JPH04117276A - Device for microbe culture medium - Google Patents

Abstract

PURPOSE:To obtain the title device capable of safely producing in simple facility and stably forming smooth and uniform slant culture medium and reduced in change of physical properties by providing a specific hydrophilic polymer layer on the surface of a device for microorganism culture medium made of a plastics. CONSTITUTION:The aimed device made of a plastics and providing a hydrophilic polymer layer consisting of a copolymer of glycosylethyl methacrylate and olefinic compound (preferably acrylic acid or styrene) thereon. The hydrophilic polymer layer is preferably formed by applying a solution of polymer consisting of the above-mentioned copolymer to the surface of plastics. Furthermore, the plastics constituting the device is preferably e.g. polycarbonate.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a microorganism growth medium device used for testing, storing, and transporting microorganisms, and more specifically, the present invention relates to a microorganism growth medium device used for testing, storing, and transporting microorganisms, and more specifically, the present invention relates to a microorganism growth medium device used for testing, storing, and transporting microorganisms. The present invention relates to a plastic microbial growth medium device.

[Conventional technology]

Conventionally, slanted media made of agar containing various components have been widely used for testing, preserving, or transporting microorganisms. Slants are generally formed in glassware such as glass test tubes.

However, (a) glass containers are easily broken during transportation, (b) it is difficult to completely remove the agar medium from glass test tubes when disposing of them, which poses problems in industrial waste disposal, and (C ) There were problems such as the slant culture medium made of agar peeling off from the glass surface during transportation, making it difficult to inoculate microorganisms.

Therefore, attempts have been made to use plastic containers instead of glass containers. However, when using a plastic container for boat fishing, the surface is hydrophobic, making it impossible to form a smooth and uniform slope when preparing a slope culture medium.As a result, bacterial growth is insufficient and accurate. It was difficult to obtain accurate test results.

On the other hand, in order to solve the above-mentioned problems with plastic containers, a method of making the inner surface of the container hydrophilic is disclosed in JP-A-1-179685 and JP-A-1-165367. That is, JP-A No. 1-179685 discloses that cell culture efficiency can be increased by applying corona discharge treatment to the inner surface of a plastic container, and JP-A No. 1-165367 discloses that the inner surface of a plastic container can be treated with corona discharge to improve cell culture efficiency. It has been disclosed that plasma treatment can be made hydrophilic and increase cell culture efficiency.

However, since high frequencies are used in corona discharge treatment and plasma treatment, there is a risk that this may have a harmful effect on workers. In addition, in order to process a large number of bottle-shaped containers at the same time, it is necessary to provide a large number of gas ejection holes.
There was also the problem that the equipment was too thick.
With hollow tube-shaped containers, it is difficult to uniformly apply corona discharge treatment or plasma treatment to the inner surface of the container.

In addition, there was also the problem that the hydrophilicity imparted by plasma treatment decreased over time.

On the other hand, as an attempt to impart hydrophilicity to plastic containers, a method is disclosed in JP-A-59-168052 in which a molded product having a hydroxyl group in the molecule is added to a resin, melted and kneaded, and then molded. has also been proposed. However, the method disclosed in JP-A-59-168052 involves kneading an aliphatic amino compound and a polystyrene resin or a polypropylene resin, and the resins that can impart hydrophilicity are limited to these. In addition, since the kneaded aliphatic amino compound has a low molecular weight, it tends to cause bleeding on the surface of the plastic container, and there is also the problem that the physical properties of the molded product deteriorate over time.

[Problem to be solved by the invention]

Therefore, the object of the present invention has been made in view of the problems of the prior art described above, and the object is to:
It can be manufactured without requiring means that have a harmful effect on the human body or large-scale equipment, and it is possible to stably form a smooth and uniform slope culture medium. It is an object of the present invention to provide a plastic microbial growth medium device that undergoes little change.

[Means to solve the problem]

The microorganism growth medium device of the present invention is constructed using plastic, and has a hydrophilic polymer layer on the surface thereof made of a copolymer of glucosylethyl methacrylate and an olefin compound copolymerizable with glucosylethyl methacrylate. It is characterized by being provided with. This hydrophilic polymer layer is made by coating the plastic surface with a polymer solution obtained by polymerizing glucosylethyl methacrylate and an olefin compound that can be copolymerized with it.
Set by drying.

