1. An abamectin coloring agent is characterized in that: the coloring agent comprises methylene blue, basic fuchsin, phenol, alcohol and distilled water.

2. A method of formulating the avermectin stain of claim 1, comprising the steps of:

(1) preparing solution A, namely putting methylene blue into a container according to a proportion, adding a certain volume of 95% alcohol, uniformly stirring, and then adding distilled water;

(2) b, preparing a solution B, namely weighing basic fuchsin, adding the basic fuchsin into 95% alcohol, and fully and uniformly mixing to prepare a basic fuchsin solution; weighing phenol, adding distilled water, and mixing uniformly to prepare a phenol solution; mixing the prepared phenol solution with the prepared basic fuchsin solution, and fully and uniformly mixing;

(3) adding the solution A into the solution B according to the proportion, fully and uniformly mixing to prepare a mixed solution, dripping a drop of the prepared mixed solution on filter paper to observe the color diffusion condition, and finely adjusting A, B liquid proportion according to the color condition until the inner ring is blue and the outer ring is red.

3. The method for preparing an abamectin coloring agent according to claim 2, wherein the method comprises the following steps: the quality of methylene blue in the solution A is as follows: 95% of the volume of the hotel essence: the volume of distilled water is 1g: 30-60 mL: 300-600 mL.

4. The method for preparing an abamectin coloring agent according to claim 2, wherein the method comprises the following steps: the mass of basic fuchsin in the B solution is as follows: the mass of phenol is as follows: volume of 95% alcohol: the volume of distilled water is 1g: 5-10 g: 20-50 mL: 80-150 mL.

5. The method for preparing an abamectin coloring agent according to claim 2, wherein the method comprises the following steps: the volume ratio of the liquid A to the liquid B in the coloring agent is 35:2 to 5.

6. A method for preparing a tablet of an abamectin stain according to claim 1, wherein the method comprises the following steps: the method comprises the following steps:

1) weighing abamectin fermentation liquor, and uniformly stirring;

2) dripping 1 drop of abamectin fermentation liquid on a glass slide, and knocking the glass slide until only one thin layer of fermentation liquid exists on the glass slide;

3) drying the slide glass with the thin layer of fermentation liquor in the step (2) on a constant-temperature heating table at 110 ℃ to fix the slide glass;

4) uniformly coloring the glass slide dried in the step (3) by using a prepared coloring agent, and standing for 3-5 seconds;

5) washing the stained glass slide with distilled water;

6) and (4) drying the slide glass punched in the step (5) on a constant-temperature heating table again at the temperature of 110 ℃, and observing under a microscope, wherein live bacteria and dead bacteria are different in color.

7. A method for preparing a tablet of an abamectin staining agent according to claim 6, wherein the method comprises the following steps: the film prepared by the method is observed under a microscope, the live thalli are blue, and the dead thalli are purple red.

Background

When the abamectin fermentation liquor is subjected to slice-making dyeing by a traditional methylene blue dyeing method using a single dyeing agent, the dyeing time is long; the use of alcohol lamp fixing pieces is not safe enough; the cost of decoloring by using hydrochloric acid alcohol is high and long, only one piece can be manufactured at a time, the quantity is small, the requirement of mass production cannot be met, and the death and the survival of infected mixed bacteria are difficult to distinguish during observation.

Disclosure of Invention

The invention aims to solve the problem that the slice-making dyeing time of the abamectin fermentation liquor by using a single dyeing agent is long and the death and activity of infectious mixed bacteria are difficult to observe and distinguish in the prior art, and provides the abamectin dyeing agent.

The invention can solve the problems by the following technical scheme: an abamectin coloring agent is prepared from methylene blue, basic fuchsin, phenol, alcohol and distilled water.

The invention also provides a preparation method of the abamectin coloring agent, which comprises the following steps: (1) preparing solution A, namely putting methylene blue into a container according to a proportion, adding a certain volume of 95% alcohol, uniformly stirring, and then adding distilled water;

(2) b, preparing a solution B, namely weighing basic fuchsin, adding the basic fuchsin into 95% alcohol, and fully and uniformly mixing to prepare a basic fuchsin solution; weighing phenol, adding distilled water, and mixing uniformly to prepare a phenol solution; mixing the prepared phenol solution with the prepared basic fuchsin solution, and fully and uniformly mixing;

(3) adding the solution A into the solution B according to the proportion, fully and uniformly mixing to prepare a mixed solution, dripping a drop of the prepared mixed solution on filter paper to observe the color diffusion condition, and finely adjusting A, B liquid proportion according to the color condition until the inner ring is blue and the outer ring is red.

