to evaluate the efficacy of the medicine. It uses about 3% agar to prepare the medium, sterilizes it, pours it into a 9 cm transparent sterile plastic petri dish, and scratches the side of the bottom of the petri dish Create a rectangular coating area of 6 cm x 1 cm. That is to say, apply fungi or bacteria to the area and then apply the same amount of fungicide to observe the germination rate or growth rate, which is a method for evaluating its efficacy. After a certain action time, the fungus or bacteria to be tested can be dragged from the platform with an inoculation loop or a suitable tool, and dragged along multiple dilution lines to the observation area outside the area for observing the antibacterial or bactericidal effect of the agent under a microscope Mechanism.

Description

Method of coating pests and fungicides on a pest culture medium platform for direct drug efficacy evaluation

The invention relates to a method for evaluating the efficacy of fungicides on germs or harmful organisms, especially a method for directly coating germs or harmful organisms on a culture medium platform and then coating agents for observation.

The research and development of fungicides, including antibiotics, generally requires the evaluation of the efficacy of the agents on the target microorganisms. In the past, the methods of drug efficacy evaluation mainly included the filter paper tablet test method in the laboratory, the medium digging test method, and the "poison food method" of medium dosing. However, these methods often require the use of a lot of consumables, and the concentration of various agents needs to be prepared to test the lowest bacteriostatic or bactericidal concentration, and it is often impossible to determine whether the tested bacteria have died or are only temporarily dormant. .

In the effective drug screening of a pathogenic bacteria, if there are many kinds of drugs for preliminary testing, the drug efficacy testing project of various concentrations is often very huge, and the manpower and material resources are very expensive. Another example is agricultural chemicals and antibiotics used in human medicine. Due to frequent repeated use in recent years, drug resistance has often occurred. For the test of drug-resistant strains, "drug efficacy test" or "control rate test" must also be carried out. If the "control rate" of a drug on the target organism is significantly reduced or reduced, it shows that the special strain has "resistance". Medicinal properties". In terms of plant diseases and insect pests, whether they are resistant to pesticides is very important for the recommendation of control prescriptions, and a rapid testing platform is also required. In terms of human pathogens, new drug-resistant strains often appear, and a rapid testing platform is also required. If the drug-resistant strains cannot be tested, the most effective prescription will not be issued. for effectively defeating such germs.

In view of this, the inventor of the present case made further research and innovation, revealing the method of coating harmful organisms and fungicides on the culture medium platform so as to evaluate the drug efficacy.

The purpose of the present invention is to provide a method for coating harmful organisms and bactericides on the culture medium platform to evaluate the efficacy of the medicine. It uses a culture medium containing about 3% agar after sterilization and pours it into a 9 cm transparent sterile plastic culture dish, and Draw a 6 cm x 1 cm rectangular coating area on one side of the bottom of the petri dish. That is to say, fungus or bacteria and other germs are applied to this area, and then the same amount of fungicide is applied to observe the germination rate or growth rate, which is a method for evaluating its efficacy. After a certain period of action, the fungus or bacteria to be tested can be dragged from the platform with an inoculation loop or a suitable tool, dragged along multiple dilution lines to the observation area outside the area, and the antibacterial or bactericidal action mechanism of the agent can be observed under a microscope By.

The testable objects of the present invention may include human pathogens and animal and plant pathogens, and may also include protozoa, nematodes and small insects or eggs. Anything that can be absorbed by a straw and moved to the "testing platform" can be tested for drug efficacy. There are no restrictions on the types of medicines, and those that can be sucked through a straw and moved to the "testing platform" can be applied for testing.

According to a method of coating harmful organisms and bactericides on a medium platform of the present invention to directly evaluate the drug efficacy, the medium uses about 3% but not limited to 3% agar as the base, with or without adding other nutrients. After sterilization, pour it into a 9 cm transparent sterile plastic culture dish. After the medium solidifies, draw (define) a rectangular coating area (1) of 6 cm x 1 cm on the bottom of the culture dish according to the template in Figure 1, and draw 5 or more dilution lines (2). That is, fungus or bacteria and other germs are spread on this "rectangular" platform, and then the same amount of fungicide or test agent is immediately spread. After a fixed period of time, the viability of the fungi or bacteria to be tested can be observed under a microscope, including germination status, growth status, and whether there are morphological changes, etc., for direct evaluation of the efficacy of the test. After the bacteria and the medicament are uniformly mixed for a certain period of time, the tested fungus or bacteria can be dragged from the platform with an inoculation loop or a suitable tool, along 5 but not limited to 5 orthogonal or oblique lines in the coating area (1 ) to the dilution line (2) to the observation area at the end of the dilution line outside the area, and observe the activity of fungi or bacteria dragged outside the area under a microscope, including germination status, growth status, and whether there are morphological changes, etc. . In the above tests, there is a control group in which a smeared bacteria is not added to the drug for comparison with the experimental group, which can be used to calculate the bacteriostatic rate or bactericidal rate of the drug, whether it is temporarily dormant, etc.

Through the above-mentioned practice, the present invention can calculate the "germination inhibition rate" or "growth inhibition rate" by comparing the experimental group and the control group (without adding medicine) in the "test platform area". The comparison between the experimental group in the "test platform area" and the "drag dilution area" can be used to understand the temporary or permanent nature of the drug. When the harmful organisms are dragged to the "drag dilution zone", if the activity cannot be restored, it means that it has a permanent bactericidal effect. When the bacteria are dragged to the "drag dilution zone", the activity can be observed, which means that it is temporarily inhibited.

According to the above method, the use of tetrachloroisobenzonitrile can completely inhibit the germination of uredospores of the fig rust fungus, and the spores outside the platform cannot recover their germination ability, indicating that tetrachloroisobenzonitrile has a "bactericidal" effect. On the contrary, when the Bordeaux mixture was tested, when it was dragged out of the platform, the germination rate increased, indicating that the Bordeaux mixture belongs to the "bacteriostatic" agent.

The method that the present invention is superior to conventional drug efficacy evaluation has the following advantages: 1. The operation is simple and can save manpower. 2. Less consumables to avoid waste of resources and benefit the environment. 3. Inspection and judgment are more accurate, which can improve the efficiency of drug efficacy evaluation.

1: coating area; 2: Dilution line.

Fig. 1 is the schematic diagram of the template for drawing the medium test platform of the present invention.

Fig. 2 is a schematic diagram of an embodiment of the prepared culture medium platform of the present invention.

1: coating area

2: Dilution line

Claims (1)

A method for coating harmful organisms and fungicides on a harmful organism culture medium platform to conduct drug efficacy evaluation, including: 1. Prepare a culture medium platform composed of transparent sterile petri dishes; 2. Use an agar medium, sterilized After that, pour it into the petri dish, and draw a coating area on the bottom surface of the petri dish, apply the germs or harmful organisms to be tested on the coating area, and then apply the fungicide, and then observe the effect of the fungicide on the petri dish. The bactericidal or bacteriostatic effect of germs or harmful organisms is to evaluate the efficacy of the bactericide; the coating area is a rectangular coating area, and multiple dilution lines are perpendicular or oblique to the rectangular coating area After the bacteria or harmful organisms and the fungicide act on the platform for a fixed period of time, the tested germs or harmful organisms are dragged from the platform with an inoculation loop or other tools, and dragged along the dilution line to the dilution line outside the area for observation Area, observe the bacteriostatic or bactericidal action mechanism of the fungicide under a microscope.

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