Hereinafter, each configuration of the present invention will be explained in detail.

In the microorganism growth medium device of the present invention, a hydrophilic polymer layer as described above is provided on the plastic surface, and this hydrophilic polymer layer is made of glucosylethyl methacrylate and an olefin compound. It is composed of a copolymer of

i) Glucosylethyl methacrylate The glucosylethyl methacrylate used to provide the hydrophilic polymer layer in the present invention is a glycoside represented by the following structural formula, and includes acrylic acid, methacrylic acid,
It can be easily copolymerized with common monomers such as methyl methacrylate, styrene, and acrylonitrile to form copolymers with various solubility.

As glucosylethyl methacrylate, it is possible to use, for example, those marketed by Nippon Fine Chemical Co., Ltd. under the trade name rsucraph-GEMAJ.

ii) Olefin Compound In the present invention, the hydrophilic polymer layer is constructed by copolymerizing the glucosylethyl methacrylate with an olefin compound. Examples of copolymerizable olefin compounds that can be used include the following.

(a) Methyl (meth)acrylate, ethyl (meth)acrylate, butyl (meth)acrylate, amyl (meth)acrylate, hexyl (meth)acrylate, octyl (meth)acrylate, 2-ethylhexyl (meth)acrylate, decyl (meth)acrylate ) Acrylate, undecyl (meth)acrylate, lauryl (meth)acrylate, stearyl (meth)acrylate, and other alkyl esters of methacrylic acid or acrylic acid; cycloalkyl esters of methacrylic acid or acrylic acid, such as cyclohexyl (meth)acrylate; styrene,
Vinyls such as vinyl acetate or vinyl propionate;
Hydrophobic olefin compounds such as nitriles such as acrylonitrile.

(b) 2-hydroxyethyl (meth)acrylate, 2
-Hydroxypropyl (meth)acrylate, 2-(dimethylamino)ethyl (meth)acrylate, 2-(diethylamino)ethyl (meth)acrylate, 3-(dimethylamino)propyl (meth)acrylate, 3-(
Hydrophilic olefinic compounds such as diethylamino)propyl(meth)acrylate, polyethylene glycol monomethacrylate, polyethylene glycol monoacrylate, acrylamide, methacrylamide, dimethylacrylamide, dimethylmethacrylamide, acrylic acid, methacrylic acid, N-vinylpyrrolidone or vinylcarbazole .

(C) Ethylene glycol di(meth)acrylate, diethylene glycol di(meth)acrylate, triethylene glycol di(meth)acrylate, vinyl(meth)acrylate, allyl(meth)acrylate, divinylbenzene, diallyl phthalate or trimethylpropane tri(meth)acrylate ) Polyfunctional olefin compounds such as acrylates, etc.

Preferably, acrylic acid, methacrylic acid, methyl methacrylate, styrene, acrylonitrile, etc. are used.

Mountain)! ! Formation of aqueous polymer layer The hydrophilic polymer layer is formed by applying a polymer solution produced by polymerizing the above-mentioned glucosylethyl methacrylate and an olefin compound onto the plastic surface and drying it.

The second polymer solution can be produced according to conventional polymerization methods such as solution polymerization, bulk polymerization, emulsion polymerization, suspension polymerization, or radiation polymerization.

- By way of example, solution polymerization is carried out in a solvent in the presence or absence of a polymerization initiator such as benzoyl peroxide, azobisisobutyronitrile or ammonium persulfate. The solvent used can be appropriately selected depending on the solubility of the copolymer to be produced. For example, a water-containing polar solvent such as isopropanol, methanol, acetone, or ethyl cellulose, or dimethyl sulfoxide, whose water content is adjusted depending on the solubility of the copolymer, is used. Preferably, an alcoholic water-containing polar solvent is used.

The polymer solution obtained as described above is diluted in advance with an alcohol-based water-containing polar solvent or the like to usually about 0.5 to 20% by weight, and then applied to the plastic surface. The coating amount is not particularly limited, but the thickness of the polymer layer after drying is 0.
．． About 1 to 500 μm, preferably 0. 3~100μ
You just need to apply it so that it becomes 50 m, and drying is usually 50 m
Although it is carried out at a temperature of ~60°C, it may also be carried out by natural drying under ventilation.