Preferably, the mass of methylene blue in the solution A is 95 percent of the volume of hotel essence, and the volume of distilled water is 1g, 30-60 mL, 300-600 mL.

Preferably, the mass of basic fuchsin in the solution B, the mass of phenol, the volume of 95% alcohol and the volume of distilled water are respectively 5-10 g, 20-50 mL and 80-150 mL.

Preferably, the volume ratio of the liquid A to the liquid B in the coloring agent is 35: 2-5.

The invention also provides a method for preparing the abamectin coloring agent, which comprises the following steps:

1) weighing abamectin fermentation liquor, and uniformly stirring;

2) dripping 1 drop of abamectin fermentation liquid on a glass slide, and knocking the glass slide until only one thin layer of fermentation liquid exists on the glass slide;

3) drying the slide glass with the thin layer of fermentation liquor in the step (2) on a constant-temperature heating table at 110 ℃ to fix the slide glass;

4) uniformly coloring the glass slide dried in the step (3) by using a prepared coloring agent, and standing for 3-5 seconds;

5) washing the stained glass slide with distilled water;

6) and (4) drying the slide glass punched in the step (5) on a constant-temperature heating table again at the temperature of 110 ℃, and observing under a microscope, wherein live bacteria and dead bacteria are different in color.

Preferably, when the slices prepared by the method are observed under a microscope, live thalli are blue, and dead thalli are purple red.

Compared with the background technology, the invention has the following beneficial effects: the method for preparing the abamectin coloring agent and flaking and dyeing the fermentation liquor greatly shortens flaking time, is safer than an alcohol lamp by using a constant-temperature heating table, has stable flaking effect, is more economic and environment-friendly than hydrochloric acid alcohol by using distilled water for decoloring, saves time, can simultaneously dye and flake a large number of flakes, fully meets production requirements, and has clear thalli, easily distinguished colors, blue live thalli and purple red dead thalli by observing the produced flakes under a microscope.

Drawings

FIG. 1 is a microscopic image of a slide made in accordance with an embodiment of the present invention;

FIG. 2 shows the result of microscope imaging of slides prepared by conventional slide staining method.

The specific implementation mode is as follows:

the technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings and embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

The preparation method of the abamectin coloring agent comprises the following steps:

1) and preparing the solution A: 1g of methylene blue is weighed and put into a container, then 30mL of 95% alcohol is added to be stirred evenly, and distilled water is added to reach 300 mL.

2) And preparing liquid B: 1g of basic fuchsin is weighed, 20mL of 95% alcohol is added, and the mixture is mixed well. Weighing 5g of phenol, adding 80-150 mL of distilled water, and uniformly mixing. The phenol solution was mixed with basic fuchsin and mixed well.

3) Preparing a mixed solution of the solution A and the solution B: adding 4mLB liquid into every 70mLA liquid, mixing evenly, dropping one drop of filter paper to observe the color diffusion condition, the inner circle is blue, the outer circle is red, and fine adjusting A, B liquid proportion according to the color condition.

The method for preparing the abamectin staining agent comprises the following steps:

stirring avermectin fermentation liquor uniformly, taking 1 drop to a glass slide, knocking the slide until only a thin layer of fermentation liquor exists on the glass slide, drying and fixing the slide on a constant temperature heating table at 110 ℃, uniformly coloring the slide by using a prepared coloring agent, standing for 3 seconds, punching by using distilled water, drying on the constant temperature heating table at 110 ℃ again, observing under a microscope, wherein the live thallus journey is blue, and the dead thallus journey is purple red.

Example 2

The preparation method of the abamectin coloring agent comprises the following steps:

1) and preparing the solution A: 1g of methylene blue is weighed and put into a container, then 60mL of 95% alcohol is added to be stirred evenly, and distilled water is added to 600 mL.

2) And preparing liquid B: first, 1g of basic fuchsin was weighed, 50mL of 95% ethanol was added, and the mixture was mixed well. 10g of phenol was weighed and mixed with 150mL of distilled water. The phenol solution was mixed with basic fuchsin and mixed well.

3) Preparing a mixed solution of the solution A and the solution B: adding 10mLB liquid into every 70mLA liquid, mixing evenly, dropping one drop of filter paper to observe the color diffusion condition, the inner circle is blue, the outer circle is red, and fine adjusting A, B liquid proportion according to the color condition.

The method for preparing the abamectin staining agent comprises the following steps:

stirring avermectin fermentation liquor uniformly, taking 1 drop to a glass slide, knocking the slide until only a thin layer of fermentation liquor exists on the glass slide, drying and fixing the slide on a constant temperature heating table at 110 ℃, uniformly coloring the slide by using a prepared coloring agent, standing for 5 seconds, punching by using distilled water, drying on the constant temperature heating table at 110 ℃ again, observing under a microscope, wherein the live thallus journey is blue, and the dead thallus journey is purple red.

Example 3

The preparation method of the abamectin coloring agent comprises the following steps:

1) and preparing the solution A: first, 1g of methylene blue is weighed and put into a container, then 50mL of 95% alcohol is added to be stirred uniformly, and distilled water is added to 500 mL.

2) And preparing liquid B: first, 1g of basic fuchsin was weighed, 30mL of 95% ethanol was added, and the mixture was mixed well. 8g of phenol was weighed and mixed with 100mL of distilled water. The phenol solution was mixed with basic fuchsin and mixed well.

3) Preparing a mixed solution of the solution A and the solution B: adding 7mLB liquid into every 70mLA liquid, mixing evenly, dropping one drop of filter paper to observe the color diffusion condition, the inner circle is blue, the outer circle is red, and fine adjusting A, B liquid proportion according to the color condition.

The method for preparing the abamectin staining agent comprises the following steps:

stirring avermectin fermentation liquor uniformly, taking 1 drop to a glass slide, knocking the slide until only a thin layer of fermentation liquor exists on the glass slide, drying and fixing the slide on a constant temperature heating table at 110 ℃, uniformly coloring the slide by using a prepared coloring agent, standing for 4 seconds, punching by using distilled water, drying on the constant temperature heating table at 110 ℃ again, observing under a microscope, wherein the live thallus journey is blue, and the dead thallus journey is purple red.

Comparative example 1

Traditional avermectin-methylene staining: the method comprises the steps of uniformly stirring abamectin fermentation liquor, dropping 1 drop of abamectin fermentation liquor onto a glass slide, knocking the glass slide until only one thin layer of fermentation liquor exists on the glass slide, drying the glass slide above an alcohol lamp (the condition that the glass slide is not scalded when hands are placed under the glass slide), enabling the glass slide to move back and forth on the alcohol lamp flame for three times, dyeing the prepared methylene blue for 1min, washing the glass slide with distilled water, decoloring the glass slide with hydrochloric acid alcohol for 30s, washing the glass slide with the distilled water, sucking the glass slide dry, observing the glass slide under.

The traditional methylene blue dyeing method has long flaking time, has potential safety hazard of fire when an alcohol lamp is used, uses hydrochloric acid for alcohol decolorization, uses hydrochloric acid which belongs to an easily toxic reagent during preparation, has potential safety hazard when the hydrochloric acid belongs to strong acid preparation, has higher cost than distilled water for flaking, can only produce one flake by using the alcohol lamp, cannot simultaneously dry and fix a plurality of flakes, has complicated operation steps compared with the method, and has no identification capability because the traditional methylene blue dyeing belongs to single dyeing.

The imaging results of the wafers produced in examples 1-3 under a microscope are shown in FIG. 1; comparative example the results of microscopic imaging of slides prepared using conventional slide staining methods are shown in figure 2.

In the attached figure 1, the blue hypha in the mass is the streptomyces avermitilis as the production bacterium, and the purple elliptical hypha is the yeast. In the attached figure 2, the blue hypha is in a dough shape and is used as the production bacteria of the streptomyces avermitilis, and the color is slightly light elliptical and is used as the yeast. The production bacteria are live bacteria when being tabletted, and the yeast is a fermentation medium component and is dead bacteria after high-temperature sterilization. Comparing the two figures, the tablets prepared by the preparation and the tabletting method of the abamectin staining agent have clear thalli, easily distinguished colors, blue live thalli and purple red dead thalli. However, the color of the slices manufactured by the traditional method is basically blue, and the death and the survival of infectious mixed bacteria are difficult to distinguish. And the method can simultaneously dye and slice a large number of pieces, thereby fully meeting the production requirement.