In the present invention, plastics that are rendered hydrophilic by providing a hydrophilic polymer layer on the surface as described above are not particularly limited, but transparent plastics may be used to observe the growth of microorganisms. Polystyrene, polypropylene, acrylic resin, vinyl chloride resin, polyethylene terephthalate, polycarbonate, etc., which have excellent properties, are preferably used.

Further, the shape of the plastic base material is not particularly limited, and may be cylindrical, box-shaped, bottle-shaped, sheet-shaped, or film-shaped. Specifically, plastic test tubes,
Examples include petri dishes, multi-hole plates, flasks, bottles, gel-supporting plates, and microscope slide plates.

The media listed in the microorganism growth medium device of the present invention include Ogawa medium, chocolate agar medium, Sabouraud medium,
Semi-solid media mainly composed of agar such as TSI medium, potato dextrose medium, MALT medium, or CZAPEC medium are exemplified, but liquid media containing malt extract, yeast extract, or nutrients required by microorganisms are also applicable. can do.

Method for initial culture of microorganisms of the present invention The microorganism growth medium device of the present invention is used, for example, as follows.

In the case of plastic test tubes, apply the above polymer solution.
After drying, a certain amount of sterilized agar medium kept at 60° C. is added, and the test tube is immediately tilted to prepare a slant medium. After cooling, the slant culture medium in the test tube is inoculated with microorganisms using a platinum loop.

[Action and effect of invention]

In the microorganism growth medium device of the present invention, the inner surface of the plastic test tube is made hydrophilic by the glucosyl groups present in the polymer layer. It can be manufactured uniformly and the reliability of test results can be maintained. Also,
Since the inner surface of the plastic utensil is made hydrophilic, accidents such as the agar medium being peeled off from the inner surface of the utensil or the utensil breaking due to impact during storage or transportation of microorganisms are prevented.

Furthermore, since the above-mentioned hydrophilic polymer layer is provided on the plastic surface, the pH of the contents during storage is difficult to change over time, and sterilization with radiation, ultraviolet rays, or ethylene oxide gas is possible; The wood-loving nature of the surface is reliably maintained even after treatment.

With the microorganism growth medium device of the present invention, there is no need to take out the agar medium after use and dispose of it separately.

That is, it can be sterilized and incinerated with the agar medium still in it. Therefore, waste disposal after the inspection can be carried out very easily.

[Explanation of Examples]

The present invention will now be elucidated by describing non-limiting examples thereof.

50% by weight aqueous solution of glucosylethyl methacrylate 3
20g, methyl methacrylate) 40g.

320g of isopropyl alcohol and 350g of distilled water
was placed in a separable flask equipped with a condenser, and the temperature was raised to 65°C after blowing in nitrogen gas.

2.2'-Azobis(2,4-dimethylvaleronitrile) (50 μm) was dissolved in 20 g of isopropyl alcohol and added thereto, followed by reaction at 65-68° C. for 8 hours under stirring in a nitrogen atmosphere. As a result, the viscosity is 1080 cp
s (rotational viscosity needle, 6 Q rpm, 25°C), and the polymerization rate at which a transparent solution was obtained was 97%, and the average molecular weight (M
W) was 9.3×10'.

The solution obtained as above was diluted approximately 10 times with a 50% by weight aqueous isopropyl alcohol solution, filled into 5 polystyrene test tubes (capacity 15 m), and after opening the entire volume by approximately 2 minutes, forced It was dried in a circulating dryer at a temperature of 60° C. for 20 minutes and then allowed to cool.

In addition, for comparison, three polystyrene test tubes that were not subjected to the above treatment were prepared.

TSI agar medium prepared separately and kept at 60°C (
4yd (manufactured by Nippon Pharmaceutical Co., Ltd.) was poured into the above-treated test tubes and a comparison test tube, a polyethylene cap was lightly attached, and the test tubes were immediately tilted.

In the test tubes treated with hydrophilic treatment, a flat slanted medium was formed. On the other hand, an uneven slope was formed in an untreated polystyrene test tube for comparison.

When both test tubes were stored at room temperature for 20 days, the color of the medium in the untreated test tube changed from orange to deep red on the second day, but no change in the color of the medium was observed in the hydrophilized test tube. There wasn't. At the end of the storage period, the pH of the medium was measured by bringing a pH measuring electrode into direct contact with the medium, and the results shown in Table 1 were obtained. In addition, the pH of the medium at the time of slope formation
was 7°3.

In addition, a polystyrene petri dish (60 mm diameter) was treated with hydrophilic treatment in the same manner, and the receding contact angle of the treated surface, that is, the contact angle when water droplets were sucked up, was measured. The results are shown in Table 2.

Table 1 Table 2; Receding contact angle (degrees) Working group I 50% by weight aqueous solution of glucosylethyl methacrylate 2
80 g of dioxane, 170 g of dioxane, 170 g of isopropyl alcohol, and 200 g of distilled water were placed in a separable flask equipped with a condenser, and the temperature was raised to 65° C. without blowing in nitrogen gas.

2. After dissolving 50 g of 2'-azobis(2,4-dimethylvaleronitrile) in 20 g of isopropyl alcohol, it was added to the solution in the above flask and reacted for 10 hours at 65 to 70'C under stirring in a nitrogen atmosphere. I let it happen. The obtained polymerization liquid had a viscosity of 760 cps (rotational viscometer, 60 cps).
rpm, 25°C). The polymerization rate was 95.
2%, average molecular weight was 8.9X10'.

The solution obtained as above was diluted about 10 times with a 33% by weight aqueous isopropyl alcohol solution, filled into 3 polycarbonate test tubes (capacity 15 d), and after about 1 hour, the entire volume was emptied. I turned it upside down and let it air dry overnight.

On the other hand, three polycarbonate test tubes that were not subjected to the above treatment were separately prepared for comparison.

The next day, prepare TSI separately and keep it warm at 60℃.
N-temperature medium (manufactured by Nippon Pharmaceutical Co., Ltd.) 4H1 was poured into polycarbonate test tubes treated as described above and comparison test tubes, the polyethylene caps were loosened, and the test tubes were immediately tilted.

A flat slanted medium was formed in the hydrophilized test tube. In the untreated polycarbonate test tube used as a control, an uneven slope was formed. Note that the pH at the time of medium preparation was 7.2.

When both test tubes were stored at room temperature for 10 days, the color of the medium in the untreated test tube changed from orange to deep red on the third day, but no change in the color of the medium was observed in the hydrophilized test tube. There wasn't. When the pH of the culture medium was measured at the end of the storage period, the results shown in Table 3 were obtained.

(Hereinafter, blank space) Table 3 The polymerization solution of Example 1 was diluted about 10 times with 50% isopropyl alcohol aqueous solution, filled into 5 test tubes made of polyethylene terephthalate, and after emptying the entire volume after about 2 hours,
It was dried for 15 minutes in a dryer at ℃ and allowed to cool. In addition, for comparison, three test tubes made of polyethylene terephthalate that had not been subjected to the above treatment were prepared.

4 m of TSI agar medium (manufactured by Nippon Pharmaceutical Co., Ltd.) prepared separately and kept at 60°C was poured into each of the above test tubes, the polyethylene cap was removed, and the test tubes were immediately tilted.

A flat slanted medium was formed in a polyethylene terephthalate test tube that had been subjected to hydrophilic treatment.

In the untreated polyethylene terephthalate test tube used as a comparison, an uneven slope was formed.

When both test tubes were stored at room temperature for 10 days, the medium color changed from orange to orange-red in the untreated test tube on the 5th day, but no change in the medium color was observed in the hydrophilic test tube. I couldn't. When the pH of the medium was measured on the 6th day of storage, the results shown in Table 4 were obtained. Note that the pH of the medium at the time of slope formation was 7°3.

In addition, a 10-diluted solution of the above polymerization solution was uniformly applied to a polyethylene terephthalate sheet (a commercially available OHP sheet was used) using a brush, and the sheet was dried for 20 minutes in a drying oven at 50''C. The results of the angle measurements are shown in Table 5 below.

(Hereafter, margin)

Claims (1)

[Claims] (1) A microorganism growth medium device made of plastic, characterized in that a hydrophilic polymer layer made of a copolymer of glucosylethyl methacrylate and an olefin compound is provided on the surface thereof. Culture medium equipment.